Rhnull phenotype in an Indian patient due to a novel c.1138 + 2 t > a mutation in the RHAG gene.

BACKGROUND: The Rh system is an extremely important RBC antigen system with over 50 antigens, 5 of which (D, C, E, c and e) are considered most clinically significant. The rare Rhnull phenotype can result from mutations in the RHD and RHCE genes or the RHAG gene that affects their expression. This is a case report of the second type. CASE REPORT: This case reports a multiparous lady who had to be evaluated for a panreactive antibody. The discrepancy was first identified at the centre she reported to. A thorough immunohematological workup was performed at a second reference laboratory. Suspecting Rhnull phenotype, a third referral (molecular typing) was requested at International Blood Group Reference Laboratory (IBGRL), Bristol. RESULTS: A novel RHAG null allele (c.1138+2t>a), causing a Rhnull phenotype was identified. The antibody was most likely an anti-Rh 29 antibody. CONCLUSION: The novel c.1138+2 t > a mutation in the RHAG gene causing the Rhnull phenotype and development of a pan reacting antibody(ies) made the patient's pregnancy challenging. Confirmation of the diagnosis, an important step in her management, required use of both serological immunohematology and molecular techniques.

Indirect Response of the Temperature, Humidity, and Rainfall on the Spread of COVID-19 over the Indian Monsoon Region.

This article examines the role of the meteorological variable in the spread of the ongoing pandemic coronavirus disease 2019 (COVID-19) across India. COVID-19 has created an unprecedented situation for public health and brought the world to a standstill. COVID-19 had caused more than 1,523,242 deaths out of 66,183,029 confirmed cases worldwide till the first week of December 2020. We have examined the surface temperature, relative humidity, and rainfall over five cities: Delhi, Mumbai, Kolkata, Bengaluru, and Chennai, which were severely affected by COVID-19. It is found that the prevailing southwest (SW) monsoon during the pandemic has acted as a natural sanitizer in limiting the spread of the virus. The mean rainfall is ~ 20-40 mm over the selected cities, resulting in an average decrease in COVID cases by ~ 18-26% for the next 3 days after the rainfall. The day-to-day variations of the meteorological parameters and COVID-19 cases clearly demonstrate that both surface temperature and relative humidity play a vital role in the indirect transport of the virus. Our analysis reveals that most COVID-19 cases fall within the surface temperature range from 24 to 30 °C and relative humidity range from 50% to 80%. At a given temperature, COVID-19 cases show a large dependency on the relative humidity; therefore, the coastal environments were more prone to infections. Wavelet transforms coherence analysis of the daily COVID-19 cases with temperature and relative humidity reveals a significant coherence within 8 days.

Heavy metal contamination in river water and sediments of the Swarnamukhi River Basin, India: risk assessment and environmental implications.

The concentration of heavy metals was analyzed each of 20 river water, suspended sediments and bed sediments along the stretch of Swarnamukhi River Basin. River water is not contaminated with heavy metals except Fe and Mn. Contamination factor in sediments shows considerable to very high degree contamination with Cr, Cu, Pb and Zn. The sources of these metals could be residential wastes, sewer outfall, fertilizers, pesticides (M-45 + carbondine) and traffic activities apart from natural weathering of granitic rocks present in the basin area. Principal component analyses indicate the interaction between metals in different media. The comparison of metals (Cu, Pb and Zn) in bed sediments of Swarnamukhi River with the Indian and world averages indicates that the values obtained in the basin are above the Indian averages and far below to the world averages. Average shale values and sediment quality guidelines point toward the enrichment and contamination of Cu, Cr, Pb and Zn to several fold leading to eco-toxicological risks in basin.

Streptomyces hyderabadensis sp. nov., an actinomycete isolated from soil.

