A multi-organ map of the human immune system across age, sex and ethnicity.

Understanding tissue biology's heterogeneity is crucial for advancing precision medicine. Despite the centrality of the immune system in tissue homeostasis, a detailed and comprehensive map of immune cell distribution and interactions across human tissues and demographics remains elusive. To fill this gap, we harmonised data from 12,981 single-cell RNA sequencing samples and curated 29 million cells from 45 anatomical sites to create a comprehensive compositional and transcriptional healthy map of the healthy immune system. We used this resource and a novel multilevel modelling approach to track immune ageing and test differences across sex and ethnicity. We uncovered conserved and tissue-specific immune-ageing programs, resolved sex-dependent differential ageing and identified ethnic diversity in clinically critical immune checkpoints. This study provides a quantitative baseline of the immune system, facilitating advances in precision medicine. By sharing our immune map, we hope to catalyse further breakthroughs in cancer, infectious disease, immunology and precision medicine.

Drip irrigation and sulphur fertilization influenced fodder yield, quality and water use efficiency of groundnut in arid region.

Availability of ample and nutritious fodder for livestock is always a challenge in arid region. Choice of crops such as groundnut that can fulfil the requirement of fodder with its crop residues along with human needs can be a viable option to bridge the gap between availability and requirement of fodder. The fodder yield and quality largely depend on soil moisture and nutrient supply especially sulphur (S), a key nutrient for improving groundnut fodder quality. However, no researchers have given emphasis on coupling effect of drip irrigation (DI) and sulphur on fodder yield, quality, digestibility and water use efficiency (WUE). Therefore, the study was conducted to determine the effects of different regimes of DI and S on productivity and quality of fodder. Results revealed that higher regimes of DI i.e. 0.8 +1.0 PE(pan evaporation) and 1.0 PE level of irrigation along with 40 kg S ha-1 significantly improved the yield, primary quality traits (crude protein, ether extract and ash), digestibility indices and significant reduction in Fibers which indicates improvement in quality of fodder. Fodder productivity was 27.0 and 25.6% higher in 1.0 PE and 0.8+1.0 PE level of irrigation, respectively, as compared to 0.6 PE level (lower water regime) of irrigation, although 0.6 PE level of irrigation recorded higher WUE and was at par with 0.8 PE and 0.6 +1.0 PE level of DI. By changing the levels of DI from 1.0 PE to 0.8+1.0 PE, considerable water can be saved without affecting the yield and quality of fodder. Similarly, crop responded to S up to 60 kg ha-1 but at par with 40 kg S ha-1 indicating that application of extra S after 40 kg did not warrant any extra benefit in terms of fodder yield, WUE and quality of fodder. Thus, adjusting the PE levels of DI for water saving and optimal S application can be a sustainable strategy to improve the productivity and quality of groundnut fodder in arid region.

Range grasses to improve soil properties, carbon sustainability, and fodder security in degraded lands of semi-arid regions.

Tropical grasses are the primary source of forage for livestock and a valuable resource for improving soil health and environmental sustainability in semi-arid regions. A study was carried out in a semi-arid region of central India to determine the short-term (6-year) impact of nine range grasses on soil physio-chemical and biological properties, carbon stock, and forage security. The experiment was carried out in a randomized block design with three replications. Results show that the majority of the grass roots were distributed in the upper soil layer (0-10 cm, 63.5-76.5 %), and then in the middle (10-20 cm, 21.3-25 %) and lower (20-30 cm, 2.2-11.5 %) layers. Perennial tussock grass (Heteropogon contortus (L.) P. Beauv. ex Roem. & Schult) had a higher root volume (2219 mm3), followed by Guinea grass [Megathyrsus maximus (Jacq.) B.K. Simon & S.W.L. Jacobs] (1860 mm3). A lower soil bulk density (BD, 1.11-1.23 g cm-3), higher gravimetric water content (GMW, 14.0-17.8 %), and soil organic carbon (0.38-0.73 %) were recorded for grass-cultivated plots compared to the barren land (1.38 g cm-3, 13.0 %, and 0.28 %, respectively). The perennial tussock grass and Guinea grass resulted in the highest soil microbial biomass carbon (SMBC, 70.1 mg kg-1 soil) and enzyme activities (dehydrogenase, 17.09 µg TPF g-1 day-1 and fluorescein diacetate activity 4.94 µg fluorescein g-1 h-1). The considerable improvement in soil properties with minimal inputs resulted in a higher sustainable yield index and carbon sustainability index in plots planted with Guinea grass (0.9 and 89.29) and perennial tussock grass (0.89 and 71.61). Therefore, the cultivation of either Guinea grass or perennial tussock grass as an intercrop or sole crop in the semi-arid environment can be an ecologically sound strategy to improve soil health, C sequestration, and fodder supply.

