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Evidence of metabolic mechanisms playing a role in multiple insecticides resistance in Anopheles stephensi populations from Afghanistan
Noor Halim Zahid Safi1,
Abdul Ali Ahmadi1,
Sami Nahzat1,
Seyyed Payman Ziapour2,3,
Seyed Hassan Nikookar2,
Mahmoud Fazeli-Dinan4,
Ahmadali Enayati ORCID: orcid.org/0000-0001-6861-07194 &
Janet Hemingway5
Malaria Journal volume 16, Article number: 100 (2017) Cite this article
Malaria is endemic in most parts of Afghanistan and insecticide-based vector control measures are central in controlling the disease. Insecticide resistance in the main malaria vector Anopheles stephensi from Afghanistan is increasing and attempts should be made to determine the underlying resistance mechanisms for its adequate management.
The contents of cytochrome P450s, esterases, glutathione S-transferases (GSTs) and acetylcholine esterase (AChE) activities were measured in the Kunar and Nangarhar populations of An. stephensi from Afghanistan and the results were compared with those of the susceptible Beech strain using the World Health Organization approved biochemical assay methods for adult mosquitoes.
The cytochrome P450s enzyme ratios were 2.23- and 2.54-fold in the Kunar and Nangarhar populations compared with the susceptible Beech strain. The enzyme ratios for esterases with alpha-naphthyl acetate were 1.45 and 2.11 and with beta-naphthyl acetate were 1.62 and 1.85 in the Kunar and Nangarhar populations respectively compared with the susceptible Beech strain. Esterase ratios with para-nitrophenyl acetate (pNPA) were 1.61 and 1.75 in the Kunar and Nangarhar populations compared with the susceptible Beech strain. The GSTs enzyme ratios were 1.33 and 1.8 in the Kunar and Nangarhar populations compared with the susceptible Beech strain. The inhibition of AChE was 70.9 in the susceptible Beech strain, and 56.7 and 51.5 in the Kunar and Nangarhar populations. The differences between all values of the enzymes activities/contents and AChE inhibition rates in the Kunar and Nangarhar populations were statistically significant when compared with those of the susceptible Beech strain.
Based on the results, the reported resistance to pyrethroid and organophosphate insecticides, and tolerance to bendiocarb in the Kunar and Nangarhar populations of An. stephensi from Afghanistan are likely to be caused by a range of metabolic mechanisms, including esterases, P450s and GSTs combined with target site insensitivity in AChE.
Malaria is endemic in Afghanistan. From a total population of 31 million, 8.5 million live in areas of high transmission and more than 15.4 million in areas of low transmission [1]. Major vectors of malaria in Afghanistan are Anopheles stephensi, Anopheles culicifacies, Anopheles superpictus, Anopheles hyrcanus, Anopheles pulcherrimus, and Anopheles fluviatilis [1], the first two being the most important in the country [2–6]. An. stephensi is widespread in different countries in the Middle East including Iran, Iraq, Bahrain, Saudi Arabia, Oman, India, Pakistan, Afghanistan, Bangladesh, South China and Myanmar (see [7]). Malaria in Afghanistan is predominantly due to Plasmodium vivax (95% of the cases) and Plasmodium falciparum (5%) in two distinct transmission seasons. The total number of confirmed cases in 2015 was 61362 [1].
As in many malaria endemic countries the main malaria control intervention was indoor residual spraying (IRS) with DDT (1950s to 1970), continued with organophosphate insecticides (OPs), such as malathion in later years, followed by insecticide-treated nets (ITNs) in the 1990s, and long-lasting insecticidal nets (LLINs) distribution from 2007 onwards [2, 3]. Deltamethrin-treated LLINs distribution to households in the main malaria-endemic provinces in Afghanistan is currently the main malaria control intervention [3]. During 2007–2014, the number of deltamethrin-treated LLIN distributed in Kunar and Nangarhar was 334,080 and 1,386,217, respectively [8]. Selection pressure from pesticides used in vector control and also in agriculture might have contributed to insecticide resistance in malaria vectors in Afghanistan especially in An. stephensi. This vector species from Nangarhar is resistant to DDT, bendiocarb, permethrin and deltamethrin, and to DDT, deltamethrin, permethrin and malathion in Kunar [8, 9]. Resistance to several insecticides including DDT, dieldrin, malathion and more recently pyrethroids have been reported in An. stephensi from the Middle East region [10–13] and in Afghanistan neighbouring countries including India [7]. The involvement of different enzymes and site insensitivity mechanisms in insecticide resistance in An. stephensi from Iran was confirmed [13–16]. An. stephensi from India had increased activities of esterases and GSTs associated with deltamethrin and permethrin resistance [17, 18]. Involvement of GSTs in insecticide resistance is evident in many insects including different mosquitoes [19]. General esterases are involved in OPs resistance in An. stephensi from Pakistan [20]. In recent years, World Health Organization (WHO) standard insecticide susceptibility bioassays have been performed on An. stephensi from Afghanistan showing resistance to organochlorines, carbamates and pyrethroid insecticides especially in the eastern provinces of Nangarhar and Kunar [4, 9, 21]. In addition, in 2014, target site insensitivity for pyrethroid insecticides known as knockdown resistance (KDR) was studied in An. stephensi from Kunar and Nangarhar. KDR is due to some single-nucleotide polymorphisms (SNP) causing voltage-gated sodium channels of the axons of the nerve cells to become insensitive to the knockdown effect exerted by pyrethroid insecticides. Although the wild type susceptible 1014L allele in the sodium channel gene was most prevalent followed by L1014S (kdr east, 21.4%) and L1014F (kdr west, 1.4%), no kdr homozygotes were detected. Only when the mutation data of kdr e and kdr w are combined, was there any significant association between kdr frequency and insecticide resistance. However, when they are separately considered, there was no significant association between kdr frequency and pyrethroid resistance. The finding that many of the bioassays survivors did not possess the kdr mutation suggests that other resistance mechanisms are present in these populations [21].
Accurate information on the underlying resistance mechanisms in An. stephensi is needed for proper management of insecticide resistance and a better management of malaria through vector control interventions. Therefore, based on the recommendations of the global plan for insecticide resistance management (GPIRM) [22], the need to develop an evidence-based malaria control plan and the suggestion that resistance mechanisms other than kdr are present in An. stephensi [21], other mechanisms operating in An. stephensi, the main malaria vector in Nangarhar and Kunar Provinces in eastern Afghanistan were investigated.
The study area was the provinces of Nangarhar (34.1718°N, 70.6217°E) and Kunar (34.8466°N, 71.0973°E) in eastern Afghanistan (Fig. 1). The sampling places were the same exact districts, villages and coordinates where collections took place for the analysis of kdr in 2014 [21]. These sampling sites were 10 villages in Behsood, Jalalabad and Kama districts in Nangarhar; and Chawkay, Nurgal and Assadabad districts in Kunar (Table 1).
Map of Afghanistan and the location of the provinces of Nangarhar and Kunar in Northeastern region of the country where sampling for Anopheles stephensi took place in October 2015
Table 1 Sampling places of Anopheles stephensi in Kunar and Nangarhar provinces, for biochemical assays of insecticide resistance in Afghanistan in 2015
Mosquito populations
Larvae of the Kunar and Nangarhar populations of An. stephensi were collected from the field (200 larvae from each district). They were reared to adults in the insectary located in Nangarhar National Malaria and Leishmaniasis Control Programme (NMLCP) field station. The adult mosquito specimens were identified to species using Glick's identification keys [23]. Two to three days old adult mosquitoes were transported in cool boxes to NMLCP of the MoPH, Kabul. Upon arrival, they were immediately placed in a freezer (−80 °C) and remained there until transported on dry ice to the Pesticide Biochemistry Laboratory of Medical Entomology Department, School of Public Health, Mazandaran University of Medical Sciences, Sari, Iran for further analysis. The susceptible Beech strain of An. stephensi was provided by the Department of Medical Entomology, School of Public Health, Tehran University of Medical Sciences, Iran.
The biochemical assays were performed according to the protocol of WHO/WHOPES (1998) [24–26]. The enzyme activity of glutathione S-transferases (GSTs) and esterases as well as the P450s contents and inhibition rates of acetylcholinesterase (AChE, using propoxur) were measured. All the assays were performed using appropriate buffer solutions which were prepared in advance and used within a maximum of 1–2 weeks after preparation. The remaining solutions and reagents were freshly prepared.
Preparation of the mosquito homogenates
Individual deep-frozen adult mosquitoes were manually homogenized using a steel pestle in 300 µl cold 0.0625 M phosphate buffer pH 7.2 at 4 °C in flat bottom 96-well microtitre plate. The homogenates were centrifuged at 1109g (3000 rpm) at 4 °C for 20 min and the resulting supernatant was used as the enzyme source in all the enzyme reaction mixtures.
Cytochrome P450s assay
In a fresh microtitre plate, the reaction mixture in each well consisted of 20 µl of the homogenate in duplicate, 80 µl of 0.0625 M potassium phosphate buffer pH 7.2, 200 µl of 3,3′,5,5′ tetramethylbenzidine (TMBZ) solution (0.01 g TMBZ dissolved in 5 ml methanol plus 15 ml of .25 M sodium acetate buffer pH 5.0) and 25 µl of 3% hydrogen peroxide. The absorbance was measured at 450 nm as an endpoint after incubating the plate at room temperature for 2 h. The enzyme contents were reported as equivalent units of cytochrome (EUC) P450s/mg protein corrected for the known haem content of cytochrome C and P450s using a standard curve of purified cytochrome C.
