Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS)-Based Biowaivers: A workshop Summary Report.

The workshop "Drug Permeability - Best Practices for Biopharmaceutics Classification System (BCS) Based Biowaivers" was held virtually on December 6, 2021, organized by the University of Maryland Center of Excellence in Regulatory Science and Innovation (M-CERSI), and the Food and Drug Administration (FDA). The workshop focused on the industrial, academic, and regulatory experiences in generating and evaluating permeability data, with the aim to further facilitate implementation of the BCS and efficient development of high-quality drug products globally. As the first international permeability workshop since the BCS based biowaivers was finalized as the ICH M9 guideline, the workshop included lectures, panel discussions, and breakout sessions. Lecture and panel discussion topics covered case studies at IND, NDA, and ANDA stages, typical deficiencies relating to permeability assessment supporting BCS biowaiver, types of evidence that are available to demonstrate high permeability, method suitability of a permeability assay, impact of excipients, importance of global acceptance of permeability methods, opportunities to expand the use of biowaivers (e.g. non-Caco-2 cell lines, totality-of-evidence approach to demonstrate high permeability) and future of permeability testing. Breakout sessions focused on 1) in vitro and in silico intestinal permeability methods; 2) potential excipient effects on permeability and; 3) use of label and literature data to designate permeability class.

Plastic waste recycling: existing Indian scenario and future opportunities.

This review article aims to suggest recycling technological options in India and illustrates plastic recycling clusters and reprocessing infrastructure for plastic waste (PW) recycling in India. The study shows that a majority of states in India are engaged in recycling, road construction, and co-processing in cement kilns while reprocessing capabilities among the reprocessors are highest for polypropylene (PP) and polyethylene (PE) polymer materials. This review suggests that there are key opportunities for mechanical recycling, chemical recycling, waste-to-energy approaches, and bio-based polymers as an alternative to deliver impact to India's PW problem. On the other hand, overall, polyurethane, nylon, and polyethylene terephthalate appear most competitive for chemical recycling. Compared to conventional fossil fuel energy sources, polyethylene (PE), polypropylene (PP), and polystyrene are the three main polymers with higher calorific values suitable for energy production. Also, multi-sensor-based artificial intelligence and blockchain technology and digitization for PW recycling can prove to be the future for India in the waste flow chain and its management. Overall, for a circular plastic economy in India, there is a necessity for a technology-enabled accountable quality-assured collaborative supply chain of virgin and recycled material. Supplementary Information: The online version contains supplementary material available at 10.1007/s13762-022-04079-x.

Angular and impact energy dependence of intensity ratio of Kα,ß x-rays to underneath bremsstrahlung radiation emitted from 15 to 25 keV electrons incident on a pure thick Cu (Z=29) target.

We present new results on angular and impact energy dependence of the relative intensity ratios of Kα and Kß characteristic (CS) x-ray lines to their respective underneath bremsstrahlung (BS) radiation produced from 15-25 keV electrons incident on a thick polycrystalline pure target of Cu (Z = 29). The angular distributions of relative intensity ratios were carried out by measuring the radiations emitted at incidence angle θi that varied from 15° to 75° in reflection geometry mode of the target. The x-rays emerging from the target surface were detected by an energy dispersive Si PIN photodiode detector. The magnitude of relative intensity ratio of Kα/BS is found to be larger than that of Kß/BS by a factor of about 5.11, 5.24 and 5.49 for 15 keV, 20 keV and 25 keV impact energy, respectively measured, for example, at θi = 15°. However, the intensity ratio of a given characteristic line to its underneath BS increases on average by a factor of about 2.2 in varying the impact energy from 15 keV to 25 keV measured at different angles. The experimental results on angular and impact energy dependence are compared with the corresponding results obtained from Monte-Carlo simulations using the PENELOPE code. The overall agreement between experiment and theory is found to be satisfactory within the uncertainty of measurements.