Adding MR Diffusion Imaging and T2 Signal Intensity to Neck Imaging Reporting and Data System Categories 2 and 3 in Primary Sites of Postsurgical Oral Cavity Carcinoma Provides Incremental Diagnostic Value.

BACKGROUND AND PURPOSE: The NI-RADS lexicon doesn't use ADC parameters and T2 weighted signal for ascribing categories. We explored ADC, DWI, and T2WI to examine the diagnostic accuracy in primary sites of postsurgical oral cavity carcinoma in the Neck Imaging Reporting and Data System (NI-RADS) categories 2 and 3. MATERIALS AND METHODS: We performed a retrospective analysis in clinically asymptomatic post-surgically treated patients with oral cavity squamous cell carcinoma who underwent contrast-enhanced MRI between January 2013 and January 2016. Histopathology and follow-up imaging were used to ascertain the presence or absence of malignancy in subjects with "new enhancing lesions," which were interpreted according to the NI-RADS lexicon by experienced readers, including NI-RADS 2 and 3 lesions in the primary site. NI-RADS that included T2WI and DWI (referred to as NI-RADS A) and ADC (using the best cutoff from receiver operating characteristic curve analysis, NI-RADS B) was documented in an Excel sheet to up- or downgrade existing classic American College of Radiology NI-RADS and calculate diagnostic accuracy. RESULTS: Sixty-one malignant and 23 benign lesions included in the study were assigned American College of Radiology NI-RADS 2 (n = 33) and NI-RADS 3 (n = 51) categories. The recurrence rate was 90% (46/51) for NI-RADS three, 45% (15/33) for NI-RADS 2, and 73% (61/84) overall. T2WI signal morphology was intermediate in 45 subjects (53.5%) and restricted DWI in 54 (64.2%). Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the American College of Radiology NI-RADS were the following: NI-RADS (75.4%, 78.3%, 90.1%, 54.5%, and 76.1%); NI-RADS A (79.1%, 81.2%, 91.9%, 59.1%, and 79.6%); and NI-RADS B (88.9%, 72.7%, 91.4%, 66.7%, and 85.1%), respectively. CONCLUSIONS: Adding MR imaging diagnostic characteristics like T2WI, DWI, and ADC to the American College of Radiology NI-RADS improved diagnostic accuracy and sensitivity.

Bacteriophage therapy of human-restricted Salmonella species-a study in a surrogate bacterial and animal model.

Salmonella Typhi has been an important human-restricted pathogen from time immemorial, and unfortunately, the indiscriminate use of antibiotics has induced the emergence of multidrug resistance in S. Typhi. Bacteriophage therapy may be a possible alternative in countering antimicrobial resistance. Therefore, this study was planned to assess the efficacy of bacteriophages in treating acute and chronic S. Typhimurium infection in the mouse as a surrogate model. We isolated bacteriophages against S. Typhimurium and selected three different bacteriophages for the in vivo experiments. The lethal dose of S. Typhimurium was decided for Swiss albino mice, and acute infection was developed. Further, bacteriophage therapy by daily intraperitoneal injection of phage cocktail was given for 14 days. While the chronic carrier state of S. Typhimurium in Swiss albino mice was developed by inoculating intraperitoneally sequential 10-fold increasing doses of the bacterium. On the successful establishment of carrier state, oral feeding of phage cocktail at a high count was given, which completely cured the carrier state within 7 days of feeding. These experiments confirmed that the phage cocktail could eradicate the S. Typhimurium from the mice in both types of infections, that is acute and chronic.