A novel actinomycete, designated strain OU-40(T), was isolated from farm soil collected from the Hyderabad region of Andhra Pradesh, southern India. The strain was found to have morphological and chemotaxonomic characteristics typical of species of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain OU-40(T) belonged to the genus Streptomyces, and was related most closely to Streptomyces pactum NBRC 13433(T) (99.0 % sequence similarity), Streptomyces olivaceus NBRC 12805(T) (99.0 %) and Streptomyces parvulus NBRC 13193(T) (98.8 %). Strain OU-40(T) could be distinguished from the type strains of its closest phylogenetic relatives based on levels of DNA-DNA relatedness and comparison of morphological and phenotypic data. It is therefore concluded that strain OU-40(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces hyderabadensis sp. nov. is proposed. The type strain is OU-40(T) (=CCTCC AA 209024(T) =PCM 2692(T)).

Identification of SNPs in crucial starch biosynthesis genes in rice.

Rice is the foremost crop catering to the major calorific requirement of the human population but has the disadvantage of having high glycaemic index (GI). The fine quality rice varieties, BPT and RNR have been recently identified as having low GI in nature and are grown mostly in southern parts of India. Starch (80%) is the major component of rice endosperm attributing to GI. The study aimed to unravel the molecular basis of low GI through targeted pooled amplicon sequencing of major starch biosynthetic genes. A total of 13 candidate genes involved in starch synthesis were amplified and pooled in equimolar proportion for sequencing. Single-nucleotide polymorphisms (SNPs) and insertions/deletions (Indels) were detected in both coding and noncoding regions. Among the genes that are under study, the highest number of variations were identified in starch synthase I (SSI) followed by starch synthase IIIA (SSIIIA) genes. Nonsynonymous SNPs with high probability of effecting gene function were validated by Sanger sequencing and molecular docking. Identified causative SNPs were mapped on 3000 rice genome database and their allele frequencies were obtained. The outcome of this study has a potential to be applied in breeding programmes to obtain low GI rice varieties with added beneficial traits.

Retrievability of calcium hydroxide intracanal medicament with three calcium chelators, ethylenediaminetetraacetic acid, citric acid, and chitosan from root canals: An in vitro cone beam computed tomography volumetric analysis.

AIM: This study compared the amount of aqueous-based and oil-based calcium hydroxide remaining in the canal, after removal with two different chelators 17% EDTA, 20% Citric acid and 0.2% Chitosan in combination with ultrasonic agitation. METHODS AND MATERIAL: Cleaning and shaping of root canals of 28 mandibular premolar was done and canals were filled either with Metapex or Ca(OH)2 mixed with distilled water. Volumetric analysis was performed utilizing cone beam-computed tomography (CBCT) after seven days of incubation. Ca(OH)2 was removed using either 17% EDTA, 20% Citric acid or 0.2% Chitosan in combination with ultrasonic agitation. STATISTICAL ANALYSIS USED: Volumetric analysis was repeated and percentage difference was calculated and statistically analyzed using Kruskal-Wallis and Mann-Whitney U test. RESULTS: All the three chelators failed to remove aqueous-based as well as oil-based Ca(OH)2 completely from the root canal. Aqueous-based Ca(OH)2 was easier to be removed than oil-based Ca(OH)2. 0.2% Chitosan in combination with ultrasonics performed better than 17% EDTA and 20% citric acid in removal of Ca(OH)2. CONCLUSION: Combination of 0.2% Chitosan and ultrasonic agitation results in lower amount of Ca(OH)2 remnants than 17% EDTA, 20% Citric acid irrespective of type of vehicle present in the mix.

Mutagenic activation of N-2-fluorenylacetamide and N-hydroxy-N-2-fluorenylacetamide in subcellular fractions from X/Gf mice.

The Salmonella mutagenesis test system was used to evaluate the in vitro mutagenic potency of N-2-fluorenylacetamide (2-FAA) and N-hydroxy-N-2-fluorenylacetamide (N-OH-2-FAA) mediated by liver and kidney subcellular fractions from X/Gf mice, a strain resistant to 2-FAA carcinogenesis. Pretreatment of the mice with the microsomal inducers 3-methylcholanthrene (MCA) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) increased the number of revertants from both liver and kidney fractions. Mutagenicity of N-OH-2-FAA mediated by liver or kidney microsomes was partially inhibited at 0.001--0.1 microM Paraxon (diethyl-p-nitrophenyl phosphate), an inhibitor of deacetylase enzyme, and the inhibition was complete (98%) in microsomes from control mice (100 microM Paraoxon). Conversely, the liver and kidney microsomal fractions from MCA- and TCDD-treated X/Gf mice were less sensitive to Paraoxon. The inhibition of kidney or liver cytosol-mediated N-OH-2-FAA mutagenicity by Paraoxon was less than that observed with the microsomal fraction (50% inhibition at 1 x 10(-7) and 1 x 10(-5) M Paraoxon, respectively). The mutagenicity of 2-FAA and N-OH-2-FAA mediated by liver or kidney subcellular fractions from X/Gf mice and its response to inducers and inhibitors of mutagenic activation processes appear similar to those observed in species both resistant (cotton rat) and sensitive (Sprague-Dawley rat, NIH Swiss mice) to 2-FAA carcinogenesis.

