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MicroRNAs (miRNAs) are involved in post-transcriptional gene regulation and single miRNAs regulating multiple mRNAs targets. Using bioinformatics approaches, this work aims to identify and characterize the miRNAs that could potentially bind with PDKCC, AP3S2 and SEC61A2 gene which regulate protein transport during heat stress. To resolve this problem, three freely available online miRNAs target finder databases were applied to identify the potential miRNAs which may target PDKCC, AP3S2 and SEC61A2 gene. The prediction results showed that a total of 115, 67 and 25 miRNAs for PDKCC, 267, 111 and 12 miRNAs for AP3S2 and 92, 120 and 31 miRNAs for SEC61A2 were identified as a possible potential candidate by TargetScan, MirTarget2 and miRanda respectively. Following the clustering process, we found eight miRNAs for PDKCC, three for AP3S2 and nine for SEC61A2s identified as possible candidates by all three databases. Five miRNAs in PDKCC, four in AP3S2 and five in the SEC61A2 gene have 100% identical sequence resemblance between human and bovine miRNAs. miRNAs binding site analysis showed that miR-34a in AP3S2 and miR-377 in SEC61A2 has the highest number of binding sites while miR-30b and miR-30c have two binding sites in 3´UTR of SEC61A2 gene whereas the rest of the miRNAs have the lowest number of the binding site in PDKCC and SEC61A2 gene respectively. Our study identified total four potential miRNAs that have different binding sites at the AP3S2 and SEC61A2 3´UTR site along with seven miRNAs which have single binding site and can modulate protein transport, translocation and other cellular pathways.

Mesoporous silica nanoparticles (MSN), multiwalled carbon nanotube (MWCNT) and zinc oxide (ZnO).

Probiotics have been suggested as alternatives to pharmacological drugs in the treatment of a variety of medical problems, including obesity management, which is often linked to low sperm production. Also, probiotic fermented products are known to boost host immune response, immunosenescence, infection tolerance, and redox homeostasis, but their direct role in male fertility has been less investigated. This study assessed the effect of two probiotic strains

Obesity has reached epidemic proportions, with major economic and health implications. The complex pathophysiology of obesity explains the difficulty provided to health policy for its clinical management. Increasing data show that obesity and metabolic abnormalities are intimately connected to differences in consumption of probiotics, its relevance to gut microbiota activity and composition. The goal of this investigation was to assess the effect of oral delivery of indigenous probiotic Lactobacillus fermentum NCDC 400 and prebiotic fructo-oligosaccharide (FOS) on obesity-associated hepatic steatosis and inflammation produced by a high-fat diet (HFD). C57BL/6 mice treated with L. fermentum NCDC 400 either independently or in conjunction with FOS demonstrated reduced body weight and abdominal obesity after 24 weeks of treatment. Also, the anti-oxidative enzyme activity went down, and the inflammatory profile got better, with less fat getting into the hepatocytes. The lipid profile changed, with HDL cholesterol going up and LDL cholesterol and triglyceride levels going down. Further, L. fermentum NCDC 400 and FOS combinations decreased fasting glucose, gHbA1c, gastric inhibitory peptide, and insulin levels in mice fed with HFD, thus improving glucose homeostasis. Overall, consumption of L. fermentum NCDC 400 alone or its combinational effects had a protective role on obesity-associated hepatic steatosis. PRACTICAL APPLICATIONS: The potential indigenous probiotic Lactobacillus fermentum NCDC 400 and prebiotic FOS had a preventive role in obesity-induced hepatic steatosis and improves anti-oxidant and anti-inflammatory properties in HFD-fed obese mice. Our finding would be helpful to prevent obesity-associated hepatic steatosis and inflammation upon supplementation of pre- and pro-biotics (synbiotics).

Nanoparticles (NPs) are important in a variety of sectors, including disease diagnostics, medicine, nutrition, and many other industries. The risk of human exposure demands an early evaluation of both the basic dynamics of NPs’ interaction with biological systems and their potential consequences. Deciphering these occurrences will provide critical information regarding the health hazards and safety advantages associated with next-generation nanoformulations in clinical practice. We examined the HepG2 cell line in a systematic manner to determine the cellular response to single-walled carbon nanotubes (SWCNTs) and zinc oxide (ZnO) NPs. With the use of high-throughput transcriptomic methods, we found that both NPs induce comparable dysregulation of the endocytic and proteasomal complex genes in liver hepatocellular carcinoma cells, at levels (> 80 percent cell viability) that do not cause over-toxicity at early incubation period (6 h). SWCNT and ZnO NPs were shown to enter cells through clathrin-mediated pathways, affecting cytoskeleton gene expression, DNA damage and repair, protein ubiquitination, and cell transcriptional machinery. Our findings indicate that early response strategies activate stress-related mechanisms. Finally, this method for studying nanomaterial–cell interactions demonstrates how changes in the transcriptome profile may predict downstream consequences even at doses that do not cause acute toxicity.

