An Unusual Presentation of Fatal Acute Haemorrhagic Pancreatitis in a Young Adult.

Acute haemorrhagic pancreatitis is a medical emergency and the most severe form of Acute Pancreatitis. It is characterized by severe epigastric pain that radiates to the back and is associated with vomiting. If not diagnosed and managed promptly, it may result in sudden, unexpected, unexplained deaths which fall within the medicolegal domain. In such cases, the role of an autopsy is of paramount importance to determine the cause of death. Here we report a young adult, who presented to the local hospital with vague abdominal discomfort and vomiting following alcohol intake and referred to our tertiary care center for further management. But he was received dead on arrival at our hospital. The diagnosis of acute haemorrhagic pancreatitis was made only after the post mortem examination. Awareness of the physicians about the unusual symptoms in acute haemorrhagic pancreatitis and the need for pancreas examination at autopsy of all sudden deaths is emphasised.

De novo assembly, transcriptome characterization and marker discovery in Indian major carp, Labeo rohita through pyrosequencing.

Labeo rohita, one of the Indian major carps, is the most popular culture species in Indian subcontinent due to its consumer preference and delicacy. A selective breeding program for harvest body weight has resulted in an average genetic gain of 17% per generation. Transcriptome resource for this species is scanty. Here, we have characterized the liver and muscle transcriptomes of rohu using Roche 454 GS-FLX next generation sequencing platform. In total, 1.2 million reads were generated, de novo assembly and clustering resulted in 4171 transcripts. Out of these, 4171 had significant blast hit against NCBI nr database, and 2130 transcripts were successfully annotated. In total, 289 SSRs were identified with an identification rate of 5.8%, and dinucleotide repeat motifs were observed to be the most abundant SSRs. Further, 2231 putative SNPs were identified with high confidence. Validation of eight putative SNPs using Sanger sequencing resulted in 100% true SNPs. Significant allelic imbalance of M1, M4 and M5 loci between growth selected and control individual were observed. Furthermore, 13 transcription factors were identified in the present study belonging to six different transcription factor families. The present study demonstrated the utility of RNAseq to develop genomics resources in non-model fish species, and the marker resources developed would support the genetic improvement program of this species.

Molecular characterization, computational analysis and expression profiling of Dmrt1 gene in Indian major carp, Labeo rohita (Hamilton 1822).

Sexual dimorphism of fish morphology, physiology and behavior is diverse and complex in nature. Doublesex and mab-3 related transcription factor (Dmrt) is a large protein family whose function is sexual development and differentiation in vertebrates. Here, we report a full-length cDNA sequence of Labeo rohita (rohu) Dmrt1 of 907 bp length having 798 bp of open reading frame encoding 265 amino acids. The molecular weight of rohu DMRT1 protein was found to be 28.74 KDa and isoelectric point was 7.53. DMRT1 protein contains 23 positively and 24 negatively charged amino acids with a GRAVY score of -0.618. A characteristic DM domain was found in DMRT1 protein, which is a novel DNA-binding domain. Phylogenetic analysis showed maximum similarity with Cyprinus carpio when compared with DMRT1 of other vertebrates. Molecular docking study identified active sites to be targeted for drug designing. Rohu DMRT1 was observed to interact with other proteins such as FOXL2, CYP19a1a, AMH and SOX9a. Differential expression study revealed higher expression in testis tissue implying its role in male sex differentiation and testicular development. The information generated in the present work could facilitate further research to resolve the issues related to gonadal maturation and reproduction of commercially important aquaculture species.

Surgical outcome in relation to duct size at the porta hepatis and the use of cholagogues in patients with biliary atresia.

