An empirical investigation on the impact of attitudes towards organ donation in India.

Background: Organ shortage is a global issue and it is imperative to take steps to bridge this gap. In the Indian context, despite its demographic dividend, the rate of organ donation has been abysmally low. This emphasizes a need to demystify the antecedents of organ donation intention among the Indian population. Methods: Using a cross-sectional research design and adopting a post-positivism research philosophy, this study identified 259 respondents by adopting a purposive sampling approach and data on knowledge of organ donation was collected using a structured, pretested questionnaire. Results: Awareness of organ donation law in India is low on specific issues and respondents from the health science & medicine discipline scored better on knowledge about organ donation. The findings show that most participants had heard about organ donation and had a favourable attitude toward it. The primary sources of information on organ donation were television and newspapers, and healthcare service providers. A complementary partial median is established (ß = .217, t = 5.889, p < .001) which implies that willingness to discuss with family significantly mediates the association between attitude towards organ & tissue donation and willingness to sign the donor card. Conclusion: This study revealed that there is a general awareness of organ and tissue donation among the Indian population, but they lack clarity on certain specific issues. Mass media has to be effectively used to build awareness campaigns revolving around enhancing knowledge on a specific issue and building acceptance of the concept of organ and tissue donation.

Biochemical characterization of a GDP-mannose transporter from Chaetomium thermophilum.

Nucleotide Sugar Transporters (NSTs) belong to the SLC35 family (human solute carrier) of membrane transport proteins and are crucial components of the glycosylation machinery. NSTs are localized in the ER and Golgi apparatus membranes, where they accumulate nucleotide sugars from the cytosol for subsequent polysaccharide biosynthesis. Loss of NST function impacts the glycosylation of cell surface molecules. Mutations in NSTs cause several developmental disorders, immune disorders, and increased susceptibility to infection. Atomic resolution structures of three NSTs have provided a blueprint for a detailed molecular interpretation of their biochemical properties. In this work, we have identified, cloned, and expressed 18 members of the SLC35 family from various eukaryotic organisms in Saccharomyces cerevisiae. Out of 18 clones, we determined Vrg4 from Chaetomium thermophilum (CtVrg4) is a GDP-mannose transporter with an enhanced melting point temperature (Tm) of 56.9°C, which increases with the addition of substrates, GMP and GDP-mannose. In addition, we report-for the first time-that the CtVrg4 shows an affinity to bind to phosphatidylinositol lipids.

The role of legal medicine professionals in preventing pregnancy and sexually transmitted infections among female victims of sexual assault.

Sexual violence can have an overwhelming impact on the victim's physical and mental health; the consequences include unintended pregnancy and sexually transmitted infections (STIs). Therefore, the examiners must assess victims for possible pregnancy and sexually transmitted infections as a part of the sexual assault examination. This article aims to orient the medico-legal examiners towards their role in preventing unintended pregnancy and sexually transmitted infections among victims of sexual assault. Prompt detection of pregnancy or STIs is critical, as any delay would adversely affect the successful administration of emergency contraception and post-exposure prophylaxis (PEP) against human immunodeficiency virus (HIV) and other sexually transmitted infections.

The effectiveness of a sexual assault nurse examiner-grounding program (SANE-GP) on knowledge, skill and practice regarding sexual assault examination (SAE) among nurses working in a tertiary care hospital in Udupi district, India: A study protocol.

The medico-legal care of victims of sexual assault is very challenging, and requires specific knowledge and skills. Professionals in the emergency departments of hospitals might not have specialised training in forensic science. Nurses have a very significant role in these settings, but they lack any formal forensic training. This study aims to develop a sexual assault nurse examiner-grounding program (SANE-GP) for Indian nurses to inculcate knowledge and skill regarding sexual assault examination. The study adopts a three-stage Delphi technique to develop the training module and uses a time-series design to evaluate the effectiveness of the program. A questionnaire on nurses' knowledge on sexual assault examination (KQSANE-I) will be developed in phase-I and subsequently used in phase-II. The protocol of SANE-GP will help the medical community to implement the program across India. The implementation of SANE-GP can also help to start a sexual assault nurse examiner network.

Early prepubertal cyclophosphamide exposure in mice results in long-term loss of ovarian reserve, and impaired embryonic development and blastocyst quality.