MicroRNAs (miRNAs) are small single-stranded non-coding RNAs that act as the master regulator of animal growth and development. RNA-RNA interaction is an important mechanism of gene regulation during biotic and abiotic stress. Heat stress (HS) is one of the most important abiotic factors which affect the growth, milk yield and reproductive health of the dairy animals. In the present investigation, we identified 387 known and 77 novel miRNAs from Tharparkar (TH) and Karan Fries (KF) cattle under HS condition. Family distribution analysis showed the identified miRNAs belong to more than 15 different families in which miR-2284 was the most abundant. We identified 42,350 targets for the known miRNAs reported in cattle. Pathway analysis of the identified targets showed most of the target genes were involved in cancer, mitogen-activated protein kinase (MAPK) signaling, calcium signaling, Ras signaling, and cAMP signaling pathways. Differential gene expression showed more than 344 miRNAs changed their expression significantly between control and HS condition. Heat map was generated for the top 20 most up and downregulated miRNAs. Ten miRNAs were validated using qRT-PCR to be heat responsive, based on read count value and differential gene expression. These novel miRNAs are new addition to the miRNA database of cattle. This study provides an overview of miRNA profile and their interaction with the target genes which leads to further understanding in deciphering the thermotolerance mechanism in cattle.

Background The cellular response to nanoparticles (NPs) for the mechanical clue and biochemical changes are unexplored. Here, we provide the comprehensive analysis of the Chinese Hamster Ovary (CHO-K1) cell line to study cell behaviour following the exposure of mesoporous silica nanoparticle (MSN), multiwall carbon nanotubes (MWCNTs), and zinc oxide (ZnO) NPs. Results Through the high-throughput proteomic study, we observed that the effect of NPs is alone not restricted to cell viability but also on cell polarisation. In the case of MSN, no drastic changes were observed in cellular morphology, but it upregulated chaperons that might prevent protein aggregation. However, MWCNT showed elongated cell appearance with numerous cytoplasmic vacuoles, and induce lamellipodia formation through actin polymerisation. The cytoskeleton remodelling was accompanied by the increased expression of Dlc-1, cofilin and Rac1 proteins. While ZnO NPs resulted in the rounded cell morphology along with nuclear abnormalities. The proteome analysis revealed that UBXN11 control cell roundness and DOCK3 leads to actin stress fibre formation and finally, loss of cell adhesion. It enhances the expression of catastrophic DNA damage and apoptotic proteins, which was unrecoverable even after 72 h, as confirmed by the colony formation assay. All three NPs trigger over-expression of the endocytic pathway, ubiquitination, and proteasomal complex proteins. The data indicate that ZnO and MSN entered into the cells through clathrin-mediated pathways; whereas, MWCNT invades through ER-mediated phagocytosis. Conclusions Based on the incubation and concentration of NPs, our work provides evidence for the activation of Rac-Rho signalling pathway to alter cytoskeleton dynamics. Our results assist as a sensitive early molecular readout for nanosafety assessment.

There is a major concern that exposure to titanium dioxide (TiO2) nanoparticles (NPs) can have degrading effects on human health as well as mammary gland because of the increased use in numerous sorts of nanotech-based health care and food merchandise. Also, there is a scarcity in NP toxicity studies on the mammary gland; therefore, the aim of the present study was to compare toxicity caused by nano- and bulk-phase TiO2 particles on the human mammary gland in vitro. In comparison to bulk-TiO2 particles, nano-TiO2 cause a significant ( p < 0.05) reduction in viability and increased reactive oxygen species generation in the human mammary epithelial cells after a dose- (1, 2, 5, 10, 20, 50, and 100 µg/mL) and time (6, 12, 24, and 48 h)-dependent exposure. Further, an increase in genotoxicity in the mammary epithelial cells was observed as percent tail DNA and comet area was increased significantly ( p < 0.05) at 12 h of exposure (10 and 100 µg/mL) with nano-TiO2. The scanning electron microscopic examination showed that a 50 µg/mL dose of both nano-TiO2 and bulk-TiO2 particles cause morphological changes and retarded growth pattern of mammary epithelial cells at 12 h. Moreover, a significant ( p < 0.05) increase in apoptosis at 10 µg/mL and necrosis at 50 µg/mL concentrations of nano-TiO2 in comparison to bulk-TiO2 was observed in mammary epithelial cells. Finally, we can conclude that the toxicity caused by nano-TiO2 particles on the human mammary gland cells was comparatively higher than the bulk-TiO2 particles.