Active Regulation of Pressure and Volume Defines an Energetic Constraint on the Size of Cell Aggregates.

We explore the relationship between the nonequilibrium generation of myosin-induced active stress within the F-actin cytoskeleton and the pressure-volume relationship of cellular aggregates as models of simple tissues. We find that due to active stress, aggregate surface tension depends upon its size. As a result, both pressure and cell number density depend on size and violate equilibrium assumptions. However, the relationship between them resembles an equilibrium equation of state with an effective temperature. This suggests that bulk and surface properties of aggregates balance to yield a constant average work performed by each cell on their environment in regulating tissue size. These results describe basic physical principles that govern the size of cell aggregates.

Applying comparative molecular modelling techniques on diverse hydroxamate-based HDAC2 inhibitors: an attempt to identify promising structural features for potent HDAC2 inhibition.

Histone deacetylase 2 (HDAC2) has been implicated in a variety of cardiovascular and neurodegenerative disorders as well as in cancers. Thus, HDAC2 has become an exclusive target for anticancer drug development. Therefore, the development of newer HDAC2 inhibitors in disease conditions is a prime goal to restrain such a scenario. Although a handful of HDAC inhibitors was accepted for the treatment of HDAC-related disease conditions, the non-selective nature of these entities is one of the major setbacks in the treatment of specific HDAC isoform-related pathophysiology. In this framework, the analyses of pre-existing molecules are essential to identify the important structural features that can fulfil the requirements for the cap and linker moieties to obtain potent and effective HDAC2 inhibition. Thus, in this study, the implementation of a combined comparative 2D and 3D molecular modelling techniques was done on a group of 92 diverse hydroxamate derivatives having a wide range of HDAC2 inhibitory potency. Besides other crucial features, this study upheld the importance of groups like triazole and benzyl moieties along with the molecular fields that are crucial for regulating HDAC2 inhibition. The outcomes of this study may be employed for the designing of HDAC2 inhibitors in future.

Bacteriophages: A possible solution to combat enteropathogenic Escherichia coli infections in neonatal goats.

Due to awareness and benefits of goat rearing in developing economies, goats' significance is increasing. Unfortunately, these ruminants are threatened via multiple bacterial pathogens such as enteropathogenic Escherichia coli (EPEC). In goat kids and lambs, EPEC causes gastrointestinal disease leading to substantial economic losses for farmers and may also pose a threat to public health via the spread of zoonotic diseases. Management of infection is primarily based on antibiotics, but the need for new therapeutic measures as an alternative to antibiotics is becoming vital because of the advent of antimicrobial resistance (AMR). The prevalence of EPEC was established using bfpA gene, uspA gene and Stx1 gene, followed by phylogenetic analysis using Stx1 gene. The lytic activity of the isolated putative coliphages was tested on multi-drug resistant strains of EPEC. It was observed that a PCR based approach is more effective and rapid as compared to phenotypic tests of Escherichia coli virulence. It was also established that the isolated bacteriophages exhibited potent antibacterial efficacy in vitro, with some of the isolates (16%) detected as T4 and T4-like phages based on gp23 gene. Hence, bacteriophages as therapeutic agents may be explored as an alternative to antibiotics in managing public, livestock and environmental health in this era of AMR.

Cell-Matrix Elastocapillary Interactions Drive Pressure-based Wetting of Cell Aggregates.

