Saturday, 28 March 2015

European Pharmacopeia updates

Some updates of interest in relation to the European Pharmacopeia:

European Pharmacopoeia 8th Edition - Supplement 8.4 (implementation date 1st April 2015)

General Chapters

The following general chapters are being revised: Materials based on plasticised poly(vinyl chloride) for containers for human blood and blood components based on plasticised poly(vinyl chloride) for tubing used in sets for the transfusion of blood and blood components

3.1.10  Materials based on non-plasticised poly(vinyl chloride) for containers for non-injectable, aqueous solutions

3.1.14  Materials based on plasticised poly(vinyl chloride) for containers for aqueous solutions for intravenous infusions

3.2.1    Glass containers for pharmaceutical use                                            

It has been decided to further emphasise the need for control of specific components that may be toxic for chronic use and for vulnerable patient groups. The statements in the definition section of Parenteral preparations (0520), in chapter 3.2.1. Glass containers for pharmaceutical use and in chapter 3.2.2. Plastic containers and closures for pharmaceutical use have therefore been supplemented.

5.12     Reference standards                                                                          

General revision to update chapter to reflect recent advances in pharmaceutical reference standards, including harmonisation with the specifications and definitions of the corresponding ISO Guides (i.e. ISO Guides 30 and 34) and introduction of:

- the definition of international standards based on WHO Technical Report Series 932,
- herbal reference standards (HRS),
- biological reference preparations (BRP) and chemical reference substances for biologicals.

Posted by Tim Sandle

Friday, 27 March 2015

Porphyrin synthesis test explained

The porphyrin synthesis test is used to identify haemin producing Haemophilus species. It avoids the problems of red cell carry over associated with tests for X and V dependence. The porphyrin test is considered to be the definitive method for the differentiation of Haemophilus species.

Haemophilus species are not readily distinguishable by their colonial morphology or gram stain appearance. Oral flora grown from routine sputum cultures often contains organisms which resemble Haemophilus species. H. influenzae, which is the principal human pathogen, can be distinguished from other Haemophilus species and oral flora by determining the need for essential factors for growth, specifically Haemin (X factor) and Nicotinamide-adenine dinucleotide (NAD/V factor). H. influenzae requires both factors for growth where as some of the other species require only one. The requirement for one or both of the growth factors nicotinamide adenine dinucleotide (NAD or V factor) and haemin (X factor) is used to characterise Haemophilus species.

Strains which produce their own haemin possess the enzyme porphobilinogen synthase which can convert d-aminolaevulinic acid (ALA) to protoporphyrin and ultimately haemin.
This test demonstrates the ability of a bacterium supplied with d-aminolaevulinic acid to synthesise and excrete porphobilinogen and other porphyrins, indicating that they are not X dependent.

In relation to the porphyrin synthesis test, Public Health England has issued a technical report, including safety information. The report can be accessed here.

Posted by Tim Sandle

Thursday, 26 March 2015

Proteins pull together as cells divide

Cell division is how new cells form, both during development and throughout an organism's life.  Successful cell division requires the formation of a dip called a cleavage furrow, a process that has remained complex. Now, researchers have found that no single molecular architect directs the cleavage furrow's formation; rather, it is a robust structure made of a suite of team players.

For information about the new research, see:

Vasudha Srivastava, Douglas N. Robinson. Mechanical Stress and Network Structure Drive Protein Dynamics during Cytokinesis. Current Biology, 2015; DOI: 10.1016/j.cub.2015.01.025

Posted by Tim Sandle

Wednesday, 25 March 2015

The first X-ray portraits of living bacteria

Researchers have captured the first X-ray portraits of living bacteria. This milestone is a first step toward possible X-ray explorations of the molecular machinery at work in viral infections, cell division, photosynthesis and other processes that are important to biology, human health and our environment.

The experiment took place at SLAC's Linac Coherent Light Source (LCLS) X-ray laser, a DOE Office of Science User Facility. The experiment focused on cyanobacteria, or blue-green algae. The cyanobacteria were passed into an ultrabright, rapid-fire LCLS X-ray pulses, producing diffraction patterns recorded by detectors.

The diffraction patterns preserved details of the living cyanobacteria that were compiled to reconstruct 2-D images. Researchers said it should be possible to produce 3-D images of some samples using the same technique.

The technique works with live bacteria and requires no special treatment of the samples before imaging. Other high-resolution imaging methods may require special dyes to increase the contrast in images, or work only on dead or frozen samples.

The technique can capture about 100 images per second, amassing many millions of high-resolution X-ray images in a single day. This speed allows sorting and analysis of the inner structure and activity of biological particles on a massive scale, which could be arranged to show the chronological steps of a range of cellular activities.

van der Schot, G. et al: For further details see: Imaging single cells in a beam of live cyanobacteria with an X-ray laser. Nature Communications, 2015; 6: 5704 DOI: 10.1038/ncomms6704

Posted by Tim Sandle

Tuesday, 24 March 2015

New insight into Yersinia pestis

Researchers discover that the accepted theory of how Yersinia pestis bacteria travel from fleabite to lymph node is off base. Most bacteria get trapped in a bottleneck and never make it to the lymph node, where infection takes root. Finding out why could lead to new ways to stop the pathogen.