Various studies in rodents have shown that nanoparticles are transferred to the breast milk. Under the present study, lactating Wistar rats were repetitively gavaged 5, 25, and 50 mg/kg bw of zinc oxide nanoparticles (ZnO-NPs) and 50 mg kg−1 bw of bulk zinc oxide (bZnO) for 19 days after parturition. The results showed that ZnO-NPs were absorbed in the small intestine of dams and distributed to the liver. Furthermore, ZnO-NPs were distributed to the intestine and liver of rat pups through dam’s milk. No significant change in body weight was observed in the dams treated with ZnO-NPs or bZnO and their offsprings as compared to the control group. The spleen weight significantly increased in the rat dams treated with 50 mg kg−1 of ZnO-NPs. ZnO-NPs were mostly excreted through feces. The levels of liver cytochrome P450 reductase and serum total antioxidant capacity significantly decreased in the rat dams treated with ZnO-NPs (50 mg kg−1) and their offsprings. The levels of serum cytokines (tumor necrosis factor-alpha and interleukin-1 beta) and liver injury marker enzymes (alanine aminotransferase and aspartate aminotransferase) significantly increased in the rat dams treated with ZnO-NPs (25 and 50 mg kg−1) and their offsprings. The level of immunoglobulin A secretion in the intestinal fluid of rat dams and their offsprings is significantly increased by increasing the dose of ZnO-NPs. Histopathology of intestine and liver of offsprings whose rat dams were treated with ZnO-NPs (50 mg kg−1) showed gross pathological changes. These results provide information for the safety evaluation of ZnO-NPs use during lactation. In conclusion, a dose-dependent postnatal transfer of ZnO-NPs is hazardous to the breastfed offsprings.

MicroRNAs (miRNAs) are small single-stranded non-coding RNAs that act as the master regulator of animal growth and development. RNA-RNA interaction is an important mechanism of gene regulation during biotic and abiotic stress. Heat stress (HS) is one of the most important abiotic factors which affect the growth, milk yield and reproductive health of the dairy animals. In the present investigation, we identified 387 known and 77 novel miRNAs from Tharparkar (TH) and Karan Fries (KF) cattle under HS condition. Family distribution analysis showed the identified miRNAs belong to more than 15 different families in which miR-2284 was the most abundant. We identified 42,350 targets for the known miRNAs reported in cattle. Pathway analysis of the identified targets showed most of the target genes were involved in cancer, mitogen-activated protein kinase (MAPK) signaling, calcium signaling, Ras signaling, and cAMP signaling pathways. Differential gene expression showed more than 344 miRNAs changed their expression significantly between control and HS condition. Heat map was generated for the top 20 most up and downregulated miRNAs. Ten miRNAs were validated using qRT-PCR to be heat responsive, based on read count value and differential gene expression. These novel miRNAs are new addition to the miRNA database of cattle. This study provides an overview of miRNA profile and their interaction with the target genes which leads to further understanding in deciphering the thermotolerance mechanism in cattle.

There is a dearth of information regarding the safety of silver nanoparticles (Ag NPs) and multiwalled carbon nanotubes (MWCNTs) with respect to their impact on human/animal health and the environment. This study aimed to determine the half-maximum inhibitory concentration (IC50) of Ag NPs and MWCNTs by employing different doses and time interval combinations in buffalo bull spermatozoa. Semen samples containing 100 million spermatozoa each were incubated with 1, 10, 25, 50, 75 and 100 µg/mL of Ag NPs and MWCNTs at 37°C for 30, 60 and 120 min. Sperm viability was monitored by the MTT assay and eosin–nigrosin staining followed by estimation of IC50 values using correlation–regression analysis. Spermatozoa treated with IC50 doses of Ag NPs and MWCNTs were also assessed for different sperm functionality parameters including oxidative stress and membrane integrity. These parameters were observed to be significantly affected in treated spermatozoa compared with the controls. We concluded that both nanomaterials showed cytotoxicity, mediated principally via oxidative stress. This work has provided valuable toxicological information that will serve as a benchmark for future studies aimed at safe use of nanomaterials.

Zinc oxide nanoparticles (ZnO NPs) are important nanomaterials with myriad applications and in widespread use. The main aim of this study was to evaluate the direct effect of ZnO NPs on steroidogenesis by considering mouse testicular Leydig cells (TM3) as an in vitro model system. The uptake, intracellular behaviour, cytotoxicity and morphological changes induced by ZnO NPs (0–200 µg/ml) in a time-dependent manner in the TM3 were assessed. A significant ( p < 0.05) decrease in TM3 viability was observed at 2 µg/ml ZnO NP after a 1-h incubation time period. Increased antioxidant enzyme activity, namely, superoxide dismutase (SOD) and catalase, was regularly observed. Not surprisingly, apoptosis also increased significantly after a 4-h exposure period. Transmission electron micrographs illustrated that ZnO NPs were taken up by Leydig cells and resulted in the formation of autophagosomes, autolysosomes and autophagic vacuoles. Concomitant real-time data indicated that ZnO NPs significantly increased the expression of steroidogenesis-related genes (steroidogenic acute regulatory protein and cytochrome P450 side-chain cleavage enzyme) and significantly ( p < 0.05) decreased antioxidant enzyme gene (SOD) expression after a 4-h incubation period. Moreover, ZnO NPs exposure significantly increased testosterone production at 2 µg/ml concentration after a 12-h incubation period. Our findings confirm the adverse effects of ZnO NPs by being cytotoxic, enhancing apoptosis, causing steroidogenic effect in Leydig cells and increasing autophagic vacuole formation possibly via alteration of antioxidant enzyme activity in TM3 cells.