Cell-matrix interfacial energies and the energies of matrix deformations may be comparable on cellular length-scales, yet how capillary effects influence tis sue shape and motion are unknown. In this work, we induce wetting (spreading and migration) of cell aggregates, as models of active droplets onto adhesive substrates of varying elasticity and correlate the dynamics of wetting to the balance of interfacial tensions. Upon wetting rigid substrates, cell-substrate tension drives outward expansion of the monolayer. By contrast, upon wetting compliant substrates, cell substrate tension is attenuated and aggregate capillary forces contribute to internal pressures that drive expansion. Thus, we show by experiments, data-driven modeling and computational simulations that myosin-driven 'active elasto-capillary' effects enable adaptation of wetting mechanisms to substrate rigidity and introduce a novel, pressure-based mechanism for guiding collective cell motion.

Spatiotemporal sensitivity of mesoderm specification to FGFR signalling in the Drosophila embryo.

Development of the Drosophila embryonic mesoderm is controlled through both internal and external inputs to the mesoderm. One such factor is Heartless (Htl), a Fibroblast Growth Factor Receptor (FGFR) expressed in the mesoderm. Although Htl has been extensively studied, the dynamics of its action are poorly understood after the initial phases of mesoderm formation and spreading. To begin to address this challenge, we have developed an optogenetic version of the FGFR Heartless in Drosophila (Opto-htl). Opto-htl enables us to activate the FGFR pathway in selective spatial (~ 35 µm section from one of the lateral sides of the embryo) and temporal domains (ranging from 40 min to 14 h) during embryogenesis. Importantly, the effects can be tuned by the intensity of light-activation, making this approach significantly more flexible than other genetic approaches. We performed controlled perturbations to the FGFR pathway to define the contribution of Htl signalling to the formation of the developing embryonic heart and somatic muscles. We find a direct correlation between Htl signalling dosage and number of Tinman-positive heart cells specified. Opto-htl activation favours the specification of Tinman positive cardioblasts and eliminates Eve-positive DA1 muscles. This effect is seen to increase progressively with increasing light intensity. Therefore, fine tuning of phenotypic responses to varied Htl signalling dosage can be achieved more conveniently than with other genetic approaches. Overall, Opto-htl is a powerful new tool for dissecting the role of FGFR signalling during development.

Effect of Cryopreservation on Semen Quality Parameters in Relation to Lipid Peroxidation and Antioxidant Profile in Indian Buffalo.

BACKGROUND: Lipid peroxidation (LPO) due to oxidative stress leads to structural and functional changes in spermatozoa. OBJECTIVE: To evaluate any association of various seminal characteristics at the pre- and post-cryopreservation stages with LPO and total antioxidant capacity (TAC) in Murrah buffalo semen samples. MATERIALS AND METHODS: Sixty-five ejaculates from seven bulls were processed for cryopreservation in liquid nitrogen. RESULTS: Only 31 (47.7%) samples were found satisfactory for inclusion in the further artificial insemination. A strong negative correlation was observed between LPO and individual progressive motility, TAC, viability, plasma membrane integrity as well as acrosome integrity of fresh spermatozoa. At the post-thaw stage, post-thaw motility, viability, plasma membrane integrity and acrosome integrity had strong positive correlation with TAC. CONCLUSION: The effort to minimize LPO and enhance TAC shall play a pivotal role in improving buffalo semen quality upon cryopreservation.

Prevalence, risk factors and morbidities of gestational diabetes among pregnant women attending a hospital in an urban area of Bhubaneswar, Odisha.

