Multi-omics analysis identifies potential microbial and metabolite diagnostic biomarkers of bacterial vaginosis.

BACKGROUND: Bacterial vaginosis (BV) is a common clinical manifestation of a perturbed vaginal ecology associated with adverse sexual and reproductive health outcomes if left untreated. The existing diagnostic modalities are either cumbersome or require skilled expertise, warranting alternate tests. Application of machine-learning tools to heterogeneous and high-dimensional multi-omics datasets finds promising potential in data integration and may aid biomarker discovery. OBJECTIVES: The present study aimed to evaluate the potential of the microbiome and metabolome-derived biomarkers in BV diagnosis. Interpretable machine-learning algorithms were used to evaluate the utility of an integrated-omics-derived classification model. METHODS: Vaginal samples obtained from reproductive-age group women with (n = 40) and without BV (n = 40) were subjected to 16S rRNA amplicon sequencing and LC-MS-based metabolomics. The vaginal microbiome and metabolome were characterized, and machine-learning analysis was performed to build a classification model using biomarkers with the highest diagnostic accuracy. RESULTS: Microbiome-based diagnostic model exhibited a ROC-AUC (10-fold CV) of 0.84 ± 0.21 and accuracy of 0.79 ± 0.18, and important features were Aerococcus spp., Mycoplasma hominis, Sneathia spp., Lactobacillus spp., Prevotella spp., Gardnerella spp. and Fannyhessea vaginae. The metabolome-derived model displayed superior performance with a ROC-AUC of 0.97 ± 0.07 and an accuracy of 0.92 ± 0.08. Beta-leucine, methylimidazole acetaldehyde, dimethylethanolamine, L-arginine and beta cortol were among key predictive metabolites for BV. A predictive model combining both microbial and metabolite features exhibited a high ROC-AUC of 0.97 ± 0.07 and accuracy of 0.94 ± 0.08 with diagnostic performance only slightly superior to the metabolite-based model. CONCLUSION: Application of machine-learning tools to multi-omics datasets aid biomarker discovery with high predictive performance. Metabolome-derived classification models were observed to have superior diagnostic performance in predicting BV than microbiome-based biomarkers.

Microbial diversity and composition in acidic sediments of freshwater finfish culture ponds fed with two types of feed: a metagenomic approach.

Microbial community profile associated with acidic pond sediments (APS) (pH = 3·0-4·5) of freshwater finfish aquaculture ponds (n = 8) was investigated. Sediment DNA extracted from the eight APS were subjected to high-throughput sequencing of V3 and V4 regions which yielded 7236 operational taxonomic units (OTUs) at a similarity of 97%. Overall results showed higher proportion of bacterial OTUs than archaeal OTUs in all the APS. Euryarchaeota (23%), Proteobacteria (19%), Chloroflexi (17%), Crenarchaeota (5·3%), Bacteroidetes (4·8%), Nitrospirae (3·2%), Nanoarchaeaeota (3%) which together constituted 75% of the microbial diversity. At the genus level, there was high preponderance of methanogens namely Methanolinea (5·4%), Methanosaeta (4·5%) and methanotrops, Bathyarchaeota (5%) in APS. Moreover, the abundant phyla in the APS were not drastically affected by the administration of chicken slaughter waste (R-group ponds) and commercial fish feed (C-group ponds), since 67% of the OTUs generated remained common in the APS of both the groups of ponds. There was a minimal difference of 24-26% of OTUs between C-group and R-group ponds, suggesting the existence of a core microbial community in these ponds driven by acidic pH over the years. This study concludes that microbial diversity in pond sediment was influenced to a lesser extent by the addition of chicken slaughter waste but was majorly driven by acidic nature of the pond.

The selenocyanate dimer radical anion in water: Transient Raman spectra, structure, and reaction dynamics.