Impact of Stress on Type 2 Diabetes Mellitus Management.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is one among the major health and socioeconomic problems worldwide. It is, however, not a somatic illness for which just symptomatic treatment will suffice. Stress is an important factor in not only causing diabetes onset or exacerbation, but also in hampering proper treatment by interfering with the treatment adherence of patients. Hence, it becomes important for physicians to acquaint themselves with the effects of stress on T2DM in order to ensure proper treatment of the latter. OBJECTIVE: Documentation of effect of stress on the management of T2DM. SUBJECTS AND METHODS: The research was a cross-sectional study on the patients attending Sri Muthukumaran Medical College, Hospital and Research Institute, Mangadu. A total of 400 people, who werepre-established diabetic patients of the hospital of age greater than 30 years, were chosen for the study. The stress levels of the patients were assessed with the Perceived Stress Scale (PSS) and treatment adherence using a questionnaire prepared exclusively for the study. Based on the data, a statistical relationship was framed between the degree of control (treatment adherence) and the stress levels of the patients. RESULTS: The FBS levels were a direct reflection of the stress levels (P<0.05). Stress had a major impact on treatment adherence among the diabetic subjects: Increased levels of stress decreased the adherence (P<0.001). The glycemic index (HbA1C level) was found to be linked to both treatment adherence and stress. Increased adherence kept it at bay (P<0.05) while stress proved abysmal to glycemic control. CONCLUSION: T2DM is the result of an interplay between various factors; environmental, psychiatric and somatic. Hence, a holistic treatment approach is required, one that involves stress management, education and mental health awareness along with pharmacological treatment, to fully control the disease.

Effects of different stocking densities on performance and activity of cattle × yak hybrids along a transhumance route in the Eastern Himalaya.

Twelve lactating cattle × yak hybrids (B. taurus × B. grunniens) were investigated on five experimental pastures (Sites 1-5), following an up- and downward transhumance route (3,200, 4,000, 4,500, 4,000 and 2,600 m a.s.l.). Hybrids were kept in replicated groups of two (low SD) and four animals per paddock (high SD). As a control, yaks (with calves) were included at low SD at Sites 2-4. Performance was described by body weight, milk yield and composition. Herbage samples as selected by the animals were analyzed. Activity patterns of the hybrids were assessed at Sites 2-4. At similar body weight change and milk composition, the individual hybrids on average produced 26% less milk at high than at low SD. However, at high SD on average still 49% more milk/m(2) of pasture area was produced. Milk fat increased with time from 5.4 to 7.1%, milk protein decreased from 4.2 to 3.3%. At high SD, the hybrids spent less time standing and more time lying. The yaks gained weight at 4,500 m a.s.l. while the hybrids lost weight (+836 and -653 g/animal/day, respectively). In conclusion, high SD reduced individual milk yield and forced hybrids to spend more time for feeding. The yaks apparently had an advantage over the hybrids at very high altitude.

Isolation and classification of a soil actinomycete capable of sulphur-specific biotransformation of dibenzothiophene, benzothiophene and thianthrene.

AIM: To isolate actinomycete spp with the ability to desulphurize sulphur-containing heterocyclic compounds present in petroleum. METHODS AND RESULTS: Enrichment cultures were set up to select and isolate sulphur heterocycle metabolizing soil micro-organisms. Screening of the microbial isolates for the desulphurization property led to isolation of R3. The isolate was characterized by PCR screening of 16S rRNA genes and classical taxonomic investigations. HPLC analysis of the desulphurization assays with R3 showed ~85% transformation of dibenzothiophene (270 µmol l(-1)), present as the sole sulphur source in basal salt medium, in 4 days. Production of the desulphurized dibenzothiophene metabolite, 2-hydroxybiphenyl, was confirmed by GC/MS analyses. GC/MS analyses also established the ability of R3 to transform benzothiophene to benzothiophene-1-oxide and benzothiophene-1, 1-dioxide, and thianthrene to thianthrene-5-oxide. PCR primers computed based on the desulphurization operon (dszABC) of Rhodococcus erythropolis IGTS8 yielded the predicted amplification products with R3 genomic DNA as template. Southern hybridization and restriction endonuclease digestion profiles indicated that R3 amplicons were homologous to dsz AB. CONCLUSIONS: The enrichment method used in this study yielded an environmental isolate with the ability to transform multiple sulphur heterocycles. The isolate R3 has taxonomic proximity to the Oerskovia sp, order Actinomycetales. The isolate R3 selectively removes sulphur from dibenzothiophene yielding 2-hydroxybiphenyl and sulphate. R3 also transforms benzothiophene and thianthrene in a sulphur-targeted manner. The desulphurization genes in R3 bear similarity to those in R. erythropolis IGTS8. SIGNIFICANCE AND IMPACT OF THE STUDY: The actinomycetes present in soil can remove sulphur from different sulphur heterocycle substrates and have potential as biodesulphurization catalysts.

Degradation of dibenzothiophene and its metabolite 3-hydroxy-2-formylbenzothiophene by an environmental isolate.