For details, see:

Rodrigo J. Gonzalez, M. Chelsea Lane, Nikki J. Wagner, Eric H. Weening, Virginia L. Miller. Dissemination of a Highly Virulent Pathogen: Tracking The Early Events That Define Infection. PLOS Pathogens, 2015; 11 (1): e1004587 DOI: 10.1371/journal.ppat.1004587## Posted by Tim Sandle

Monday, 23 March 2015

Italian cemetery provides clues on cholera's evolution

A team of archaeologists and other researchers hope that an ancient graveyard in Italy can yield clues about the deadly bacterium that causes cholera.

Researchers are excavating the graveyard surrounding the abandoned Badia Pozzeveri church in the Tuscany region of Italy. The site contains victims of the cholera epidemic that swept the world in the 1850s, said Clark Spencer Larsen, professor of anthropology at The Ohio State University and one of the leaders of the excavation team.

Archaeologists have spent the past four summers excavating remains in a special section of the cemetery used for cholera victims. Finding traces of the pathogen that caused cholera among the human remains could reveal details about how people lived -- and died -- in this region of Europe.

The bodies of the cholera victims were hastily buried and covered in lime, which hardened like concrete around the bodies. Researchers suspect residents were trying to keep the disease from spreading.

The researchers are of the view that if they find the DNA we could see how cholera has evolved and compare it to what the bacteria is like today. That's the first step to possibly finding a cure.

For further details see: Ohio State University.

Posted by Tim Sandle

Sunday, 22 March 2015

Bio Products Laboratory at 60

The U.K.'s Bio Products Laboratory (BPL) recently celebrated 60 years of operating. You can read about the plasma fractional plant's history here.

Posted by Tim Sandle

Saturday, 21 March 2015

The remarkable Zymomonas mobilis

The bacterium Zymomonas mobilis can convert atmospheric nitrogen gas into ammonium, researchers at the University of Indiana have found. Their results were published in PNAS this week (February 2).

“It was known already that the Zymomonas genome contains all the genes that are needed, but nobody had checked whether they really are able to fix nitrogen,” said Uldis Kalnenieks of the University of Latvia who was not involved in the research.

“The genes might have been there and annotated in the past, but this [study] now enables perhaps the industrial side to develop it,” said Steve Brown of Oak Ridge National Laboratory in Tennessee who also was not involved in the work. “This opens the door to further studies in this system so I think that’s a pretty exciting result.”

For further details, see The Scientist

Posted by Tim Sandle

Friday, 20 March 2015

Optochin test explained.

Susceptibility to optochin is a simple and reliable method of differentiating Streptococcus pneumoniae from other alpha-haemolytic streptococci.

Optochin is a chemical, ethylhydrocupreine hydrochloride and is completely soluble in water. The optochin test detects an organism’s susceptibility to the chemical optochin. The chemical tests the fragility of the bacterial cell membrane and causes S. pneumoniae to lyse due to changes in surface tension.

The optochin test is widely used in the form of filter paper discs, impregnated with ethylhydrocupreine hydrochloride, which are applied directly to inoculated plates before incubation.

The optochin test is less time-consuming than the bile solubility test.

In relation to the optochin test, Public Health England has issued a technical report, including safety information. The report can be accessed here.

Posted by Tim Sandle

Thursday, 19 March 2015

ISO 11133:2014

A new ISO standard of interest to those involved with culture media use:

ISO 11133:2014  - Microbiology of food, animal feed and water -- Preparation, production, storage and performance testing of culture media

ISO 11133:2014 defines terms related to quality assurance of culture media and specifies the requirements for the preparation of culture media intended for the microbiological analysis of food, animal feed, and samples from the food or feed production environment as well as all kinds of water intended for consumption or used in food production. These requirements are applicable to all categories of culture media prepared for use in laboratories performing microbiological analyses.

ISO 11133:2014 also sets criteria and describes methods for the performance testing of culture media. It applies to producers such as:
  • commercial bodies producing and/or distributing ready-to-use or semi-finished reconstituted or dehydrated media;
  • non-commercial bodies supplying media to third parties;
  • microbiological laboratories preparing culture media for their own use.

The revised standard replaces ISO/TS 11133-1:2009.#

Posted by Tim Sandle

Wednesday, 18 March 2015

Link found in how cells start process necessary for life

Researchers have found an RNA structure-based signal that spans billions of years of evolutionary divergence between different types of cells, according to a new study. The finding could alter the basic understanding of how two distinct life forms -- bacteria and eukaryotes -- begin the process of protein synthesis.