The selenocyanate dimer radical anion (SeCN)2 •-, prepared by electron pulse irradiation of selenocyanate anion (SeCN)- in water, has been examined by transient absorption, time-resolved Raman spectra, and range-separated hybrid density functional (ωB97x and LC-ωPBE) theory. The Raman spectrum, excited in resonance with the 450 nm (λmax) absorption of the radical, is dominated by a very strong band at 140.5 cm-1, associated with the Se-Se stretching vibration, its overtones and combinations. A striking feature of the (SeCN)2 •- Raman spectrum is the relative sharpness of the 140.5 cm-1 band compared to the S-S band at 220 cm-1 in thiocyanate radical anion (SCN)2 •-, the difference of which is explained in terms of a time-averaged site effect. Calculations, which reproduce experimental frequencies fairly well, predict a molecular geometry with the SeSe bond length of 2.917 (±0.04) Å, the SeC bond length of 1.819 (±0.004) Å, and the CN bond length of 1.155 (±0.002) Å. An anharmonicity of 0.44 cm-1 has been determined for the 140.5 cm-1 Se-Se vibration which led to a dissociation energy of ∼1.4 eV for the SeSe bond, using the Morse potential in a diatomic approximation. This value, estimated for the radical confined in a solvent cage, compares well with the calculated gas-phase energy, 1.32 ± 0.04 eV, required for the radical to dissociate into (SeCN)• and (SeCN)- fragments. The enthalpy of dissociation in water has been measured (0.36 eV) and compared with the value estimated by accounting for the solvent dielectric effects in structural calculations.

Transient Raman spectra, structure, and thermochemistry of the thiocyanate dimer radical anion in water.

Time-resolved resonance-enhanced Stokes and anti-Stokes Raman spectra of the thiocyanate dimer radical anion, (SCN)2•-, prepared by pulse radiolysis in water, have been obtained and interpreted in conjunction with theoretical calculations to provide detailed information on the molecular geometry and bond properties of the species. The structural properties of the radical are used to develop a molecular perspective on its thermochemistry in an aqueous solution. Twenty-nine Stokes Raman bands of the radical observed in the 120-4200 cm-1 region are assigned in terms of the strongly enhanced 220 cm-1 fundamental, weakly enhanced 721 cm-1, and moderately enhanced 2073 cm-1 fundamentals, their overtones, and combinations. Calculations by range-separated hybrid density functionals (ωB97x and LC-ωPBE) support the spectroscopic assignments of the 220 cm-1 vibration to a predominantly SS stretching mode and the features at 721 cm-1 and 2073 cm-1 to CS and CN symmetric stretching modes, respectively. The corresponding bond lengths are 2.705 (±0.036) Å, 1.663 (±0.001) Å, and 1.158 (±0.002) Å. A first order anharmonicity of 1 cm-1 determined for the SS stretching mode suggests a convergence of vibrational states at an energy of ∼1.5 eV, using the Birge-Sponer extrapolation. This value, estimated for the radical confined in solvent cage, compares well with the calculated gas-phase energy of 1.26 ± 0.04 eV required for the radical to dissociate into SCN• and SCN- fragments. The enthalpy of dissociation drops to 0.60 ± 0.03 eV in water when solvent dielectric effects on the radical and its dissociation products upon S-S bond scission are incorporated in the calculations. No frequency shift or spectral broadening was observed between light and heavy water solvents, indicating that the motion of solvent molecules in the hydration shell has no perceptible effect on the intramolecular dynamics of the radical. The Stokes and anti-Stokes Raman frequencies were found to be identical within the experimental uncertainty, suggesting that the frequency difference between the thermally relaxed and spontaneously created vibrational states of (SCN)2•- in water is too small to be observable.

The nature of the CO2 (-) radical anion in water.