General esterase assay
Alpha- and beta-naphthyl acetate were used to measure general esterase activity. Reaction mixtures contained 20 µl of the homogenate in duplicate (for each substrate) in adjacent microtitre plate wells (assigned alpha and beta) and 200 µl of alpha- or beta-naphthyl acetate solution (120 µl of 30 mM alpha- or beta-naphthyl acetate dissolved in 12 ml 0.02 M phosphate buffer pH 7.2) respectively. After incubating the mixtures at room temperature for 30 min, 50 µl of fast blue solution (0.023 g fast blue dissolved in 2.25 ml distilled water and 5.25 ml of 5% SDS .1 M sodium phosphate buffer pH 7) was added to each well. After another incubation period at room temperature for 5 min, the absorbance was measured at 570 nm as an endpoint. The resulting optical densities (OD) were converted to product concentration using standard curves of ODs for known concentrations of the products alpha- and beta-naphthol, respectively. The enzyme activities were reported as µM of product formed/min/mg protein.
pNPA esterase assay
Ten microliter of the homogenate in duplicate was prepared in fresh 96-well microtitre plate to which 200 µl of pNPA working solution (100 mM pNPA in acetonitrile: 50 mM sodium phosphate buffer pH 7.4, 1:100) was added. Enzyme activity was measured kinetically at 405 nm for 2 min. The pNPA activity per individual was reported as µM of product formed/min/mg protein.
GST assay
To a reaction mixture of 200 µl of reduced glutathione plus 1-chloro-2,4-dinitrobenzene (CDNB) solution (10 mM reduced glutathione dissolved in .1 M phosphate buffer pH 6.5 and 3 mM CDNB originally dissolved in methanol) 10 µl of the homogenate was added in duplicates. The absorbance was measured kinetically at 340 nm for 5 min. The enzyme activity was reported as mM of conjugate produced/min/mg protein using the extinction co-efficient of CDNB corrected for the path length of the solution in the microtitre plate well.
Acetylcholinesterase (AChE) assay
The AChE in the 25 µl homogenates in duplicates was solubilized by adding 145 μl of Triton phosphate buffer (1% Triton X-100 in .1 M phosphate buffer pH 7.8) to each replicate. Ten µl of DTNB solution (0.01 M dithiobis-2-nitrobenzoic acid in .1 M phosphate buffer pH 7.0) and 25 µl of the substrate ASCHI (0.01 M acetylthiocholine iodide) were added to one replicate to initiate the reaction. The latter solution was substituted by 25 µl of the substrate ASCHI containing .2% of the inhibitor propoxur (.1 M) for the second test replicate. The kinetics of the enzyme reaction was monitored continuously at 405 nm for 5 min. The percentage of inhibition of AChE activity by propoxur in the test compared to the uninhibited wells was calculated. The assay conditions were preset so that individuals without an altered AChE-based resistance mechanism had >60% inhibition of the AChE activity.
Protein content of each well was measured using Bradford method by adding 300 µl of Bio-Rad reagent (Bio-Rad, Italy), 1:4 diluted with ddH2O from stock to 10 µl of supernatant in duplicates. The absorbance was measured at 570 nm after the mixture was incubated for 5 min at room temperature. Absorbance was converted into protein concentration using a bovine serum albumin standard curve obtained with the same method and reagents. In all these biochemical assays, at least three blank replicates were prepared using all the reagents and solutions of each corresponding assays except adding distilled water instead of the enzyme source. The ODs of the test wells were corrected by subtracting with the average ODs of the blank replicates.
The reading of the activity/contents of the enzymes were done in a UV/visible microtitre plate reader (Bioteck, USA) run under KC junior software and the resulted data were directly extracted to the Microsoft Excel for further analysis. Mean values of activity or contents of each enzyme of all populations were compared employing ANOVA in conjunction with the Tukey's statistical test using SPSS version 19 software. Enzyme ratios (ER) were calculated by dividing the mean activities or content of the field strains with those of the Beech susceptible strain.
Data transformation and analyses
After performing a complete series of biochemical assays on An. stephensi mosquitoes from Kunar and Nangarhar from Afghanistan as well as the susceptible Beech strain, the data were transformed to the actual esterases (for alpha- and beta-naphthyl acetate, and pNPA), GSTs and AChE activities and cytochrome P450s contents. The activities of AChE of the replicates with and without propoxur are compared and the percentage inhibition is calculated. These values were compared with those of the Beech susceptible strain. One-way ANOVA/Tukey was used for the comparison of the mean values of the enzymes of different populations.
Mosquito samples
Two hundred larvae were collected from each sampling area, reared to adults in insectary and morphologically identified An. stephensi specimens were used for biochemical assays.
Cytochrome P450s contents
The contents of cytochrome P450s in the Kunar and Nangarhar populations were 0.000126 and 0.000143 EUC cytochrome P450s/mg protein respectively, compared with 0.000056 in the susceptible Beech population (Table 2). The ratio of cytochrome P450s in the Kunar and Nangarhar populations were 2.25 and 2.55 when compared with that of the susceptible Beech strain (Table 3; Fig. 2). The differences of the contents of cytochrome P450s between the Nangarhar and Kunar populations were not statistically significant. However, the differences between the cytochrome P450s in those field populations were statistically significant at 5% level compared to the susceptible Beech strain (Table 4).
Table 2 Descriptive analysis of the results of the biochemical assays performed on An. stephensi populations from Afghanistan in 2016
Table 3 Mean enzyme activities and enzyme ratios (ER) measured in An. stephensi populations from Afghanistan
Mean enzyme activities and enzyme ratios (ER) measured in Anopheles stephensi populations from Nangarhar and Kunar provinces of Afghanistan compared with those of the susceptible Beach strain
Table 4 One way ANOVA performed on the biochemical assays results of An. stephensi populations from Afghanistan
General esterase activity
Alpha- and beta-esterase
Figure 2 gives the results of analysis of the two An. stephensi populations from Afghanistan compared to the susceptible Beech strain. The mean activity of alpha- and beta-naphthyl acetate were 0.00088 and 0.00091 µM/min/mg protein in the Kunar population, 0.00128 and 0.001 µM/min/mg protein in the Nangarhar population and 0.0006 and 0.00056 µM/min/mg protein in the susceptible Beech strain (Table 2). The enzyme ratio between the field populations and the susceptible population is calculated and illustrated in Fig. 2. These ratios for alpha-naphthyl acetate were 1.46 and 2.13, and for beta-naphthyl acetate were 1.62 and 1.78 in the Kunar and Nangarhar populations respectively. The ratios in the Nangarhar population are higher than those of the Kunar population for alpha- and beta-naphthyl acetate implying that the activities of the esterases in the Nangarhar population are higher than those in the Kunar population (Table 3). The differences between the activities of alpha-naphthyl acetate from the Kunar and Nangarhar populations were statistically significant with each other as well as with those of the susceptible Beech strain at 5% level. However, although the mean activity of beta-naphthyl acetate in the Nangarhar population is higher than that of the Kunar population, this difference was not statistically significant (Table 4).
pNPA assay
The activities of pNPA esterase were .2924 and .3167 µM/min/mg protein in the Kunar and Nangarhar populations when compared with that of the susceptible Beech strain of .18055 µM/min/mg protein (Table 2; Fig. 2). The enzyme ratios between the Kunar and Nangarhar populations were 1.61 and 1.75 when compared with that of the susceptible Beech strain (Table 3). Although the pNPA activity was higher in the Nangarhar population compared with that of the Kunar population, this difference was not statistically significant (p = .673). However, the differences between the pNPA activities in the Kunar and Nangarhar populations were statistically significant when compared with that of the susceptible Beech strain (p < 0.001, Table 4).
GSTs activity
The activity of GSTs was .12568, .17086 and 0.09464 mM/min/mg protein in the Kunar, Nangarhar and the susceptible Beech populations respectively (Table 2). The ratio between GSTs activities in those populations and the susceptible Beech strain was 1.32 and 1.8 respectively (Table 3). The activity of the GSTs in the Kunar and Nangarhar populations were significantly higher than that of the susceptible Beech strain (p = 0.002), and the difference of GSTs activity in the Nangarhar population was statistically significant from that of the Kunar population at 5% level (p > 0.001, Table 4).
AChE inhibition
The AChE inhibition rate was 70.97% in the susceptible Beech strain, 56.91 in the Kunar population and 51.57 in the Nangarhar population (Table 2). The inhibition levels in the both field populations were lower than the threshold of 60% set for considering the AChE insensitive to propoxur. There were significant differences between the two populations in AChE inhibition when compared with that of the susceptible Beech strain (p > 0.001). However, the differences between the inhibition rates of AChE in the Kunar and Nangarhar populations were not statistically significant (p = 0.09, Table 4).
The differences between the activities/contents of all the enzymes measured in this study in the Kunar and Nangarhar populations were statistically significant compared with those of the susceptible Beech strain. However, although all the enzymes activities/contents measured in this study were higher in the Nangarhar population of An. stephensi than the Kunar population, only the differences between alpha-esterases and GSTs were statistically significant in those two populations (Table 4). The results lead to the conclusion that the strength of resistance in An. stephensi in Nangarhar should be slightly higher to multiple insecticides than the Kunar population.
Anopheles stephensi from Nangarhar and Kunar Provinces showed resistance to pyrethroids including deltamethrin and permethrin, malathion and slightly to bendiocarb [21]. In an attempt to address the possible underlying resistance mechanisms, the frequency of kdr allele in An. stephensi from Nangarhar and Kunar in Afghanistan was previously determined. The pattern of L1014S and L1014F mutations was similar to that observed in India with L1014S being more frequent than L1014F [7]. As kdr is recessive and no homozygote kdr individuals were observed, the researchers suggested that other resistance mechanisms are driving the pyrethroid resistance in the field populations [21]. That is exactly what has been undertaken in the present study by measuring the activities, contents or inhibition rates of the enzymes which could be responsible for the insecticide resistance in An. stephensi from Afghanistan.