The emerging field of nanoscience and nanotechnology is potentially important for the growth of the economy, but it is also imperative to think through its possible environmental and health hazards. There is an urgent need to adopt a robust strategy for quick safety evaluation of Nanomaterials (NM’s) across a wide range of biochemical responses. These NM’s are new age materials which are having dimensions in nanoscale region viz, 1 to 100 nm. High-throughput screening (HTS) and high-content analysis (HCA) for nano risk assessment allows multiparametric testing for various NMs on different types of model systems at different concentrations that efficiently saves time and cost. The mechanism based high-throughput (HT) in vitro analysis approach are increasing efficacy for making safety decisions by replacing laborious animal studies from pathway-based testing in cell lines. HTS/HCA approaches describe a technique by which multiple endpoints can be analyzed at a time that facilitates classification of important biological markers at the nano-bio interface. Herein we will discuss various HT approaches that have been developed in respect to study safety aspects of NMs such as omics based methods (transcriptomics, proteomics, and metabolomics), HT flow cytometry, impedance-based monitoring and genotoxicity through HT comet assay and image-based HCS. Insights gained from HTS/HCS and should aim to deepen our fundamental understanding of NMs hazards as well as to assist in developing the safer by design approach. Moreover, it is necessary to gain insights into the large dataset to decipher the biological meaning and for this purpose, we have discussed various data analysis software employed to make predictions and the improvement of structure activity relationship for concerned NMs.

The increasing use of nanomaterials has naturally caused heightened concerns about their potential risks to human and animal health. We investigated the effect of zinc oxide nanoparticles (ZnO NPs) and mesoporous silica nanoparticles (MSN) on steroidogenesis in the corpus luteum (CL) of pregnant mice and testis of male offspring. Pregnant albino mice were exposed to ZnO NPs and MSN for 2 days on alternate days, gestation days 15–19. Hepatic injury marker enzymes increased in the higher concentration of NM-exposed mother mice, but histological examination revealed no changes in the placenta of pregnant mice, whereas testis of male offspring showed gross pathological changes. The expression pattern of progesterone biosynthesis-related genes was also altered in the CL of NP-exposed pregnant mice. In utero exposure of ZnO NPs increased the relative expression of StAR in 100 mg/kg body weight (BW) ZnO NP-treated and bulk ZnO-treated groups and P450 side-chain cleavage enzyme (P450scc) in 50 mg/kg BW ZnO NP-treated and 100 mg/kg of bulk ZnO-treated male offspring. Serum testosterone concentration significantly increased in the 100 mg/kg of bulk ZnO-treated group and decreased in the 250 mg/kg of MSN-treated group and a single dose of 300 mg/Kg BW of ZnO NPs caused miscarriages and adversely affected the developing foetus in mice.

Nanomaterials have revolutionized the drug delivery and therapeutic industry due to their unique physical characteristics, which render them extremely manipulative at nano-scale. One such category of nanomaterials is mesoporous silica nanoparticles. Due to their small size and rigid honeycomb-like structure, they are highly conducive for packaging of drugs, dyes, antibodies, etc. In addition, they show excellent biocompatibility. These new generation nanomaterials can be further functionalized by incorporating surface modifications, thus increasing their acceptability as carriers for drugs and molecules. In this chapter, a brief and comprehensive review covering various aspects of the recent advancements in synthesis of mesoporous nanomaterials and post-synthesis strategies for functionalization has been presented. Further, it also sheds light on how efficiently these smart nano-carriers are involved in transport and site-specific delivery of highly toxic drugs, like chemotherapeutic agents for cancer treatment, and their biocompatibility evaluation from a biosafety point of view.

Emetic and diarrheal toxins produced by endospore-forming Bacillus cereus causes a huge loss to dairy industry. Milk is the suitable medium for the growth of B. cereus. The pathogen is of particular concern in the dairy industry because in spite of aggressive cleaning practices performed by the dairy industry, it is impossible to destroy these hydrophobic spores which adhere to the pipelines of the dairy-processing plant and further these spores may also form biofilms in the milk. The reporting rate of illness caused by B. cereus is vague, usually due to the short duration of (

A rapid technique to measure the mechanical properties of pharmaceutical tablets is presented. The Young's modulus of elasticity and Poisson ratio are determined by fitting the measured transient force, generated by the impact of tablets with a dynamic load cell, by the theoretical force predicted from a Hertzian spring-dashpot model. Results of the method applied to a series of pharmaceutical compacts having different porosities and prepared from different excipient blends, demonstrate good agreement between the calculated force and the measured force. In particular, the Young's modulus and Poisson ratio calculated for a 22% porous microcrystalline cellulose compact are 3.5 GPa and 0.34, respectively. As the total measurement time is less than 100 μs, the technique can be used as an in-line process measurement for manufacturing process control.