The reductive conversion of CO2 into industrial products (e.g., oxalic acid, formic acid, methanol) can occur via aqueous CO2 (-) as a transient intermediate. While the formation, structure, and reaction pathways of this radical anion have been modelled for decades using various spectroscopic and theoretical approaches, we present here, for the first time, a vibrational spectroscopic investigation in liquid water, using pulse radiolysis time-resolved resonance Raman spectroscopy for its preparation and observation. Excitation of the radical in resonance with its 235 nm absorption displays a transient Raman band at 1298 cm(-1), attributed to the symmetric CO stretch, which is at ∼45 cm(-1) higher frequency than in inert matrices. Isotopic substitution at C ((13)CO2 (-)) shifts the frequency downwards by 22 cm(-1), which confirms its origin and the assignment. A Raman band of moderate intensity compared to the stronger 1298 cm(-1) band also appears at 742 cm(-1) and is assignable to the OCO bending mode. A reasonable resonance enhancement of this mode is possible only in a bent CO2 (-)(C2v/Cs) geometry. These resonance Raman features suggest a strong solute-solvent interaction, the water molecules acting as constituents of the radical structure, rather than exerting a minor solvent perturbation. However, there is no evidence of the non-equivalence (Cs) of the two CO bonds. A surprising resonance Raman feature is the lack of overtones of the symmetric CO stretch, which we interpret due to the detachment of the electron from the CO2 (-) moiety towards the solvation shell. Electron detachment occurs at the energies of 0.28 ± 0.03 eV or higher with respect to the zero point energy of the ground electronic state. The issue of acid-base equilibrium of the radical, which has been in contention for decades, as reflected in a wide variation in the reported pKa (-0.2 to 3.9), has been resolved. A value of 3.4 ± 0.2 measured in this work is consistent with the vibrational properties, bond structure, and charge distribution in aqueous CO2 (-).

Interleukin-1 gene cluster variants in hemodialysis patients with end stage renal disease: An association and meta-analysis.

We evaluated whether polymorphisms in interleukin (IL-1) gene cluster (IL-1 alpha [IL-1A], IL-1 beta [IL-1B], and IL-1 receptor antagonist [IL-1RN]) are associated with end stage renal disease (ESRD). A total of 258 ESRD patients and 569 ethnicity matched controls were examined for IL-1 gene cluster. These were genotyped for five single-nucleotide gene polymorphisms in the IL-1A, IL-1B and IL-1RN genes and a variable number of tandem repeats (VNTR) in the IL-1RN. The IL-1B - 3953 and IL-1RN + 8006 polymorphism frequencies were significantly different between the two groups. At IL-1B, the T allele of - 3953C/T was increased among ESRD (P = 0.0001). A logistic regression model demonstrated that two repeat (240 base pair [bp]) of the IL-1Ra VNTR polymorphism was associated with ESRD (P = 0.0001). The C/C/C/C/C/1 haplotype was more prevalent in ESRD = 0.007). No linkage disequilibrium (LD) was observed between six loci of IL-1 gene. We further conducted a meta-analysis of existing studies and found that there is a strong association of IL-1 RN VNTR 86 bp repeat polymorphism with susceptibility to ESRD (odds ratio = 2.04, 95% confidence interval = 1.48-2.82; P = 0.000). IL-1B - 5887, +8006 and the IL-1RN VNTR polymorphisms have been implicated as potential risk factors for ESRD. The meta-analysis showed a strong association of IL-1RN 86 bp VNTR polymorphism with susceptibility to ESRD.

Effect of direct loading of phytoestrogens into the calcium phosphate scaffold on osteoporotic bone tissue regeneration.

3D porous calcium deficient hydroxyapatite (CDHA) scaffolds with phytoestrogens were fabricated for osteoporotic bone tissue regeneration through a combination of 3D printing techniques and cement chemistry as a room temperature process. Quercetin, one of the major phytoestrogens isolated from onions and apples, was directly incorporated into CDHA for local administration in place of bisphosphonates (BPs) (which are recognized as standard treatment in osteoporosis), to avoid drug side effects. The CDHA scaffolds with quercetin (QC-CDHA) showed favorable mechanical properties (compressive strength < 21 MPa) as well as pore morphology. Quercetin was steadily released with the biodegradation of CDHA scaffolds in vitro without any initial burst. The QC-CDHA scaffolds greatly influenced both osteoblast and osteoclast cell activities. The QC-CDHA scaffolds significantly increased pre-osteoblast cell (MC3T3-E1) proliferation, differentiation, and mineralization, whereas osteoclast cell (RANK treated RAW 264.7) proliferation and differentiation were dramatically suppressed. The influence of quercetin on bone tissue regeneration was superior to alendronate, which is one of the most commonly administered BPs. All results indicated that quercetin in CDHA scaffolds plays an important role in both enhancing bone formation and suppressing bone resorption. Consequently, this technology promises great potential in osteoporotic bone tissue regeneration.