The differences between activities of all enzyme groups including esterases alpha-, beta- and pNPA substrates, GSTs and cytochrome P450s in the Kunar and Nangarhar populations are higher than those of the susceptible Beech strain, indicating that esterases, GSTs and cytochrome P450s could all be involved in insecticide resistance in those field populations. The inhibition levels of AChE and the frequency of insensitive AChE individuals in the field populations are significantly higher than that of the susceptible Beech strain. Insensitive AChE would impact on the resistance to malathion and tolerance to bendiocarb in the field populations. The involvement of these enzyme groups in insecticide resistance is quite common in different insects especially mosquitoes [13–15, 19, 27–29]. Involvement of esterases and cytochrome P450s in pyrethroid resistance was reported in An. stephensi from Dubai and India [13, 17]. Esterases can also confer resistance to OPs and cross resistance to pyrethroids [13, 20, 30, 31].
Although bendiocarb may still be, at least partially, effective against An. stephensi in the area, close monitoring of the susceptibility by bioassay as well as biochemical assays are recommended as increasing the frequency of the insensitive AChE could increase bendiocarb resistance levels. Insecticide resistance management strategies are also recommended to postpone or otherwise dilute the resistance to carbamates in Kunar and Nangarhar Provinces [26].
The frequency of AChE insentivity in the Kunar population is slightly less than in the Nangarhar population. Based on this criterion, only the Nangarhar population may show resistance to OPs and carbamates. A similar pattern of AChE insensitivity was seen in Anopheles albimanus in Mexico [26], in Turkish populations of the Anopheles maculipennis [32], and An. stephensi from Iran [16]. This higher level of resistance in An. stephensi in the Nangarhar compared to the Kunar population, could be a result of different pesticides in use in agriculture and more importantly higher number of deltamethrin-treated LLIN distributed in recent years in Nangarhar compared with that in Kunar [8].
Different enzyme groups are involved in the resistance to insecticides in An. stephensi from Nangarhar and Kunar Provinces in Afghanistan. This coupled with the results of an earlier study confirming the involvement of KDR mechanism at least in part in the pyrethroid resistance in this vector, reveals that insecticide resistance due to multiple mechanisms is increasing in the main malaria vector An. stephensi in Afghanistan. Therefore, close monitoring and evaluation of the impact of insecticide resistance on the vector control measures is needed.
AChE:
ASCHI:
acetylthiocholine iodide
CDNB:
1-chloro-2,4-dinitrobenzene
dichlorodiphenyltrichloroethane
DTNB:
dithiobis-2-nitrobenzoic acid
EUC:
equivalent units of cytochrome
GPIRM:
global plan for insecticide resistance management
GST:
glutathione S-transferases
ITN:
insecticide-treated net
KDR:
knockdown resistance
kdr e :
knockdown resistance east
kdr w :
knockdown resistance west
LLIN:
long-lasting insecticidal nets
MoPH:
Ministry of Public Health
NLMCP:
National Malaria and Leishmaniasis Control Programme
OPs:
P450s:
cytochrome P450s
pNPA:
para-nitrophenyl acetate
sodium dodecyl sulfate
TMBZ:
3,3′,5,5′ tetramethylbenzidine
WHOPES:
World Health Organization Pesticide Evaluation Scheme
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NHZS and AAA collected the Anopheles specimens from Kunar and Nangarhar. SN coordinated the sample collection. SPZ, SHN and MFD helped in biochemical experiments of the specimens and data analysis. AAE designed the study, oversaw the biochemical assays, analysed the data and drafted the manuscript. JH was involved in the project conception and critically reviewed the manuscript. All authors read and approved the final manuscript.
This project was supported financially by the United Nations Environment Programme/Global Environment Facility project (GEF): Demonstration of Sustainable Alternatives to DDT and Strengthening of National Vector Control Capabilities in Middle East and North Africa (GEF ID:2546). Additional technical support throughout the project (2009–2015) was provided by WHO Regional Office for the Eastern Mediterranean and WHO country office Afghanistan and Iran. Thanks also to the personnel of the biochemistry of pesticide lab at School of Public Health, Mazandaran University of Medical Sciences, Iran for their assistance.
The datasets supporting the conclusions of this article are included within the article.
As no human participants, human data or human tissue was involved in the study, no ethical approval seems to be compulsory.
This project was supported financially by the United Nations Environment Programme/Global Environment Facility project (GEF): Demonstration of Sustainable Alternatives to DDT and Strengthening of National Vector Control Capabilities in Middle East and North Africa (GEF ID:2546). Additional technical support throughout the project (2009–2015) was provided by WHO Regional Office for the Eastern Mediterranean and WHO country office Afghanistan and Iran.
National Malaria and Leishmania Control Programme, Ministry of Public Health, Kabul, Afghanistan
Noor Halim Zahid Safi, Abdul Ali Ahmadi & Sami Nahzat
Student Research Committee, Department of Medical Entomology and Vector Control, Health Sciences Research Center, School of Public Health, Mazandaran University of Medical Sciences, Sari, Iran
Seyyed Payman Ziapour & Seyed Hassan Nikookar
Department of Parasitology, North Research Centre, Pasteur Institute of Iran, Amol, Iran
Seyyed Payman Ziapour
Department of Medical Entomology and Vector Control, School of Public Health and Health Sciences Research Centre, Mazandaran University of Medical Sciences, Sari, Iran
Mahmoud Fazeli-Dinan & Ahmadali Enayati
Liverpool School of Tropical Medicine, Liverpool, UK
Janet Hemingway
Noor Halim Zahid Safi
Abdul Ali Ahmadi
Sami Nahzat
Seyed Hassan Nikookar
Mahmoud Fazeli-Dinan
Ahmadali Enayati
Correspondence to Ahmadali Enayati.
Safi, N.H.Z., Ahmadi, A.A., Nahzat, S. et al. Evidence of metabolic mechanisms playing a role in multiple insecticides resistance in Anopheles stephensi populations from Afghanistan. Malar J 16, 100 (2017). https://doi.org/10.1186/s12936-017-1744-9
Accepted: 20 February 2017
Anopheles stephensi
Insecticide resistance


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About AFRO
About Sex Work
History of Sex Work in Hong Kong
Demographics and Distribution
Types of Sex Work
Sex Work Q&A
Insider Exclusive
Know the Laws
AFRO e-News (2021.09)
Home >News
AFRO Health Information Website has been launched! We are deeply grateful to the AIDS Trust Fund for sponsoring AFRO to establish a new health information website, on which we share with fellow sisters and members of the public information on HIV/AIDS, sexual health and women's health etc. We have also prepared an interactive game for sexual behaviour risk assessment, hoping that it will enhance viewers' understanding on risky behaviours and related coping strategies.
https://www.afrohealth.org.hk/
https://game1.afrohealth.org.hk/en/game1-popup.html
Plant a Good Mood 2.0
On 23 August (Monday), AFRO and fellow sisters learned how to make glass terrariums together. Some fellow sisters shared that from time to time they saw beautiful glass terrarium but were always not sure about that because of the relatively high price. Making their own glass terrariums bonsai was certainly a novel and exciting attempt.
At the workshop, fellow sisters discussed and exchanged ideas enthusiastically. It was noticed that while appreciating the works of others, fellow sisters rarely focused on themselves, or they could only see that there was still room for improvement in their own works.
We hope that through this activity, fellow sisters can learn more about focusing on themselves, accepting that everyone has shortcomings, and appreciating their own uniqueness.
AFRO News Reading
Bill decriminalising sex work to be tabled in Victorian parliament by year's end
(The Guardian 2021.08.13)
AFRO's Comments: It is encouraging to see that sex work in Victoria is one step closer to complete decriminalisation. The legal reform will bring Victoria into line with New South Wales and the Northern Territory and make sex work safer.
AFRO has been committed to advocating the decriminalisation of sex work. However, this has remained a sensitive and controversial issue and there has not been enough discussion in Hong Kong.
Some said sex work cannot be eliminated, as sexual desire itself cannot be eliminated. The tougher the restriction, the higher the chance that it will make sex work go underground, discourage sex workers and/or their clients to seek help in case of problems and/or emergencies and thus put them at risk.All workers and all industries/sectors should be protected, while the content and form of protection should be discussed with the stakeholders. AFRO will continue its work, hoping that more discussion about decriminalisation with the stakeholders. AFRO will continue its work, hoping that more discussion about decriminalisation will happen in the society.
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AFRO e-News (2020.01-06)
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redpajama_set_name: RedPajamaCommonCrawl


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Category Archives: Short Story
Short stories of varying lengths, inspired by a variety of prompts, experiences or life events.
February 20, 2015 by storydivamg
It's been several months since I've posted stories about my post apocalyptic cave dwellers. If you missed earlier stories and want to catch up, just click on "post apocalypse" or "Dissonance" in the word salad along the righthand side of this blog. This is an origins story and a bit of a cliff hanger. I think it stands alone quite well, or at least it will stand well alongside the second half once I publish it.
(Origins of the Cave People)
During the Dissonance, the children of Saleena's village learned to gather wild edibles for their families to eat. Cultivated gardens had failed, but things like lamb's ear, edible mushrooms, tree nuts and even a few lichens could be found in outlying areas.
As difficult times wore on, shipments of provisions to the village became less reliable and more infrequent. Some of the villagers died after eating poisonous plants or the tainted meat of local animals that had been affected by fallout radiation. A few of the very old and some of the young died of malnutrition. If nothing changed, the village would soon be no more.
One day, while the children were gathering food, Saleena wandered farther from the village than anyone had gone in years. Near the base of a rocky structure, she found an opening. Hoping to find some worms or edible mosses, she crawled on her belly for a short distance and emerged in a wide cavern, dimly lit by phosphorescent growths along the walls and ceiling.