BACKGROUND: Small ductules communicating with the bile ducts have been described at the porta hepatis in extrahepatic biliary atresia (EHBA) and these form the basis for hepatic portoenterostomy. The use of cholagogues like dehydrocholic acid (DHC) and ursodeoxycholic acid (UDCA) to enhance bile flow postoperatively has been reported. AIMS: This communication describes our experience with the use of cholagogues following surgery in EHBA and attempts to correlate the outcomes with the diameter of the ductules. MATERIAL AND METHODS: Fifty five EHBA patients treated by the Kasai procedure form the basis of this study; 35 patients treated during 1979-1986 and administered DHC (3-5 mg/kg) postoperatively and 20 patients treated during 1999-2002 and administered UDCA (15 mg/kg) postoperatively. The diameter of ductules was measured using an optical micrometer on 5 microm serial sections; the ducts were classified as type I (no demonstrable ducts, n = 14), type II (< 50 microm, n=22) and type III (> 50 microm, n = 19). The clinical outcome was categorized as 1 (jaundice free survival at 5 years follow-up, n = 7), 2 (initial good response but deteriorated after one year, n = 27) and 3 (expired within one year following surgery, n = 21). The response to surgery was monitored using biochemical liver function tests (LFT), hepatobiliary scintigraphy (HIDA scan) and occurrence of cholangitis. RESULTS: Age did not affect the size of ducts in both DHC and UDCA groups but patients in the DHC group were older than those treated with UDCA (mean age DHC: 105.22 +/- 33.53 days, UDCA: 74.68 +/- 23.73 days; p = 0.009). There was no statistically significant difference between duct size and postoperative LFT in both groups (DHC p = 0.1, UDCA p = 0.5). Bile excretion on HIDA scan was significantly better with larger ducts (DHC p = 0.003, UDCA p = 0.025); overall UDCA showed significantly better bile excretion (p = 0.003) but this was not reflected in the surgical outcome. There was no significant difference in the surgical outcome of those treated with DHC or UDCA but a significantly higher incidence of cholangitis was seen with smaller ducts in the UDCA group (p = 0.02). CONCLUSIONS: There was no correlation between duct diameter and postoperative LFT but type III ducts were associated with better bile flow on HIDA scan. Cholangitis was seen more often with type I and II ducts in both DHC and UDCA groups. UDCA administration seemed to be beneficial in patients with type III ducts in increasing bile flow and reducing cholangitis.

Capturing transformation of flood hazard over a large River Basin under changing climate using a top-down approach.

Existing flood modeling studies over coastal catchments involving different combinations of model chain setup imparting complex information fails to entail the needs of policy or decision-makers. Thus, a comprehensive framework that pertains to the requirements of practitioners and provides more perspicuous flood hazard information is required. In this paper, a novel approach translating complex flood hazard information in the form of decision priority maps derived using a rational combination of models (physical and statistical) is elucidated at the finest administrative scale. The proposed methodology is illustrated over a highly flood-prone deltaic region in Mahanadi River Basin, India, to characterize impacts of climate change for a 1:100 years return period flood event under future conditions (2026-2055). The modeled flood events are further analyzed to capture the transformation dynamics of flood hazard classes (FHCs) in near-future, for prioritizing areas with greater hazard potential. Interestingly, the results capture a high transformation characteristic from low to high FHCs in agriculture-dominated areas, which are significantly greater than the areas experiencing flood hazard reduction. The results show a significant increase of 12.5% and 27.35% in areas with high FHCs under RCP4.5 and RCP8.5 scenarios, respectively. Moreover, a notable climate change response is indicated under both climate change scenarios, with approximately 22% (RCP4.5) and 25% (RCP8.5) in villages showing a drastic increment in flood hazard magnitude. The results thus highlight the importance of identifying and prioritizing the areas for flood adaptation where a relative change in flood hazard potential is higher due to climate change. Therefore, we conclude that this study can provide an insight into the implication of new approaches for effective communication of flood information by bridging the gaps between scientific communities and decision-makers in appraisal for better flood adaptation measures.

Population structure and genetic diversity of hatchery stocks as revealed by combined mtDNA fragment sequences in Indian major carp, Catla catla.

