Monday, 31 July 2017

The pathologic appendix

Antonella Chesca and Tim Sandle have written an article looking at the pathologic appendix.

Here is the abstract:

Appendix pathology brings together different forms and is found in different age segments. Most susceptible to disease are children. In this context, the children and young people Appendix pathology may be complicated or can be accompanied by damage to adjacent organs, located in the abdominal cavity. This morphological study presents the structural aspects of normal appendix and the ulcerated appendicitis. For observation of structural aspects, has been used classic stainings. It also shows the appendix using immunohistochemical technique.

Here is the reference:

Cheşcă A., Ciomeica A., Sandle T. Structural aspects on pathologic appendix versus normal appendix, Medicine and Ecology, 80 (3): 120-123

The publication can be accessed here.

Posted by Dr. Tim Sandle

Sunday, 30 July 2017

Battling infections with bioelectricity

Is the answer to fighting pathogens connected with bioelectricity? Promising new research suggest this is possible by using drugs to changing electrical charge of cells. Studies have been performed in frogs.
What is remarkable about the studies is the fact they have used existing medications. The drugs, which are approved for use in humans, were used to eliminate an Escherichia coli infection in frogs. The action of the drugs triggered a reversal of the electrical charge in the cells of the frigs, wiping out the pathogen.

For the study drugs that either depolarize (positively charge) or hyperpolarize (negatively charge) cells were tested. It was found that the negative charge effect is of significance. The drug that achieved this in an optimal way was one called ivermectin. Ivermectin is a medication effective against many types of parasites, such as head lice, scabies, river blindness, strongyloidiasis, and lymphatic filariasis.
Wiping out bacteria
Analyzing what's happening, Tufts University biologists discovered that administering specific drugs leads to the interior of cells becoming more negatively charged. In tadpoles (of the frog Xenopus laevis), this strengthened the innate immune response to E. coli infection and injury. Hence the researchers have found a hitherto unknown and novel aspect of the immune system. This has been termed "regulation by non-neural bioelectricity." The finding could be significant, possibly opening up a new approach for clinical applications of the technique in human medicine.

As the lead researcher, Dr. Michael Levin explains: "All cells, not just nerve cells, naturally generate and receive electrical signals." He adds that by "being able to regulate such non-neural bioelectricity with the many ion channel and neurotransmitter drugs that are already human-approved gives us an amazing new toolkit to augment the immune system's ability to resist infections."
Bioelectricity refers to electric potentials and currents produced by or occurring within living organisms. These potential vary between one to a few hundred millivolts. Perhaps the best known example is with the electric eel.
Such electric fish produce their electrical fields from a specialized structure called an electric organ. This is made up of modified muscle or nerve cells, which became specialized for producing bioelectric fields stronger than those that normal nerves or muscles produce.
The pathogen killing effect also rests with the immune system. All vertebrates possess two kinds of immunity: the adaptive immune system, which relies on the memory of previous exposure to a specific pathogen; and the innate immune system, which is the first line of defense against pathogens. It is the innate immune system that is being studied in relation to bioelectricity.
Immune system in action
The reason why this is important relates to the role this facet of the immune system plays with tissue repair and regeneration. Also important to the process is the neurotransmitter serotonin, which acts as an intermediary between voltage and immune response. Serotonin is found in all bilateral animals, where it mediates gut movements and the animal's perceptions of resource availability (primarily to food).
The combination of ivermectin and bioelectricity led to the proportion of embryos the bacterial increasing average 32 percent. This was compared tadpoles not given the drug. Here, with the untreated tadpoles, the mortality in untreated rate was 50 to 70 percent.
It is hoped the developed will lead to a new understanding of innate immunity can lead to new developments for fighting pathogens, especially new ones from which there is no adaptive immune system memory. The longer-term aim is to develop a treatment for people by manipulating the bioelectric microenvironment through new combinations of depolarizing drugs. Before doing so a greater understanding is required of how and why bacteria respond to the changes to cell environment and why they find these changes inhospitable and potentially destructive.
At present it is though the electrical variations alter the membrane potentials of the bacterial cell wall
The research has been published in the journal npj Regenerative Medicine, under the title "Bioelectric regulation of innate immune system function in regenerating and intact Xenopus laevis."
In related news, researchers are looking at bioelectric effects further, such as evaluating the antibacterial efficacy of a bioelectric dressing containing silver and zinc against various wound pathogens. With this, the microbial killing is possibly enhanced first by bioelectric field attracted microbes carrying a negative charge to the positive pole containing silver and zinc.