The well documented source for adult multipotent stem cells is spermatogonial stem cells (SSCs) of mammalian testis. It is foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. SSCs isolation from mammalian testis is difficult because of their scarcity and the lack of well characterized cell surface markers. Thus, the isolation of SSCs is of great interest for exploration of spermatogonial physiology and therapeutic approaches for fertility preservation. CD9 is a surface marker expressed in mouse and rat male germline stem cells. In this study, CD9 positive SSCs were successfully isolated from the goat testis using enzymatic digestion followed by three step purification: Differential plating, Percoll discontinuous density gradient followed by Magnetic activated cell sorting (MACS). Percoll discontinuous density gradient showed significant differences in the percentage of CD9+ SSCs across individual fraction. The fraction 36% and 40% gave the highest percentage of CD9+ SSCs i.e. 82% ± 1.2 and 9.2% ± 1.3 respectively. Magnetic activated cell sorting of CD9+ cells in the magnetic fraction of goat testes was in the range of 15% - 18% which is upto threefolds. CD9+ SSCs were further recovered with appreciable efficiency after immunomagnetic isolation by using various bead: cells ratio in which 4:1 ratio gave the highest yield of 69.06 × 105 with 18% of CD9+ SSCs. Magnetic activated cell sorting using anti-CD9 antibodies provides an efficient and fast approach as compared to conventional approaches such as differential plating and percoll discontinuous density gradient for enrichment strategy for spermatogonial stem cells from goat testes for undertaking research on basic and applied reproductive biology.

Contents Chromosomal fragmentations or damage in sperm DNA has considerable value in determination of semen quality. However, rapid and/or simple method to assess sperm DNA integrity in buffalo has apparently not been reported. In the present study, SCD was used for the first time in buffalo bulls for assessment of sperm DNA fragmentation. A modified SCD protocol, under bright field microscope was developed and validated by comparison with other routine tests which can be used for processing of samples. The DNA fragmentation index (DFI) from SCD was correlated with semen quality parameters viz. viability (r = −0.68, p

Heat shock proteins are ubiquitously expressed intracellular proteins and act as molecular chaperones in processes like protein folding and protein trafficking between different intracellular compartments. They are induced during stress conditions like oxidative stress, nutritional deficiencies and radiation. They are released into extracellular compartment during necrosis. However, recent research findings highlights that, they are not solely present in cytoplasm, but also released into extracellular compartment during normal conditions and even in the absence of necrosis. When present in extracellular compartment, they have been shown to perform various functions like antigen presentation, intercellular signaling and induction of pro-inflammatory cytokines. Heat shock proteins represents as dominant microbial antigens during infection. The phylogenetic similarity between prokaryotic and eukaryotic heat shock proteins has led to proposition that, microbial heat shock proteins can induce self reactivity to host heat shock proteins and result in autoimmune diseases. The self-reactivity of heat shock proteins protects host against disease by controlling induction and release of pro-inflammatory cytokines. However, antibodies to self heat shock proteins haven been implicated in pathogenesis of autoimmune diseases like arthritis and atherosclerosis. Some heat shock proteins are potent inducers of innate and adaptive immunity. They activate dendritic cells and natural killer cells through toll-like receptors, CD14 and CD91. They play an important role in MHC-antigen processing and presentation. These immune effector functions of heat shock proteins are being exploited them as therapeutic agents as well as therapeutic targets for various infectious diseases and cancers.

Carbon nanotubes (CNTs) are viewed as a class of nanomaterials with high potential for many biomedical applications due to their unique properties. One of the prospective bio-application of CNTs is in the area of tissue engineering as a scaffold for cell culture. Supporting the use of CNTs as scaffold in tissue engineering a few cell types have been successfully grown on it but no report for any germline cell. In the present study, the in vitro maintenance of spermatogonial cells on carbon nanotube scaffold has been evaluated for the first time. Protein adsorption on the CNT scaffolds along with other substrates has also been evaluated because it is very likely that adsorbed proteins in serum influence the properties for the cell growth. Microscopic analysis (Light and SEM) showed that cells maintain their proper shape and adhered on all CNT scaffolds during in vitro culture. It was observed that the cells were maintained for at least 21 days on all CNT scaffolds compared to the positive control (Sertoli feeder layer). The results express a degree of biocompatibility between spermatogonial cells and CNTs and the possibility for CNTs to be used as substratum for in vitro growth of these cells.

Contents Spermatogonial stem cells transplantation provides a unique approach for studying spermatogenesis. Initially developed in mice, this technique has now been extended in farm animals and provides an alternative means to preserve valuable male germ line and to produce transgenic animals. The aim of this study was to enrich type A spermatogonial cells amongst the isolated cells from goat testis, to cryopreserve these enriched populations of cells and their subsequent transplantation in unrelated recipient goats under ultrasound guidance. The cells were isolated enzymatically and enriched by differential plating and separation on discontinuous percoll gradient. Ultrasound guided injection of trypan blue dye into rete testis resulted in 20–30% filling of the seminiferous tubules. Prior to transplantation, the cells were labelled with a fluorescent dye to trace donor cells in recipient seminiferous tubules after transplantation. The fluorescent-labelled cells were observed up to 12 weeks after transplantation.

