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The Medical Minute: The Disease Behind the Ice Bucket Challenge
September 5, 2014 12:30 am·
Author: exploreJefferson
The ice-bucket challenge that was all over social media a few weeks ago raised about $100 million for ALS, a devastating, fatal motor neuron disease. But how much do those who participated and donated really know about the condition, diagnosis and treatment?
"The challenge was important in terms of the awareness and money it has raised," said Dr. Zachary Simmons, professor of neurology and director of the Penn State Hershey ALS Center. "In my experience most individuals understand it is a bad disease, but they don't know much more than that."
Amyotrophic lateral sclerosis, also known as Lou Gehrig's disease for the famous baseball player who died from it, is a condition that causes loss of voluntary muscle strength, causing weakness in the arms and legs, and creating difficulty with speech and swallowing.
While the number of diagnoses hasn't changed much since the disease was first identified in 1869, the amount of research has progressively increased. The challenge is that scientists still don't know what causes the condition.
"We know that certain processes don't work right, but we don't know why that is or what is the initial trigger," Simmons said. "Until we know that, it is going to be very difficult to identify treatments."
The only drug approved for treating ALS is riluzole, which prolongs the life span of affected individuals for three months on average. "We don't really know whether it is effective in everyone who takes it," Simmons said. "We know it can't stop the disease in its tracks or reverse it."
The first symptoms of ALS usually include asymmetric weakness in the limbs — a weak hand, a foot drop, an arm that can't be raised. In about a third of patients, slurred speech or difficulty swallowing is the first sign.
Because the symptoms of ALS look much the same as symptoms of more common diseases, it often takes about a year from the onset of symptoms until medical professionals can make a definitive diagnosis.
"There is no one specific test for it," Simmons said. "A diagnosis is made by combining a number of different factors — history, a physical exam, electromyography and nerve conduction studies — with blood tests, MRIs and sometimes muscle biopsies and spinal taps to rule out other conditions."
The average life span for someone with ALS is two or three years after the first onset of symptoms. After five years, the survival rate is 20 percent. "It is devastating because it involves progressive loss of movement and paralysis," Simmons said.
Before death, those affected often lose the ability to speak and swallow. The cause of death is usually respiratory failure. Simmons said, "It poses enormous challenges for those affected and the people caring for them, and we have no effective medical treatments."
In some of those affected, the disease appears to have a genetic component. The Penn State Hershey Institute for Personalized Medicine is collecting saliva samples from ALS patients and as many family members as possible to better understand how mutations in some genes can lead to the death of motor neurons.
Simmons said people are always asking whether stem cell research can help. Trials are underway at a handful of places around the country to determine the safety of such treatments and to determine the proper dosages. He said no one knows yet whether a cure will result: "The hope is if you introduce stem cells and they develop into healthy motor neurons, or if they provide a healthier environment for existing motor neurons, we might be able to change the natural history of the disease."
See the dean/CEO of Penn State Hershey complete the ice bucket challenge.
The Medical Minute is a weekly health news feature brought to you by Penn State Milton S. Hershey Medical Center. Articles feature the expertise of Penn State Hershey faculty physicians and staff, and are designed to offer timely, relevant health information of interest to a broad audience.


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January 1982–March 1982
Cello Hiki no Gauche (1982)
January 23, 1982 JST
The story concerns Goshu, a professional cellist. During rehearsals for a performance, he is scolded by the conductor because his playing is not good enough. His timing is off and he seems to have no "feel" for the music.
Goshu returns to his lonely cottage and starts practicing. Then a cat enters who tricks him into understanding the inner meaning of the music. The importance of practice is shown by a cuckoo, rhythm by a badger and tenderness by a mouse. In four days he learns the true meaning and feeling of music and develops into a great musician.
Laced with popular classical music this special combines fun and inspiration for all ages and audiences.
[Source: AnimeNfo]
Doraemon Meets Hattori the Ninja
January 3, 1982 JST
No synopsis has been added for this series yet.
Click here to update this information.
Sentou Mecha Xabungle
February 6, 1982 JST
Zora was an outlaw planet where they had a rule that one could own stolen things if he could run away keeping it for three days.
When Motorcycle gang, Sand Rat, was driving on the planet, they were attacked by a walker machine. Then, an injured boy, Jiron, helped them. Lag, the leader of Sand Rat was interested in Jiron because he was going to the bazaar to steal a walker machine. That bazaar was held by Charring Cargo who was a courier of Blue Stone, and there must be the guards of Carrying and rowdy fellows at the bazaar. Furthermore, to be surprised, it was Carrying's latest walker machine, Xabungle, that he wanted to steal.
Wagahai wa Neko de Aru
February 17, 1982 JST
"I" is a cat without a name who lives with an English teacher, Mr. Kushami (Mr. Sneeze). "I" can't believe how stupid human beings are and chronicles some of their most incredible foibles. In particular, he is fascinated by Kushami's student Mizushima who falls in love with Haruko, the only daughter of businessman Kaneda.
Based on Natsume Souseki's comical novel.
[Source: The Anime Encyclopedia]

redpajama_set_name: RedPajamaCommonCrawl


UCLA gymnast on her electrifying floor routines: "What you see is literally how I feel"
UCLA gymnastics star Katelyn Ohashi scored another perfect 10 over the weekend for the floor routine that made her an internet sensation. But it is her smile, not the score, that is important to Ohashi and her coach, Valorie Kondos Field. Together, they overcame Ohashi's physical and emotional scars so she could rediscover a love of the sport and herself.
Ohashi's electrifying tumbles are punctuated with joy.
"I think competing is one of my favorite things ever. And so what you see is literally how I feel every time I step on the floor," Ohashi told CBS News correspondent Jamie Yuccas.
Six times this year, her floor routine earned a perfect 10. But Ohashi laughed at the move getting so much attention where she hits the floor in splits. She recalled the first time she did it.
"I remember you could probably hear me screaming across the gym. I'm like 'AH!' coming down. And it just kind of happened where I like bounced up because of the springs in the floor. And it just kind of stuck after that," she said.
The excitement has caught on. The UCLA gymnastics team is setting attendance records with sell-out crowds of nearly 13,000.
But behind that smile is a legacy of overcoming pain.
Ohashi literally grew up in a gym. By the age of 12, she made the national team and became the target of body shaming.
"We take it in and believe a lot of the things we hear. So to hear that you're not good enough, to hear you're too fat, to hear just a lot of hurtful and verbal abuse, you start to think that that's normal and that's what you deserve," Ohashi said.
Like many elite gymnasts, Ohashi developed an eating disorder.
"Me and my friends would create these like jokes almost, or like games and stuff. And like we would try to eat the least amount of food," Ohashi said. She acknowledged that wasn't normal, but it was accepted.
"I remember one time I couldn't make a floor routine 'cause I didn't have the energy. I'm like, 'Well, I only had raspberries today.' And they're like, 'OK, haha, so?'" Ohashi said.
As a way to cope, she kept a journal, writing: "I cry myself to sleep most of the time now... I've been told I looked like I swallowed an elephant or a pig."
Her body began to break down. In 2013, Ohashi scored better than Simone Biles in the American Cup. But while Biles went on to win four Olympic gold medals, Ohashi suffered a fractured back and torn shoulders.
"I really was so unhappy and like hated the sport. The joy just got ripped away so soon," Ohashi said.
She credits her coach at UCLA, Kondos Field, with bringing back the passion and the fun.
"I don't come to athletics with this burning desire to win at all cost 'cause that's not in my DNA," Kondos Field said. "I get to develop superheroes through the sport of gymnastics. So I get to help everyone of them become the best them they can be. And let's go take that out into the world and make it a better place."
Ohashi worked with a sports psychologist and a dietitian to develop a healthy relationship with food and body image. But as she turned the corner, she faced a new battle: ulcerative colitis, an auto-immune disease that causes the colon to bleed and robs the body of needed nutrition.
"So I was diagnosed with ulcerative colitis two years ago. And the first thing I hear is, 'Oh my God! You look so amazing! Like, how you did you lose so much weight so fast?' Chronically ill," Ohashi said. "And people are just talking about how good you look because you weigh a certain amount."
That sent Ohashi back to her pen and paper to process her new world. She started a blog called "Behind the Madness." In it she writes, "There is no normal, we just don't talk about the abnormal."
Ohashi bravely takes it all in stride. As her gymnastics career comes to a close next month, she hopes her next chapter includes a book deal. But first, her team is trying to repeat as national champs with Ohashi showing off her signature moves.
"Miss Val always goes, 'Booty! Booty!' whenever I do it," Ohashi said with a laugh.
"Do you love that?" Yuccas asked.
"Yeah. So much fun. And like having the team behind me is just like amazing," Ohashi said.
What's amazing for everyone else applauding a woman who can stand taller than most of us at just 4-foot-10.