BACKGROUND: Due to improved treatment, there is an increasing focus on the reproductive potential of survivors of childhood cancer. Cytotoxic chemotherapy accelerates the decline in the number of primordial follicles within the mammalian ovary at all ages, but effects on the developmental potential of remaining oocytes following prepubertal cancer treatment are unclear. OBJECTIVES: To investigate whether cyclophosphamide (CY) exposure in the prepubertal period in female mice influences ovarian function and the functional competence of oocytes in adulthood. METHODS: This study used Swiss albino mice as the experimental model. Female mice were treated with 200 mg/kg CY on either postnatal day 14 (CY14), 21 (CY21) or 28 (CY28) i.e at a prepubertal and 2 young postpubertal ages. At 14 weeks of life, ovarian function, functional competence of oocytes, and embryo quality were assessed. RESULTS: The number of primordial follicles decreased significantly in CY14 and CY21 groups compared to control (p < 0.01). The number of oocytes from superovulated was 8.5 ± 1.4, 24.1 ± 2.9 and 26.8 ± 2.1 in CY14, CY21 and CY28 respectively which was significantly lower than control (50.2 ± 3.2; p < 0.001). In vitro culture of CY14 embryos demonstrated only 55.4% blastocyst formation (p < 0.0001) and reduced ability of inner cell mass (ICM) to proliferate in vitro (p < 0.05) at 120 and 216 h post insemination respectively. On the other hand, ICM proliferation was unaltered in 2 young postpubertal ages. CONCLUSION: Our results indicate long-term effects on the developmental competence of oocytes exposed to CY in early but not adult life. These data provide a mechanism whereby long-term fertility can be impaired after chemotherapy exposure, despite the continuing presence of follicles within the ovary, and support the need for fertility preservation in prepubertal girls before alkylating agent exposure.

Structural and functional characterization of CMP-N-acetylneuraminate synthetase from Vibrio cholerae.

Several pathogenic bacteria utilize sialic acid, including host-derived N-acetylneuraminic acid (Neu5Ac), in at least two ways: they use it as a nutrient source and as a host-evasion strategy by coating themselves with Neu5Ac. Given the significant role of sialic acid in pathogenesis and host-gut colonization by various pathogenic bacteria, including Neisseria meningitidis, Haemophilus influenzae, Pasteurella multocida and Vibrio cholerae, several enzymes of the sialic acid catabolic, biosynthetic and incorporation pathways are considered to be potential drug targets. In this work, findings on the structural and functional characterization of CMP-N-acetylneuraminate synthetase (CMAS), a key enzyme in the incorporation pathway, from Vibrio cholerae are reported. CMAS catalyzes the synthesis of CMP-sialic acid by utilizing CTP and sialic acid. Crystal structures of the apo and the CDP-bound forms of the enzyme were determined, which allowed the identification of the metal cofactor Mg2+ in the active site interacting with CDP and the invariant Asp215 residue. While open and closed structural forms of the enzyme from eukaryotic and other bacterial species have already been characterized, a partially closed structure of V. cholerae CMAS (VcCMAS) observed upon CDP binding, representing an intermediate state, is reported here. The kinetic data suggest that VcCMAS is capable of activating the two most common sialic acid derivatives, Neu5Ac and Neu5Gc. Amino-acid sequence and structural comparison of the active site of VcCMAS with those of eukaryotic and other bacterial counterparts reveal a diverse hydrophobic pocket that interacts with the C5 substituents of sialic acid. Analyses of the thermodynamic signatures obtained from the binding of the nucleotide (CTP) and the product (CMP-sialic acid) to VcCMAS provide fundamental information on the energetics of the binding process.

Crystal structures and kinetics of N-acetylneuraminate lyase from Fusobacterium nucleatum.

N-Acetyl-D-neuraminic acid lyase (NanA) catalyzes the breakdown of sialic acid (Neu5Ac) to N-acetyl-D-mannosamine (ManNAc) and pyruvate. NanA plays a key role in Neu5Ac catabolism in many pathogenic and bacterial commensals where sialic acid is available as a carbon and nitrogen source. Several pathogens or commensals decorate their surfaces with sialic acids as a strategy to escape host innate immunity. Catabolism of sialic acid is key to a range of host-pathogen interactions. In this study, atomic resolution structures of NanA from Fusobacterium nucleatum (FnNanA) in ligand-free and ligand-bound forms are reported at 2.32 and 1.76 Šresolution, respectively. F. nucleatum is a Gram-negative pathogen that causes gingival and periodontal diseases in human hosts. Like other bacterial N-acetylneuraminate lyases, FnNanA also shares the triosephosphate isomerase (TIM)-barrel fold. As observed in other homologous enzymes, FnNanA forms a tetramer. In order to characterize the structure-function relationship, the steady-state kinetic parameters of the enzyme are also reported.