BACKGROUND: Gestational diabetes mellitus (GDM) causes several maternal and neonatal complications. AIMS: This exploratory study was conducted to estimate the prevalence, determine the risk factors and morbidities among pregnant women. METHODS: In this prospective study, 1557 pregnant women attending the Gyn. & Obs. clinic of a hospital in an urban area of Bhubaneswar were enrolled. Various socio-demographic factors and clinical profiles were assessed. We used a Glucometer for the diagnosis of GDM. RESULTS: More younger pregnant women residing in slums, sedentary and overweight were having diabetes. A large percentage of pregnant women living in rural areas and slums visit the government hospitals as they are benefitted by the State govt.'s scheme, Mamata. Pregnant women residing in the urban areas prefer to go for ante-natal check-ups in private Nursing homes/Clinics owing to the crowd and prolonged waiting hours. In this study, body mass index (BMI) and family history of the pregnant women appeared to be the significant risk factors for the gestational diabetes. Out of 1557 pregnant women, 154 were having diabetes, the prevalence being 9.89%. This is low when compared to the studies reported from other regions of the country. CONCLUSIONS: Gluco-One is suitable for screening gestational diabetes using the optimal threshold capillary glucose level of 140 mg/dl. As the pregnant women find it difficult to come the next day just to collect the results, this facilitated in getting the test results promptly and appropriate consultation by Doctor the same day. Glucometer can be used for accurate screening of gestational diabetes mellitus. Pregnant women with screening values not normal were identified on the spot and followed up at regular intervals. Screening for diabetes among pregnant women would result in early case detection indirectly resulting in better outcomes of treatment and prevention of complications.

Polymorphism distribution of RYR1, PRKAG3, HFABP, MYF-5 and MC4R genes in crossbred pigs.

This study was designed to screen the crossbred pigs for SNPs in five candidate genes, associated with pork quality traits and to differentiate their genotypes by PCR-RFLP. The results indicated that genotypes of crossbred pigs were NN (90%) and Nn (10%) for RYR1; RR (83%) and QR (17%) for PRKAG3; HH (98%), Hh (1%) and hh (1%) for HFABP; DD (99%) and CD (1%) for MYF-5; and AG (57%), GG (26%) and AA (17%) for MC4R SNPs, respectively. Allelic frequencies for five SNPs {RYR1 (1843C>T), PRKAG3 (c.599G>A), HFABP (c.1322C>T), MYF-5 (c.1205A>C) and MC4R (c.1426A>G)} were 0.95 and 0.05 (N/n), 0.08 and 0.92 (Q/R), 0.99 and 0.01 (H/h), 0.00 and 1.00 (C/D) and 0.45 and 0.55 (A/G), respectively. The effect of RYR1 (1843C>T) SNP was significant on pH45 (P < 0.05), pH24 (P < 0.05) and protein % (P < 0.05). The PRKAG3 (c.599G>A) and MC4R (c.1426A>G) SNP had significant association with dressing percentages. The results revealed that RYR1, PRKAG3 and MC4R SNPs may be used in marker associated selection for pork quality traits in crossbred pigs.

Identification of mega-environments and rice genotypes for general and specific adaptation to saline and alkaline stresses in India.

In the present study, a total of 53 promising salt-tolerant genotypes were tested across 18 salt-affected diverse locations for three years. An attempt was made to identify ideal test locations and mega-environments using GGE biplot analysis. The CSSRI sodic environment was the most discriminating location in individual years as well as over the years and could be used to screen out unstable and salt-sensitive genotypes. Genotypes CSR36, CSR-2K-219, and CSR-2K-262 were found ideal across years. Overall, Genotypes CSR-2K-219, CSR-2K-262, and CSR-2K-242 were found superior and stable among all genotypes with higher mean yields. Different sets of genotypes emerged as winners in saline soils but not in sodic soils; however, Genotype CSR-2K-262 was the only genotype that was best under both saline and alkaline environments over the years. The lack of repeatable associations among locations and repeatable mega-environment groupings indicated the complexity of soil salinity. Hence, a multi-location and multi-year evaluation is indispensable for evaluating the test sites as well as identifying genotypes with consistently specific and wider adaptation to particular agro-climatic zones. The genotypes identified in the present study could be used for commercial cultivation across edaphically challenged areas for sustainable production.

Genetic diversity, population structure and association analysis in linseed (Linum usitatissimum L.).

