Search for Antideuterons of Cosmic Origin Using the BESS-Polar II Magnetic-Rigidity Spectrometer.

We searched for antideuterons (d[over ¯]'s) in the 4.7×10^{9} cosmic-ray events observed during the BESS-Polar II flight at solar minimum in 2007-2008 but found no candidates. The resulting 95% C.L. upper limit on the d[over ¯] flux is 6.7×10^{-5} (m^{2} s sr GeV/n)^{-1} in an energy range from 0.163 to 1.100 GeV/n. The result has improved by more than a factor of 14 from the upper limit of BESS97, which had a potential comparable to that of BESS-Polar II in the search for cosmic-origin d[over ¯]'s and was conducted during the former solar minimum. The upper limit of d[over ¯] flux from BESS-Polar II is the first result achieving the sensitivity to constrain the latest theoretical predictions.

Molecular characterization of N-methyl-d-aspartate receptor from Bemisia tabaci.

The N-methyl-d-aspartate receptors (NMDARs) are ionotropic ligand gated channels that are highly permeable to calcium ions. In insects, NMDARs are associated with glutamatergic neurotransmission governing diverse physiological and biological processes like vitellogenesis and ovarian development. Therefore, NMDAR may act as attractive target for insect pest control. In present study, we performed structural and functional characterization of NMDARs in Bemisia tabaci, a highly invasive crop pest and potent virus vector. We identified that NMDAR consists of three subunits each encoded by single gene in whiteflies which are highly conserved among different insect orders. Expression analysis suggests that subunit 1 (BtNR1) and subunit 2 (BtNR2) are the main functional units. External supplementation of NMDAR ligand or BtNRs silencing was lethal to insects, which suggested that NMDAR function is highly balanced in whiteflies.

Source of Energetic Protons in the 2014 September 1 Sustained Gamma-ray Emission Event.

We report on the source of > 300 MeV protons during the SOL2014-09-01 sustained gamma-ray emission (SGRE) event based on multi-wavelength data from a wide array of space- and ground-based instruments. Based on the eruption geometry we provide concrete explanation for the spatially and temporally extended γ -ray emission from the eruption. We show that the associated flux rope is of low inclination (roughly oriented in the east-west direction), which enables the associated shock to extend to the frontside. We compare the centroid of the SGRE source with the location of the flux rope's leg to infer that the high-energy protons must be precipitating between the flux rope leg and the shock front. The durations of the SOL2014-09-01 SGRE event and the type II radio burst agree with the linear relationship between these parameters obtained for other SGRE events with duration ≥ 3 hrs . The fluence spectrum of the SEP event is very hard, indicating the presence of high-energy (GeV) particles in this event. This is further confirmed by the presence of an energetic coronal mass ejection with a speed > 2000 km s - 1 , similar to those in ground level enhancement (GLE) events. The type II radio burst had emission components from metric to kilometric wavelengths as in events associated with GLE events. All these factors indicate that the high-energy particles from the shock were in sufficient numbers needed for the production of γ -rays via neutral pion decay.

Studies on drying and storage of chilgoza (Pinus gerardiana) nuts.

Present studies were undertaken with the aim of screening a suitable mode of drying and packaging material for storage of chilgoza nuts. A temperature of 55 °C was found most suitable for the drying of nuts in cabinet drier. Cabinet drier was found the best drying mode among four for drying of chilgoza nuts on the basis of quality characteristics such as moisture, water activity and sensory attributes. Further, out of five packaging materials selected in the study, glass jar followed by aluminium laminate pouch was found to be suitable for the packing and storage of dried nuts in ambient conditions for 6 months on the basis of retention of better physico-chemical and sensory attributes.

Bacterial isolates and its antibiotic susceptibility pattern in NICU.

BACKGROUND: Neonatal sepsis is one of the major causes of morbidity and mortality among the newborns in the developing world. OBJECTIVES: To determine the common bacterial isolates causing sepsis in neonatal intensive care unit and its antibiotic susceptibility pattern. METHODS: A one year discriptive prospective study was conducted in neonatal intensive care unit to analyse the results of blood culture and to look into the sensitivity of the commonly used antibiotics. RESULTS: The blood culture yield by conventional method was 44.13% with nosocomial sepsis accounting for 10.79%. 84.08% were culture proven early onset sepsis and 15.95% were late onset sepsis. Klebsiella infection was the commonest organism isolated in early, late and nosocomial sepsis but statistically not significant. Gram positive organisms were 39.36% in which Staphylococcus aureus was the leading microorganism followed by coagulase negative staphylococcus areus. Gram negative organisms were 60.64% amongst them Klebsiella was the most often encountered followed by Pseudomonas. The most common organism Klebsiella was 87.5% and 78.3% resistance to ampicillin and gentamycin respectively. Among gram negative isolates 87.5% and 77.2% were resistance to ampicillin and gentamycin respectively. Among gram positive isolates 58.5% and 31.5% resistance were noted to ampicillin and gentamycin respectively. Resistance to cefotaxim to gram negative and gram positive isolates were 87.34% and 59.35% respectively. CONCLUSION: Klebsiella is most common organism which is almost resistance to first line antibiotics. Resistance to both gram negative and gram positive isolates among firstline antibiotics and even with cefotaxim is emerging and is a major concern in neonatal intensive care unit.