Crystal structures and kinetic analyses of N-acetylmannosamine-6-phosphate 2-epimerases from Fusobacterium nucleatum and Vibrio cholerae.

Sialic acids are nine-carbon sugars that are found abundantly on the cell surfaces of mammals as glycoprotein or glycolipid complexes. Several Gram-negative and Gram-positive bacteria have the ability to scavenge and catabolize sialic acids to use as a carbon source. This gives them an advantage in colonizing sialic acid-rich environments. The genes of the sialic acid catabolic pathway are generally present as the operon nanAKE. The third gene in the operon encodes the enzyme N-acetylmannosamine-6-phosphate 2-epimerase (NanE), which catalyzes the conversion of N-acetylmannosamine 6-phosphate to N-acetylglucosamine 6-phosphate, thus committing it to enter glycolysis. The NanE enzyme belongs to the isomerase class of enzymes possessing the triose phosphate isomerase (TIM) barrel fold. Here, comparative structural and functional characterizations of the NanE epimerases from two pathogenic Gram-negative bacteria, Fusobacterium nucleatum (Fn) and Vibrio cholerae (Vc), have been carried out. Structures of NanE from Vc (VcNanE) with and without ligand bound have been determined to 1.7 and 2.7 Šresolution, respectively. The structure of NanE from Fn (FnNanE) has been determined to 2.2 Šresolution. The enzymes show kinetic parameters that are consistent with those of Clostridium perfringens NanE. These studies allowed an evaluation of whether NanE may be a good drug target against these pathogenic bacteria.

Conformational transitions of the sodium-dependent sugar transporter, vSGLT.

Sodium-dependent transporters couple the flow of Na+ ions down their electrochemical potential gradient to the uphill transport of various ligands. Many of these transporters share a common core structure composed of a five-helix inverted repeat and deliver their cargo utilizing an alternating-access mechanism. A detailed characterization of inward-facing conformations of the Na+-dependent sugar transporter from Vibrio parahaemolyticus (vSGLT) has previously been reported, but structural details on additional conformations and on how Na+ and ligand influence the equilibrium between other states remains unknown. Here, double electron-electron resonance spectroscopy, structural modeling, and molecular dynamics are utilized to deduce ligand-dependent equilibria shifts of vSGLT in micelles. In the absence and presence of saturating amounts of Na+, vSGLT favors an inward-facing conformation. Upon binding both Na+ and sugar, the equilibrium shifts toward either an outward-facing or occluded conformation. While Na+ alone does not stabilize the outward-facing state, gating charge calculations together with a kinetic model of transport suggest that the resting negative membrane potential of the cell, absent in detergent-solubilized samples, may stabilize vSGLT in an outward-open conformation where it is poised for binding external sugars. In total, these findings provide insights into ligand-induced conformational selection and delineate the transport cycle of vSGLT.

Structural and functional analysis of the DOT1L-AF10 complex reveals mechanistic insights into MLL-AF10-associated leukemogenesis.

The mixed-lineage leukemia (MLL)-AF10 fusion oncoprotein recruits DOT1L to the homeobox A (HOXA) gene cluster through its octapeptide motif leucine zipper (OM-LZ), thereby inducing and maintaining the MLL-AF10-associated leukemogenesis. However, the recognition mechanism between DOT1L and MLL-AF10 is unclear. Here, we present the crystal structures of both apo AF10OM-LZ and its complex with the coiled-coil domain of DOT1L. Disruption of the DOT1L-AF10 interface abrogates MLL-AF10-associated leukemic transformation. We further show that zinc stabilizes the DOT1L-AF10 complex and may be involved in the regulation of the HOXA gene expression. Our studies may also pave the way for the rational design of therapeutic drugs against MLL-rearranged leukemia.

Automation aided optimization of cloning, expression and purification of enzymes of the bacterial sialic acid catabolic and sialylation pathways enzymes for structural studies.

The process of obtaining a well-expressing, soluble and correctly folded constructs can be made easier and quicker by automating the optimization of cloning, expression and purification. While there are many semiautomated pipelines available for cloning, expression and purification, there is hardly any pipeline that involves complete automation. Here, we achieve complete automation of all the steps involved in cloning and in vivo expression screening. This is demonstrated using 18 genes involved in sialic acid catabolism and the surface sialylation pathway. Our main objective was to clone these genes into a His-tagged Gateway vector, followed by their small-scale expression optimization in vivo. The constructs that showed best soluble expression were then selected for purification studies and scaled up for crystallization studies. Our technique allowed us to quickly find conditions for producing significant quantities of soluble proteins in Escherichia coli, their large-scale purification and successful crystallization of a number of these proteins. The method can be implemented in other cases where one needs to screen a large number of constructs, clones and expression vectors for successful recombinant production of functional proteins.