Posted by Dr. Tim Sandle

Saturday, 29 July 2017

Bacterial pathogens interact to cause disease

New methodology allowed researchers to more easily investigate mechanisms of infection and provide new insight into how pathogens can work together to cause disease. Using the new tool, researchers confirmed a safer model for study of Brucella species, which cause a potentially debilitating infectious disease in humans and cattle.

Brucellosis is an infectious disease of livestock that may be transmitted to farm workers or consumers of unpasteurized dairy products. Easy to spread and hard to detect, the bacteria that cause the illness, Brucella species, are considered potential bioterror weapons. Yet, precisely because Brucella species are so dangerous to handle, research on this important disease-causing agent, or pathogen, has lagged behind that of other infectious diseases.

Using an innovative method they developed to study the infectious process, investigators at Beth Israel Deaconess Medical Center (BIDMC) established a safer way to study Brucella. In an early test of the model, the research team observed a surprising and previously undocumented interaction during the infectious process. The presence of another pathogen appeared to improve the infectious potential of Brucella.

The researchers' new technique creates the light-emitting bacteria by introducing genes for fluorescent proteins into their genomes. The concept itself is not new, but the genetic "tool kit" developed by Kirby and Kang greatly streamlines the process by using easy-to-manipulate genes called transposons -- sometimes called jumping genes -- to quickly and safely label the bacteria.

Read more:

Yoon-Suk Kang, James E. Kirby. Promotion and Rescue of Intracellular Brucella neotomae Replication during Coinfection with Legionella pneumophila. Infection and Immunity, 2017; 85 (5): e00991-16 DOI: 10.1128/IAI.00991-16

Posted by Dr. Tim Sandle

Friday, 28 July 2017

Aseptic and Sterile Processing: Control, Compliance and Future Trends

The Aseptic and Sterile Processing: Control, Compliance and Future Trends is the most important text discussing aseptic and sterile manufacturing that has been published in the last decade! This text looks at both today and tomorrow in regard to these two vital processing procedures.

The book is edited by Tim Sandle and Ed Tidswell, for details and a special offer for ordering please see the flyer below.

Posted by Dr. Tim Sandle

New approach to battle drug-resistant bacteria

Researchers at University of Utah Health have developed a rapid screening method to identify beneficial pairs of existing FDA-approved drugs to combat multi-drug resistant (MDR) bacterial infections. The microbiologists analyzed a bacterial dataset consisting of 4,000 E. coli mutants grown in the presence of 100 FDA-approved drugs. Each mutant is missing a specific gene and interacts with each drug in a specific way, producing a unique chemical genetic signature.

The approach identified 14 drugs that could be paired synergistically. The most promising synergistic pairing in this study combined azidothymidine (AZT), one of the earliest drugs prescribed to treat HIV-AIDS, with floxuridine, a cancer drug with a similar chemical genetic signature as a commonly prescribed antibiotic ¾ trimethoprim.

AZT blocks DNA replication, while floxuridine, like the antibiotic trimethoprim, prevent bacteria from repairing their DNA. While many bacteria have developed resistance to trimethoprim, floxuridine remains effective, even against trimethoprim-resistant bacteria, because it works by another mechanism.

Several of the drugs identified in this study are not currently used as antibiotics, but produce an effective response against MDR bacteria. The synergistic drug pairing also has the potential to slow the evolution of resistance to the drug therapies.


Morgan A. Wambaugh, Viplendra P. S. Shakya, Adam J. Lewis, Matthew A. Mulvey, Jessica C. S. Brown. High-throughput identification and rational design of synergistic small-molecule pairs for combating and bypassing antibiotic resistance. PLOS Biology, 2017; 15 (6): e2001644 DOI: 10.1371/journal.pbio.2001644

Posted by Dr. Tim Sandle

Thursday, 27 July 2017

EMA issues guidance document in preparation for Brexit

The European Medicines Agency (EMA) and the European Commission have published guidance to help pharmaceutical companies prepare for the United Kingdom’s withdrawal from the European Union.