Measurement of the cosmic-ray antiproton spectrum at solar minimum with a long-duration balloon flight over antarctica.

The energy spectrum of cosmic-ray antiprotons (p's) from 0.17 to 3.5 GeV has been measured using 7886 p's detected by BESS-Polar II during a long-duration flight over Antarctica near solar minimum in December 2007 and January 2008. This shows good consistency with secondary p calculations. Cosmologically primary p's have been investigated by comparing measured and calculated p spectra. BESS-Polar II data show no evidence of primary p's from the evaporation of primordial black holes.

Search for antihelium with the BESS-Polar spectrometer.

In two long-duration balloon flights over Antarctica, the Balloon-borne Experiment with a Superconducting Spectrometer (BESS) collaboration has searched for antihelium in the cosmic radiation with the highest sensitivity reported. BESS-Polar I flew in 2004, observing for 8.5 days. BESS-Polar II flew in 2007-2008, observing for 24.5 days. No antihelium candidate was found in BESS-Polar I data among 8.4×10(6) |Z|=2 nuclei from 1.0 to 20 GV or in BESS-Polar II data among 4.0×10(7) |Z|=2 nuclei from 1.0 to 14 GV. Assuming antihelium to have the same spectral shape as helium, a 95% confidence upper limit to the possible abundance of antihelium relative to helium of 6.9×10(-8)} was determined combining all BESS data, including the two BESS-Polar flights. With no assumed antihelium spectrum and a weighted average of the lowest antihelium efficiencies for each flight, an upper limit of 1.0×10(-7) from 1.6 to 14 GV was determined for the combined BESS-Polar data. Under both antihelium spectral assumptions, these are the lowest limits obtained to date.

Status of glaciers and climate change of East Karakoram in early twenty-first century.

This study aims to assess the glaciers change status in Shyok basin, East Karakoram in terms of area and mass balance, and to ascertain if glaciers in this part of Karakoram also display similar anomaly like central or western counterparts. The spatio-temporal monitoring of glaciers during the time frame of 24 years (1990/2000/2014) suggests major percentage of stable glaciers and insignificant retreat in the total glaciated area. The percentage of retreating glaciers has increased after year 2000. Area change analysis of glaciers having size >1 km2 (569 glaciers) suggests the number of fluctuated glaciers have substantially increased i.e. overall 56% glaciers remained stable, 35% retreated and 9% advanced during 1990-2014. The geodetic based findings using SRTM-C (2000) and Cartosat-1 (2010/2011/2014) stereo-imageries for 201 glaciers suggest the mass loss at the rate of -0.10 ± 0.07 m w.e. a-1. To better apprehend the status of glaciers changes in the region, climatic studies using in-situ observations as well as reanalysis dataset (ERA-I) were also undertaken of past 30 years. Therefore, this study is also a maiden attempt to ascertain if along with Karakoram anomaly, a climatic anomaly exists in the Eastern parts of Karakoram or not. A long term field collected snow-meteorological data of East Karakoram region suggests overall warming trend in annual temperature and no trend for snowfall during 1985-2015. The statistically significant increased rates of warming and decreased snowfall after year 2000 support the spatial variations in glaciers of East Karakoram and marginal mass loss. The observations of the marginal mass loss along with warming temperatures indicate that no Karakoram or climatic anomaly is existent over the East Karakoram region. Our study further refutes the prevalence of the elevation-dependent warming (EDW) over East Karakoram region vis-à-vis North-West Himalayan ranges. The impact of hiatus in global warming was also not observed over studied region.

Bottom-Up Nanofabrication with Extreme-Ultraviolet Light: Metal-Organic Frameworks on Patterned Monolayers.