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Ben Chaplin: We had a giggle filming raunchy scenes for Apple Tree Yard
Ben Chaplin in Apple Tree Yard (Credit: BBC)
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Ben Chaplin is a commitment-phobe. It's the nature of the job, he tells me when we catch up ahead of his latest role in BBC1's four-part adaptation of Louise Doughty's best-selling psychological thriller Apple Tree Yard. "Being an actor has an in-built flake effect, which means you can't commit to things too far in advance," he explains, saying that, often, he has to turn down invitations to friends' weddings. "I have to be non-committal, but it's my job."
Commitment problems are central to the narrative of Apple Tree Yard, whose plot hinges around an affair between geneticist Yvonne Carmichael (Emily Watson) and Chaplin's charismatic Mark Costley – an impulsive relationship that leads to a life-changing act of violence and the pair of them ending up in a Crown Court trial.
In person, Chaplin is as charming as his character first appears to be. He talks about his job with an infectious enthusiasm and speaks with ease about what it was like filming raunchy scenes with Watson. "We had a giggle," he says of the sex scenes. "You couldn't wish to do it with a nicer, more professional, more thoughtful, more clever woman. We laughed about it over the phone before we started."
"We had a giggle. You couldn't wish to do it with a nicer, more professional, more thoughtful, more clever woman. We laughed about it over the phone before we started."
Chaplin admits he hadn't read the novel before he took the part of the mysterious Costley. He finds, he says, that blissful ignorance of the original text makes it easier to get inside a character's head.
"It's such a powerful, personal experience reading a book because you have a personal relationship with it, you've set everything in your mind, so everything is as perfect as your imagination wants it to be," he says. "Even if you've read the book years before and then get asked to do a screen adaptation, it's very difficult to separate what's in the screenplay that you have to work with from what was in the book. And they're different animals, really." Naturally the book informs what you do as an actor, he says, "but sometimes it informs you down a blind alley".
"It's such a powerful, personal experience reading a book because you have a personal relationship with it, you've set everything in your mind, so everything is as perfect as your imagination wants it to be."
Is he nervous that people will compare the BBC adaptation with the book? Maybe if he were the producer or the director, he claims. "But as an actor I'm not too worried. My impact is limited to my contribution to the character."
As for that character, Chaplin feels very protective towards him, as he does with all his roles. "It's very easy for a character like that to come across as sleazy," he says. "This is going to sound pretentious and self-aggrandising, but my main concern was that, as an actor, you're the guardian of a character. So you mustn't comment on them, you must be inside them and care about them, whoever they are. If you don't care about them, it will show."
Ben Chaplin and Emily Watson in Apple Tree Yard (Credit: BBC)
Expelled from drama school
After 26 years in the industry, Chaplin is well versed in how these things work. He first became interested in acting at school before attending London's Guildhall School of Music and Drama – from where he was expelled before being allowed back in. "I was told the second term would be my last. But then on the last day of the second term, I was told I could stay, after I'd said goodbye to everybody," he says, jovially. "They gave me a stay of execution.
"I have quite a rebellious family. They're quite mouthy. That's probably why I got kicked out of drama school. I hope I'm not rude."
"I don't think I learned much about acting [there] other than how to deal with massive rejection," he says, admitting that he's always been rather nonconformist. "I have quite a rebellious family. They're quite mouthy. That's probably why I got kicked out of drama school. I hope I'm not rude."
I'm surprised by how level-headed Chaplin sounds. There was a moment in his career, I broach, when he could have become a poster boy for British acting, another Hugh Grant – he was living in America, working on a number of Hollywood films. Was it a deliberate decision on his part not to go down that route? "No, not at all," he says. "But what has been deliberate is that I've never wanted to play the same role twice."
"I've never wanted to play the same role twice."
From head footman Charlie in his big-screen debut, 1993's The Remains of the Day, to the lascivious agoraphobe Matthew in 1990s TV series Game On, Private Bell in Terrence Malick's war epic The Thin Red Line to a bank clerk ordering a Russian bride in Birthday Girl, erratic property tycoon Alvo in Mad Dogs to Robert Tibbo, the Canadian who hid Edward Snowden in last year's Snowden, typecasting has certainly never been a problem for Chaplin.
Treading the boards
His next project is on stage, in a play called Consent by Nina Raine at the National. It's been four years since he was last in a play, and he's excited to tread the boards once again. "It's been too long," he says. "It's like going to the gym for actors – it's your training."
Does he still love acting as much as ever? Yes, he says, although he jokes that he might have Stockholm syndrome. "Most people don't enjoy their job that much, do they? I've got 26 years out of loving it, so that's not bad."
'Apple Tree Yard' begins at 9pm on Sunday on BBC1
Apple Tree Yard