Lower extremity trauma is common. Epidemiology studies have shown that open long bone fractures occur at a rate of 11.5 per 100,000 persons per year, and many of these are open tibial shaft fractures1-3. Type III open fractures are the most severe, involving extensive soft tissue trauma and complex fracture patterns4. Many of these injuries can be managed with salvage involving either local muscle flaps or microvascular free tissue transfer for wound coverage. The alternative to reconstruction for such injuries is amputation. Over the past decades, many studies have sought to delineate the advantages and disadvantages of these two principal options for managing severe lower extremity trauma. A recent systematic review of all available literature demonstrated that, despite the abundance of evidence, neither option is superior with respect to clinical outcomes5. Because microvascular reconstruction is technically demanding, time-consuming, and fraught with substantial risk of complications, we must look beyond clinical outcomes to evaluate the costs and utilities afforded to patients by this option. Studies have shown that even with successful limb salvage, many patients are still not ambulatory after two years; persistent pain and disability are not uncommon after this type of injury6, 7. Despite the residual problems with reconstruction, however, some studies suggest that patients with salvaged lower limbs have a better quality of life and impose lower costs on the health care system8-10. As surgeons, we want the best for our patients, but as stewards of modern medicine who work with limited healthcare resources, we are also charged with making evidence-based, fiscally-sound decisions. When dealing with complex operations of uncertain effectiveness, cost-utility analyses can help provide the scientific evidence upon which these decisions can be based. Although costs10 and utilities11 have been measured, a detailed cost-utility analysis with robust sensitivity analysis, which is needed to consider the wide-range of assumptions in clinical decision-making, has not been conducted. This study presents a cost-utility analysis for management of severe open tibial fractures, and will serve to guide surgeons and patients in deciding between the two competing management options.

A rapidly advancing nanotechnology is focusing on producing better, efficient, and safer molecules to be used for several biomedical and diagnostic applications. Hybrid nanomaterials have been produced encompassing a plethora of applications in the areas of medicine and therapeutics. These new generation materials are proving to be much better than their earlier counterparts because of their excellent biomimetic properties, which are reflected in their healthy interactions with cells and tissues. However, without a comprehensive assessment of their properties and associated effects, these cannot be marked as completely safe. In this context, it becomes relevant to realize their cytocompatibility, adverse effects, and experimental evaluation by in vivo and in vitro testing. So here, we have tried to provide a brief and comprehensive review highlighting biosafety and/or biocompatibility features of nanocomposites and nanoceramics.

Traumatic articular cartilage lesions have a limited capacity to heal. We tested the hypothesis that overexpression of a human insulin-like growth factor I (IGF-I) cDNA by transplanted articular chondrocytes enhances the repair of full-thickness (osteochondral) cartilage defects in vivo. Lapine articular chondrocytes were transfected with expression plasmid vectors containing the cDNA for the Escherichia coli lacZ gene or the human IGF-I gene and were encapsulated in alginate. The expression patterns of the transgenes in these implants were monitored in vitro for 36 days. Transfected allogeneic chondrocytes in alginate were transplanted into osteochondral defects in the trochlear groove of rabbits. At three and 14 weeks, the quality of articular cartilage repair was evaluated qualitatively and quantitatively. In vitro, IGF-I secretion by implants constructed from IGF-I-transfected chondrocytes and alginate was 123.2+/-22.3 ng/10(7) cells/24 h at day 4 post transfection and remained elevated at day 36, the longest time point evaluated. In vivo, transplantation of IGF-I implants improved articular cartilage repair and accelerated the formation of the subchondral bone at both time points compared to lacZ implants. The data indicate that allogeneic chondrocytes, transfected by a nonviral method and cultured in alginate, are able to secrete biologically relevant amounts of IGF-I over a prolonged period of time in vitro. The data further demonstrate that implantation of these composites into deep articular cartilage defects is sufficient to augment cartilage defect repair in vivo. These results suggest that therapeutic growth factor gene delivery using encapsulated and transplanted genetically modified chondrocytes may be applicable to sites of focal articular cartilage damage.

The fragility of the CAPM has led to a resurgence of research that frequently uses trading strategies based on sorting procedures to uncover relations between firm characteristics (such as ?value? or ?glamour?) and equity returns. We examine the propensity of these strategies to generate statistically and economically significant profits due to our familiarity with the data. Under plausible assumptions, data snooping can account for up to 50 percent of the in-sample relations between firm characteristics and returns uncovered using single (one-way) sorts. The biases can be much larger if we simultaneously condition returns on two (or more) characteristics

The zona pellucida (ZP) in a majority of species consists of three antigenic families with wide inter species cross reactivity. The glocoproteins of heat solubilized goat zona pellucida (gZP) were isolated into three families by high performance liquid chromatography on a gel filtration column. Their molecular identities were 180, 95 and 75 kDa for gZP1, gZP2 and gZP3 respectively. However under reducing conditions, four zona pellucida fractions of 135, 95, 75 and 42 kDa were observed. The monoclonal antisera Mab-5H4 against porcine ZP4 (a 23 kDa glycoprotein) easily recognized the 180 and 42 kDa gZP fractions. It was also found that the 5H4 antisera strongly inhibited homologous goat sperm-zona binding. Finally, using immunofluorescent staining the goat occytes reacted positively with the 5H4 antisera. The fluorescence was uniform at 1 : 10 antisera dilution but tended to fade at higher dilution’s. Results suggest that the gZP has an antigen epitope correlative to the 5H4 antisera which is involved in sperm-zona interaction and could be a good candidate for the development of a contraceptive vaccine. # 2000 Elsevier Science B.V. All rights reserved.