Objective structured practical examination (OSPE) in Forensic Medicine: students' point of view.

The purpose of this study was to assess the attitudes of undergraduate medical students towards the objective structured practical examination (OSPE) in Forensic Medicine, in a medical college in Nepal. Participants included 59 undergraduate medical students of the 7th semester. Findings indicated that the OSPE was an acceptable tool considering the conduct of practical examination in Forensic Medicine at the undergraduate level. The overall mean attitude score was towards the favourable side. Students strongly agreed that the OSPE tested a wide range of skills. They also strongly agreed that it was a good form of examination as well as a learning experience. The introduction of the OSPE replacing the conventional method of practical examination in Forensic Medicine is a step in the right direction taken to objectively assess undergraduate medical students.

Composition, recruitment and regulation of the PRC2 complex.

The PRC2 histone methyltransferase complex is an important regulator of gene expression programs in metazoans. Gene expression regulation by the PRC2 complex is critical for development and cell differentiation. Several recent studies have begun to shed light on the molecular basis for the physiological function of the PRC2 complex. Here, we discuss some of these results and how they provide new insights into the composition, recruitment and regulation of the PRC2 complex.

Sexual dimorphism of the index to ring finger ratio in South Indian adolescents.

Sex determination from prepubertal human remains is a challenge for forensic experts and physical anthropologists worldwide as definitive sexual traits are not manifested until after the full development of secondary sexual characters that appear during puberty. The research was undertaken in 350 South Indian adolescents to investigate sexual dimorphism of the index and ring finger ratio. The index finger length (IFL) and the ring finger length (RFL) were measured in millimeters in each hand and the index and ring finger ratio was computed by dividing the index finger length by the ring finger length. Mean RFL was greater than mean IFL in both males and females. Mean RFL was significantly higher in males. The index and ring finger ratio showed a statistically significant difference between males and females (p

Wounds due to a modified shot gun (home-made): a case report.

In a case of firearm fatality, the autopsy surgeon is required to opine as to the range of fire in addition to the cause of death which will help in reconstruction of the events. Problems may arise in estimating the range of fire based on wound ballistics when there is an alteration or modification in the internal ballistics. We encountered such a case in the department of Forensic Medicine, Kasturba Medical College, Manipal, which is discussed.

Sex differences from fingerprint ridge density in the Indian population.

Determination of individuality is one of the prime concerns in forensic investigation. The study of fingerprints is widely used in providing a clue regarding identity. The present study was undertaken to study the sex differences in fingerprint ridge density in the Indian population. The study done on 100 males and 100 females revealed that significant sex differences occur in the fingerprint ridge density. The present study shows a statistically significant difference in fingerprint ridge densities of male and female fingerprints in people of Indian origin. A mean fingerprint ridge density of 12 ridges/25 mm(2) or less is found to be more likely to be of males and a mean ridge count of more the 12 ridges/25 mm(2) is more likely to be of female origin.

Sex differences from fingerprint ridge density in Chinese and Malaysian population.

The fingerprints are very typical for a human being. The present study was undertaken to study the gender differences in fingerprint ridge density in Chinese and Malaysian population. The study done on 200 subjects (100 males and 100 females) of Chinese origin and 100 subjects (50 males and 50 females) of Malaysian origin revealed that significant gender differences occur in the finger ridge density. Fingerprint mean ridge density of 12 ridges/25 mm(2) or less is found to be more likely to be of males and a mean ridge count of more the 13 ridges/25 mm(2) is more likely of female origin in Chinese subjects. Fingerprint mean ridge density of 11 ridges/25 mm(2) or less is found to be more likely to be of males and a mean ridge count of more the 13 ridges/25 mm(2) is more likely of female origin in Malaysian subjects.

A rare case of myocardial infarction due to parathion poisoning.

Organophosphorus poisoning occurs very commonly in southern India where farmers form a significant proportion of the population who commonly use organophosphorus compounds like parathion as insecticides. Awareness of the complications caused by parathion poisoning is essential for proper monitoring and treatment. We report a case of suicidal parathion poisoning in a farmer who was treated with atropine and oximes. He subsequently developed myocardial infarction, to the treatment of which, he showed no response and succumbed. In conclusion, myocardial infarction is a rare complication of parathion poisoning. Awareness of this complication of parathion poisoning, proper monitoring and treatment could prevent fatalities.