The fourth edition of Biological Safety: Principles and Practices, 4th Edition continues the format of the previous edition, focusing closely on infectious and toxic biological agents and their identification and control. Several major events have had an impact on the fields of biosafety and biosecurity since the publication of the third edition, notably the anthrax mailings of 2001 and the promulgation regulations for select agents. This newest edition examines significant developments throughout the field and discusses current regulations including those handed down from the Centers for Disease Control and Prevention and the U.S. Department of Agriculture.
Written by authorities with decades of experience in the field, the book is divided into five main sections that comprehensively cover the identification, assessment, and management of biological hazards. Chapters outline the human, animal, and agricultural considerations of a wide range of specific biohazards, from pathogenic organisms, viruses, prions, and cell cultures, to toxins and allergens. Numerous chapters detail practical systems for biohazard control. A brand-new chapter details critical safety considerations in a maximum containment (BSL 4) laboratory. Appropriate updates have been made to chapters carried over from the third edition, and a host of new contributors offer fresh perspectives on topic such as packaging and shipping of biological materials.
This book presents the essentials for a comprehensive biological safety program in venues ranging from the basic research laboratory to agricultural, pharmaceutical, educational, and commercial laboratories. Biological Safety: Principles and Practices, 4th Edition, is an indispensable resource for those involved in biological safety, including program managers, biological safety professionals, members of biohazard review committees, principles investigators, administrators, and students.
Electronic only, 622 pages, illustrations, index.
The terms normal microbial flora, normal commensal flora, and indigenous flora are synonymous and are used to describe microorganisms that are frequently found in particular anatomic sites in healthy individuals. Microbial flora is associated with the skin and mucous membranes of every human from shortly after birth until death and represents an extremely large and diverse population of microorganisms. The normal microbial flora for various anatomic sites is reviewed in this chapter. The microbial factors that contribute to the virulence of a microorganism can be divided into three major categories: (i) those that promote colonization of host surfaces, (ii) those that evade the host's immune system and promote tissue invasion, and (iii) those that produce toxins that result in tissue damage in the human host. Most infections are initiated by the attachment or adherence of the microbe to host tissue, followed by microbial replication in order to establish colonization. Organisms within biofilms are more resistant to antibiotics than individual bacteria and are partially protected from phagocytes as well. Some mucosal surfaces, such as the mouth, stomach, and small intestine, are protected from microbial colonization because they are constantly being washed with fluids. Toxins produced by certain microorganisms during growth may alter the normal metabolism of human cells with damaging and sometimes deleterious effects on the host. Many pathogenic organisms produce extracellular enzymes such as hyaluronidase, proteases, DNases, collagenase, elastinase, and phospholipases which are capable of hydrolyzing host tissues and disrupting cellular structure.
This chapter focuses on the zoonotic diseases caused by some indigenous agents of common laboratory animals which may pose an occupational hazard to animal handlers. The intent is to inform those working in animal facilities, including clinical and other research scientists and biological safety personnel, about zoonotic pathogens associated with animals used in laboratory research. Potential zoonotic hazards are associated with many laboratory animals, but the actual transmission of zoonotic disease has become uncommon due to the increased use of animals specifically bred for research over many generations. The majority of small laboratory animals (e.g., mouse, rat, and rabbit) used in research in the United States have been produced commercially in highly controlled environments under the oversight of veterinary care programs. The chapter addresses the intrinsic agents of potential significance in zoonotic diseases associated with eight animals: all of the animals from the primary category (dogs, macaques, mice, pigs, rats, rabbits) along with cats and sheep from the secondary category. It also provides some basic information on zoonotic diseases from common laboratory animals.
This chapter focuses on risk characterization and mitigation of worker exposure during culturing, inoculation of plants, and diagnosis of plant pathogens known to affect human health. There are increasing numbers of plant-pathogenic microbial organisms associated with human diseases or maladies. By the standard that agents are not associated with disease in healthy adult humans, almost all the plant pathogens and plant-associated microorganisms are in NIH risk group 1, for which biosafety level 1 (BL1) is usually recommended. By-products of plant pathogens in food, such as mycotoxins, can also cause illness. Healthy adults are not normally at risk of being infected by plant-associated microorganisms, but allergic reactions may occur. Good laboratory practices and some specific suggestions for dealing with plant-associated microorganisms are found in the methods manuals previously cited and the NIH guidelines. Samples of plant material obtained for isolation of pathogens or biocontrol agents are protected from contamination by using aseptic techniques. In the laboratory or greenhouse, autoclaving cultures and pathogen-infected material, or otherwise rendering them biologically inactive, is routine. Chemicals are not available or cost-effective for controlling many plant pathogens, and biocontrol agents are few. Other management practices to decrease the inoculum, and thus decrease exposure, are crop rotation, planting of resistant varieties (where available), planting date, and plowing under infected or infested plant material. These practices decrease inoculum by the process of competition with other microorganisms in the soil, where many plant pathogens are poor survivors.
This chapter examines the extent of documented laboratory-associated infections (LAIs) classified as viral, parasitic, bacterial, fungal, including rickettsial infection by reviewing those reported over the past 75 years, the incidence of LAIs, the means by which workers are exposed, and the contributions to exposure made by host and environmental factors. Epidemiology is concerned with the extent and types of illnesses and injuries in groups of people and with the factors that influence their distribution. Interventions can be defined and control measures, procedures and practices that may prevent the occurrence or recurrence of the event, can be implemented. This chapter focuses on the application of epidemiological methods to LAIs to provide tools for identifying and preventing health problems and disease processes. It also reviews both historical data and information accumulated in the last 26 years, pointing out observations that can provide the tools to control and prevent these LAIs. Laboratory studies of potential sources of infection have focused on hazards associated with aerosols produced from routine microbiological techniques. Regarding occupational exposures to pathogenic microorganisms, the worker is pivotal in controlling the safe outcome of any operation. The most effective strategy for the prevention or minimization of LAIs is to make certain that only approved procedures are consistently utilized. The risks associated with work involving infectious agents can be minimized if appropriate attention is given to biological safety.
This chapter focuses on agent- and activity-based risk assessments. An appropriately trained professional is needed to assess the risk associated with the agent. The European Union and the United States limit their lists of pathogens to those causing disease in healthy adult humans, the workers at risk. The protocols or standard operating procedures being developed for the specific tasks and equipment involving the etiologic agents of human disease can be assessed to identify the need for special containment practices or protective equipment. Work activities, such as centrifugation, homogenization and sonication must be assessed. A risk assessment is especially important for new procedures accompanying technological advances in related fields, such as the creation of infectious virus from its genetic blueprint. The National Research Council (NRC) recommended that risk assessment of work involving environmental release of recombinants be based on the nature of the organism and the environment into which it is introduced, and not on the method by which it is produced. A more thorough risk evaluation of the agent-host-activity triad is required to develop appropriate containment for actual work with etiologic agents. After the agent and activity are assessed for risk, the remainder of the risk assessment involving the host is not the purview of the biosafety professional. The risk assessment needs to be kept current and relevant to the work in progress to control any potential increase in risk. This chapter talks about acceptability of the risk of work with biological hazards, and risk prioritization.
Author: Joseph H. Coggin, Jr.
This chapter gives adequate information about bacterial pathogens and their host relationships and provides a general understanding of their biological traits and how they can cause work-related infections, especially laboratory-acquired infections (LAIs), when mishandled. It presents a synopsis of pathogenic bacteria that infect humans in their workplace, whether in the laboratory, industrial, or health care setting. The chapter discusses mechanisms that cause damage in bacterial infections. Some pathogenic bacteria, such as Shigella spp., which produce human dysentery, have evolved to trigger premature or unscheduled apoptosis in the host cells they infect. Knowledge of the mechanism of action of toxinogenic bacterial toxins helps in understanding certain disease processes of pathogenic bacteria. Compliance with recombinant-DNA guidelines is required when pathogenic bacteria or their genes or gene products are used. The chapter talks about classic human pathogens. Although Bacteroides spp. are members of normal gut and oral florae, work in the laboratory with these bacteria should be at biosafety level (BSL)-2 because they are known to cause human disease and can carry antibiotic resistance markers. Cutaneous anthrax is the most commonly acquired form in humans. The pathogenic bacteria Yersinia, Francisella, and Pasteurella have animal reservoirs as their natural hosts and produce serious diseases in humans.
This chapter educates laboratorians, biosafety personnel, and health care workers about the potential hazards of working in settings in which exposures to viable parasites could occur. It provides information about parasites that have caused or could cause accidental infections in laboratorians and health care workers. Factors that influence whether infection and disease develop after accidental exposures are also provided in this chapter. The chapter focuses on the protozoa that cause leishmaniasis, malaria, toxoplasmosis, Chagas' disease (American trypanosomiasis), and African trypanosomiasis. Summary data about 180 occupationally acquired cases of infection with the protozoa that cause these diseases are provided in this chapter. Blood and tissue protozoa of potential relevance to laboratorians and health care workers are also discussed in this chapter. Few laboratory-acquired helminthic infections have been reported. Even if laboratorians became infected by ingestion of infective eggs or through penetration of skin by infective larvae, they typically would have low worm burdens and few, if any, symptoms because most helminths do not multiply in humans. The fact that some of the persons who acquired parasitic infections did not recall discrete exposures suggests that subtle exposures (e.g., contamination of unrecognized microabrasions and exposure through aerosolization or droplet spread) can result in infection.