Acclimation of brackish water pearl spot (Etroplus suratensis) to various salinities: relative changes in abundance of branchial Na(+)/K (+)-ATPase and Na (+)/K (+)/2Cl (-) co-transporter in relation to osmoregulatory parameters.

The present study was conducted to elucidate the osmoregulatory ability of the fish pearl spot (Etroplus suratensis) to know the scope of this species for aquaculture under various salinities. Juvenile pearl spot were divided into three groups and acclimated to freshwater (FW), brackish water (BW) or seawater (SW) for 15 days. The fish exhibited effective salinity tolerance under osmotic challenges. Although the plasma osmolality and Na(+), K(+) and Cl(-) levels increased with the increasing salinities, the parameters remained within the physiological range. The muscle water contents were constant among FW-, BW- and SW-acclimated fish. Two Na+/K+-ATPase α-isoforms (NKA α) were expressed in gills during acclimation in FW, BW and SW. Abundance of one isoform was up-regulated in response to seawater acclimation, suggesting its role in ion secretion similar to NKA α1b, while expression of another isoform was simultaneously up-regulated in response to both FW and SW acclimation, suggesting the presence of isoforms switching phenomenon during acclimation to different salinities. Nevertheless, NKA enzyme activities in the gills of the SW and FW individuals were higher (p < 0.05) than in BW counterparts. Immunohistochemistry revealed that Na(+)/K(+)-ATPase immunoreactive (NKA-IR) cells were mainly distributed in the interlamellar region of the gill filaments in FW groups and in the apical portion of the filaments in BW and SW groups. The number of NKA-IR cells in the gills of the FW-acclimated fish was almost similar to that of SW individuals, which exceeded that of the BW individuals. The NKA-IR cells of BW and SW were bigger in size than their FW counterparts. Besides, the relative abundance of branchial Na(+)/K(+)/2Cl(-) co-transporter showed stronger evidence in favor of involvement of this protein in hypo-osmoregulation, requiring ion secretion by the chloride cells. To the best of our knowledge, this is the first study reporting the wide salinity tolerance of E. suratensis involving differential activation of ion transporters and thereby suggesting its potential as candidate for fish farming under different external salinities.

The nature of the superoxide radical anion in water.

Vibrational properties of the superoxide radical anion (O2(-●) in liquid water have been experimentally investigated for the first time. The stretching frequency, its shift from the gas-phase to aqueous solution, anharmonicity constant, and the Raman bandwidths provide an insight into the radical-water interactions and the hydration cage. In view of the spectroscopic information obtained in this work, the structural models based on molecular dynamics simulation in solution and gas-phase infrared studies of the water molecules bound to O2(-●) are critically examined.

The early events in the OH radical oxidation of dimethyl sulfide in water.

The time-resolved Raman observation of a prototype of the hetero-atom three electron bonds (-X-OH) that often form on encounter of the OH radical with chemical species in water is reported. In spite of their wide chemical and biochemical importance, no experimental structural information exists, thus far, on any such bond in solution or in the gas phase. The nature of the >S-O bond formed on the reaction of the OH radical with dimethyl sulfide in water, investigated in the present work, would necessitate a reexamination of the existing reaction mechanisms in related biological systems and development of the appropriate computational methods.

Identification of a novel heterozygous mutation in exon 50 of the COL1A1 gene causing osteogenesis imperfecta.