The question-and-answer document contains information around company location in the context of centralised procedures and certain activities, including the location of orphan designation holders, qualified persons for pharmacovigilance (QPPVs), and companies’ manufacturing and batch release sites. EMA is preparing a series of further guidance documents relating to the subject of Brexit that will be published on its website in due course.

See: EMA

(News via European Pharmaceutical Review)

Posted by Dr. Tim Sandle

Wednesday, 26 July 2017

Moist-heat sterilization of blood bags

The most suitable type of autoclaves for sterilizing blood bag systems are autoclaves in which sterilizing medium is a mixture of steam and air. This method, one of the counter pressure types of sterilization, is also called ‘air-over-steam’. It allows controlling the sterilization pressure independently of the temperature: this independence is impossible with conventional pure-steam autoclaves, as the temperature and pressure of pure saturated steam are linked by a one-to-one relation.

An article of interest: “This article provides basic information on the sterilization of blood bags systems by moist-heat. Problems of pressure compensation and steam penetration into the system parts without water inside, and the process choice between single and double autoclaving are discussed.”

For further details, see: Sterilize.IT

Posted by Dr. Tim Sandle

Tuesday, 25 July 2017

High prevalence of CRE in Washington, D.C.

Carbapenem-resistant Enterobacteriaceae (CRE), a family of highly pathogenic antibiotic-resistant organisms, are endemic across Washington, D.C. healthcare facilities, with 5.2 percent of inpatients testing positive for the bacteria, according to new research.

As part of the Healthcare Antibiotic Resistance Prevalence -- DC (HARP-DC) project, the three agencies partnered to conduct a multi-center study of CRE in D.C. Sixteen facilities voluntarily participated, including eight acute care hospitals, two long-term acute care hospitals, one inpatient rehabilitation facility, and five skilled nursing facilities. Each facility conducted bacterial colonization surveillance over a one-to-three-day interval from January to April, 2016. The researchers used the Centers for Disease Control and Prevention CRE surveillance definition and tested all cultures at a single laboratory to ensure consistency.

Of 1,022 completed tests, 53 samples tested positive for CRE, which corresponds to a prevalence rate of 5.2 percent, confirming that CRE has become endemic in healthcare facilities in Washington, D.C. The median prevalence rate by facility was 2.7 percent, with one facility measuring as high as 29.4 percent of tested patients, indicating the potential for hyperendemicity. Male patients had a significantly higher prevalence of CRE compared to females (7.1 percent vs. 3.7 percent). Adults ages 20-39 (8 percent) also showed higher prevalence than any other age group.

Of the positive samples, 18 were determined to share genetic similarity with at least one other sample. This revealed the potential transmission of CRE within and between facilities. The ability to determine similarity of strain profiles from culture and molecular testing also enabled the researchers to detect an ongoing outbreak in one facility, further demonstrating the utility of these laboratory techniques for surveillance programs.


Jacqueline Reuben, Nancy Donegan, Glenn Wortmann, et al Healthcare Antibiotic Resistance Prevalence – DC (HARP-DC): A Regional Prevalence Assessment of Carbapenem-Resistant Enterobacteriaceae (CRE) in Healthcare Facilities in Washington, District of Columbia. Infection Control & Hospital Epidemiology, 2017; 1 DOI: 10.1017/ice.2017.110

Posted by Dr. Tim Sandle

Monday, 24 July 2017

Chronic obstructive pulmonary disease

Antonella Chesca and Tim Sandle have written an article looking at chronic obstructive pulmonary disease.

Here is the abstract:

The present study refers to the exploration of the respiratory function of patients who presented acute symptoms of chronic obstructive pulmonary disease. In the medical specialty units, examination was performed using a standard chest X-ray imaging investigation; followed by a spirometry test, according to the patient's severity of symptoms, using the betamimetics test. Both the X-ray result and the imaging investigation of spirometry showed changes. Changes varied according to the patients from different investigate disease groups in relation to chronic obstructive pulmonary disease.

Here is the reference:

Cheşcă A., Cheşcă S. A., Sandle T. (2016) An approach on chronic obstructive pulmonary disease, Medicine and Ecology, 80 (3): 116-119

The publication can be accessed here.