Traditional distinctions regarding inherent virulence and portal of infection provide a useful starting point for considering the pathogenic fungi. Discussion of fungal agents of human disease in this chapter is organism based rather than disease based. This discussion is not exhaustive for all documented fungal pathogens, but instead seeks to address those fungi that are seen with regularity in clinical and environmental mycology laboratories. The Public Health Security and Bioterrorism Preparedness and Response Act of 2002 required establishment of regulations regarding the possession, use, and transfer of select biological agents and toxins. Only one fungal genus, Coccidioides, contains species pathogenic for humans and other animals and is classified as requiring Biosafety level (BSL)-3 containment precautions. Coccidioides immitis recently was split into two species based on DNA analysis. The two species, C. immitis and Coccidioides posadasii, have identical potential for causing infection, disease, and death in humans and other animals. Containment and biosafety issues are more problematic for molds than for yeasts because molds have evolved the capacity to form airborne spores routinely as a dispersal mechanism. Dematiaceous fungi can cause infection when traumatically inoculated to skin and subcutaneous tissue, and disease from inhaled spores is possible as well. In contrast to fomite specimens, medical specimens pose little hazard of airborne fungal infection. Routine containment practices for specimen handling and processing provide sound protection for laboratory personnel as well as protection of specimens and cultures from extraneous contamination.
This chapter focuses on certain organisms for which the airborne route is the predominant means of transmission to humans. The in vivo tissue forms of the fungi are yeasts or spherules and are not readily transmissible to other humans, either by direct contact or by the airborne route. Efficient means of production of droplet nuclei in nature are sneezing, coughing, and vibration of the larynx, all of which introduce energy that subdivides fluids into tiny droplets. Qualitative risk assessment is encompassed in the biosafety levels (BSLs) established in Biosafety in Microbiological and Biomedical Laboratories. The current laboratory safety-oriented biosafety classification of microorganisms reflects what is known regarding the tendency for an organism to be transmitted by an airborne route, if there is effective treatment available for infection, and whether a vaccine is available. Mycobacterium tuberculosis must be inhaled deep into the lung and reach the alveoli as the first step in a successful infection of a new host. Hazard assessment in the laboratory should focus critical attention on the manipulation of fluids. All opening of tubes, pipetting, transfers, sonication, vortex mixing, etc., should be carefully contained. Many bacteria that have caused laboratory acquired infections in humans include Brucella, Francisella tularensis, and Burkholderia pseudomallei. The greatest problem encountered in the laboratory safety arena is generally not those involving decision patterns for known problems. Rather, it is more often the unknown safety precautions required for a particular agent that causes the laboratorian to fear infection.
The early use of continuous cell lines (CCLs) for the manufacture of biological products is represented by the manufacture of foot-and-mouth disease vaccine in the Syrian hamster cell line baby hamster kidney (BHK), the production of interferon from the B-lymphoblastoid cell line Namalwa, and the introduction of monoclonal antibodies from hybridoma cells. Long-standing experience has shown that contamination with pathogenic agents is the most important hazard and merits careful assessment of safety precautions. The culture types, in order of decreasing risk, are primary cell cultures; CCLs; and intensively characterized cells, including human diploid fibro-blasts. Most national guidelines recommend that human and other primate cells be handled using biosafety level 2 practices and containment and that all work be performed in a biosafety cabinet (BSC). For airfreight, cell cultures have been classified as ''diagnostic specimens'' under the International Air Transport Association (IATA) regulations. In many cases gene therapy concepts are based on viral vectors (e.g., adenoviruses and retroviruses). This chapter describes safety measures for large-scale production of biologicals, and focuses on characterization of cell lines. PCR is now commonly used to identify cell lines and to detect both cross contamination by other cell lines and contamination by adventitious agents or in combination with other methods. Safety testing of cells used in the manufacture of biologicals is usually performed with procedures accredited to good laboratory practice.
This chapter deals with laboratory animal allergy (LAA) as a common and important occupational hazard with different biological systems. Higher levels of allergen exposure correlate well with both the development and severity of symptoms. The most important risk factor in the development of LAA is the level of exposure to laboratory animal allergens. The most common symptom of LAA is allergic rhinoconjunctivitis, which consists of nasal congestion, clear nasal discharge, sneezing, and itchy, watery eyes. The diagnosis of suspected LAA can be confirmed with the use of skin tests or radioallerabsorbent tests (RASTs), which test for the presence of IgE antibodies to specific allergens. The major allergens from species including gerbils, hamsters, such as cow allergen, have been identified as members of the lipocalin family. The first step in prevention of allergy is identifying which workers may be more susceptible to the development of LAA. Epidemiological studies have also shown that the greater the exposure to animal allergens, the more likely will one become sensitized and have symptoms related to work. For an animal facility worker with suspected animal allergy, the diagnosis is largely made on the history of clinical symptoms associated with exposure. The allergens can be carried on small airborne particles and can remain airborne for long periods. By understanding the etiology, pathophysiology, prevention, and management of LAA, hopefully the necessary measures can be implemented to control and prevent the disease.
This chapter deals with basic biomedical and clinical laboratories at biosafety level 2 (BSL-2) and with containment laboratories, with the main emphasis on BSL-3 and their enhancements. In BSL-4 cabinet laboratories the focus is on enhanced primary containment by working with viable agents in a class III biological safety cabinets (BSCs). Primary barriers are specialized items designed for capture or containment of biological agents, e.g., BSCs, chemical fume hoods (CFHs), and animal cage dump stations. Secondary barriers are facility-related design features that separate the laboratory from nonlaboratory areas or from the outside. An administrative area, physically separated from all hazardous aspects of laboratory work, should be planned near the main entry to each building or floor. Caging systems as primary containment are an important consideration in the risk assessment and design of animal BSL-3 (ABSL-3) facilities. Historically, laboratories had fairly simple static heating, ventilation, and air-conditioning (HVAC) systems. Air change rates vary depending on specific needs, types of HVAC systems, number of exhausted containment devices per laboratory, and cooling requirements of rooms. Laboratories often have two types of drainage systems: sanitary and laboratory waste. A plan must be developed to provide a clear method for distribution of HVAC, plumbing, and electrical systems to the facility to allow for ease of operation and maintenance. Clinical laboratories are one of the most successful users of flexible casework. Cabinets for flammable and acid storage should be provided in each laboratory where chemicals are used and stored.
Primary barriers range from a basic laboratory coat to a biological safety cabinet (BSC). This chapter addresses some of the more common primary containment devices and personal protective equipment and a variety of equipment-associated hazards. The history of laboratory-acquired illnesses amply demonstrates how important primary barriers are and how equally important it is to select appropriate primary barriers and use them correctly. Personal protective equipment includes all clothing and other work accessories designed to serve or be worn as a barrier against workplace hazards. Employers and employees must be conscious of the fact that personal protective equipment alone does not eliminate the hazard. If the primary containment fails or is insufficient, personal protective equipment often becomes an important barrier against exposure. This chapter illustrates the four biosafety levels (BSLs) and their corresponding primary barriers. The Occupational Safety and Health Administration (OSHA) standards outline the specific provisions for the various types of personal protective equipment. The choice of style and fabric should be based on the job tasks to be performed, and the material or hazards to which the wearer may be exposed. The selection and use of the appropriate primary barriers constitute just one important component of the overall laboratory safety program. Laboratory-acquired infections have occurred due to the lack of or misuse of primary containment devices and personal protective equipment. Personal protective equipment should be chosen carefully and utilized appropriately.
Authors: David G. Stuart, David C. Eagleson, Charles W. Quint, Jr.
In a review of laboratory-associated infections (LAIs), the case is made that there is renewed interest in biosafety in laboratories and health care facilities because of the emergence of new infectious agents (human immunodeficiency virus), continuing problems with known agents (hepatitis B virus), and reemergence of old agents (Mycobacterium tuberculosis). Maximum-containment glove boxes, now called class III biological safety cabinets (BSCs), were developed during the 1940s, and partial-containment fume hood-like class I BSCs made their appearance in the mid-1950s. The first publication of microbiological testing of the performance of "laminar flow biological safety cabinets," as they were called then, was in 1968. The term "minute quantities" means that such chemicals will not be weighed out and diluted in the BSC. These activities are to be performed in appropriate equipment such as fume hoods or glove boxes. Type A1 cabinets meet or exceed the requirements of the microbiological tests, but they are not suitable for use with volatile hazardous chemicals. The microbiological aerosol tracer test is not suitable for performance testing of BSCs in the working laboratory because of the spores used as the tracer. A safety professional should be consulted to perform a risk assessment before selecting or using a glove box. If the risk assessment results in a requirement for a primary barrier that is not satisfied by standard models of available equipment, modifications of existing designs or special designs can often solve the problem.
This chapter discusses the subject of respiratory protection to individuals working in the field of microbiology. Respirators can be divided into two general classes: atmosphere-supplying respirators and air-purifying respirators. Particles may be captured on a filter by both mechanical and electrostatic mechanisms. Four mechanical mechanisms contribute to particle deposition: interception, diffusion, gravity, and inertial impaction. There are very few recommendations for use of respiratory protection to control exposures to specific aerosolized microorganisms, or bioaerosols as they are sometimes called. Assigned protection factors (APFs) are designated by type of respirator. In simplified terms, the APF is the factor by which a properly selected and fitted respirator will reduce contaminant exposures. If an APF is 10, then the concentration of contaminant that reaches the wearer's lungs will be reduced by a factor of 10. A qualitative method has been proposed for selecting respirators for protection in infectious aerosol environments which is based on the National Institute for Occupational Safety and Health (NIOSH) respirator decision logic. Respirator selection will be based on the best available practices, most current knowledge, and professional judgment and reemphasizes the need to utilize engineering and administrative controls when possible.