Effect of microsomal enzyme inducers on the urinary excretion pattern of mutagenic metabolites of the carcinogen 2,4-toluenediamine.

2,4-Toluenediamine [(TDA) CAS: 95-80-7] was administered to rats pretreated with the microsomal enzyme inducers phenobarbital (PB), beta-naphthoflavone (beta NF), or 3-methylcholanthrene (MCA). The 24-hour urines of male F344 rats were examined for their mutagenic potency by means of the Salmonella assay, with the Aroclor 1254-pretreated rat liver S-9 fraction as an activating system. No revertants were found with TDA or its urinary metabolites in the absence of the S-9 fraction. In the presence of S-9, the number of revertants increased as the concentration of TDA or its urinary metabolites increased. The urinary metabolites, generated after the microsomal enzyme inducers (PB, beta NF, MCA), had increased mutagenic activity as compared with the controls (saline, corn oil). In the presence of beta-glucuronidase (beta G), increased numbers of TA98 revertants were noted in the urine of rats pretreated with PB, saline, or corn oil. Addition of sulfatase did not alter the number of TA98 revertants. Conversely, beta G treatment of urine from rats pretreated with MCA or beta NF led to a decrease in the number of TA98 revertants as compared to levels in urine without beta G. Addition of known urinary metabolites of TDA, such as 4-acetylamino-2-aminobenzoic acid or 2,4-diacetylaminobenzoic acid, to beta NF-pretreated rat urine had no inhibitory effect on the mutagenicity in the absence of beta G. However, in the presence of beta G, the inhibitory effect was similar to that noted with beta NF-pretreated rat urine. Upon separation of urinary metabolites (beta NF-pretreated rat urine) into free, conjugated, and water-soluble forms, the maximum number of TA98 revertants was associated with the free ethyl acetate-extractable fraction, which accounted for the total mutagenic activity associated with the original volume of urine. Conjugated metabolites showed much less mutagenic activity, and an inhibitory principle was associated with the water-soluble fraction.

Hepatocarcinogenic and promoting action of a choline-devoid diet in the rat.

Male Fisher 344 rats were solely fed a choline-supplemented diet for 65 to 105 weeks or a choline-devoid diet for 24 to 102 weeks. Hepatocellular carcinomas developed in the latter animals, beginning at 24 weeks. Other groups of rats were given a single dose of 20 mg diethylnitrosamine/kg, 18 h after a partial hepatectomy and were fed, 4 weeks thereafter, either a choline-supplemented, or a choline-devoid diet for up to 48 weeks. In rats fed the choline-supplemented diet, the only relevant lesion observed was a small transect number of foci of enzyme-altered hepatocytes. On the other hand, a significant number of foci, of preneoplastic nodules, and of hepatocellular tumors developed in rats fed the choline-devoid diet. The results obtained are consistent with those previously reported by others, indicating that diets devoid of choline, or of choline and methionine, are carcinogenic. The diets appear to act as complete carcinogens, since they are also efficient promoters of chemical hepatocarcinogenesis, as shown again, in the present study, by the results obtained in the diethylnitrosamine-pretreated rats.

Free energy component analysis for drug design: a case study of HIV-1 protease-inhibitor binding.