Posted by Dr. Tim Sandle

Sunday, 23 July 2017

New antibiotic effective against drug-resistant bacteria

Scientists have discovered a new antibiotic effective against drug-resistant bacteria: pseudouridimycin. The new antibiotic is produced by a microbe found in a soil sample collected in Italy and was discovered by screening microbes from soil samples. The new antibiotic kills a broad spectrum of drug-sensitive and drug-resistant bacteria in a test tube and cures bacterial infections in mice.

Pseudouridimycin inhibits bacterial RNA polymerase, the enzyme responsible for bacterial RNA synthesis, through a binding site and mechanism that differ from those of rifampin, a currently used antibacterial drug that inhibits the enzyme. Because pseudouridimycin inhibits through a different binding site and mechanism than rifampin, pseudouridimycin exhibits no cross-resistance with rifampin, functions additively when co-administered by rifampin and, most important, has a spontaneous resistance rate that is just one-tenth the spontaneous resistance rate of rifampin.

Pseudouridimycin functions as a nucleoside-analog inhibitor of bacterial RNA polymerase, meaning that it mimics a nucleoside-triphosphate (NTP), the chemical "building block" that bacterial RNA polymerase uses to synthesize RNA. The new antibiotic binds tightly to the NTP binding site on bacterial RNA polymerase and, by occupying the NTP binding site, prevents NTPs from binding.

Pseudouridimycin is the first nucleoside-analog inhibitor that selectively inhibits bacterial RNA polymerase but not human RNA polymerases.


Sonia I. Maffioli, Yu Zhang, David Degen, Thomas Carzaniga, Giancarlo Del Gatto, Stefania Serina, Paolo Monciardini, Carlo Mazzetti, Paola Guglierame, Gianpaolo Candiani, Alina Iulia Chiriac, Giuseppe Facchetti, Petra Kaltofen, Hans-Georg Sahl, Gianni Dehò, Stefano Donadio, Richard H. Ebright. Antibacterial Nucleoside-Analog Inhibitor of Bacterial RNA Polymerase. Cell, 2017; 169 (7): 1240 DOI: 10.1016/j.cell.2017.05.042

Posted by Dr. Tim Sandle

Saturday, 22 July 2017

Bacteria from hot springs solve mystery of metabolism

Combustion is often a rapid process, like fire. How can our cells control the burning process so well? The question has long puzzled researchers. Using bacteria from hot springs, researchers from Stockholm University now have the answer.

Light a match and place it near a candle. You would see a fire and feel the heat when the stearin is burning, while consuming oxygen from air and converting the fuel to carbon dioxide and water. However, when our body burns fat, sugar or protein containing the same amount of energy, we do not vanish into fire and smoke, but use the energy for moving our muscles or for thinking. How does our body control the burning process so well? Researchers at Stockholm University have finally been able to monitor the process and to uncover the mechanism.
"We have shown how oxygen is combusted after it has been transported by blood to our cells. We have also shown how the combustion of oxygen provides energy, for example, for muscle contraction or to generate electricity in our nerve cells," says Peter Brzezinski, Professor at the Department of Biochemistry and Biophysics, Stockholm University.

The combustion of oxygen in our cells takes place in the so-called respiratory chain that carefully controls the process. Electrons, which come from digestion, are transferred to the oxygen we breathe. The oxygen molecules bind to an enzyme in our mitochondria, the cellular power plant. However, the bound oxygen is not immediately combusted to form water, like in an uncontrolled fire, but it is converted to water gradually in a carefully controlled process. Up until now we only had a very basic knowledge about the mechanism of this process, since the reaction is too rapid too be studied using available techniques. One possibility would be to follow the reactions at low temperatures, at about -50 degrees Celsius, where they would be sufficiently slow. However, this is not practically possible.

In this project, researchers Federica Poiana and Christoph von Ballmoos studied oxygen combustion in a bacterium that lives in hot springs -- they thrive in almost boiling water. When the research group performed their studies at 10 degrees, the bacterium found it extremely cold -- as if our mitochondria were exposed to minus 40 degrees. The reactions then became sufficiently slow to allow studies using available instruments. By combining their experimental studies with theoretical calculations, the researchers could translate their observations to the equivalent processes in human cells.

"In addition to just being curious and wanting learn how the process works, our studies are also motivated by trying to understand the so-called Mitochondrial diseases, caused by malfunction in oxygen combustion," says Peter Brzezinski.