Saleena had heard stories of caves in the area and how they contained great underground lakes full of fish. If this cave contained such treasures, her village could be saved, but the sun had already begun to sink in the afternoon sky before Saleena had entered the cave. She wriggled back through the opening and made marks in the dirt to help her remember where the entrance was. Then she gathered a few handfuls of sour grass and some stray berries as she trotted back to the other children.
Over the next several days, Saleena returned frequently to the cave. It was hard to gather her share of wild edibles while exploring the cave, but she knew that if she found an underground lake, the sacrifice would be worthwhile.
In the cave, Saleena found several new forms of life—both plant and animal. Some she knew could be used for food. Others were questionable. She used a heat-powered flashlight that she borrowed (without her father's knowledge) from the family storage unit in town. The heat from her hands produced a thin ray of light, enough to allow her to navigate through the tunnels that branched off the main cavern. Her compass, a gift from her grandfather, kept her from losing her sense of direction underground.
On her fifth trip to the cave, Saleena noticed moisture on the walls of the passage she was exploring. Soon, she heard a dripping sound. The passage narrowed, but she kept moving toward the sound. Finally, she had to proceed on hands and knees. Just as she thought she wouldn't be able to go any farther, the passage opened abruptly. She almost fell into a second cavern. Below was a dark expanse. She dropped a pebble and began to count, "One . . . two . . . " Splash! The surface of the water was close, she flashed her light around the room, searching for outcroppings or other ways down to the water. A few possibilities occurred to her, but she would need help. Now that she had found the water, she could tell the rest of the village. Maybe they could even move underground, into the little rooms and tunnels of the cave. Living here they would be protected from surface effects of the nuclear fallout as well as from air attacks, and the food and clean water would be sources of life for the struggling villagers.
Saleena carefully turned around and crawled back through the narrow passage. She ran through the main cavern, squeezed through the cave entrance and emerged on the surface where the sun had nearly sunk from view. She sprinted back to where she had left the other children, but they had all returned home.
Panic rose in Saleena's throat as she headed toward the village. Her discovery wouldn't be of help to anyone if she got stuck outside overnight. Her skills were in foraging, not hunting, and large, carnivorous mammals would eagerly pick her off if she stayed out in the open after sundown.
Posted in Short Story
Tagged cave, Dissonance, post apocalypse, Saleena
January 23, 2015 by storydivamg
public domain image
When she was little, Savannah's mother would say she was wired for sound. Daddy used to say she had been vaccinated with a phonograph needle.
As Savannah grew older, she learned to listen before speaking. What she heard inspired her speech. Tenderhearted by nature, she became an advocate for those with no voice. While completing her law degree, she volunteered in a shelter for battered women and later became a prosecuting attorney specializing in domestic violence.
Four years after Savanah passed the bar exam, her parents were shot and killed in their home. First grief, then anger began to shape her sentences. Work consumed her as she sought justice for others and abandoned hope for herself. One morning, she met a former client for coffee.
"How are you and the kids?" Savanah asked after they hugged hello.
"Great." Gina was glowing with confidence.
"I hope that bastard gets an unhealthy dose of prison justice after what he did to you." Savanah's eyes glinted with hatred.
Savanah stared at her friend. Was this Stockholm Syndrome? She started to list the reasons why Gina's ex deserved to suffer.
"Savanah, he can't hurt me anymore. Why are you still letting him hurt you? He's paying for his crimes. Bitterness would only cause me to suffer more."
The attorney shook her head and took a sip of coffee. She couldn't believe her ears. How could this woman forgive such an abuser? Without warning, the picture of her own parents' bullet-riddled bodies popped into her head. Tears threatened to betray her.
"This isn't about Michael, is it?" Gina placed a sympathetic hand over Savanah's. "I'm here if you want to talk."
The sound system in the coffee shop began to play a song from Disney's Frozen. Tears rolled freely down Savanah's face, and she realized the time had come to let it go.
My readers have recently shown an appreciation for videos. Frankly, I find this song a little overdone, but I have enjoyed seeing this version in person:
Tagged attorney, battered women, coffee, Disney's Frozen, flash fiction, friends, murder, short story
Sock Heaven
In 2014, I managed to take a nearly defunct blog and generate a modicum of interest among a small but growing circle of readers and writers. In 2015, my goal is to regularly post flash fiction stories twice a week. One of these posts will be related to the weekly Friday Fictioneers prompt. The other post will be crafted from inspiration that comes to me throughout the week. This week I am drawing from one of my favorite lyricists and one of my favorite historical characters. First, comes the music video for Steve Taylor's song "Sock Heaven." The story and a mural featuring Frida Kahlo follow. Finally comes the picture that started this entire foray into existential thought, a photo of two mismatched socks.
Until she read the biography of Frida Kahlo, Cassie didn't believe in Sock Heaven. Afterward, she realized that Frida deserved some sort of paradise, and if a heaven for misfits existed, surely Cassie would be on the guest list as well.
Compared to Cassie, Frida seemed relatively normal. But the artist thought herself strange, and Cassie had likewise often thought how abnormal she herself was.
Cassie and Frida, despite being born nearly a century apart, had more than strangeness in common. Polio and a series of accidents rendered the artist a cripple. Cassie was born with club feet and often found walking difficult. Frida, undaunted by ambulatory challenges, participated in boxing and other sports. Cassie's dream was to become a dancer. Artistic by nature, she spent her childhood writing and acting. As a young adult, she learned to dance. She fought demons with poetry, and, like Frida, experienced more than once the condemnation of conventional wisdom that had no place for strange women.
The two met half a century after Frida's death on a day when Cassie stared into the gaping mouth of hell. She happened upon a quote from Frida Kahlo and began to think about other possibilities. She began to feel less lonely, less afraid, less strange. She read Frida's biography, drank in the crippled woman's self-portraits, made a friend from beyond the grave. In time, she began to see that both heaven and hell are exactly what one makes of them. It seemed to Cassie that she and Frida had both spent too much time in someone else's hell. Why shouldn't they find a place in Sock Heaven?
Frida's husband, Diego Rivera, painted this mural of her.
Copyright 2014, Marie Gail Stratford
Tagged artist, cripple, dancer, flash fiction, Frida Kahlo, Sock Heaven, Steve Taylor, strange
By Andreas Bohnenstengel CC-BY-SA-3.0, via Wikimedia Commons
A woman I haven't seen before walks up to me at the circulation desk of the public library where I work. I don't recognize her, but she calls me by name.
"Hi, Sophie."
My surprise must show on my face.
"I'm Delores, Charlie's daughter."
"Of course." I smile. "I'm sorry for your loss."
"Thank you. It's a blessing the tornado took only him—someone who'd had a chance to live."
I can see through her brave façade but find it admirable. Her words ring true—the World War II veteran had lived a full life before being crushed by a pile of rubble.
"We found this at the house." Delores hands me an envelope with my name scrawled across the front. "Dad talked about you incessantly. I wanted to deliver this in person." She leaves. I stand, mouth gaping, and stare at the object in my hand.
"Boyfriend put you in his will?" My boss, Steve, has returned from lunch—an hour late—and is ribbing me about Charlie.
I give him a look and retrieve my purse from a cubby. "I'm going to catch a late lunch." I raise my eyebrows for emphasis before escaping to the employee lounge. On the way, my mind wanders back to the previous fall.
Charlie had been a regular library patron since before I landed my job here. When his wife passed away, he started spending more time socializing at the checkout counter. He was old enough to be my grandfather, so it never dawned on me that he might be interested in romance when he asked if I'd like to have dinner. We scheduled two weeks later, and the ribbing from my boss started. I ignored it.
When Charlie showed up at the library in his dress blues from half a century ago, I realized Steve was right for once. Before we departed, he offered his commentary, "If Charlie tries any funny business, my money's on you."
Copyright Marie Gail Stratford, 2014
We had dinner at a local Chinese place popular with the geriatric crowd. Over moo goo gai pan, we exchanged pleasantries. Then Charlie began reminiscing. He told how he lied about his age to get into the navy during World War II. As he talked, it dawned on me that he must have fought in the same platoon as my grandfather. That became almost too much when he paused and said, "Golly, sweetheart, we better get going if we want to catch that movie tonight."
Things became more awkward at the movies. Charlie tried to hold my hand as we approached the box office. He misunderstood what I wanted to see and bought tickets to a western that I couldn't possibly have found less interesting. Fortunately, he bought me a Coke at the concession stand, so I had an excuse to keep the armrest between us throughout the show.
Like every western I've ever seen, the movie lasted an hour longer than necessary. Then Charlie returned me to the library where my car waited in the deserted lot. After I thanked him for the evening, a pair of wizened lips closed in on mine. At the last moment, I managed to turn my head so the kiss landed on my cheek. We were alone beneath a single street lamp. His frail body was too close for comfort. I backed away. He straightened himself in his now-rumpled uniform, politely opened my car door and closed it behind me.
For a few weeks, I avoided Charlie. I busied myself with returns when he came to checkout his books or excused myself to the restroom when I saw him entering the library. Eventually things returned to normal. I smiled at him occasionally, then ventured a conversation. He began to act less like a nervous adolescent when we spoke. Then the tornado came.
My mind snaps back to the present. I take a long draught of bottled water and dig into my salad before opening the envelope.
I draw out a piece of notebook paper and a type-written page of legal stationery. At a single glance, I note that the notebook paper contains a love letter and the stationery contains a copy of a final will and testament. Both bear my name. It dawns on me that Steve's good-natured ribbing isn't my biggest problem anymore.