A theoretically rigorous and computationally tractable methodology for the prediction of the free energies of binding of protein-ligand complexes is presented. The method formulated involves developing molecular dynamics trajectories of the enzyme, the inhibitor, and the complex, followed by a free energy component analysis that conveys information on the physicochemical forces driving the protein-ligand complex formation and enables an elucidation of drug design principles for a given receptor from a thermodynamic perspective. The complexes of HIV-1 protease with two peptidomimetic inhibitors were taken as illustrative cases. Four-nanosecond-level all-atom molecular dynamics simulations using explicit solvent without any restraints were carried out on the protease-inhibitor complexes and the free proteases, and the trajectories were analyzed via a thermodynamic cycle to calculate the binding free energies. The computed free energies were seen to be in good accord with the reported data. It was noted that the net van der Waals and hydrophobic contributions were favorable to binding while the net electrostatics, entropies, and adaptation expense were unfavorable in these protease-inhibitor complexes. The hydrogen bond between the CH2OH group of the inhibitor at the scissile position and the catalytic aspartate was found to be favorable to binding. Various implicit solvent models were also considered and their shortcomings discussed. In addition, some plausible modifications to the inhibitor residues were attempted, which led to better binding affinities. The generality of the method and the transferability of the protocol with essentially no changes to any other protein-ligand system are emphasized.

Hepatic vitamin A status of rats during feeding of the hepatocarcinogen 2-aminoanthraquinone.

Dietary administration of the dyestuff intermediate 2-aminoanthraquinone (AAQ) at a 2% level for periods of 2, 4, 8 or 12 weeks decreased the total liver vitamin A significantly (P less than 0.001) at all time periods in both male and female rats, compared to controls and pair-fed controls, respectively. A decrease in the body weights of AAQ-fed rats was much more pronounced in the females than male rats.

Macromolecular adduction by trichloroacetonitrile in the Fischer 344 rat following oral gavage.

Male Fisher 344 rats were administered 1- or 2-[14C]trichloroacetonitrile (TCAN) by oral gavage. DNA was isolated from the liver, kidneys and stomach and several protein fractions (globin, albumin and globulins) were isolated from blood. TCAN binds to both the DNA and the blood proteins in a dose-related manner. More radiolabel was associated with the DNA when the carbon at C2 position was labeled, than at C1 position. However, the position of the radiolabel did not influence the levels of radioactivity associated with the blood proteins. The stomach exhibited the highest level of DNA binding, followed in order by the liver and kidney. TCAN binding level was higher in DNA isolated from rats killed at 24 h than at 4 h after administration. In contrast, the three blood proteins showed similar binding levels, regardless of the exposure time. Radioactivity associated with DNA was not incorporated into the nitrogen bases (i.e. via de novo synthesis) and a covalent binding index (mumol chemical bound/mol nucleotide phosphate per mmol/kg body wt. of chemical administered) of 30-120 was observed for various tissues. Most of the radioactivity (60-80%) associated with globin could be released and separated from the protein by the treatment with concentrated ammonium hydroxide and precipitation of protein by organic solvent. Three peaks were observed in the HPLC elution profiles of the radioactivity released from the globin. Trichloroacetic acid co-eluted with one of these released products (peak II), however, the chemical identity of the material under the major peak (peak III) and peak I are still uncharacterized.

Effects of dietary choline deficiency on the mutagenic activation of chemical carcinogens by rat liver fractions.

Salmonella mutagenesis assays were used to evaluate the mutagenicity of several chemical carcinogens as mediated by liver S-9 fractions from rats fed a choline-supplemented (CS) or choline-devoid (CD) diet. The liver S-9 fraction from CD diet-fed rats was found to have a significantly decreased ability to activate 2-acetylaminofluorene (2-AAF), 2-aminoanthracene (2-AA) and 6-aminochrysene (6-AC), but not N-hydroxy-2-acetylaminofluorene (HO-N-2-AAF) and dimethylnitrosamine (DMN). The same liver S-9 fraction was also less effective in deactivating N-methyl-N1-nitro-N-nitrosoguanidine (MNNG) but not methylnitrosourea (MNU). A decrease (20%) in the cytochrome P-450 content was found in liver microsomes of CD diet-fed rats. Although it has been shown that feeding a CD diet to rats enhances chemical hepatocarcinogenesis, the data presented here suggest that CD diet does not increase the activation of the chemical procarcinogens tested.