This chapter reviews the risks associated with the blood-borne pathogens of major concern for work-places handling human clinical specimens, and the evolution and efficacy of prevention methods developed to reduce exposures and transmission of infection. It is important to review the documented cases in detail in order to emphasize rational precautions for work with human specimens. The major premise involved the careful handling of all human blood and certain body fluids as if all were contaminated with human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) or other blood-borne pathogens. Standard microbiological practices form the basis for BSL-2, with additional protection available from personal protective equipment (PPE) and biological safety cabinets (BSCs) when appropriate. The National Institute for Occupational Safety and Health (NIOSH) guidelines establish criteria, such as the need to install container openings at a height of 52 to 56 in. to provide the ergonomically correct position for 95% of all adult female workers. The NIOSH guidelines also provide evaluation tools to help select the most appropriate container for the facility. Any defective personal protective equipment (PPE) must be replaced, and reusable protective clothing must be laundered and maintained by the institution. Finally, all laboratory workers must be instructed in the proper use of PPE and its location. The ultimate indication of compliance with standard (universal) precautions is the reduction in workplace exposures and infection with blood-borne pathogens.
This chapter focuses on worker protection; however, the use of prudent biosafety practices can also protect coworkers, and the local community, from infection and protect the work, product, and the environment from contamination. The National Research Council (NRC) Committee on Hazardous Biological Substances in the Laboratory recommended seven basic prudent biosafety practices to avoid exposure to infectious agents. These practices provide barriers against the known routes of exposure for most diseases and are recommended for work with any biohazardous agent. The chapter discusses criteria for use of containment levels for human pathogens. Indigenous and exotic agents vary by country and within regions of some countries; thus, there must be some flexibility for the assignment of containment levels. Using the voluntary guidelines and mandated regulations, work practices are selected or developed to provide appropriate barriers to prevent exposure and subsequent infection in the trained, healthy adult worker. The chapter talks about examples of barriers that can be used to block the known points of entry and routes of transmission of infection. Pipetting was one technique associated with laboratory infections in the past. To manage the risk of working with biohazards and to actually reduce the risk of exposure, one must identify and interpret the recommended practices. Immunocompromised individuals need extra precautions to prevent exposure to microbial agents, even at biosafety level 1 (BSL-1).
This chapter describes basic strategies for decontaminating surfaces, items, and areas in laboratories to eliminate the possibility of transmission of infectious agents to laboratory workers, the general public, and the environment. Emphasis is placed on the general approaches to decontamination practices. The principles of sterilization and disinfection are discussed and compared in the context of decontamination procedures used in laboratories. The definitions of sterilization, disinfection, antisepsis, decontamination, and sanitization are reviewed in the chapter to avoid misuse and confusion. The definitions and implied capabilities of each inactivation procedure are discussed with an emphasis on achieving and, in some cases, monitoring each stage of microbial reduction. Decontamination in the microbiology laboratory requires great care. It may entail disinfection of work surfaces or decontamination of equipment so that it is safe to handle. CD gas sterilization can be used for decontamination of laboratory rooms, equipment, glove boxes, and incubators. Recommendations for the decontamination of items and areas contaminated with Bacillus anthracis (anthrax) spores are based on two historical sources. The first is the industrial setting, where animal hides and hairs are processed. The second is the laboratory setting, where biological safety protocols have been developed to address decontamination of high concentrations of anthrax spores after spills in the laboratory. Anthrax is unique among the agents of bioterrorism because the etiologic agent is a bacterial spore that is more resistant than other pathogens. Appropriate materials and methods for decontamination, disinfection, and sterilization are required for the safe conduct of work with biohazardous agents.
This chapter provides practical guidance to facilitate compliance with current national and international regulations that govern the packing and shipping of hazardous materials and dangerous goods. Topics in this chapter include terminology, classification and naming of diagnostic specimens and infectious substances, marking and labeling packages, packaging material, documentation, training and certification of personnel, practical suggestions for classifying diagnostic specimens and infectious substances, and resources for additional information. Medical waste which is reasonably believed to have a low probability of containing infectious substances must be packed and shipped as Medical Waste. Department of Transportation (DOT) regulations, International Air Transport Association (IATA) requirements, and IATA packing instructions (PI) describe the minimum standards for the safe transport of various biological materials. The marking and labeling on the outer container communicate essential information regarding the shipper and consignee of the package, the nature and weight of the contents of the package, the potential hazard of the substance, how the substance is packed, and information to be used in case of an emergency. There are several simple but extremely important points which must be regularly and strongly emphasized to persons who pack and ship diagnostic specimens and infectious substances.
A biological safety program management system, following the process identified by ISO 14001, is an organized and documented approach to managing biosafety issues within an organization. This chapter describes six key steps to an effective management system. Hazard identification requires the organization to identify, evaluate, and prioritize, in a systematic manner, the full set of biological hazards associated with its activities, products, and services. The biosafety program must have a system to keep its analysis of hazards up-to-date. All legal constraints imposed on an organization to control its biological materials should be identified. These include federal, state (or provincial), and local laws and regulations, permits, registrations, orders, and consent decrees. Establishing objectives and targets requires taking the program policy and the results from the significance criteria assessment and establishing more specific goals with regard to biosafety. Communication from nonregulatory external parties should be handled in a consistent and responsive manner. A biosafety manual is the most common means used to capture written information (paper or electronic) that describes core elements of the biosafety program. Corrective and preventive action provides the framework for identifying and correcting problems in the overall system, keeping the program on track relative to its goals and objectives, and for investigating and addressing any nonconformance with the defined biosafety program. Establishing a formalized, documented biosafety program management system is not an easy task, but the benefits are well established.
This chapter provides an overview of the regulations and guidelines for handling biohazards and recombinant DNA (rDNA) in workplaces around the world. The Genetic Manipulation Advisory Committee (GMAC) oversees the development and use of innovative genetic manipulation techniques in Australia so that risks to the safety of workers or potential hazards to the community or environment associated with the genetics of manipulated organisms are identified and can be managed. Most biosafety issues are addressed at the provincial level under transportation of dangerous goods legislation, occupational health and safety legislation, general public health and safety legislation, and federal and provincial Workplace Hazardous Materials Information System (WHMIS) legislation. A bill must be passed by both the U.S. House of Representatives and the U.S. Senate before it becomes a Public Law. The International Civil Aviation Organization (ICAO) has developed regulations for the safe transport of dangerous goods by air. These regulations have been incorporated by the International Air Transport Association's (IATA) into a set of internationally accepted regulations which cover the packaging and transportation of dangerous goods (including infectious materials) internationally.
The principal goal for an occupational medicine program is to promote a safe and healthy workplace through the provision of work-related medical services. In a biomedical research setting that involves biohazardous materials, those services should include a preplacement interview and counseling, a practical plan for responding to suspected exposures and infections, and occasionally the provision of medical surveillance for suspected health hazards in the work environment. A preplacement medical evaluation is recommended for individuals who may be exposed to potential human pathogens, including zoonotic agents. This chapter focuses on medical care for occupational injuries and illnesses. Medical surveillance is an important component of occupational medical support services. The practice of occupational medicine is unique in that the practitioner is responsible to both the employee-patient and the worker's employer. Participation in a serum storage program is generally voluntary, although it should be strongly recommended by the health care provider. Sera should be preserved for the workers' benefit at -20°C or lower in a non-self-defrosting freezer. The importance of the basic principles for designing medical support services for a workplace—a proper risk assessment and thoughtful advance planning for work-related medical needs—cannot be overemphasized when the workplace is a maximum containment laboratory. Specialized medical surveillance programs and immunization regimens may be necessary. Strict adherence to postexposure monitoring protocols is needed, and provisions for adequate medical facilities in the event of an exposure and/or subsequent infection should be made prior to undertaking biosafety level 4 work.
This chapter provides mechanisms for the evaluation of the components of a biosafety program and gives examples of tools that can be used for this process. A biosafety program may include blood-borne pathogens, infectious materials, diagnostic specimens, recombinant-DNA (rDNA) oversight, management of biosafety cabinet (BSC) certification, and/or the select agent program. The most common method used to evaluate a biosafety program is an audit of the laboratory. Biosafety manager, self-inspection, safety generalist, outside consultant and regulatory agency are explained in this chapter. One logical way to evaluate a component of the biosafety program is through review of records and documentation rather than a site visit. In many institutions the biosafety program manages the BSC certification program. The chapter discusses annual reverification of biosafety level 3 (BSL-3) laboratories. Inspection should be scheduled well in advance to ensure that laboratory experiments are not in progress, since the annual reverification focuses on facility operations and is not a review of laboratory procedures. A BSL-3 facility should be reverified on an annual basis to ensure that it continues to meet the BSL-3 criteria stated in Biosafety in Microbiological and Biomedical Laboratories (BMBL).