The fabrication of integrated circuits with ever smaller (sub-10 nm) features poses fundamental challenges in chemistry and materials science. As smaller nanostructures are fabricated, thinner layers of materials are required, and surfaces and interfaces gain a more important role in the formation of nanopatterns. We present a new bottom-up approach in which we use the high optical resolution offered by extreme ultraviolet (EUV) lithography to print patterns on self-assembled monolayers (SAMs). Upon radiation, low-energy electrons induce chemical changes in the SAM so that the projected image is transferred to the substrate surface. We use the chemical differences between exposed and unexposed regions to promote a selective growth of hybrid structures that can act as an etch-resistant layer for further pattern transfer or can be used as functional nanostructures. The EUV doses required to promote selective growth on exposed areas are close to industrial requirements. Furthermore, this method allows for the independent tuning of different steps in the EUV lithography process (photo-induced chemistry, spatially resolved chemical contrast, and formation of nanopatterns), an advantage over current resists, in which the same material plays all roles.

The M10 locus of murine gammaherpesvirus 68 contributes to both the lytic and the latent phases of infection.

Murine gammaherpesvirus 68 (MHV-68) is closely related to Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus (KSHV) and provides a small-animal model to study the pathogenesis of gammaherpesvirus (gammaHV) infections. According to the colinear organization of the gammaHV genomes, the M10 locus is situated at a position equivalent to the K12 locus of KSHV, which codes for proteins of the kaposin family. The M10 locus of MHV-68 has been predicted to code for three overlapping open reading frames (M10a, M10b, and M10c [M10a-c]) with unknown function. In addition, the M10 locus contains a lytic origin of replication (oriLyt). To elucidate the function of the M10 locus during lytic and latent infections, we investigated, both in vitro and in vivo, the following four recombinant viruses which were generated using MHV-68 cloned as a bacterial artificial chromosome: (i) a mutant virus with a deletion which affects both the coding region for M10a-c and the oriLyt; (ii) a revertant virus in which both the M10a-c coding region and the oriLyt were reverted to those of the wild type; (iii) a virus with an ectopic insertion of the oriLyt, which restores the function of the oriLyt but not the M10a-c coding region; and (iv) a mutant virus with a deletion in the oriLyt only. While the mutants were slightly attenuated with regard to lytic replication in cell culture, they showed severe growth defects in vivo. Both lytic replication and latency amplification were strongly reduced. In contrast, both the revertant virus and the virus with the ectopic oriLyt insertion grew very similarly to the parental wild-type virus both in vitro and in vivo. Thus, we provide genetic evidence that mutation of the oriLyt, and not of putative protein coding sequences within the M10a-c region, is responsible for the observed phenotype. We conclude that the oriLyt in the M10 locus plays an important role during infection of mice with MHV-68.

Standardization of pre-treatments for the preparation of dried arils from wild pomegranate.

Pre-treatments for drying of wild pomegranate arils were standardized to check discolouration of dried arils. Steam blanching of arils for 30 s followed by sulphur fumigation at 0.3% for 60 min was found suitable as they took minimum time to dry a given tray load, and had minimum non-enzymatic browning, furfural, hydroxyl methyl furfural and moisture contents. The dried arils of the standardized pre-treatment recorded maximum scores for sensory characteristics like colour, texture, taste, aroma and overall acceptability.

An epistatic interaction between pre-natal smoke exposure and socioeconomic status has a significant impact on bronchodilator drug response in African American youth with asthma.

BACKGROUND: Asthma is one of the leading chronic illnesses among children in the United States. Asthma prevalence is higher among African Americans (11.2%) compared to European Americans (7.7%). Bronchodilator medications are part of the first-line therapy, and the rescue medication, for acute asthma symptoms. Bronchodilator drug response (BDR) varies substantially among different racial/ethnic groups. Asthma prevalence in African Americans is only 3.5% higher than that of European Americans, however, asthma mortality among African Americans is four times that of European Americans; variation in BDR may play an important role in explaining this health disparity. To improve our understanding of disparate health outcomes in complex phenotypes such as BDR, it is important to consider interactions between environmental and biological variables. RESULTS: We evaluated the impact of pairwise and three-variable interactions between environmental, social, and biological variables on BDR in 233 African American youth with asthma using Visualization of Statistical Epistasis Networks (ViSEN). ViSEN is a non-parametric entropy-based approach able to quantify interaction effects using an information-theory metric known as Information Gain (IG). We performed analyses in the full dataset and in sex-stratified subsets. Our analyses identified several interaction models significantly, and suggestively, associated with BDR. The strongest interaction significantly associated with BDR was a pairwise interaction between pre-natal smoke exposure and socioeconomic status (full dataset IG: 2.78%, p = 0.001; female IG: 7.27%, p = 0.004)). Sex-stratified analyses yielded divergent results for females and males, indicating the presence of sex-specific effects. CONCLUSIONS: Our study identified novel interaction effects significantly, and suggestively, associated with BDR in African American children with asthma. Notably, we found that all of the interactions identified by ViSEN were "pure" interaction effects, in that they were not the result of strong main effects on BDR, highlighting the complexity of the network of biological and environmental factors impacting this phenotype. Several associations uncovered by ViSEN would not have been detected using regression-based methods, thus emphasizing the importance of employing statistical methods optimized to detect both additive and non-additive interaction effects when studying complex phenotypes such as BDR. The information gained in this study increases our understanding and appreciation of the complex nature of the interactions between environmental and health-related factors that influence BDR and will be invaluable to biomedical researchers designing future studies.