In this article we use a single unifying framework to analyze the sources of profits to a wide spectrum of returnbased trading strategies implemented in the literature. We show that less than 50% of the 120 strategies implemented in the article yield statistically significant profits and, unconditionally, momentum and contrarian strategies are equally likely to be successful. However, when we condition on the return horizon (short, medium, or long) of the strategy, or the time period during which it is implemented, two patterns emerge. A momentum strategy is usually profitable at the medium (3to 12-month) horizon, while a contrarian strategy nets statistically significant profits at long horizons, but only during the 19261947 subperiod. More importantly, our results show that the cross-sectional variation in the mean returns of individual securities included in these strategies plays an important role in their profitability. The cross-sectional variation can potentially account for the profitability of momentum strategies and it is also responsible for atten

The authors test whether the reaction of international stock markets to oil shocks can be justified by current and future changes in real cash flows and/or changes in expected returns. They find that, in the postwar period, the reaction of U.S. and Canadian stock prices to oil shocks can be completely accounted for by the impact of these shocks on real cash flows alone. In contrast, in both the United Kingdom and Japan, innovations in oil prices appear to cause larger changes in stock prices than can be justified by subsequent changes in real cash flows or by changing expected returns. Copyright 1996 by American Finance Association.

The present study has examined the effect of different concentrations (1 μg/ml, 10 μg/ml and 100 μg/ml) of titanium oxide (TiO2) nanoparticles (NPs) (2 NP albeit at lower concentrations. In DNA fragmentation assay, there was dose-dependent increase in the DNA fragmentation ( r = 0.96). Ultrathin cross-sections revealed TiO2 NPs inside head and plasma membrane of the buffalo spermatozoa as assessed by TEM. These studies suggest that TiO2 NPs may have cytotoxic effect on buffalo spermatozoa by affecting sperm functionality and causing high amount of DNA fragmentations.

Summary The aim of this study was to isolate and subsequently enrich type A spermatogonia from pre-pubertal buffalo testis. Two-step enzymatic digestion was used to isolate spermatogonia from 10 to 14 months pre-pubertal buffalo calves, resulting in maximal release of spermatogonia from the seminiferous tubules. After enzymatic digestion, the type A spermatogonia were subsequently enriched by differential plating and Percoll gradient centrifugation. The identity of type A spermatogonia was determined by light microscopy and further characterised by Dolichos biflorus agglutinin, a specific marker for bovine type A spermatogonia by fluorescence-activated cell sorting analysis. After enzymatic isolation, the cell suspension contained about 27% of type A spermatogonia, which was enriched up to 71% with >70% cell viability. Further flow cytometric analysis showed the presence of THY1+ cells (cells expressing thymocyte differentiation antigen 1), suggesting that THY1 is a conserved marker of the undifferentiated spermatogonial cells in buffalo. The isolation of the enriched type A spermatogonia from buffalo testis opens ways to study the further biochemical characteristics of this important class of germ cells in this species.

Mechanical integrity and properties of drug tablets may adversely affect their therapeutic and structural functions. An embedded ultrasound monitoring system for tablet mechanical property monitoring during compaction and a non-contact/non-destructive off-line air-coupled technique for determining the mechanical properties of coated drug tablets are presented. In the compaction monitoring system, the change of ToF and the reflection coefficient for the upper-punch surface interface as a function of compaction pressure has been studied. In the air-coupled measurement approach, air-coupled excitation and laser interferometric detection are utilized and their effectiveness in characterizing the mechanical properties of a drug tablet by examining its vibrational resonance frequencies is demonstrated. An iterative computational procedure based on the finite element method and Newton's method is developed to extract the mechanical properties of the coated tablet from a subset of its measured resonance frequencies. The mechanical properties characterized by this technique are compared to those obtained by a contact ultrasonic method.

We show that the positive volatility-volume relation documented by numerous researchers actually reflects the positive relation between volatility and the number of transactions. Thus, it is the occurrence of transactions per se, and not their size, that generates volatility; trade size has no information beyond that contained in the frequency of transactions. Our results suggest that theoretical research needs to entertain scenarios in which (1) both the frequency and size of trades are endogenously determined, yet (2) the size of trades has no information content beyond that contained in the number of transactions. Article published by Oxford University Press on behalf of the Society for Financial Studies in its journal, The Review of Financial Studies.

We examine the effects of trading and information flows on the short-run behavior of stock prices by comparing the behavior of stock return volatility during trading and nontrading periods. We define nontrading periods as periods when exchanges and businesses are open but traders endogenously choose not to trade. After correcting for the bid/ask bounce and stickiness in quotes, we find that a large proportion of daily stock return volatility occurs without trades, especially for large firms. Furthermore, we provide new evidence that public (versus private) information is the major source of short-term return volatility.