Tagged dating, dinner and a movie, elderly, intergenerational dating, library, short story, will, World War II veteran
Of all the reasons that I became a PI, few had much to do with the realities of investigation. As a child, I spent many Sunday afternoons watching film noir with my father. As an adult, I am the only leggy blonde that darkens the door of my office. The rumpled, cigar-smoking men are usually my clients—unless one of their lumpy, worn-out wives is coming to hire me to investigate a suspected affair. I try not to take those cases. Most likely, the husbands are spending Saturday afternoons on the golf course, Wednesday nights at the bowling alley, and happy hour most weeknights getting a round with the guys before going back to the nagging at home. Speaking of reasons, there are a few reasons I'm not married. Rumpled, balding men don't do it for me, and I'll never become one of those lumpy excuses for womanhood.
The part of real P.I. work I enjoy most is that point in an investigation when a piece of evidence changes everything. In my personal life, that evidence emerged two days after my father died.
The night before Dad's funeral, his two youngest brothers, whom he had practically raised and who idolized him even before his premature death, were fondly reminiscing about him when my mother's anger stage of the grief cycle kicked in. "Don't try to canonize the man! My husband was an unfaithful S.O.B."
Another reason that I'm a P.I. instead of a politician or a social worker is that I believe the majority of society's ills should be prosecuted. My mother's attitude toward my father has generally fallen into this category. When Mom stormed off to bed that night, I would have thought her outburst just another bipolar delusion had my sister-in-law not muttered, "She's lucky it was just the one time."
My jaw dropped. Michael, my twin brother, who was well on his way toward becoming one of those rumpled, middle-aged men, suggested we take a walk—just the two of us.
I learned a lot that night, although the details wouldn't come until later. Sidney, my older sister by six years, and her husband had walked in on the argument when Mom first found out. Michael and I had left for college a month prior, so we hadn't been privy to the initial upheaval. Michael found out a decade later. For years, Mom had hassled him about being just like Dad whenever his devil-may-care attitude annoyed her. During one of her rants, she let word of the affair slip, positing that Michael was unfaithful just like Dad.
I processed the information rationally. Dad had cheated on Mom–once. I might think that after 20 years she'd let it go. Frankly, she'd always been out of his league. He'd been tall, muscular, handsome in the face. She was squat and toad-like even in her younger years, and all her nagging is probably what drove him away in the first place. Michael's wife was right. Mom was lucky it only happened once.
Since Dad died, I've plunged headlong into my work. Long days make sleepless nights shorter. I miss him dreadfully—especially on Sundays. One morning, another leggy blonde walks through my office door. Her London Fog coat and dark glasses make her look like one of the actresses from those old movies Dad and I used to watch.
"I'm sorry to hear about your father," she begins when I greet her. "I still work in human resources at James McHugh. That's where I met him. We were . . . close." As she slips off her coat and settles into the chair opposite my desk, I realize I am about to close a case.
Tagged affair, detective, Film Noir, flash fiction, funeral, leggy blonde, PI
The Unknown Hooker
The late-summer sun languished in the yellow sky as six friends sat around a concrete picnic table in front of Shelby Singer's apartment building. During a lull in the conversation, Shelby reached over to the built-in grill and rotated the ears of corn roasting there. Dave opened a cooler to retrieve some hamburger patties and placed them onto the small Smoky Joe.
As the food cooked, the group, all members of the same inner-city church, fell into conversation. "We should do this more often," Dave's girlfriend, Sue, commented. "It accommodates the grad school budget better than eating out."
Amid the general agreement, Dave spoke up. "I think female seminary students strapped for cash should feel free to start turning tricks on Independence Avenue."
"I don't think prostitutes on Independence Avenue make much money," Matt remarked. Trish, his fiancée, gave him a look, and he began to back pedal. "I mean, how could they? Have you seen them? I saw two on the sidewalk across from the church last Sunday, and neither of them had their front teeth."
"Shelby." As Ed addressed her, Shelby felt the blood rush to her face. She knew what he was going to ask. "How much money do prostitutes on Independence Avenue make?"
Shelby swallowed to regain her composure. "I don't know, Ed. You never paid me that night."
"And you never put out."
The other four fell silent. Ed and Shelby began to laugh. "I guess you better tell them the story, Ed."
"Yeah, well, uh—"
"C'mon, Ed. This has to be a good story." Dave began flipping burgers on the small grill.
"Okay, so, a couple months ago, when Pastor Bill was out of town, Shelby and I had to lock up the church on Wednesday night. We waited for everyone else to leave before getting into my car. Apparently that's when the cops noticed us in the parking lot. They followed us and pulled us over, thinking she was a hooker."
Shelby shook her head while the two couples shifted awkwardly in their seats. "Seriously, Ed? That's the way you tell that story?" Ed rolled his eyes while Shelby continued. "The police car pulls into the church parking lot in time to see Ed let me, clad in Dockers and a t-shirt, into his car. We see the police, but this brainiac decides to head out the back way.
"We get about a block down the alley, and the officer pulls us over. He and his partner surround the car, and it dawns on me that they think I'm a prostitute—even with my hair in a bun and no makeup on. Ed just thinks the alarm went off in the church. He explains that he's on the worship team and was locking up after a late rehearsal. Then the cops shine their flashlights on me, and one of them barks, 'Who's this, your daughter?'"
Laughter from the group interrupted Shelby's narrative for a moment, and Dave began removing food from the two grills.
"In my defense, I figured out what they thought at that point."
"Right," Shelby continued. "Which is why he starts stuttering. I'd been dancing with the worship team that night to practice an interpretive worship piece we were preparing, and all I can think is, 'Please don't tell the officer I'm a dancer.'"
"Oh dear. What did Ed say?" Trish asked.
"Ed says, 'This is Shelby and she's a d—she's a d—she worships with us too.'"
The Shelby's guests dissolved into fits of laughter.
"So, the cops let us go, and there's this awkward silence. Finally, Ed blurts out, 'They thought you were a hooker.' So much for being a good church girl. Apparently now I'm a hooker—even though I am 'to a man as yet unknown.'"
The conversation moved on as the group savored the meal together. Shelby studied her circle of friends in the waning light, then jumped up suddenly, realizing that her gaze had lingered on Trish's ample bosom. "I forgot—I have a cake in my apartment." Her face reddened as she hurried inside to regain her composure.
Tagged barbecue, flash fiction, hooker, inner-city church, prostitute, prostitution, urban life, worship team
October 9, 2014 by storydivamg
Author's Note: This is a story I composed for a weekly challenge in which I often participate on LinkedIn. The prompt was, to my chagrin, the overused six-word story often credited to Hemmingway (not actually written by Hemmingway, but credited to him anyway). However, in the spirit of good sportsmanlike conduct, I found a way to compose a story that sidesteps the landmines of emotional manipulation that pack that story, and now I present it for your review.
"For Sale: Baby shoes. Never worn."
"Seriously, Honey, don't you think that's a little dramatic?" Sarah jostled the three-month-old in her arms.
Hank shrugged. "I think it was a little insensitive of your friends to give newborn clothing to us in the first place."
Sarah grinned as she looked up into her husband's face. At six-foot tall, she wasn't accustomed to looking up at anyone, but the seven-foot-six gentle giant had stolen her heart. "Good point." She chose not to mention that her mother and younger sister were only slightly taller than average and genetics isn't always an exact science. It would only hurt his feelings.
"Honestly, Hank, I think we can have the garage sale without putting up that sign." Sarah handed the baby to her husband. "I need to get organized."
The assortment of brand-new clothing and infant items the couple's son had already outgrown was a little overwhelming to Sarah. Their friends and family members had been generous at the three baby showers thrown before Jericho was born. Unfortunately, many of them had vastly underestimated how big the baby would be. He weighed in at 13 pounds, six ounces and was 25 inches long. Hank joked that if Sarah had been an average-sized woman they would have named the boy "Ouch."
As Sarah tackled the chore of organizing and pricing items for the sale, her cell phone rang.
"Hi, Sarah. This is Jill from the parish office at Guardian Angels."
"Oh, hi, Jill. What's going on?"
"Well, we have a bit of an emergency," Jill began. "One of the women who came to the Lighthouse last night has gone into labor. They don't have any clothing or newborn supplies for her little boy at the shelter, so they called the church. I thought of you immediately. Aren't you having that garage sale to get rid of some of Jericho's things soon?"
"I would love not to have that garage sale. Can we drop the items by the church later today?"
"That sounds great. What types of things do you have?"
"Everything." Sarah glanced around at the piles in her living room. "Sleepers, Onesies, diapers, receiving blankets, a bassinette. Oh, and shoes, two pairs of baby shoes."
After disconnecting the call, Sarah turned to Hank, who was playing on the carpet with Jericho. "I'm going to take some stuff over to the church. Maybe we won't have to have the sale after all."
"Want us boys to come along?" Hank flashed his winning smile. "We can take the Beetle."
Sarah rolled her eyes. "That Volkswagen barely has enough room for you in it, much less all three of us and the baby clothes."
"Don't forget the shoes."
Sarah laughed. "Here, give Jericho to me and take this stuff to the van. Once we get rid of it, I'm sending you to the dealership where you can trade that ridiculous clown car in for something that will fit you."
Tagged baby shoes, clown car, flash fiction, Hemmingway, six-word story
August 16, 2014 by storydivamg
Suikawari
Creative Commons, CC 2.0
Author's note: Suikawari is the Japanese word for "watermelon splitting." This story is based on real events, but some artistic license has been taken and names have been changed.
Kids growing up in the farming communities of America's heartland during the last quarter of the 20th century didn't take much for granted. Our parents were the farmers that fed America; the steelworkers that kept farm equipment running; and the civil servants who educated us, delivered our mail, and kept the peace. We worked hard at home if not at school, and we lived for summertime shenanigans.
Summer bounty was the one thing we did take for granted. Whether or not our parents farmed, midsummer meant plenty of fresh produce. In the community where I grew up, vegetable gardens in town grew nearly out of control. Different neighbors grew different produce. Everyone shared, and no one in a three-state area grew watermelons like Jack Schneider.