UNLABELLED: A 19-year-old woman was diagnosed with osteogenesis imperfecta (OI). She had sustained numerous low-trauma fractures throughout her childhood, including a recent pelvic fracture (superior and inferior ramus) following a low-impact fall. She had the classical blue sclerae, and dual energy X-ray absorptiometry (DEXA) bone scanning confirmed low bone mass for her age in the lumbar spine (Z-score was -2.6). However, despite these classical clinical features, the diagnosis of OI had not been entertained throughout the whole of her childhood. Sequencing of her genomic DNA revealed that she was heterozygous for the c.3880_3883dup mutation in exon 50 of the COL1A1 gene. This mutation is predicted to result in a frameshift at p.Thr1295, and truncating stop codon 3 amino acids downstream. To our knowledge, this mutation has not previously been reported in OI. LEARNING POINTS: OI is a rare but important genetic metabolic bone and connective tissue disorder that manifests a diverse clinical phenotype that includes recurrent low-impact fractures.Most mutations that underlie OI occur within exon 50 of the COL1A1 gene (coding for protein constituents of type 1 pro-collagen).The diagnosis of OI is easily missed in its mild form. Early diagnosis is important, and there is a need for improved awareness of OI among health care professionals.OI is a diagnosis of exclusion, although the key diagnostic criterion is through genetic testing for mutations within the COL1A1 gene.Effective management of OI should be instituted through a multidisciplinary team approach that includes a bone specialist (usually an endocrinologist or rheumatologist), a geneticist, an audiometrist and a genetic counsellor. Physiotherapy and orthopaedic surgery may also be required.

Impact of alphamethrin on biochemical parameters of Channa punctatus.

Impact of alphamethrin (synthetic pyrethroid) on profiles of lactate dehydrogenase (LDH), catalase (CAT), DNA, RNA and protein in liver, brain, gill and skeletal muscle of the freshwater food fish Channa punctatus were investigated. Exposure of sublethal concentration of alphamethrin (0.018 ppm for 14 days) increased the activity of LDH in liver (1.8 fold), brain (1.4 fold), gill(1.6 fold), and skeletal muscle (2.2 fold) of the fish. However, it significantly decreased the activity of CAT in the tissues of liver (54%), skeletal muscle (52%), gill (51%) and brain (49%) of the fish. Similarly, DNA (skeletal muscle (36%), liver (30%), brain (28%) and gill (25%)) RNA (liver (42%), brain (32%), gill (35%) and skeletal muscle (45%)) and protein content (45%), brain (42%), gill (36%), and skeletal muscle (27%)) declined in different tissues of the fish exposed to alphamethrin. Maximum increase in the level of LDH was in skeletal muscle (2.2 fold) and minimum in brain (1.4 fold). Maximum reduction in CAT profile was in liver (54%), and minimum in brain (49%). Declines in DNA was maximum in skeletal muscle (36%) and minimum in gill (25%) whereas RNA and protein content were maximum in liver (42% and 45% respectively) and minimum in skeletal muscle (45% and 27% respectively). Alphamethrin was toxic to the freshwater fish due to its inducing effect on anaerobic enzyme (LDH) and inhibitory effect on antioxidant enzyme (CAT), DNA, RNA and protein. This reflected alphamethrin associated increase in anaerobiosis and decrease in oxidative defense and impairment in protein synthesizing capacity of C. punctatus. Further, induction in LDH and reduction in CAT and protein profile may be used as biomarker of alphamethrin toxicity in fish.