Federica Poiana et al. Splitting of the O–O bond at the heme-copper catalytic site of respiratory oxidases. Science Advances, June 2017 DOI: 10.1126/sciadv.1700279

Posted by Dr. Tim Sandle

Friday, 21 July 2017

New health and safety standard

A new International Standard due out early next year – ISO 45001. This is ISO’s first standard for occupational health and safety management systems.

According to ISO: “ISO 45001, Occupational health and safety management systems – Requirements with guidance for use, will provide the requirements for implementing a management system and framework that reduces the risk of harm and ill health to employees.

The standard is being developed by a committee of occupational health and safety experts and will follow in the footsteps of ISO’s other management systems approaches, such as ISO 14001 (environment) and ISO 9001 (quality). It will take into account other International Standards in this area including OHSAS 18001, the International Labour Organization’s ILO-OSH Guidelines, various national standards and the ILO’s international labour standards and conventions.”

For further details see: ISO

Posted by Dr. Tim Sandle

Thursday, 20 July 2017

European Pharmacopoeia revises Biological Indicator Chapter

The general chapter in the European Pharmacopoeia (5.1.2) relating to biological indicators has undergone a significant revision. The new version has been issued with the release of supplement 9.2 to the 9th edition of the pharmacopoeia (1). The changes to the chapter are signaled by the revision to the title, which now becomes “Biological indicators and related microbial preparations used in the manufacture of sterile products”. Previously the chapter was headed “Biological indicators of sterilization.” The change of title reflects the adaption of the chapter to take into account microbial preparations used for sterilization grade filtration. This article assesses the key changes.

The reference is:

Sandle, T. (2017) European Pharmacopoeia revises Biological Indicator Chapter, GMP Review, 16 (1): 4-6

For a copy, please contact Tim Sandle

Posted by Dr. Tim Sandle

Wednesday, 19 July 2017

What's that smell?

Did you know that bacteria make us humans and other animals smell the way we do? On top of that, animals use that smell to communicate with each other.

The “fermentation hypothesis of chemical recognition” says that bacteria in the scent glands of mammals generate metabolites with specific odors that animals use to communicate with each other. What’s more, this hypothesis explains how variations in these chemical signals are actually due to variations in those populations of bacteria.

News from Laboratory Roots:

Let’s start with humans, Corynebacterium is responsible for that distinct “body odor” that emanates from our armpits. That odor is caused by 3-methyl-2-hexenoic acid (3M2H) and 3-hydroxy-3-methylhexanoic acid (HMHA). It turns out that the precursors for these chemicals are cleaved by a zinc-dependent bacterial aminoacylase.

In one study, researchers isolated bacteria from the human axilla. They identified 19 strains of Corynebacterium and 25 strains of Staphylococcus. Curiously, only isolates of Corynebacterium, not Staphylococcus, produced 3M2H or HMHA from the precursors 3M2H-Gln or HMHA-Gln, indicating that these strains produced an Nα-acylglutamine aminoacylase.

So, what’s the use of human body odor? It’s safe to say that most people think body odor smells bad - it’s definitely not attractive to other humans. Some researchers think that may be the point - or at least it was at some point in our evolution. Body odor may have been used by early humans as a way to assert dominance or repel rivals.

One of these odor chemicals is trimethylamine, which is specific for the olfactory receptor TAAR5. Trimethylamine synthesis requires two steps, one of which involves bacteria. We’ve all smelled trimethylamine before, it’s the stinky smell of bad breath and spoiled food!

In humans, trimethylamine is the byproduct of bacteria metabolizing dietary choline, and researchers wanted to know if the same held true for mice. To figure things out, they collected urine from mice that were fed a choline/methionine-free diet or that were treated with an antibiotic. Sure enough, these mice produced less trimethylamine in their urine than control mice.

Likewise, urine from wild type mice contained trimethylamine that activated its receptor TAAR5 (assayed with a reporter gene), but urine from mice on the choline-free diet or treated with antibiotics did not activate TAAR5. These findings suggest that bacteria produce trimethylamine from dietary choline.

While trimethylamine is produced by commensal bacteria, pathogens produce their own array of odors. Mice have receptors in their vomeronasal organ that recognize formylated peptides that are produced by tissue damage or bacterial infection. There are also chemosensory cells in the respiratory epithelium that detect bacterial quorum sensing molecules called acyl-homoserine lactones.