Balantidiasis in an Asiatic elephant and its therapeutic management.

A 14 years old female Asiatic elephant was presented to the hospital with a history of mucoid watery diarrhea, inappetence and lethargy. Clinical examination revealed normal body temperature (98.2 °F), tachycardia (42 bpm), eupnoea (14/min), congested mucous membrane and dehydration. Haemato-biochemical parameters are well within the range. Microscopic examination of faecal sample revealed presence of live, motile and pear shaped ciliated Balantidium coli protozoa. Based on clinical and laboratory examination, the condition was diagnosed as balantidiasis. The animal was treated with Tab. Metronidazole (10 mg/Kg, PO, BID) for 5 days. Supportive treatment was done with antacids, hepatoprotectants and multivitamin supplements. An uneventful recovery was noticed after 5 days of treatment.

Genetic characterization of Bactrocera fruit flies (Diptera: Tephritidae) from Northeastern India based on DNA barcodes.

The Northeastern region of India, one of the mega biodiversity hot spots has enormous potential for the production of fruits and vegetables. Fruit flies of the genus Bactrocera Macquart are important pests of fruits and vegetables, and one of the limiting factors in successful production of these commodities. The relationship among some of the species is unclear due to their high molecular and morphological similarities. Moreover, due to the significant morphological resemblance between fruit fly species, reliable identification is very difficult task. We genetically characterized 10 fruit fly species of the genus Bactrocera by using standard DNA barcoding region of COI gene. The characterization and identification of eight species were straight forward. This study was unable to establish the molecular identity of Bactrocera sp. 2. Within the 547 bp region of partial COI gene, there were 157 variable sites of which 110 sites were parsimony informative, 153 were synonymous substitutions and 4 were non-synonymous substitutions. The estimate of genetic divergence among the ten species was in the range of 0-21.9% and the pairwise genetic distance of Bactrocera. (Bactrocera) dorsalis (Hendel) with B. (B.) carambolae was only 0.7%. Phylogenetic analysis formed separate clades for fruit and vegetable infesting fruit flies. B. (B.) aethriobasis Hardy, B. (B.) thailandica and B. (B.) tuberculata (Bezzi) have been reported for the first time from the Northeastern India. The information generated from this study would certainly have implications for pest management, taxonomy, quarantine and trade.

Laser Raman spectroscopy of THF clathrate hydrate in the temperature range 90-300 K.

In situ Raman and Fourier transform infrared (FTIR-NIR) spectroscopic studies on tetrahydrofuran (THF-C(4)H(8)O) clathrate hydrate (CH) were reported. The Raman results in lattice (64 cm(-1)), ring breathing and C-H stretching mode regions are in conformity with earlier reports, while the FTIR (NIR) studies in second order mode region were reported for the first time. Comparison of the results indicate that the band assigned to ring breathing mode around 922 cm(-1) (in Raman) and corresponding second order mode in NIR around 4295 cm(-1) broadens and shifts in enclathrated THF. The ring breathing mode at lower temperatures (T<120 K) is highly asymmetric and splits into two and are due to different host-guest interactions at lower temperatures.

Large proximal ureteral stones: Ideal treatment modality?