Microbial degradation of dibenzothiophene (DBT) beyond 3-hydroxy-2-formylbenzothiophene (HFBT), a commonly detected metabolite of the Kodama pathway for DBT metabolism, and the catabolic intermediates leading to its mineralization are not fully understood. The enrichment cultures cultivated from crude oil contaminated soil led to isolation of ERI-11; a natural mixed culture, selected for its ability to deplete DBT in basal salt medium (BSM). A bacterial strain isolated from ERI-11, and tentatively named A11, degraded more than 90 % of the initial DBT (270 µM), present as the sole carbon and sulfur source, in 72 h. Gas chromatography-mass spectrophotometry (GC-MS) analyses of the DBT degrading A11 culture medium extracts led to detection of HFBT. The metabolite HFBT, produced using A11, was used in degradation assays to evaluate its metabolism by the bacteria isolated in this study. Ultra violet-visible spectrophotometry and high-performance liquid chromatography analyses established the ability of the strain A11 to deplete HFBT, present as the sole sulfur and carbon source in BSM. GC-MS analyses showed the presence of 2-mercaptobenzoic acid in the HFBT degrading A11 culture extracts. The findings in this study establish that the environmental isolate A11 possesses the metabolic capacity to degrade DBT beyond the metabolite HFBT. The compound 2-mercaptobenzoic acid is an intermediate formed on HFBT degradation by A11.

Contamination of potable water by enterotoxigenic Escherichia coli: qPCR based culture-free detection and quantification.

Tourists visiting to endemic zones may acquire Enterotoxigenic Escherichia coli (ETEC) infection resulting into diarrhea due to consumption of contaminated potable waters. In this study, a qPCR assay (SYBR Green), targeting LT1 and ST1 genes was designed to quantify ETEC in potable waters derived from civic water supply. The assay could detect lowest 1CFU/PCR targeting LT1/ST1 gene from ten-fold diluted culture of the reference strain (E. coli MTCC 723) and is ten-fold more sensitive than the conventional PCR. The quantification of the ETEC in potable waters collected from civic supply of a major city of the northern India exhibiting high flow of tourists reveals that all the sites that ran along sewage line were contaminated by the ETEC. Contamination was due to percolation of sewage. The assay could be used for the regular monitoring of potable water in places exhibiting heavy flow of tourists to prevent ETEC induced diarrhea.

Study of K-line radiation of thick titanium produced in collisions of keV electrons.

The characteristic K-line yields Y(E0) of a pure thick titanium (Z=22) element target are measured for 8-18 keV electron impact and compared with the simulation calculations using PENELOPE code. A fair agreement between experiment and simulation results is found within the existing experimental uncertainty of measurements. The ratio F of indirectly produced characteristic Ti K X-ray yield to its total (directly+indirectly) yield is determined by employing an approximate analytical formulation of Hanson and Cowan (Hanson, H.P., Cowan, D.J., 1961. Phys. Rev. 124, 22-26). It is found that F changes strongly with impact energy E0 for normal angle of incidence in contrast to a mild change predicted by the simulation calculations. Furthermore, experimental and simulation results for peak to effective continuum ratio R of Ti K-line are compared and discussed for the energy range of impact of the present investigation.

Culture-free detection and enumeration of STEC in water.

Shiga toxin-producing Escherichia coli (STEC) causes worldwide outbreaks of food and waterborne diseases. Rapid identification of causative agents is critical for early intervention in the case of widespread diarrheal epidemics to prevent mortality. In this study, a Molecular-Beacon targeting stx2 gene (highly associated with human illness) was designed to develop a culture-independent real-time PCR assay for detection and quantification of STEC in water samples. The assay could detect lowest 10 genomic equivalent (GE) of the reference strain (E. coli I.T.R.C.-18) per PCR or 100 GE/mL. The presence of 10(6)CFU/mL of non-pathogenic E. coli DH5α has no impact on sensitivity of the assay. The assay could successfully enumerate STEC in surface water (collected from a sewage impacted river) and potable water samples collected from Lucknow city without prior enrichment. The assay will be useful in pre-emptive monitoring of surface/potable waters to prevent waterborne outbreaks caused by STEC.

Metabolic profiles and phylogenetic diversity of microbial communities from chlorinated pesticides contaminated sites of different geographical habitats of India.