Prions are unprecedented transmissible pathogenic agents that cause a group of invariably fatal neurodegenerative diseases, including scrapie (the prototype prion disease in sheep and goats), bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in deer and elk, and Creutzfeldt-Jakob disease (CJD) in humans. A complete list of currently known animal and human prion diseases is provided in this chapter. The biosafety issues related to prions have been addressed in several guidelines published by health authorities over the years, with recommendations to limit the potential risk associated with prion contamination in laboratory studies as well as in foods and medicinal products. In the chapter, the authors strive to sort out the truth from the myth and the legitimate from the unreasonable in relation to the biosafety issues, and provide a factual basis for concerns as well as an informed rationale for actions to be implemented. The authors also explore the potential consequences of underreaction as well as overconcern with respect to these issues. The potential transmission of prions from human to human by blood transfusion or through administration of blood components or plasma derivatives has long been a cause for concern among health professionals throughout the world. The chapter also discusses the epidemiological data regarding classical CJD and blood, and deals separately with variant of CJD (vCJD) and its implications for the safety of blood products.
This chapter addresses some of the basic safety issues and risk assessment considerations for those individuals who will be affiliated with biosafety level 4 (BSL-4) laboratory operations. It also discusses basic laboratory design and engineering considerations for reducing daily operational risks in these unique laboratories. The class III biological safety cabinet was designed for work at BSL-4 with microbiological agents, and it offers the highest degree of personnel and environmental protection from infectious aerosols, as well as protection of research materials from microbiological contaminants. The laboratory-specific BSL-4 safety and operations manual and its comprehensive protocols should be read by trainees and used for refresher training of experienced personnel. A supervised apprenticeship is completed when the trainer is satisfied that the trainee fully understands all of the principles of BSL-4 operations, and has satisfactorily demonstrated the requisite skills and temperament to work in the BSL-4 environment. Factors influencing the external containment envelope include clothing change room, personal shower and chemical disinfectant shower. Factors influencing the inside containment envelope include laboratory, microbiological security and communication training. The increase in construction of BSL-4 laboratories is the result of the need to study new and emerging diseases associated with high morbidity and mortality, as well as the concern that bioterrorists may use weaponized versions of exotic disease agents.
This chapter provides a general overview of the construction, safety features, and suggested containment level of viral gene transfer vectors in common laboratory use or under development. The discussion of each vector focuses on similarities of the biosafety features and issues arising from the construction and use of these gene transfer systems. The safety assessments and suggested containment levels presented in the chapter can be used to inform the discussions of local institutional biosafety committees (IBCs), investigators, and biosafety professionals, and assist in the assessment of the risks inherent in the use of these vectors in the laboratory. Characteristics of viral systems commonly used for recombinant gene transfer have been provided in the chapter. Replication-defective vector genomes persist in one of two forms in the infected cell: either as an episome or integrated into the host cell genome. In this discussion, an episomal viral vector means that the vector genome remains intact yet is separable from the host cell DNA by physical methods.
As a general rule, teaching laboratories tend to be densely populated with large numbers of individuals with limited experience in the hazards of a science laboratory. A section of this chapter focuses on college and university microbiology teaching laboratories where hazardous organisms are handled. Numerous common procedures conducted in the microbiology teaching laboratory may create aerosols, including improper sterilization of inoculating loops, centrifugation and microcentrifugation, use of bead beaters or shearing blenders, pipetting, and handling of contaminated animal bedding. Each biosafety level (BSL) is based on the accepted potential hazard of the agent, as well as the general operations of the laboratory. Increasing attention has been focused on the potential for microbiology teaching laboratories to be exploited by terrorists as a means of acquiring agents or knowledge to conduct acts of bioterrorism. While animal and plant pathogens may represent potential threats for misuse as bioterrorism agents, at the same time many of these organisms are invaluable tools in the microbiology teaching laboratory. The selection of appropriate microorganisms and their toxins for instructional purposes in the microbiology teaching laboratory is integral to creating a biologically safe learning environment. The strategic choice of organisms and toxins suited to introductory, intermediate, or advanced course levels complements instructor efforts to increase student awareness of potential biosafety risks, and educates students in good microbiological laboratory practice to minimize those risks. By making safety training a part of every microbiology laboratory, and by providing careful counseling, risks such as exposure to biohazardous microorganisms can be minimized.
This chapter addresses the biosafety challenges commonly experienced in cultivating recombinant and pathogenic microbes and the use of mammalian cells for the production of therapeutic proteins and viruses. It summarizes the place and application of viruses, bacteria, and fungi in pharmaceutical research and development. Recombinant viral agents have become a significant starting material for both gene therapy and vaccine production. Recombinant bacteria are frequently used to produce pharmacological proteins, enzymes, and plasmid DNA. The two bacterial species most frequently employed are Escherichia coli and Bacillus subtilis. Containment for fungi and yeasts can vary from biosafety level 1 (BSL-1) for organisms such as Saccharomyces cerevisiae through BSL-3 for Histoplasma species. Containment for known fungi should follow the recommendations provided by national guidelines, such as in Biosafety in Microbiological and Biomedical Laboratories (BMBL). Mammalian cells may be used for the direct generation of pharmacological proteins, usually created by recombinant DNA techniques. The biosafety regulations utilized at a specific company are frequently dictated by the home country for the organization. Clinical assays can present challenge for the biosafety professional within the pharmaceutical industry. The global harmonization of biosafety practices would allow benchmarking to determine the best practices for this industry.
This chapter highlights the selection and design of equipment and facilities to achieve a safe work environment. There are few absolutes that can be applied across the board, since the decisions to be made are dependent on the risk assessment of the organism and the processes used. Fortunately, similar equipment and facility design criteria are used, so there are common biosafety principles that can be utilized which are discussed herein. Primary containment is provided by the equipment utilized and the use of appropriate biosafety practices, administrative practices, and personal protective equipment. In general, all of the standard and special practices and safety equipment identified in Biosafety in Microbiological and Biomedical Laboratories (BMBL), along with the recommendations in the NIH recombinant-DNA guidelines, are applicable to large-scale processes. Bioreactors used for growth of microorganisms and those used for cell culture can share many attributes. A biological safety cabinet (BSC) can be utilized for smaller equipment that does not generate much turbulence. In some instances, BSC manufacturers, or other specialty equipment fabricators, can make specialized containment devices for specific equipment. The facility design and construction provide the secondary containment that protects people outside of the immediate work area, both in other parts of the facility and in the community at large.
This chapter describes the facility requirements and work practices of biosafety level 3 agriculture (BSL-3 Ag). This BSL, unique to agriculture, was developed to protect the environment from economic, high-risk pathogens in situations where studies employ large animals or other situations where facility barriers, which normally serve as secondary barriers, must serve as primary containment. In agriculture, special biocontainment features are required for certain types of research involving high-consequence livestock pathogens, in animal species or other research where the room provides primary containment. The descriptions and requirements for BSL-3 Ag studies are based on the use of high-risk organisms in animal systems or other types of agriculture research where the facility barriers, usually considered secondary barriers, now act as primary barriers. There are circumstances where certain agents, typically handled at BSL-3 Ag for work with agricultural animals, may be studied in an enhanced BSL-3 laboratory or enhanced ABSL-3 for work with small animals for which primary containment devices can be utilized. Filter testing is intended to be completed in a manner consistent with industry standards for certification of HEPA filters in biological safety cabinets (BSCs). An alternate procedure may be used as outlined in the USDA-ARS Facility Design Standards (USDA-ARS, 2002).
Written by recognized authorities with decades of experience in the biological safety field, the third edition of this best-selling volume is an essential resource as well as an excellent text for courses in biosafety. The epidemiology of laboratory-associated infections, including some previously unreported cases, is thoroughly covered. Detailed chapters cover hazard assessments of the wide range of pathogens and biological toxins encountered in biomedical laboratories as well as other occupational settings. All facets of hazard control, from personal protective equipment to institution wide provisions and policies, are discussed.
As the subtitle indicates, the editors seek to address both principles of control and the practice of biosafety with this book.
The fourth edition broadens the original scope of the book in the wake of recent untoward events, e.g., the 2002 anthrax letters. It also addresses Select Agent regulations disseminated by the United States Centers for Disease Control and Prevention. Notwithstanding other good journal articles and useful websites, this book is a complete syllabus of pertinent information.
The editors indicate that it should be used as a resource by biosafety professionals, those who teach, and those who work with pathogenic agents in research, production, and teaching. More than 50 authors contribute the 33 chapters.
in the pharmaceutical industry, large-scale production of microorganisms, special considerations for agriculture pathogen biosafety and regulatory impacts).
There is little doubt that there is a considerable demand for this material. The field of likely coverage, e.g., teaching laboratories, the pharmaceutical industry, large-scale production facilities, and those involving agricultural pathogens is similarly vast. Many of us have at least seen earlier editions, if not used them. Broadly written books devoted to microbiology, infectious disease, or even occupational health are certainly not a good substitute for this book. I think it will find its way into nearly every regulated facility (and most are these days!). I can envision it will be read in its entirety by biohazard specialists, but be of use to a much broader panel of professionals, including physicians, microbiologists, veterinarians, and industrial hygienists, both in the U.S. and abroad.