Next are the meerkats. These members of the mongoose family are native to South Africa. They produce a smelly paste from a pouch beneath their tails. They use this paste to mark their territories, applying it to plants, rocks, and their meerkat pals. Researchers found over 1,000 types of bacteria and some 220 odorous chemicals in the stinky paste. The key finding was that specific odors are produced by specific microbial communities - a specific meerkat family smells the way it does because of its own specific microbes.

The group detected five main phyla of bacteria in the meerkats’ anal pouch - Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, and Fusobacteria. Specific genera of bacteria found in the paste were Porphyromonas, Fusobacterium, Anaerococcus, and Campylobacter.

The researchers also found that subordinate and dominant male meerkats had different bacterial communities in their pouches. However, there were no significant differences among dominant and subordinate females.

Finally, like meerkats, hyenas, deposit a scented paste from anal scent pouches to communicate with other hyenas. In one study, researchers used scanning electron microscopy to look for bacteria in the scent pouches of both spotted and striped hyenas (two groups that differ rather significantly in their lifestyles and behavior).

They found that the bacterial communities differed between spotted and striped hyenas, but all communities were largely made up of fermentative anaerobes - specifically, species from the order Clostridiales. For the spotted hyenas, they identified Clostridiales from the genera Anaerococcus, Clostridium, Fastidiosipila, Finegoldia, Murdochiella, Peptoniphilus, and Tissierella. The bacterial communities differed, however, based on sex and female reproductive status.

Tuesday, 18 July 2017

Genetic Data for 1000 Microbes Released

Microbial organisms are a major component of our environment, but scientists have only begun to study their genetic composition in depth. Using current genetic technologies, researchers with the U.S. Department of Energy Joint Genome Institute (DOE JGI) have begun to learn more, and have released their findings. The team has reported 1,003 phylogenetically reference genomes of bacterial and archaeal organisms in Nature Biotechnology.

Genome sequencing and analysis data for 1,003 genomes is now available through the Integrated Microbial Genomes with Microbiomes (IMG/M) system. The DOE JGI is aiming to provide interested scientists with a trove of new sequence data. It could aid in the characterization of biotechnologically relevant secondary metabolites or reveal more about enzymes that act under certain conditions.

According to the Nature paper:

"We present 1,003 reference genomes that were sequenced as part of the Genomic Encyclopedia of Bacteria and Archaea (GEBA) initiative, selected to maximize sequence coverage of phylogenetic space. These genomes double the number of existing type strains and expand their overall phylogenetic diversity by 25%. Comparative analyses with previously available finished and draft genomes reveal a 10.5% increase in novel protein families as a function of phylogenetic diversity. The GEBA genomes recruit 25 million previously unassigned metagenomic proteins from 4,650 samples, improving their phylogenetic and functional interpretation. We identify numerous biosynthetic clusters and experimentally validate a divergent phenazine cluster with potential new chemical structure and antimicrobial activity. This Resource is the largest single release of reference genomes to date. Bacterial and archaeal isolate sequence space is still far from saturated, and future endeavors in this direction will continue to be a valuable resource for scientific discovery."

Posted by Dr. Tim Sandle

Monday, 17 July 2017

Pathological appendix versus normal appendix

Antonella Chesca and Tim Sandle have written a paper of interest to those working in pathology or medicine. It concerns the pathological appendix compared with the normal appendix.

Here is the abstract:

This article makes reference to certain structural issues, that can be found during the appendix surgery and which are examined microscopically. For the study permanent preparations resulting from appendectomies, charged for removal in cases of gangrenosum appendicitis, were examined microscopically. For comparison and analytically, describable microscopic permanent preparations with gangrenosum appendix, were compared with normal appendix. Stains used in this structural study are Hematoxylin-Eosin, van Gieson and Gomori silver impregnation. Preparations were examined using Nikon microscope and magnifying lenses with powers x10, x20 and x40.

The reference is:

Chesca, A. and Sandle, T. (2017) A new approach related to structural aspects of pathological appendix versus normal appendix, Medicine and Ecology, 82 (1): 115-118

The article can be accessed here.

Posted by Dr. Tim Sandle