Prior work with competitive rational expectations equilibrium models indicates that there should be a positive relation between trading volume and differences in beliefs or information among traders. We show that this result is sensitive to whether and how transaction costs are modeled. In a specialist market with endogenous transaction costs we show that trading volume can be negatively related to the degree of informational asymmetry in the market. Our analysis highlights the dependence of volume on market structure, and our results suggest that the "volume effects" of corporate or macroeconomic events reflect a decrease, rather than an increase, in heterogeneity of beliefs or asymmetry of information. PRIOR RESEARCH ON RATIONAL expectations equilibrium models of competitive markets indicates that trading volume is generated "primarily" by differences in beliefs which arise because agents possess different information and noise in the market prevents prices from perfectly aggregating private information (see, for example, Milgrom and Stokey (1982), Pfleiderer (1984), Grundy and McNichols (1989), Holthausen and Verrecchia (1990), and Kim and Verrecchia (1991a, 1991b)). The common conclusion of previous studies is that volume is (a) directly related to (some measure of) asymmetric information or differences in beliefs; and (b) this relationship is unambiguously positive. For example, Kim and Verrecchia (199 ib) state that; "The presence of differential precision [of private information] thus causes differential belief revisions among investors which, in turn, creates volume. When there are no differences in precision across investors... there is no volume."1

ATP-binding cassette (ABC) transporters utilize the energy present in cellular ATP to drive the translocation of structurally diverse set of solutes across the membrane barriers of eubacteria, archaebacteria and eukaryotes. In bacteria, these transporters are considered to be important virulence factors because they play role in nutrient uptake and in the secretion of toxins. The advances in structural determination and functional analysis of bacterial transporters have greatly increased our understanding of the mechanism of transport of these ABC transporters. Although progress in the field of structural biology has been made with the prokaryotic family members, it is likely that eukaryotic transporters will utilize the same mechanisms for translocation process. In this review, we summarize the function of the known MsbA ABC transporters in E. coli and mechanistic insights from structural and possible flippase mechanism studies.

This paper presents data on the emission of helium fragments due to break up of 200 A GeV 32S ions in collision with emulsion nuclei. A total of 499 minimum bias events have been studied in an emulsion stack and 97 events in emulsion chambers. Emission angles of He fragments have been measured with high precision. This data has been compared with data of breakup of heavier as well as lighter projectiles and we observe that the value of the momentum spread, σ(p), increases with increasing projectile mass. The bulk part of the helium emission is consistent with a single source for helium fragments with an average excitation energy of (9.1±0.9) MeV. A tail of a few (

We show that the returns to the typical long-term contrarian strategy implemented in previous studies are upwardly biased because they are calculated by cumulating single-period (monthly) returns over long intervals. The cumulation process not only cumulates "true" returns but also the upward bias in single-period returns induced by measurement errors. We also show that the remaining "true" returns to loser or winner firms have no relation to overreaction. This study has important implications for event studies that use cumulative returns to assess the impact of information events. RECENT RESEARCH HAS UNCOVERED substantial predictability in both shortterm (Conrad and Kaul (1988, 1989) and Lo and MacKinlay (1988)) and long-term (Fama and French (1988) and Poterba and Summers (1988)) stock returns. An increasingly popular interpretation of return predictability emphasized by a number of researchers is that the stock market consistently overreacts to new information. The "stock market overreaction" hypothesis asserts that stock prices take temporary swings away from their fundamental values due to waves of optimism and pessimism (see, for example, DeBondt and Thaler (1985, 1987), Lehmann (1990), and Shefrin and Statman (1985)). Compelling evidence in favor of long-term overreaction was first provided by DeBondt and Thaler (1985). They show that losers and winners, determined by their performance relative to the aggregate stock market over the past three to five years, consistently outperform and underperform the market in subsequent three- to five-year periods. For example, they find that the arbitrage (zero investment) portfolio of losers and winners earns an

Regulation at the level of translation in eukaryotes is feasible because of the longer lifetime of eukaryotic mRNAs in the cell. The elongation stage of mRNA translation requires a substantial amount of energy and also eukaryotic elongation factors (eEFs). The important component of eEFs, i.e. eEF2 promotes the GTP-dependent translocation of the nascent protein chain from the A-site to the P-site of the ribosome. Mostly the eEF2 is regulated by phosphorylation and dephosphorylation by a specific kinase known as eEF2 kinase, which itself is up-regulated by various mechanisms in the eukaryotic cell. The activity of this kinase is dependent on calcium ions and calmodulin. Recently it has been shown that the activity of eEF2 kinase is regulated by MAP kinase signalling and mTOR signalling pathway. There are also various stimuli that control the peptide chain elongation in eukaryotic cell; some stimuli inhibit and some activate eEF2. These reports provide the mechanisms by which cells likely serve to slow down protein synthesis and conserve energy under nutrient deprived conditions via regulation of eEF2. The regulation via eEF2 has also been seen in mammary tissue of lactating cows, suggesting that eEF2 may be a limiting factor in milk protein synthesis. Regulation at this level provides the molecular understanding about the control of protein translocation reactions in eukaryotes, which is critical for numerous biological phenomenons. Further the elongation factors could be potential targets for regulation of protein synthesis like milk protein synthesis and hence probably its foreseeable application to synthetic biology.