BACKGROUND AND PURPOSE: Ideal treatment modality for patients with large impacted proximal ureteral stone remains controversial. We compared laparoscopic transperitoneal ureterolithotomy (Lap-TPUL) and semirigid ureteroscopy for large proximal ureteric stones to evaluate their efficacy and safety. PATIENTS AND METHODS: From November 2012 to December 2014, we enrolled 122 patients with large (≥1.5 cm) proximal ureteral stone in the study. Patients were randomly divided into two groups: Group A (60 patients), retrograde ureteroscopic lithotripsy using a semirigid ureteroscope; Group B (62 patients), transperitoneal LU (Lap-TPUL). RESULTS: The overall stone-free rate was 71.6% and 93.5% for Group A and Group B respectively (P = 0.008). Auxiliary procedure rate was higher in Group A than in Group B (27.3% vs. 5.6%). The complication rate was 11.2% in Group B versus 25% in Group A. Mean procedure time was higher in laparoscopy group as compared to ureterorenoscopy (URS) groups (84.07 ± 16.80 vs. 62.82 ± 12.71 min). Hospital stay was 4.16 ± 0.67 days in laparoscopy group and 1.18 ± 0.38 days in URS group (P < 0.0001). CONCLUSION: Laparoscopic transperitoneal ureterolithotomy is a minimally invasive, safe and effective treatment modality and should be recommended to all patients of impacted large proximal stones, which are not amenable to URS or extracorporeal shock-wave lithotripsy or as a primary modality of choice especially if patient is otherwise candidate for open surgery.

Complete mitochondrial genome sequence of a phytophagous ladybird beetle, Henosepilachna pusillanima (Mulsant) (Coleoptera: Coccinellidae).

Ladybird beetles are generally considered as agriculturally beneficial insects, but the ladybird beetles in the coleopteran subfamily Epilachninae are phytophagous and major plant feeding pest species which causes severe economic losses to cucurbitaceous and solanaceous crops. Henosepilachna pusillanima (Mulsant) is one of the important pest species of ladybird beetle. In this report, we sequenced and characterized the complete mitochondrial genome of H. pusillanima. For sequencing of the complete mitochondrial genome, we used the Ion Torrent sequencing platform. The complete circular mitochondrial genome of the H. pusillanima was determined to be 16,216 bp long. There were totally 13 protein coding genes, 22 transfer RNA, 2 ribosomal RNA and a control (A + T-rich) region estimated to be 1690 bp. The gene arrangement and orientations of assembled mitogenome were identical to the reported predatory ladybird beetle Coccinella septempunctata L. This is the first completely sequenced coleopteran mitochondrial genome from the beetle subfamily Epilachninae from India. Data generated in this study will benefit future comparative genomics studies for understanding the evolutionary relationships between predatory and phytophagous coccinellid beetles.

Modulatory role of leptin on ovarian functions in water buffalo (Bubalus bubalis).

The aim of the present study was to demonstrate the modulatory role of leptin on bubaline granulosa cells (GCs) and luteal cells (LCs) functions using an in vitro cell culture system and to establish a cross talk between leptin and insulin-like growth factor-1 (IGF-1). GCs were collected from group IV follicles (>13 mm size) and LCs from mid-luteal phase corpus luteum and were grown in serum-containing media supplemented with leptin at three different dose rates (0.1, 1, and 10 ng/mL) and time durations (24, 48, and 72 hours). We evaluated the production and secretion of estradiol (E2) and progesterone (P4) using RIA and the mRNA expression of steroidogenic acute regulatory protein (STARD1), cytochrome P450 cholesterol side-chain cleavage (CYP11A1), 3ß-hydroxysteroid dehydrogenase (3ß-HSD), cytochrome P450 aromatase (CYP19A1), sterol regulatory element-binding protein 1 (SREBP1), steroidogenic factor-1 (SF1), anti-apoptotic gene PCNA, pro-apoptotic gene caspase 3 and endothelial cell marker, Von Willebrand factor (vWF), using quantitative real-time polymerase chain reaction. The results depicted a direct inhibitory action of leptin on GCs steroidogenesis in a time-dependent manner (P < 0.05), whereas in the presence of IGF-1 the inhibitory effect was reverted. Furthermore, leptin augmented both cellular proliferation (PCNA) and apoptosis (caspase 3). On the other hand, in LCs, leptin alone showed an apparent stimulatory effect on steroidogenesis (P < 0.05); however, in the presence of IGF-1, an antagonistic effect was witnessed. Moreover, leptin had an inhibitory effect on apoptosis while promoted cellular proliferation and angiogenesis. These findings were further strengthened by immunocytochemistry. To conclude, these observations for the first time reported that in buffaloes leptin has a direct dose-, time-, and tissue-dependent effect on ovarian steroidogenesis, angiogenesis, and cytoprotection, and furthermore, it can regulate the effect of systemic factors like IGF-1. Hence, this in vitro study provides an insight into the putative roles of leptin alone and its interactions in vivo.

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 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.