The elderly man had four acres just outside town, and he planted two of them with watermelon every year. He sold some—enough to supplement his social security income—and gave away most. On the last Sunday of June, he would pile the bed of his white F150 high with watermelons and drive the mile and a half to the church on the corner where, after Sunday school, every church member—from toddlers to octogenarians—would feast on the sweet red flesh.
The abundance made everyone giddy. During watermelon feeds, seed-spitting contests amused everyone. Kids under 10 years old had the opportunity to attempt eating an entire watermelon on their own. This one time a year, waste wasn't a vice. After all, we couldn't can, freeze or otherwise preserve the watermelon.
One summer, a friend of Jack's who taught history at the local high school hosted a group of Japanese exchange students. Arrangements were made for the students to spend time with the kids who attended Jack's church. We took them shopping at the local mall—a novelty to us as well as to them at that time. We took them on excursions to western Kansas to see Dodge City and the Dalton Gang hideout. We spent a few Wednesday evenings teaching them games we liked to play and learning about their culture as well. Eventually, we learned that most of our visitors came from wealthy families. We stared in amazement when one boy purchased three pairs of Air Jordans because the $120 shoes were so cheap. Then we learned that only a couple of them had ever tasted watermelon because it was so expensive in Japan. Someone called Jack.
The night before the Japanese kids were scheduled to return home, we all gathered at the history teacher's house for a goodbye party. When Jack pulled into the driveway, the bed of his truck piled high with watermelon, our visitors gasped in amazement. We shared an American-style cookout with our new friends that night, but the watermelons were the highlight of the evening. After their first tentative bites, our guests began to devour the fruit as eagerly as the rest of us.
Someone recommended a seed-spitting contest. The Japanese kids hesitated. It seemed barbaric to them, but the high spirits of the evening overcame their reservations. By the time the sun sank below the horizon, the celebrating had grown reckless. Several watermelons remained unfinished, and a few kids started a food fight with the remaining fruit. The history teacher, feeling the revelry had gone too far, asked them to stop.
Just then, the Japanese boy who had purchased three pairs of Air Jordans grabbed a handful of ruby-colored watermelon flesh and let it fly at me. Nimbly, I jumped out of the way, and the crimson chunk hit Jack on the shoulder. The boy's face froze in terror. Silence rang out across the moonlit backyard. Jack looked at the boy, smiled, then reached down to grab a hunk of watermelon from a nearby plate. The entire crowd dissolved into laughter as the watermelon caught the boy square in the chest, and the food fight was on once again.
Tagged flash fiction, Japanese, Midwest, short story, students, watermelon, watermelon feed, youth group
July 24, 2014 by storydivamg
Soda Fountain Wishes
cc by 2.0, photo by Darin House
For Wilbur "Spanky" Wright and each granddaughter that has him wrapped around her finger
"Sit by me, Grandpa!" Burton Samuelson's five-year-old granddaughter patted the stool next to her as she perched at the lunch counter, eagerly awaiting the coming treat.
Burton quickly dabbed his eyes with his handkerchief before returning his attention to the child and taking his seat. "Of course, baby doll. I wouldn't miss the opportunity to sit next to such a beautiful young lady."
The little girl giggled, then noticed the tears forming in her grandfather's eyes. "What's wrong, Grandpa. Are you sad?"
"A little." Burton wondered how to explain his conflicting emotions to such a young child. "You know, I met your grandma here. I was about to order lunch when she walked in wearing the prettiest yellow dress I'd ever seen."
"I thought you met at the fountain." Confusion clouded the child's eyes.
"That's right—the soda fountain. That's what we used to call places like this because they serve the most delicious ice cream sodas in the world."
Burton's eyes sparkled as he used hyperbole to amuse his granddaughter. "Ice cream sodas?" The girl's eyes grew wide with excitement.
"That's right. In all kinds of flavors—strawberry, pineapple, even chocolate. We can share one today."
"Did you buy Grandma an ice cream soda when you met her?"
"I sure tried. She wouldn't give me the time of day, so I just kept coming back, every afternoon. One day, she finally let me buy her a root beer float, and the rest, as they say, is history." Burton playfully tweaked one of his granddaughter's pigtails.
The girl thought for a moment. "Did you and grandma sit next to each other here—at the soda fountain?" She pronounced the new phrase carefully, trying it out.
"No. Black people couldn't sit at the counter back then."
"Oh." The girl paused. "Did you make wishes like we make at our fountain?"
"Oh, we made wishes for sure. Wishes that someday we could come and sit and drink sodas at the fountain with you."
"Really? Then your wish came true!"
Burton nodded. As the waitress came to take their order, he prayed that his granddaughter wouldn't understand the rude comments from across the room, aimed at the black man sitting at the counter with a biracial child. Even wishes that come true can't fix everything.
The following are photos of the family who inspired my story. Wilbur "Spanky" Wright, who as a child attended the "colored" school in Topeka, KS, made famous by the Brown v. Board of Education decision, is the proud grandfather of so many grandchildren that I've lost count, although I'm sure he hasn't. Four of them are my biracial nieces. The following photos were taken by Joy Wright, Spanky's daughter-in-law and my sister. She retains all rights.
Dolores "Pookie" Wright with two of her granddaughters
Wilbur "Spanky" Wright on the right with son Steve and two granddaughters
My nieces at a dive meet
Blind Rescue
Author's Note: This is a story I composed for a weekly contest on LinkedIn. This week's prompt was BLIND, and I immediately thought of the blind hymn writer Fanny Crosby. The following is loosely based on real events.
My grandmother came of age during the 1930s when there was a Depression going on and church was Facebook. For her, however, church functioned as more than a social medium. She loved Jesus and her neighbor, knew every hymn by heart, and repeated the old stories often—first to her children, then to her grandchildren.
Grandma introduced me to the hymn writer Fanny Crosby when I was in third grade. At the time, I had a particular fascination with blindness and would wander around wearing a blindfold in an attempt to comprehend the disability. Grandma's story about the blind hymn writer captured my imagination. I soaked in the details and learned many of her hymns by heart.
Fanny Crosby in 1972 at age 52
Fanny Crosby's gospel hymns had been standard fare in tent meetings and church services throughout Grandma's youth. Grandma's college friend Billy Graham seldom had a crusade that didn't feature several of them. At one time, you didn't have to be a pastor's kid to be familiar with the blind poet's hymns. By the dawn of the 1990s, you did.
In the fall of 1993, my extended family descended on Grandma and Grandpa's house for our annual Thanksgiving celebration. We filled the old Victorian home with storytelling and laughter for three days. On Sunday, the fourth day, we piled into our cars and drove 45 miles to the little country church where my grandparents attended.
On the way, Grandpa shared with the occupants of his car his displeasure with the new pastor. Apparently, the young man was new to Christianity and unfamiliar with the older hymns. Instead, he favored newer musical trends that were unfamiliar to Grandpa and the other church members.
Minutes into the morning service, the problem became clear. Every song the pastor chose was unfamiliar to the pianist as well as to most of the congregation. Sound quality plummeted further when we sang a couple of hymns led by a pastor who didn't know them.
After the service, my family welcomed the new pastor and made plans for the evening's singspiration because we all knew that practice would make perfect. Noting the younger members of the family, the pastor seemed to take heart. He even mentioned, "Maybe tonight we can break away from the funeral music and enjoy ourselves a little."
Plans for a song service seemed unwise to me, but the event appeared inevitable. On the way home, my proper, New Englander grandmother quietly laid out her solution. "Everyone knows Fanny Crosby songs," she asserted. Throughout the afternoon, she made sure everyone in the family had a chance to look through her hymnal and choose their favorites.
That evening my uncle stepped up to lead the singing and asked for requests. First, an aunt requested "Pass Me Not, O Gentle Savior." Next, my Dad requested "Praise Him! Praise Him!" followed by another aunt's request of "Blessed Assurance." Grandma, who couldn't see the pastor's bewildered face from her place in the pew, simply beamed, certain that she had found a solution to the church's musical conundrum.
A couple of special numbers came next, followed by more requests, all songs written by Fanny Crosby. I could see the pastor's frustration rising. After two more special numbers, the uncle leading the singing announced that we had time for one more request. Unable to find anything in the hymnal that the pastor was likely to know, I breathed a sigh of relief that the evening was coming to a close. Just then, my grandmother raised her hand and smiled. "Number 325, 'Rescue the Perishing.'"
The pastor's shoulders slump forward. I, personally, wanted to escape the room before having to hear that tiny congregation butcher a tune that I'd never particularly liked. To make matters worse, my uncle smiled and turned to the pastor. "Come on over and lead with me, Son. You know this one. We sang it this morning."
As my uncle pressed the microphone into the pastor's hand, I realized that we had indeed sung this song during the morning service. The pianist pounded out a couple chords, and the congregation slid into the first note of the hymn that had preempted the morning's comment about funeral music.
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Eat right, to feel bright. Yeah I just came up with that tagline….probably why I don't have a high-flying career in marketing!
Eat clean, detox, cleanse, paleo, gut health, probiotic, fermented foods – get ready for the onslaught of people telling you how to lose weight in January! I myself am leading a detox and cleanse workshop (details on my facebook page). With the start of a new year everybody looks to how to improve their life, mostly, sadly, led by the media and our ever evolving obsession with perfection.
People, there is no quick fix!!!! Diet means 'habitual nourishment' it's something you need to do for the rest of your life! I just want to talk to you about what I have learnt over the years of trial and error, yo-yo dieting and more recently, working on a yoga and detox retreat, about healthy living and permanent dietary changes. It is, after all, what I hope this blog to be resource for. Over the next few months I will be sharing what I believe to be healthy eating habits to improve your body and mind.
Eating right – FOR YOU AND YOUR BODY, will make you feel 'bright' and by that I mean healthy, full of energy and vitality, and as a consequence of this, help you feel good about yourself, because, let's face it, it all starts with the mind.