AIMS: To study the microbial communities in three sites contaminated with chlorinated pesticides and evaluation of dehydrodechlorinase (linA) gene variants involved in gamma-hexachlorocyclohexane (γ-HCH, lindane) degradation. METHODS AND RESULTS: Using a culture-independent method, 16S rRNA genes were amplified from microbial communities occurring in contaminated soils. From 375 clone libraries analysed, 55 different restriction fragment length polymorphism phylotypes were obtained. Dehydrodechlorinase (linA) gene, which initiates the γ-HCH degradation, was directly amplified by PCR from the DNA extracted from soils. Deduced amino acid sequences of eight variant genotypes of linA showed few amino acid changes. All the variants of linA had mutations of F151L and S154T, and one of the genotype carried 12 amino acid changes when compared to a linA of Sphingomonas sp. reported from the same soil. CONCLUSIONS: The microbial communities displayed complex and diverse groups similar to bacteria involved in biodegradation. The presence of biodegradative genes like linA indicates the presence of communities with capacity to biodegrade the persistent pesticide HCH. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insights to evaluate the presence of catabolic genes and assessing the bioremediation potential of the industrial soils contaminated by chlorinated pesticides.

Eventration with diaphragm perforation leading to secondary diaphragmatic hernia and intestinal strangulation.

We report a rare occurrence of a previously asymptomatic eventration that presented with intestinal obstruction followed by respiratory distress. The thinned out diaphragm had a nontraumatic perforation with herniation of the small bowel through the narrow defect. The herniated gut became strangulated and dilated inside the thorax, resulting in respiratory compromise. The rare occurrence of this vicious cycle of obstruction and respiratory failure leading to a sudden clinical deterioration in a previously stable patient is described.

Enterotoxigenic Escherichia coli in sewage-impacted waters and aquatic weeds: quantitative PCR for culture-independent enumeration.

AIM: To develop quantitative PCR for culture-independent enumeration of enterotoxigenic Escherichia coli (ETEC) in sewage-impacted waters and aquatic weeds. METHODS AND RESULTS: Two fluorescent probes (TaqMan and FRET) based on two different real-time PCR chemistries were designed in highly conserved region of LT1 gene encoding heat labile enterotoxin. Both the assays could detect 2 CFU ml(-1) from serially diluted (two-fold and ten-fold) culture of reference strain (E. coli MTCC 723). FRET performed better in terms of CT value and PCR efficiency than TaqMan. The presence of 10(6) CFU ml(-1) of nonpathogenic E. coli reduced the detection limit two-fold with both the probes. However, the performance for two chemistries in various environmental samples was significantly (student's t-test, P<0.05) different. CONCLUSION: It could be inferred from this study that real-time PCR chemistries (TaqMan and FRET) could detect very few copies of target DNA in pure cultures, but may give varied response in the presence of nonspecific DNA and natural inhibitors present in environmental sample matrices. SIGNIFICANCE AND IMPACT OF THE STUDY: The assays can be used for pre-emptive monitoring of aquatic weeds (a potential nonpoint source), surface and potable waters to prevent waterborne outbreaks caused by ETEC.

Determination of antimicrobial resistance and virulence gene signatures in surface water isolates of Escherichia coli.

AIMS: To determine the occurrence of Escherichia coli harbouring virulence markers of shiga- or entero-toxins and resistance to antimicrobials in surface waters. METHODS AND RESULTS: Surface water samples were collected at six locations of the river Gomti. E. coli isolates (n = 90) were characterized for their pathogenic potential using polymerase chain reaction to detect virulence genes as well as their sensitivity to antimicrobial agents using disc diffusion methods. In this study, 57.8% of E. coli isolates exhibited resistance to three or more antimicrobial agents. Sensitivity to cephotaxime, gentamicin and norfloxacin was observed in 7.8%, 48.9% and 77.8% of isolates, respectively. Both stx1 and stx2 genes were present in 15.6% of isolates while remaining isolates had either stx1 (17.8%) or stx2 (6.7%). The stx1 gene (33.3%) was more prevalent than stx2 (22.2%). The results indicate that the LT1 and ST1 genes were positive in 21.2% of isolates. CONCLUSIONS: The presence of multi-drug resistance and virulence genes in E. coli isolated from surface water being used for domestic and recreational purposes may result in waterborne outbreaks. SIGNIFICANCE AND IMPACT OF THE STUDY: The data will be useful in monitoring surface waters for forecasting and management of waterborne outbreaks.

Isolation of hexachlorocyclohexane-degrading Sphingomonas sp. by dehalogenase assay and characterization of genes involved in gamma-HCH degradation.