Temperature-dependent alterations in metabolic enzymes and proteins of three ecophysiologically different species of earthworms

The effects of varying temperatures (12 - 44° C) on the specific activity of cytoplasmic malate dehydrogenase ((cMDH), mitochondrial malate dehydrogenase (mMDH) and lactate dehydrogenase (LDH) of some earthworms (Metaphire posthuma, Perionyx sansibaricus and Lampito mauritii) were studied. The effects of different temperatures on supernatant and mitochondrial protein contents were also investigated. The specific activities of cMDH, mMDH and LDH of the earthworms decreased gradually as a function of increasing temperature from 12 to 44°C. Higher metabolic energy was needed to maintain the activity at low temperatures. Hence, the earthworms showed increased enzyme specific activity at low temperatures. However, the protein content increased upto 28°C. Afterwards, with the increase in the temperature from 28 to 42°C, the proteins in the earthworms showed a significant decrease. The temperature-associated changes in the protein content could be explained by the fact that protein synthesizing capacity was hampered above and below the optimum temperature range. The most pronounced effects of varying temperatures were on P. sansibaricus. It might be due to the epigeic nature of the earthworm species. Then minimum effect was on the endogeic earthworm M. posthuma. Virtually, the differences in the enzymes physiology were associated with the differences in the ecological categories of the earthworms. This clearly demonstrate a possible link between the physiology and ecology at aerobic (cMDH, mMDH) and anaerobic (LDH) levels in the tropical earthworms.

Association of angiotensin-converting enzyme gene I/D polymorphism with steroid responsiveness in childhood nephrotic syndrome.

The aim of the study was to study the distribution of angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism, and its association with steroid responsiveness in children with idiopathic nephrotic syndrome (INS). One hundred twenty-five children with INS were classified into two groups: steroid-sensitive nephrotic syndrome (SSNS: n = 90) and steroid-resistant nephrotic syndrome (SRNS: n=35). The control group consisted of 150 unrelated healthy children. Genomic DNA was extracted from peripheral leucocytes by the standard salting-out method. ACE genotyping was performed and ACE genotypes DD, ID, and II were compared between different groups. The frequency distribution of the DD genotype was significantly increased in children with INS compared to control subjects (P = 0.0012) while the difference was not significant (P = 0.071) between SSNS and control subjects. The frequency distribution of the DD genotype was significantly high in the SRNS group compared to control subjects (P < 0.0001). The distribution of the DD genotype was high in SRNS compared to SSNS group patients (P = 0.016). In conclusion, the presence of the DD genotype may predict risk for steroid resistance in childhood INS.

Glucose oscillations, more than constant high glucose, induce p53 activation and a metabolic memory in human endothelial cells.

AIMS/HYPOTHESIS: Damage persists in HUVECs exposed to a constant high glucose concentration long after glucose normalisation, a phenomenon termed 'metabolic memory'. Evaluation of the effects of exposure of HUVECs to oscillating high glucose on the induction of markers of oxidative stress and DNA damage (phospho-γ-histone H2AX and PKCδ) and onset of metabolic memory, and the possible role of the tumour suppressor transcriptional factor p53 is of pivotal interest. METHODS: HUVECs were incubated for 3 weeks in 5 or 25 mmol/l glucose or oscillating glucose (24 h in 5 mmol/l glucose followed by 24 h in 25 mmol/l glucose) or for 1 week in constant 5 mmol/l glucose after being exposed for 2 weeks to continuous 25 mmol/l high glucose or oscillating glucose. Transcriptional activity of p53 was also evaluated in the first 24 h after high glucose exposure. RESULTS: High constant glucose upregulated phospho-γ-histone H2AX and protein kinase C (PKC)δ compared with control. Oscillating glucose was even more effective than both normal and constant high glucose. Both constant and oscillating glucose resulted in a memory effect, which was more pronounced in the oscillating condition. Transcriptional activity of p53 peaked 6 h after glucose exposure, showing a predicted oscillatory behaviour. CONCLUSIONS/INTERPRETATION: Exposure to oscillating glucose was more deleterious than constant high glucose and induced a metabolic memory after glucose normalisation. Hyperactivation of p53 during glucose oscillation might be due to the absence of consistent feedback inhibition during each glucose spike and might account for the worse outcome of this condition.

Reparation of chlorpyrifos-induced impairment by thyroxine and vitamin C in fish.