Induction of nuclear anomalies in the gastrointestinal tract by polycyclic aromatic hydrocarbons.

A selective list of polycyclic aromatic hydrocarbons (PAH) with varied carcinogenic and mutagenic potencies, which are identified as common contaminants at industrial sites and which often contaminate the neighboring ground water, are investigated for their ability to induce nuclear anomalies (NA) in the mouse gastrointestinal (G.I.) tract. These studies examined the hypothesis that a relationship between NA induction and carcinogenic potency of these PAH exists. Among the PAH tested, 7,12-dimethylbenzanthrene (DMBA) was most effective inducer of NA in all G.I. tract tissues examined, with the relative potency in duodenum of DMBA much much greater than benzo[a]pyrene (B[a]P) much greater than benzo[b]fluoranthene (B[b]F). The induction of NA by benzo[a]anthracene (B[a]A), pyrene (PY) and benzo[e]pyrene (B[e]P) was not different from that elicited by vehicle controls. MNU, a known potent inducer of NA in the mouse G.I. tract, yielded a high level of NA in duodenum and proximal colon but was less effective than DMBA in the forestomach. The data suggest that induction of NA by DMBA and B[a]P PAH are in approximate accordance with their relative carcinogenic potency in the gastrointestinal tract. When binary mixtures of some PAH were administered the yield of NA was less than that expected by simple additivity and closer to that expected by averaging the activities of the two PAH comprising the mixture. Thus, this short-term in vivo assay may be useful as a predictor of the genotoxic or carcinogenic strength of individual PAH and/or mixtures of these compounds.

Site-specific modulation of carcinogen-induced gastrointestinal tract nuclear anomalies in B6C3F1 mice by chloroform.

Chloroform (CHCl3) is an established rodent carcinogen and a prevalent contaminant of chlorine-disinfected drinking water. Thus in the United States CHCl3, along with other trihalomethanes, is regulated not to exceed 100 ppb in potable water. Recently, several studies have shown that CHCl3 also has anti-cancer properties as it inhibits tumor growth in mouse liver and in the gastrointestinal tract of the rat. In this paper we show that CHCl3 also inhibits the propensity for three gastrointestinal tract carcinogens, benzo(a)pyrene (BAP), 1,2-dimethylhydrazine (DMH) and methylnitrosourea (MNU), to induce nuclear anomalies in the proximal colon of the B6C3F1 mouse. For example, in mice pre-adapted to 1800 ppm CHCl3 for 30 days prior to the carcinogen administration the level of nuclear anomalies induced in the proximal colon by BAP was reduced by four-fold (0.9 +/- 0.7 v. 3.6 +/- 1.0 anomalies/10 crypts; p less than 0.001) and two-fold for MNU (2.4 +/- 1.0 v. 4.6 +/- 1.6; p less than 0.001) and DMH (0.9 +/- 0.9 v. 1.7 +/- 0.8; p = 0.03). In the duodenum CHCl3 was effective at inhibiting unclear anomalies only for MNU (45.3 +/- 4.6 v. 30.4 +/- 3.5; p = 0.02). The inhibitory effect of CHCl3 does not extend to nuclear anomalies of the forestomach. The anti-cancer properties of CHCl3 are discussed in light of its cancer causing potential and possible application to human risk assessment.

Mutagenicity of urine from rats after administration of 2,4-diaminoanisole: the effect of microsomal enzyme inducers.