AIM: To screen and identify bacteria from contaminated soil samples which can degrade hexachlorocyclohexane (HCH)-isomers based on dechlorinase enzyme activity and characterize genes and metabolites. METHODS AND RESULTS: Dechlorinase activity assays were used to screen bacteria from contaminated soil samples for HCH-degrading activity. A bacterium able to grow on alpha-, beta-, gamma- and delta-HCH as the sole carbon and energy source was identified. This bacterium was a novel species belonging to the Sphingomonas and harbour linABCDE genes similar to those found in other HCH degraders. Gamma-pentachlorocyclohexene 1,2,4-trichlorobenzene and chlorohydroquinone were identified as metabolites. CONCLUSIONS: The study demonstrates that HCH-degrading bacteria can be identified from large environmental sample-based dehalogenase enzyme assay. This kind of screening is more advantageous compared to selective enrichment as it is specific and rapid and can be performed in a high-throughput manner to screen bacteria for chlorinated compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: The chlorinated pesticide HCH is a persistent and toxic environmental pollutant which needs to be remediated. Isolation of diverse bacterial species capable of degrading all the isomers of HCH will help in large-scale bioremediation in various parts of the world.

Effect of pressure on molecular and ionic motions in ultraviscous acetaminophen-aspirin mixture.

Effect of pressure and temperature on molecular motions and dc conductivity in ultraviscous, 50 wt% acetaminophen-aspirin melt has been studied by dielectric relaxation spectroscopy. The spectra obtained over the pressure range, 5-300 MPa and temperature range, 295-320 K show a distribution of relaxation times greater than found in pure acetaminophen. The equilibrium dielectric permittivity and relaxation time, tau, of the melt increase with increase in pressure and decrease in temperature and the dc conductivity, sigma(dc), decreases. The pressure and temperature variation of the limiting high frequency permittivity shows significant contribution from infrared polarization. The volumes of activation for sigma(dc) and tau vary with both the pressure and temperature, indicating that there is also a structural effect that determine sigma(dc). This effect would add a non-linear term to the Debye-Stokes-Einstein equation for variation of sigma(dc) with tau. The ultraviscous liquid remains stable with time, and with change in temperature and pressure, suggesting that a more stable glassy state of a pharmaceutical may be obtained by mixing a second component.

Doppler effect in fluorine K-Auger line produced in electron-induced core ionization of SF6.

An experimental evidence is reported on the observation of the Doppler effect in fluorine K-Auger line emitted from a core-ionized SF6 molecule under an impact of 16 keV electrons. The emitting source of the Auger line is found to acquire a kinetic energy of 4.7+/-0.3 keV. We propose that such large energy is released from the Coulomb repulsion taking place between F+ and SF5+ fragment ions under influence of an intense focusing field of the incident electrons. In the presence of the Coulomb field of these ions, the Auger line obtains a polarization P = 76%+/-7%.

Modulation of acute cadmium toxicity by Emblica officinalis fruit in rat.

The efficacy of Emblica officinalis in modifying the acute cytotoxicity of cadmium in male rats was evaluated. Oral administration of Emblica fruit juice (500 mg/kg, b.w.) for 8 days followed by a single toxic dose of Cd as CdCl2 (3 mg/kg,b.w. ip), considerably reduced the mortality in rats as well as prevented to some extent the cadmium induced histopathological damage in testis, liver and kidneys. Biochemical investigation also revealed reduced levels of Cd induced serum glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and gamma glutamyltranspeptidase. The enhanced levels of Cd and lipid peroxidation in liver, kidney, and testes and metallothionein and total sulphydryl in liver and kidney by Cd were significantly reduced by Emblica pretreatment. These results suggest cytoprotective potential of Emblica fruit in acute cadmium toxicity which could be due to its multiple role in biological system.

Pilot study of expedited HIV-1 testing of women in labor at an inner-city hospital in New York City.

To evaluate the feasibility of offering voluntary counseling and expedited human immunodeficiency virus (HIV) testing to women in labor, and to assess the characteristics of two rapid HIV assays compared with results from an expedited standard enzyme immunoassay (EIA), with Western blot confirmation, as indicated, we undertook a pilot study immediately prior to enactment of New York State regulations (August 1999) requiring expedited testing of laboring women (or newborns) with undocumented HIV status. From June 9, 1999 through July 2, 1999, we offered HIV counseling and testing (C&T) to all medically stable women in active labor, 106 of 125 (85%) of whom accepted. One woman was confirmed HIV-1 seropositive. Rapid assay sensitivity and specificity were: SUDS 100 and 98%, and Multispot 100 and 100%, respectively in comparison with 100 and 99% for the standard EIA. The positive predictive values (PPV) were SUDS 33%; Multispot 100%; and EIA 50%. While our sample size was small, it appears that the accuracy of rapid and expedited HIV assays may be improved by requiring two different reactive assays before informing women of HIV-seropositive results or initiating antiretroviral treatment.