The organophosphate pesticide chlorpyrifos significantly decreased the specific activities of catalase (CAT) and lactate dehydrogenase (LDH) in brain, liver, gill and skeletal muscle of the freshwater catfish, Heteropneustes fossilis. It may be due to binding of the pesticide or its metabolite(s) with the enzyme molecules or affecting the synthesis and/or degradation of the enzymes. RNA and protein content also declined significantly in different tissues of the fish in response to chlorpyrifos, which may be due to alterations in turnover of macromolecules involved in protein synthesis. Administration of thyroxine (T4) and ascorbic acid (vitamin C) separately or conjointly to chlorpyrifos-exposed fish brought CAT, LDH, RNA and protein up to their control levels. The protective effect may be attributed to hormone, vitamin and pesticide interaction. Therefore, chlorpyrifos-induced impairment in antioxidative, anaerobic and protein synthesizing capacity can be repaired by ascorbic acid and T4 in fish.

Formulation and In Vitro evaluation of pH sensitive oil entrapped polymeric blended gellan gum buoyant beads of clarithromycin.

BACKGROUND AND THE PURPOSE OF THE STUDY: A gastroretentive pH sensitive system has been a frontier approach to release the drug in controlled manner in stomach and duodenum. The aim of this study was to develop buoyant beads of gellan based, wherein, the oil was entrapped, blended with hydroxypropyl methyl cellulose or carbopol 934 in order to evaluate its potential for targeted sustained delivery of clarithromycin in the gastric region. METHODS: Buoyant beads of gellan was developed by inotropic gelation technique using calcium carbonate as gas forming agent and the drug polymer dispersion was emulsified with mineral oil. The oil was entrapped and blended with hydroxypropyl methyl cellulose or carbopol 934. The developed beads were evaluated in terms of diameter,% floating, encapsulation efficiency, In vitro drug release, In vivo gastric residence efficacy and clarithromycine concentration in the mucosa of the experimental animal model. RESULTS: The scanning electron microscope photograph indicated that the prepared beads were spherical in shape and buoyancy, encapsulation efficiency and drug content obtained from all batches were satisfactory. Particle size and percentage buoyancy of the gel beads increased by raising the concentration of calcium carbonate. The formulation exhibited sustained release profile and was best fitted in the Peppas model with n<0.45. Subsequent coating of microbeads exhibited zero-order sustained pattern of the drug release up to 8 hrs. Batch B(4) showed comparatively better residence and the drug concentration in the gastric mucosa of the treated animals. CONCLUSION: The result provides evidence that the prepared optimized formulation may be used effectively for pH sensitive gastric targeted antibiotic such as clarithromycin.

Genetic association of phase I and phase II detoxification genes with recurrent miscarriages among North Indian women.

Allelic variants of the detoxification genes that have impaired biotransformation functions may increase susceptibility to reproductive toxicity leading to endometriosis, recurrent miscarriage (RM) or poor pregnancy outcome. In the present study, we have investigated CYP1A1, CYP2D6, GSTT1, GSTP1 and GSTM1, which are involved in the phase I and phase II detoxification systems, in relation to their role in the etiology of unexplained RMs. In a case-control study, we have investigated 200 females with RM and 300 age and ethnically matched healthy controls with successful reproductive history from North India. The frequencies of phase I wild-type genotypes of CYP1A1 and CYP2D6 in RM cases were 0.56 and 0.60, whereas in controls these were 0.68 and 0.65, respectively (both P < 0.05). The GSTM1 null-genotype frequencies were 0.66 and 0.84 among RM cases and controls, respectively, the GSTT1 null-genotype frequencies were 0.52 and 0.45 (P < 0.005) and the GSTP1 variant allele frequencies were 0.23 and 0.20, respectively. In conclusion, we observed significant protective effects of phase I wild-type genotypes and association of the GSTT1 null genotype with RM. Through combined analyses we have highlighted the importance of the balance of phase I/phase II detoxification systems, in the etiology of RM.