Scientists have long thought that the molecular signals that initiate protein synthesis in bacteria and eukaryotes are mutually exclusive. Scientists in Kieft's lab explored whether a structured RNA molecule from a virus that infects eukaryotic cells could function in bacteria. Surprisingly, they found that it could initiate protein syntheses, a process necessary for life.

For details see:

Timothy M. Colussi, David A. Costantino, Jianyu Zhu, John Paul Donohue, Andrei A. Korostelev, Zane A. Jaafar, Terra-Dawn M. Plank, Harry F. Noller, Jeffrey S. Kieft. Initiation of translation in bacteria by a structured eukaryotic IRES RNA. Nature, 2015; DOI: 10.1038/nature14219

Posted by Tim Sandle

Tuesday, 17 March 2015

Potassium Hydroxide Test explained

Many Bacillus and Clostridium species organisms that have lost some of the integrity of their cell wall, appear Gram negative on staining resulting in possible misidentification.

The potassium hydroxide test may aid in differentiation between Gram positive and Gram negative organisms and is a useful complement to the Gram stain and the antibiotic disc test. Like the Gram stain reaction, the test is based on differences in the chemistry of the bacterial cell wall.

In the presence of potassium hydroxide, Gram negative cell walls are broken down, releasing viscid chromosomal material which causes the bacterial suspension to become thick and stringy. Gram positive organisms remain unaffected. Hence the alternative name for this procedure, the “String Test”.

In relation to the potassium hydroxide test, Public Health England has issued a technical report, including safety information. The report can be accessed here.

Posted by Tim Sandle

Monday, 16 March 2015

Dracunculiasis and the Long Decline of an Ancient Disease

The disease dracunculiasis (guinea worm disease) has ravaged human populations for thousands of years (reference to the disease is documented in the Egyptian medical Ebers Papyrus, dating from around 1550 BC.) Current indications suggest that global incidences of the disease have been rapidly declining due to the concerted efforts of national and international health agencies. Here, only 148 dracunculiasis cases were reported worldwide in 2013 (which represents the lowest annual total ever recorded) and only four endemic countries remain: Chad, Ethiopia, Mali and South Sudan. With these countries, the majority of the cases occur in South Sudan . Nonetheless across these regions the number of endemic villages has declined from the peak of 23,735 in 1991 to 79 in 2013.

Guinea-worm disease is caused by the parasitic worm Dracunculus medinensis or "Guinea-worm". This worm is the largest of the tissue parasite affecting humans. The adult female, which carries about 3 million embryos, can measure 600 to 800 mm in length and 2 mm in diameter. The parasite migrates through the victim's subcutaneous tissues causing severe pain especially when it occurs in the joints. The worm eventually emerges (from the feet in most of the cases), causing an intensely painful oedema, a blister and an ulcer accompanied by fever, nausea and vomiting.
To find out more about this disease and why a decline has occurred, Tim Sandle has written a review paper for the journal Journal of Ancient Diseases & Preventive Remedies.

The reference is:

To view a copy, please contact Tim Sandle.

Sunday, 15 March 2015

ICH Q3D Guideline on Elemental Impurities

The ICH Q3D Guideline on Elemental Impurities reached Step 4 of the ICH Process in December 2013 and now enters the implementation period (Step 5). This new guidance has been developed to provide a global policy for limiting metal impurities qualitatively and quantitatively in drug products and ingredients. The existing ICH Q3A Guideline classifies impurities as organic, inorganic, and residual solvents. The Q3A and Q3B Guidelines effectively address the requirements for organic impurities. An additional Guideline Q3C was developed to provide clarification of the requirements for residual solvents. The new Q3D Guideline would provide similar clarification of the requirements for metals, which are included in the ICH inorganic impurities classification.

For further details, see: Q3D guideline

Posted by Tim Sandle

Saturday, 14 March 2015

FDA - cGMP requirements for combination products

The FDA has issued a new draft guidance.

This guidance describes and explains the final rule on CGMP requirements for combination products. Prior to issuance of the final rule, although CGMP regulations were in place to establish requirements for drugs, devices, biological products, and Human Cells, Tissues, and Cellular and Tissue-Based Products (HCT/Ps), there were no regulations to clarify and explain the application of these CGMP requirements to combination products. The final rule was intended to provide such clarification and specify how compliance with applicable CGMP requirements may be demonstrated.

Section II of the guidance provides the definition of a combination product, an overview of the final rule, and the role of the lead center and other agency components with respect to combination product CGMP issues.

Section III addresses certain general considerations for CGMP compliance for combination products, and Section IV presents the purpose and content of specific CGMP requirements addressed in the final rule, and Section V analyzes hypothetical scenarios intended to clarify how to comply with certain CGMP requirements addressed in the final rule. Throughout this guidance, the agency also refers to existing guidance and additional sources of information that address CGMP requirements for drugs, devices, biological products, and HCT/Ps, to further inform combination product manufacturers on how to comply with CGMP requirements.

For further details, see FDA.

Posted by Tim Sandle