Ring 14C-labelled 2,4-diaminoanisole disulfate was administered to rats pretreated with the microsomal inducers phenobarbital (PB), beta-naphthoflavone (BNF), 3-methylcholanthrene (MC) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The 24-h urine from rats pretreated with PB showed a 2-fold increase in revertant rate over the corresponding control as measured by the Ames Salmonella test system. Pretreatment of rats with BNF, MC or TCDD decreased the mutagenicity of urine by about 70% when an activating system was used. However, in the absence of an activating system, the urine from rats induced with BNF, MC or TCDD showed a significant (P < 0.001) degree of mutagenicity compared with urine from controls or urine from phenobarbital-induced rats. Release of conjugates by beta-glucuronidase increased the mutagenicity of urine even in the absence of an activating system, but the number of revertants was almost doubled in the presence of an activating system. The urine from rats treated only with the 4 inducers did not show any mutagenicity. 2,4-Diaminoanisole itself was mutagenic only in the presence of an activating system. alpha-Naphthoflavone (ANF) (0.1 mM) inhibited by 85--90% the in vitro mutagenicity of urine, mediated by Aroclor 1254, MC or TCDD induced rat-liver microsomes. The mutagenicity mediated through PB-induced rat-liver microsomes was, however, inhibited only by 16%. Similarly, 0.1 mM metyrapone (MP) inhibited the mutagenicity of urine by Aroclor 1254, MC or TCDD induced rat-liver microsomes by 13--18%. For the same MP concentration a 50% inhibition of the mutagenicity mediated through PB-induced rat-liver microsomes was observed. The mutagenicity pattern for urine in vitro was shown to be similar with liver S9 from rats induced either with Aroclor 1254 or with MC.

Streptomyces osmaniensis sp. nov., isolated from soil.

A novel actinomycete, designated strain OU-63T, was isolated from garden soil collected on the campus of Osmania University in Hyderabad, southern India. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain belonged to the genus Streptomyces, and was related most closely to Streptomyces chartreusis NBRC 12753T (98.6% similarity). However, the mean level of DNA-DNA relatedness between the two strains was only 25+/-1.73%. Based on DNA-DNA relatedness, morphological and phenotypic data, strain OU-63T could be distinguished from the type strains of phylogenetically related species. It is therefore considered to represent a novel species of the genus Streptomyces, for which the name Streptomyces osmaniensis sp. nov. is proposed. The type strain is OU-63T (=CCTCC AA209025T =PCM 2690T).

Effect of copper sulfate on hematology, blood chemistry, and hepato-somatic index of an Indian catfish, Heteropneustes fossilis (Bloch), and its recovery.

The present communication deals with the effect of copper sulfate (0.25 ppm) on hematology, blood chemistry, and hepato-somatic indices of an Indian catfish, Heteropneustes fossilis (Bloch) and its recovery from intoxication. Analysis of various parameters was made after 24 hr, and 5, 10, 20, and 30 days of CuSO4 exposure. Subsequently, the fish were allowed recovery periods of 24 hr and 5 days in normal water. Changes in various parameters from normal values during exposure and recovery were recorded and probable reasons for alterations have been discussed in detail.

Endogenous hepatic growth-modulating factors and effects of a choline-devoid diet and of phenobarbital on hepatocarcinogenesis in the rat.

The activities of an endogenous inhibitor and of a stimulator of cell proliferation were assayed in the livers of sham-operated (SO) or partially hepatectomized (PH) adult rats; rats fed a choline-supplemented (CS) or a choline-devoid (CD) diet; the same diets followed by acute CCl4 intoxication; the same diets supplemented with phenobarbital (PHB); or a CD diet containing DL-ethionine (ETH). The inhibitor and the stimulator were semipurified by fractional ethanol precipitation of a liver cytosolic fraction, and their activities were assessed by means of bioassays in vitro. The livers of SO rats and of rats fed the CS diet contained only inhibitor activity. Following PH, a CD diet, or CCl4 intoxication the inhibitor activity was suppressed, and there was a simultaneous appearance of a stimulator activity. Thus, PH, a CD diet, and CCl4 intoxication cause similar cellular (loss and regeneration) and humoral-homeostatic changes in adult rat livers. We propose that these changes constitute a basic attribute of the mechanism whereby the three conditions affect similarly hepatocarcinogenesis in the rat, especially in the case of a CD diet, because the changes it induces are chronic rather than acute. PHB, another promoter of chemical hepatocarcinogenesis, affected neither the inhibitor nor the stimulator activity. Thus, PHB seems to be acting by a different mechanism than that of the other three agents. ETH did not modify the shift in the balance of the growth-modulating factors induced by a plain CD diet. This shift may account for the marked stimulation of carcinogen-induced oval cell proliferation exerted by a CD diet. The significance of these results is discussed in the context of known effects of a CD diet and of PHB on hepatocarcinogenesis in rats.