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He's been mentoring business owners in Perth, around Australia and overseas for over 15 years now. And, he's helped business and companies generate over $100M in additional sales. Most of all, David has a huge heart and loves helping people. He knows what he is doing and has is very successful at it.
I have been receiving his weekly business improvement newsletter for years now and I thought of you because you've shared that you've been struggling with your business and I was hoping this introduction could support you.
David is quite busy, but has offered to give you a free consultation. If you're still interested in improving your business then I recommend giving David Buttsworth from ABC Business Improvement a call.
His website is www.abcbusinessimprovement.com.au And his work phone number is 1300 858 836.
I also copied him in on this email so you can respond to him via email if you prefer.
I recommend you give him a call right away, because his schedule in February is filling up fast.
← It's time to strike, whilst the iron's hot!
It's time to strike, whilst the iron's hot!


Pink tweed dresses are a perennial favorite. The very first dress that I can remember buying was at age 10.
Remarkably the dress that I feature here today is quite similar! !
Trimmed with metallic and grosgrain ribbon, this classic even has pockets !
Now this sheath dress is knitted and is so cozy and soft. Light grey tights are thick and warm for a cold day. I met a friend for lunch at Bergdorf Goodman.
Indeed, it takes a lot to get me up and out in this frigid weather. Dear friends are the only reason I would make such an effort. This particular friend is in fact, quite dear to my heart. Therefore, I do hope to see her soon!
Punctually is a must for me, so I arrived early. While there, I took two snaps which will SNAP you out of the winter doldrums! Ready?
Eye candy dresses that are red carpet ready!
Quite a difference from the pink tweed dresses!
Below another snap of my tweedy number, taken at home by my friend!
Near Christmas you saw me pair these necklaces! One is from KCSignatures and the other, more colorful piece, is one I made years ago.
Red added a pop of color with my pearl headband.
Crimson red also matched the ribbon trim of the dress and shoes!
See the trim finishing and the fun pointy pumps that I wore that day. After the lunch I relaxed!
Nostalgia hits me whenever I see the cameo vase that was my mother's!
Pink tweed dresses can pop for Winter!
TIP: Always add accessories, and add them liberally when it is dull outside! The sparkle will lift your mood.
Shopping for pink tweed dresses can be challenging this time of year. However, please do try to scout out this year round color. The pink can be muted with warm peach tones like mine, or bold in vivid Dahlia! For instance, here is a dress that is very well priced from Saks. Pease try to shop local and consider second hand shopping as well. Ebay and Etsy are always good sources.
What color are you gravitating towards this season!
Are you also dealing with very cold weather?
If you are in Chicago, or anywhere else affected by extreme weather, I wish you safety and warmth!
Pink for winter? Yes or no?
Let me know. You know how!
Oh I love pink tweed dresses! And pink is fine with me in winter. Wore a pink dress a few days ago!
I am sure it looked fabulous on you Patti!
thanks, yes it is a fun color and so soft!
Thanks, I can feel the warmth !
I love pink in the winter and tweed is a classic. I think your white tights are very groovy. We are watching Mad Men again and they often wear white tights in the 60s.
We were hit with a massive snow storm that dumped about 2 ft of snow on us 3 days ago and now it is -20 C here today. I HATE it! Canadians keep going about their days though, we have to.
Oh that awful snow there! I don't know how you cope! Please take care my friend! I would just hate that!
Tweed is near and dear to my heart too. I'm not sure why, but I'm always drawn to it even though I have so much of it already in my closet. I think it's the mix of textures and different colors all in one material.
And good for you for getting out. Friends make it worth the while!!
Tweed is so classic but looks so fun updated with bold accessories !
My friend sent me a link on facebook to some of the cold temps in the US at the moment – hope you are staying warm! It's crazy, we never see anything that cold here at all!
Hope that you are having a great week! We finally have someone coming over to fix the aircon today, yey! Has been no fun in the heatwave without it! So very different from the temps you are having, ha!
oh, I am so sorry about the heatwave. Such extreme weather..
Thanks so very much Mica, stay cool!
Pink for every season! I love this fun tweed dress – you look just stunning, Elle, all dripping with your jewels. What a fun outfit. Thanks for the lovely shots of those dresses – the second one really rings my bell. Wishing you a lovely weekend, and maybe some warmer temps, my dear!
Yes to year round rose! I do love the pile it on! lol!
I love tweed! I just purchased a pre loved tweed skirt, I searching for long to find a beautiful tweed skirt! You are wearing a fabulous dress and I always adore your choice of accessories!
Thanks! I am sure you will find whatever it is you want. I know you will style it well, too!
You look lovely! It takes a lot to get me outside in this weather — yesterday, I braved it to walk the streets of NYC to visit my doctor.
Thanks you very much ! It does take a lot to go out in the bitter cold.
I do hope you are well!
Perfectly said once again my friend ! The shoes are comfy !
I love reading about your daily adventures! And you look especially lovely in your elegant pink dress, these gorgeous shoes and your pretty accessories. I agree with your advice – fun accessories and colors do lift up my mood in cold or gray winter days!
Yes, you are the queen of fun accessories and bold pattern mix. I do admire your style!
Ah yes that tweed dress is traditionally chic and on trend. And that feather dress, oh la la eye candy for sure! Love it. And it was great seeing you the other night I had a wonderful time.
Oh yes the dress at Bergdorfs!
Yes, it was wonderful to see you!
Love love those pointed shoes! I admired them in one of your other posts with the thick buckle detail. Your hair is beautiful up Elle and I love the headband. You are such a fan of details, like myself! Those dresses are so fun and would certainly brighten up my mood, love the feathered one. Your tweed dress is beautiful and I love the thick leather belt.
Thanks so much sweet friend! These pointy shoes are quite comfy actually!