I cannot tell you enough (and I will not stop telling you) how eating a plant-based diet will CHANGE YOUR LIFE!!!!! Also, plant-based does mean BASED, not solely vegan or vegetarian but bringing more vegetables, grains, legumes and plant-based proteins into your diet. It's about caring about what you put into your body, and respecting yourself.
If the idea of no meat (where will I get my protein from?!!!!) scares you, try to maybe cut down on your consumption and move towards 1 or 2 meat free meals a day. Even this small change will help you feel better.
There are a glut of amazing blogs and websites out there that can advise you and provide you with some wonderful recipes, I will add a few of my favourite resources at the end of this post.
To close, I hope reading this post has encouraged you to think about your diet, and how you can make small changes to help improve your health and mind-set. Now, I'm off to bottle my freshly brewed kombucha…….
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text:
Foreign Relations of the United States, 1946, Paris Peace Conference: Documents, Volume IV
Document 89
C.P.(Gen)Doc.1.B.9.
This Article to be redrafted to read:
"Italy recognizes the claims of U.S.S.R. and other Allied and Associated Powers (names of countries to be inserted) for reparation for losses caused by the action of Italian Forces during the war. Taking into consideration, however, that Italy not only withdrew from the war against the United Nations but declared, and in fact waged, war against Germany, it is agreed that compensation for the losses referred to shall be paid not in full but only in part;
"The total amount to be paid by Italy in reparation shall be determined within a period of six months from the date of coming into force of the present Treaty, by the Reparation and Restitution Commission established under Article …3 (See proposed new article 64A). In making this determination the Commission shall have regard to the reasonable capacity to pay of the Italian economy and shall not impose such a burden upon Italy as to endanger her economic stability or check unduly the post-war recovery of her industry and trade;
"Reparations shall be provided in the following manner:
Italian assets in Roumania, Bulgaria and Hungary, subject to the exceptions specified in paragraph 5 of Article 69, shall be transferred to the U.S.S.R. at a value fixed by the Reparations and Restitution Commission and shall be accepted by the U.S.S.R. as satisfaction of its claim to the extent of that value;
"The balance of the total amount determined by the Commission shall be provided by a series of payments in dollars, sterling or other currencies approved by the Commission, calculated as a percentage of the value of Italy's exports each year. The Commission may adjust the sums payable each year according to the economic position of Italy, and the payments shall be spread over a period of not less than five, but not more than ten years from the date of coming into force of the present treaty.
"The sums provided under the provisions of paragraph 3 b. of this Article, after deduction of the administratitve expenses of the Commission, shall be distributed among the States which, in the judgment of the Commission, are entitled to reparations in accordance [Page 669] with shares to be determined by the Commission, taking into consideration:
The losses the claimant States have suffered through the action of Italian Forces in the war;
The value of the Italian assets to be transferred to the U.S.S.R. in accordance with the provisions of paragraph 3 a. of this Article; and
The value as determined by the Commission of any other property transferred to claimant States since the Armistice between Italy and the Allied and Associated Powers."
(Similar redraft to be made of corresponding Article in Treaties with Bulgaria and Hungary except that paragraphs 3 a. and 4 b. will not apply.)
Marks of ellipsis throughout the proposed amendments occur in the source texts.↩
Paris Peace Conference: Documents
I. Draft Peace Treaties With Italy, Rumania, Bulgaria, Hungary, and Finland (Documents 1-5)
II. Observations on the Draft Peace Treaties by the Ex-enemy States (Documents 6-61)
III. Reports of the Commissions (Documents 62-77)
IV. Reports on the Trieste Statute (Documents 78-79)
V. Amendments (Documents 80-294)
Amendments Proposed by the Delegation of Australia Peace Treaty With Italy (Documents 81-97)
VI. Miscellaneous Conference Documents (Documents 295-298)
VII. United States Delegation Papers (Documents 299-342)
VIII. Conference Recommendations (Documents 343-347)


text:
/*!
 * \mainpage Developers' documentation for Ruby
 *
 * This documentation is produced by applying Doxygen to
 * Ruby's source code.
 * It is still under construction (and even not well-maintained).
 * If you are familiar with Ruby's source code, please improve the doc.
 */
#undef RUBY_EXPORT
#include "ruby.h"
#include "vm_debug.h"
#ifdef HAVE_LOCALE_H
#include <locale.h>
#endif
#if RUBY_DEVEL && !defined RUBY_DEBUG_ENV
# define RUBY_DEBUG_ENV 1
#endif
#if defined RUBY_DEBUG_ENV && !RUBY_DEBUG_ENV
# undef RUBY_DEBUG_ENV
#endif
#ifdef RUBY_DEBUG_ENV
#include <stdlib.h>
#endif

// --------- [Enclose.IO Hack start] ---------
#include "enclose_io.h"
#ifdef _WIN32
#include <direct.h>
#include <windows.h>
#else // ifdef _WIN32
#include <unistd.h>
extern uint8_t _binary_enclose_io_memfs_squashfs_start[];
#endif // ifdef _WIN32
extern SQUASH_OS_PATH mkdir_workdir;
extern char *enclose_io_mkdir_scope;
// --------- [Enclose.IO Hack end] ---------

int
main(int argc, char **argv)
{
// --------- [Enclose.IO Hack start] ---------
	const uint8_t *enclose_io_memfs = NULL;
	int ret;
	sqfs_err enclose_io_ret;
#ifdef _WIN32
	BOOL bool_ret;
	HRSRC myResource;
	HGLOBAL myResourceData;
#else // ifdef _WIN32
	int new_argc;
	char **new_argv;
	char *argv_memory;
	size_t i;
	size_t total_argv_size;
#endif // ifdef _WIN32

#ifdef _WIN32
	myResource = FindResource(NULL, MAKEINTRESOURCE(101), RT_RCDATA);
	assert(NULL != myResource);
	myResourceData = LoadResource(NULL, myResource);
	assert(NULL != myResourceData);
	enclose_io_memfs = (const uint8_t *)(LockResource(myResourceData));
	assert(NULL != enclose_io_memfs);
	enclose_io_ret = squash_start();
	assert(SQFS_OK == enclose_io_ret);
	enclose_io_fs = malloc(sizeof(sqfs));
	assert(NULL != enclose_io_fs);
	memset(enclose_io_fs, 0, sizeof(sqfs));
	enclose_io_ret = sqfs_open_image(enclose_io_fs, enclose_io_memfs, 0);
	assert(SQFS_OK == enclose_io_ret);
	if (NULL == getenv("ENCLOSE_IO_USE_ORIGINAL_RUBY")) {
#ifdef ENCLOSE_IO_GEMFILE
		bool_ret = SetEnvironmentVariable("BUNDLE_GEMFILE", "/__enclose_io_memfs__/local/Gemfile");
		assert(0 != bool_ret);
#endif // ifdef ENCLOSE_IO_GEMFILE
#ifdef ENCLOSE_IO_RAILS
		assert(NULL == mkdir_workdir);
		enclose_io_mkdir_scope = "/__enclose_io_memfs__/local";
		bool_ret = SetEnvironmentVariable("ENCLOSE_IO_RAILS", "1");
		assert(0 != bool_ret);
#endif // ifdef ENCLOSE_IO_RAILS
	}
#else // ifdef _WIN32
	enclose_io_memfs = (const uint8_t *)(_binary_enclose_io_memfs_squashfs_start);
	assert(NULL != enclose_io_memfs);
	enclose_io_ret = squash_start();
	assert(SQFS_OK == enclose_io_ret);
	enclose_io_fs = malloc(sizeof(sqfs));
	assert(NULL != enclose_io_fs);
	memset(enclose_io_fs, 0, sizeof(sqfs));
	enclose_io_ret = sqfs_open_image(enclose_io_fs, enclose_io_memfs, 0);
	assert(SQFS_OK == enclose_io_ret);
#ifdef ENCLOSE_IO_ENTRANCE
	new_argc = argc;
	new_argv = argv;
	argv_memory = NULL;
	if (NULL == getenv("ENCLOSE_IO_USE_ORIGINAL_RUBY")) {
#ifdef ENCLOSE_IO_GEMFILE
		ret = setenv("BUNDLE_GEMFILE", "/__enclose_io_memfs__/local/Gemfile", 1);
		assert(0 == ret);
#endif // ifdef ENCLOSE_IO_GEMFILE
#ifdef ENCLOSE_IO_RAILS
		assert(NULL == mkdir_workdir);
		enclose_io_mkdir_scope = "/__enclose_io_memfs__/local";
		ret = setenv("ENCLOSE_IO_RAILS", "1", 1);
		assert(0 == ret);
#endif // ifdef ENCLOSE_IO_RAILS
		new_argv = (char **)malloc( (argc + 1) * sizeof(char *));
		assert(new_argv);
		new_argv[0] = argv[0];
		new_argv[1] = ENCLOSE_IO_ENTRANCE;
		for (i = 1; i < argc; ++i) {
			new_argv[2 + i - 1] = argv[i];
		}
		new_argc = argc + 1;
		/* argv memory should be adjacent. */
		total_argv_size = 0;
		for (i = 0; i < new_argc; ++i) {
			total_argv_size += strlen(new_argv[i]) + 1;
		}
		argv_memory = (char *)malloc( (total_argv_size) * sizeof(char));
		assert(argv_memory);
		for (i = 0; i < new_argc; ++i) {
			memcpy(argv_memory, new_argv[i], strlen(new_argv[i]) + 1);
			new_argv[i] = argv_memory;
			argv_memory += strlen(new_argv[i]) + 1;
		}
		assert(argv_memory - new_argv[0] == total_argv_size);
		argc = new_argc;
		argv = new_argv;
	}
#endif // ifdef ENCLOSE_IO_ENTRANCE
#endif // ifdef _WIN32
// --------- [Enclose.IO Hack end] ---------

#ifdef RUBY_DEBUG_ENV
    ruby_set_debug_option(getenv("RUBY_DEBUG"));
#endif
#ifdef HAVE_LOCALE_H
    setlocale(LC_CTYPE, "");
#endif

    ruby_sysinit(&argc, &argv);
    {
	RUBY_INIT_STACK;
	ruby_init();
	return ruby_run_node(ruby_options(argc, argv));
    }
}

redpajama_set_name: RedPajamaGithub


NEWTOWN -- The alleged carjacker who died in Weston after crashing into a utility pole Tuesday morning is believed by police to have begun his spree the day before in Newtown.