The present investigation aimed to explore the level of genetic diversity, determine the population structure in a larger set of germplasm of linseed using microsatellite marker and identify linked markers through association mapping. A total of 168 accessions of linseed were evaluated for major agro-economic traits and SSRs markers deployed for diversity assessment. A total of 337 alleles were amplified by 50 SSRs ranging from 2 to 13 with an average of 6.74 ± 2.8 alleles per loci. The neighbor joining based clustering grouped all the accessions into three major clusters that were also confirmed by scatter plot of PCoA. While model based clustering determined four sub-populations (K = 4). Further, analysis of molecular variance analysis considering three population showed that maximum variation (79%) was within the population. We identified one putative SSR marker (Lu_3043) linked with days to 50% flowering through both GLM and MLM analysis of association mapping. The results of this preliminary study revealed genetic diversity, population structure in linseed and linked marker which could be utilized in future breeding program.

Expression dynamics of HSP70 during chronic heat stress in Tharparkar cattle.

Six male Tharparkar cattle aged 2-3 years were selected for the study. The animals were acclimatized in the psychrometric chamber at thermoneutral zone (TNZ) for 15 days and then exposed to 42 °C temperature up to 23 days followed by 12 days of recovery period. Physiological responses were estimated, and peripheral blood mononuclear cells (PBMCs) were isolated at TNZ on day 1, day 5, and day 12; after 6 h of heat stress exposure on day 16 to day 20, day 25, day 30, day 32, day 34, day 36, and day 38; and a recovery period on day 45 and day 50. The PBMCs were cultured to study the effect of thermal challenge on HSP70 messenger RNA (mRNA) expression pattern at different temperature-time combinations. The mRNA and protein expression of HSP70 in PBMCs along with serum extracellular HSP70 (eHSP70) was increased (P < 0.05) and showed two peaks on day 17 and day 32 (2nd and 17th days of thermal challenge, respectively). The HSP70 mRNA expression was increased (P < 0.05) in a temperature- and time-dependent manner in heat stress challenge treatment as compared to control in cultured PBMCs. HSP70 expression was found to be higher (P < 0.05) after 10 days of heat exposure (corresponds to chronic heat stress) as compared to the first 5 days of heat stress (corresponds to short-term heat stress) and control period at TNZ. The present findings indicate that HSP70 is possibly involved in heat stress adaptive response in Tharparkar cattle and the biphasic expression pattern may be providing a second window of protection during chronic heat stress.

Expression and localization of fibroblast growth factor (FGF) family in buffalo ovarian follicle during different stages of development and modulatory role of FGF2 on steroidogenesis and survival of cultured buffalo granulosa cells.

The present study investigated the expression and localization of FGF and its functional receptors in the follicle of buffalo and the treatment of FGF2 on mRNA expression of CYP19A1 (aromatase), PCNA, and BAX (BCL-2 associated X protein) in cultured buffalo granulosa cells (GCs). Follicles were classified into four groups based on size and E2 level in follicular fluid (FF): F1, 4-6mm diameter, E2<0.5ng/ml of FF; F2, 7-9mm, E2=0.5-5ng/ml; F3, 10-13mm, E2=5-40ng/ml; F4, >14mm, E2>180ng/ml. The qPCR studies revealed that the mRNA expression of FGF1, FGF2 and FGF7 were maximum (P<0.05) in theca interna (TI) whereas the transcripts of FGFR1, FGFR2, FGFR2IIIB and FGFR2IIIC were up-regulated (P<0.05) in GCs of F4 follicles. Protein expression of most members were maximum (P<0.05) in F4 follicles except FGFR3 and FGFR4. All members were localized in GC and TI with a stage specific immunoreactivity. Primary culture of GCs with treatment of FGF2 at different dose-time combinations revealed that the mRNA expression and immunoreactivity of CYP19A1 and PCNA were maximum (P<0.05) whereas BAX was minimum (P<0.05) with 200ng/ml at 72h of incubation. The findings indicate that FGF family members are expressed in a regulated manner in buffalo ovarian follicles during different stages of development where FGF2 may promote steroidogenesis and GC survival through autocrine and paracrine manner.