text:
Q: Issue with labels I am writing a program such that a user can enter his details one window and a new window comes showing the output. But, for some reason, the labels in the second window are not being printed.
from tkinter import *

window = Tk()
window.title("Login")
window.geometry("500x500")
window.resizable(0,0)
l = Label(window, text="Name: ", font="Roman 20 bold",padx=10,pady=10)
l.grid(row=0,column=0)
l = Label(window, text="Password: ", font="Roman 20 bold",padx=10,pady=10)
l.grid(row=1,column=0)
e1 = Entry(window, font="Roman 20 bold")
e1.grid(row=0,column=1)
e2 = Entry(window, font="Roman 20 bold",show="*")
e2.grid(row=1,column=1)

def test():
    a= e1.get()
    b= e2.get()
    mess = Tk()
    mess.title("Output")
    mess.geometry("500x500")
    mess.resizable(0,0)

m= StringVar()
m.set("Name: "+a)
l = Label(mess, textvariable=m, font="Roman 20 bold",padx=10,pady=10)
l.grid()

n= StringVar()
n.set("Password: "+b)
l = Label(mess, textvariable=n, font="Roman 20 bold",padx=10,pady=10)
l.grid(row=1,column=0)
mess.mainloop()

b = Button(window,text="Click",font="Roman 15 bold",width=15,command= test)
b.grid(row=2,column=0)

window.mainloop()

I don't think it is an issue with the get() method as I am able to print the values obtained on the Python shell screen. So, I think it might be an issue with the label(), though I might be wrong.

A: You aren't updating the labels with l.configure(text="Password: " + c") so it takes the value that was set at the start which is none.
Also you defined b as the button so you have to define the entry get as another.
Also I deleted the string car because I found it unnecessary.
Updated code:
from tkinter import *

window = Tk()
window.title("Login")
window.geometry("500x500")
window.resizable(0,0)
l = Label(window, text="Name: ", font="Roman 20 bold",padx=10,pady=10)
l.grid(row=0,column=0)
l = Label(window, text="Password: ", font="Roman 20 bold",padx=10,pady=10)
l.grid(row=1,column=0)
e1 = Entry(window, font="Roman 20 bold")
e1.grid(row=0,column=1)
e2 = Entry(window, font="Roman 20 bold",show="*")
e2.grid(row=1,column=1)

def test():
    a = e1.get()
    c = e2.get()
    mess = Tk()
    mess.title("Output")
    mess.geometry("500x500")
    mess.resizable(0,0)
    l2 = Label(mess, font="Roman 20 bold",padx=10,pady=10)
    l2.grid()

    l = Label(mess, font="Roman 20 bold",padx=10,pady=10)
    l.grid(row=1,column=0)
    l.config(text="Password: " + c)
    l2.config(text="Name: " + a)
    mess.mainloop()

b = Button(window,text="Click",font="Roman 15 bold",width=15,command= test)
b.grid(row=2,column=0)

window.mainloop()

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