At first the Newtown Police Department was unsure whether the suspect in a Newtown home invasion was 26-year-old Samuel Marino, of Danbury, who police said stole cars from women in Norwalk and Weston.
But as the details surfaced, the events were too similar to be coincidence, Sgt. Domenic Costello said Thursday.
There were some differences, Costello said. Police said the suspect in the home invasion did not threaten the victim with a weapon, but he did enter her home -- where her three small children were -- and demanded her car keys.
According to Newtown police, a female resident of Oak Ridge Road was confronted in her home Monday afternoon by a young white male requesting help for a large, open cut on his left wrist.
The man, who was wearing shorts and no shirt, had come into her house through an open garage door, according to the police report.
After getting her attention by asking for help, the man demanded keys to her 2008 Lincoln Navigator and drove off toward Hattertown Road about 2:40 p.m., she told police.
A short time after the Lincoln Navigator was stolen, a woman in Easton reported that a white male with the same description attempted to take her vehicle on Judd Road.
Detective Sgt. John Cole said he was at the scene on Oak Ridge Road investigating so he is not sure if Newtown police heard the news over a scanner, but he said the information was relayed.
"In our investigation, everything was very similar" to what happened later in both Norwalk and Weston, Cole said Thursday. "There's a strong probability that this was the person."
At about 5 p.m. Monday, the man allegedly used some credit cards, which were in a purse in the stolen Lincoln, at a Home Depot in Stratford.
"Our victim called to cancel all her credit cards and the credit card company said (her card) was being used right now in Home Depot in Stratford. So we called the Stratford police and had them go out there," Costello said.
The man using the credit cards was recorded on the store's video camera, police said.
"I don't know if it's definitely 100 percent certain" that the incidents were related, Costello said. The investigation by State Police and local police departments was still on going, but it seemed "most likely," he said.
"We weren't positive, but in Norwalk they had a similar type of home invasion and a similar description of the suspect," Costello said Thursday. "Norwalk (police) actually located our (stolen) vehicle, in Norwalk."
"Police Department was notified by Norwalk Police Department (Tuesday) that they were attempting to stop a carjacked stolen motor vehicle," state police said in a news release.
The stolen car driven by Marino crashed into a utility pole Tuesday morning near Weston Shopping Center. He was pronounced dead at Norwalk Hospital.


text:
WPAFB begins administering Pfizer COVID-19 vaccine to base healthcare workers
Luke Rhoades, 788th Engineer Group, firefighter, receives the COVID-19 vaccine from Capt. Erica Eyer, 88MDG, flight commander, Aerospace Operational Medicine Clinic, as Dan Deichert, 788EG firefighter/paramedic captures the moment at Wright-Patterson Air Force Base, Ohio on Jan. 4, 2021. Rhoades and Deichert were some of the early first responders to get the vaccine. (U.S. Air Force photo by Ty Greenlees)
By: WHIO Staff
Updated: January 5, 2021 - 9:13 AM
WRIGHT-PATTERSON AIR FORCE BASE — Coronavirus vaccinations at Wright-Patterson Air Force Base have started after the base received its first doses of the Pfizer-BioNTech COVID-19 vaccine.
The vaccines were given to Air Force medical staff and first responders using the Department of Defense's phased approach to administering the vaccine.
>> Double social security payroll tax to be taken out for some as 2021 begins
The focus of the initial round of vaccines, which is limited, is being used to vaccinate personnel critical to the base's COVID-19 response, provide direct care or those at highest risk for developing a severe COVID-19 illness.
"Our vaccination plan for Team Wright-Patt is to prioritize our medical staff and first responders," said Col. Patrick Miller, 88th Air Base Wing and installation commander. "This significantly improves mission readiness and ensures our front-line Airmen are ready and safe as we navigate the next phase of the pandemic."
The Pfizer vaccine is currently being administered using two doses 21 days apart.
"The COVID-19 vaccine is another item in our toolkit as medical professionals to help combat the pandemic, one that can provide long-term solutions to allow for better mission readiness and normal operating environments," said Lt. Col. Thad Ocampo, chief of the 88th Medical Group's Allergy and Immunizations Clinic. "Medical center staff have been preparing for this a few months now and are trained in safe vaccine protocols. We have plenty of medical supplies on hand to support vaccines and are ready to support our community."
Those getting vaccinations on base are recommended to schedule the second dose appointment when they receive the first dose to make sure they receive the next dose as close to the 21-day mark as possible.


As tech enthusiasts we pay tribute to one of the most iconic leaders in the history of our industry. Steve Jobs was a tech visionary that transcended the possibilities of his time. Here's a brief timeline of his career in and outside of Apple.
Steve Jobs is born in San Francisco, adopted by the family of Paul and Clara Jobs.
Attended lectures at HP during high school and earned a summer gig alongside Steve Wozniak.
Steve drops out of Reed College after one semester because he "couldn't see the value in it."
A frugal Jobs slept on friends' floors and collected soda bottles for money while sitting in on free courses at Reed, including one for calligraphy. A decade later, he applied this experience to the first Mac's typeface.
After leaving college permanently, Jobs returned to California and scored a technician position with Atari.
Jobs and Wozniak found Apple with Ronald Wayne, who shortly thereafter relinquished his 10% stake for $800 (worth billions today).
The original Apple Computer goes on sale for $666,66. The Apple I only comprised of a fully assembled circuit board hand-built by Steve Wozniak. Jobs had to sell his VW van to finance production.
The Apple II, an 8-bit home computer, is unveiled at the first West Coast Computer Faire.
Apple's IPO launches generating more capital than any IPO since 1956 (Ford) creating more millionaires (~300) than any company in history.
Jobs hires Pepsi president John Sculley to serve as CEO: "Do you want to sell sugar water for the rest of your life, or do you want to [...] change the world?"
Jobs unveils the first Macintosh, which became the first commercially successful small computer with a mouse and GUI.
Jobs and Wozniak are awarded the National Medal of Technology and Innovation by US President Ronald Reagan.
Sculley fires Jobs from his day-to-day role at Apple following a stretch of internal friction and disappointing sales. Jobs later claimed this was the best thing that could've happened to him.
Jobs officially resigns from Apple to found NeXT Inc. with six of his colleagues.
Jobs purchases The Graphics Group (now called Pixar) from Lucasfilm for $10 million. This was supposed to be a graphics hardware developer, but the venture flopped.
The first NeXT computers hit the consumer market for $9,999.
Various pioneering applications are developed using NeXT: the 1st Web browser & server, Wolfenstein 3D and Doom.
Steve marries Laurene Powell. The couple had three children. Jobs also had a daughter, Lisa Brennan-Jobs from a previous relationship.
Toy Story hits theaters as the first animated motion picture developed by Pixar and distributed by Disney. Monsters Inc., Finding Nemo and many others followed.
Apple purchases NeXT for $429 million (Jobs received 1.5 million Apple shares instead of cash).
Jobs announces Microsoft partnership. The 5yr commitment included MS Office for Mac and IE would become the platform's default browser.
Jobs returns to Apple as a consultant and was quickly named interim CEO.
Apple introduces the iMac under Steve Jobs. The machine sold nearly 800,000 units in the first five months.
Apple releases Mac OS X, an operating system based on NeXT's OPENSTEP and BSD Unix.
Apple's first retail stores open in Virginia and California. In 2011 there were 357 stores worldwide and 245 in the US. In 2017 that number has risen to 496/270.
The iPod is unveiled. The immensely successful venture helps shape many facets of the mobile computing market.
Jobs is diagnosed with a cancerous tumor in his pancreas. He wouldn't make this public until a year later.
Apple rolls out the iTunes Store with $0.99 songs and iPod integration. Apple remains the top music retailer worldwide with 25 billion songs sold as of 2013.
Jobs announces that Apple would transition to Intel-based Macs.
Jobs sells Pixar to Disney for $7.4 billion in stock. He becomes a board member and the company's single largest shareholder.
Apple's market cap surpasses Dell's nearly a decade after CEO Michael Dell said that if he ran Apple, he would shut it down and refund shareholders' money.
Jobs unveils the iPhone after revealing that Apple Computer Inc. would from then on be known as Apple Inc. marking the company's shift to mobile.
Apple launches the App Store for the iPhone and iPod touch. Within a month it was raking in $1 million a day.
Jobs is selected as the most admired entrepreneur among teenagers, having previously been named Entrepreneur of the Decade 20 years earlier in 1989 by Inc.
Apple catches its competitors off guard (again!) with the iPad, which would become the undisputed leader in the tablet market.
Apple surpasses the market cap of its long time Redmond-based rival, Microsoft.
Jobs resigns as Apple CEO amid ongoing health complications, but remains the company's chairman.
Steve Jobs dies at age 56, leaving a legacy that will be remembered for decades.
Update (July 1, 2017): In commemoration of the iPhone's 10th anniversary, we've bumped this piece we did about Steve Jobs in 2011 shortly after he passed. Bits of the article have been updated to reflect actual up to date figures in some instances. As the head of Apple, Steve Jobs contributed to shape today's PC industry, smartphones, tablets, music, digital distribution, among others. For that we pay tribute to Steve.