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis, angiogenesis and survivability of cultured buffalo luteal cells.

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P4 ) secretion and mRNA expression of phosphotidylinositide-3kinase-protein kinase B (PI3K-AKT), phosphoinositide-dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real-time RT-PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P4 secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P4 secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P4 secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.

Expression and localization of angiopoietin family in buffalo ovarian follicles during different stages of development and modulatory role of angiopoietins on steroidogenesis and survival of cultured buffalo granulosa cells.

The present study investigated the expression and localization of angiopoietin (ANPT) family members in buffalo ovarian follicles of different size. It also looked at the role of ANPTs in estradiol secretion and mRNA expression of phosphoinositide-3-kinase-protein kinase B signaling pathway cellular proliferation (phosphoinositide-dependant kinase and protein kinase B [AKT]) and proapoptotic (BAD) factors with caspase 3 in cultured buffalo granulosa cells (GCs). The mRNA and protein expression of ANPT-1 was greatest (P < 0.05), whereas ANPT-2 was reduced (P < 0.05) in preovulatory follicles as compared to F1 follicle. Tyrosine kinase with immunoglobulin-like and EGF-like domains 1 transcripts and protein expression did not change in all follicular groups, whereas tyrosine kinase with immunoglobulin-like and EGF-like domains 2 mRNA was highest (P < 0.05) in theca interna but not GC layer of preovulatory follicle. All members of ANPT family were localized in GC and theca interna showing a stage specific immunoreactivity. Cultured GCs were treated with ANPT-1 and ANPT-2 separately at doses of 1, 10, and 100 ng/mL and in combination at 100 ng/mL for three incubation periods (24, 48, and 72 hours). Estradiol secretion was highest (P < 0.05) at 100 ng/mL at 72 hours of incubation when GCs were treated with either protein alone. The mRNA expression of phosphoinositide-dependant kinase and AKT was highest (P < 0.05), and BAD with caspase 3 was lowest (P < 0.05) at 100 ng/mL at 72 hours of incubation, when cultured GCs were treated separately with each protein or in combination. The immuoreactivity of AKT, pAKT, and pBAD were maximal, whereas BAD was minimal with 100 ng/mL at 72 hours when cultured GCs treated with either protein alone. The findings indicate that ANPTs are expressed in a regulated manner in buffalo ovarian follicle during different stages of development where they may promote steroidogenesis and GC survival through autocrine and paracrine actions.

Development of Crystalline Cellulosic Fibres for Sustained Release of Drug.

Natural quinoline alkaloid camptothecin (CPT) is used for the treatment of colon, lung, breast and ovarian cancers still facing challenges due to low solubility in aqueous and biological fluids. Its lactone form easily converts into a toxic carboxylic form at slightly basic pH, typical in blood and tissue fluid has rapid clearance from systemic administration. We report a new approach based on micro crystalline cellulose (MCC) and nano crystalline cellulose (NCC) isolated from natural sources such as Cymbopogan flexuosus to stabilize and regulate the release kinetics of CPT in physiological solution. Langmuir and Freundlich isotherm studies approve that degree of crystallinity i.e. ratio of amorphous and crystalline cellulose regulate the adsorption of CPT. The freeze dried celluloses of Cymbopogan flexuosus origin (MCC and NCC) further were optimized for drug delivery with a mimicked physiologically relevant solution. Both carriers can significantly extend the release of drug as compared to reported values, however, NCC showed better results. Not only the crystallinity but crystal size and hydrogen bonding play critical role in drug release. Free diffusion of drug into physiological solution follows the Ritger- Peppes kinetic model. The coefficient of the model signifies the Fickian diffusion mechanism of release. The investigation indicates that NCC cellulosic matrix can act as a better carrier of CPT for its sustained release formulation.