Catla catla is the second most important Indian major carp due to high growth rate and acceptance to consumers for food value. It is widely cultured in the Indian subcontinent as monoculture or polyculture. In the present study, genetic diversity among hatchery stocks (total 218 samples of catla) collected from different geographical regions of India was examined using mtDNA fragment sequence of Cyt b (306 bp) and D loop (710 bp). High numbers (57) of population specific haplotypes were observed in the present study. The results revealed significant genetic heterogeneity for the sequence data (FST = 0.27546, p < .05). Analysis of molecular variance revealed significant genetic differentiation among different catla populations. The information generated in present study could be useful to develop broad genetic base populations of catla.

Profile of trauma related mortality at Manipal.

BACKGROUND: The main objective of the present study is to analyse the pattern of death related to trauma/injuries at Manipal in Udupi district, Karnataka state, India. MATERIAL AND METHODS: Retrospective data (January 2001 to December 2003) were collected from autopsy reports maintained in the department of forensic medicine, and the inquest reports from the police. Out of the total (n=470) autopsies, 344 victims had traumatic fatality. The demographic data, type of trauma, duration of survival, body areas involved was recorded. RESULTS: 470 autopsies were performed during the period of study. Of these 344 (73%) were traumatic fatalities. Males were preponderant (71%). More than half of the victims were between 1-40 years. The youth age of 21-30 years were affected maximum due to all type of trauma except for falls and assault in this area of the country (India). The leading type of trauma was Road Traffic Accident (RTA) (52%) followed by burn (24%). Females were predominant with 74% in trauma due to burns. In majority of cases where the death was reported in the hospital, among them more than 50% of victims were survived in hospital for more than 24 hours. The victims those expired within 3 hours were in majority from RTA and those who survived for more than 24 hours were in majority from burns. Majority (258) of the victims had head injuries followed by thoracic injuries (169), and abdomen (125). Twenty four percent of the victims died on the spot or before reaching hospital. CONCLUSION: In this area of the country the trauma due to RTA is a leading cause of death among the traumatic death followed by burns.

Role of inflammatory and hemostatic biomarkers in Alzheimer's and vascular dementia - A pilot study from a tertiary center in Northern India.

INTRODUCTION: A reliable plasma biomarker in differentiating between Alzheimer's disease (AD) and Vascular dementia (VaD) is the need of the hour, in most memory clinics. Even though there is no disease modifying treatment, it is important to know the type of dementia for both symptomatic treatment and prognostication. METHODS: Neuropsychological assessment, MRI brain, FDG-PET brain and CSF biomarkers of AD (Aß42 and total tau) were used for establishing the diagnosis of Mild Cognitive Impairment (MCI), AD or VaD. RESULTS: 68 diagnosed patients of AD/MCI/VaD were included. FDG PET brain, plasma fibrinogen, d dimer, IL6 and CRP were done in all 68 patients while 48 patients underwent CSF biomarker analysis. Sixteen patients had MCI, of which 11 were MCI-AD and 5 were MCI-VaSC. There were 41 patients with AD (Mild AD-9, Mod AD-23, Severe AD-9) and 11 patients with VaD. Alzheimer group (MCI-AD and AD) and Vascular group (MCI VaSC & VaD) consisted of 52 and 16 patients respectively. Alzheimer and Vascular groups did not exhibit significant difference in IL6 and CRP levels. Plasma fibrinogen levels were significantly higher in VaD and vascular group as compared to Alzheimer group. But MCI-VaSC was not significantly different from MCI-AD. Plasma d dimer levels were significantly higher in all vascular subgroups compared to Alzheimer subgroups except between MCI-VaSC and MCI-AD. CONCLUSION: Hemostatic biomarkers were higher in Vascular group compared to Alzheimer group whereas there was no difference in inflammatory biomarkers. But the sensitivity and specificity of fibrinogen and d-dimer were not high enough for routine clinical use. Further studies in a larger sample are required to confirm these results.

Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin.

Wound dressings that can be formed in situ offer several advantages over the use of preformed dressings such as conformability without wrinkling or fluting in the wound bed, ease of application and improved patient compliance and comfort. Here we describe such an in situ forming hydrogel wound dressing from gelatin, oxidized alginate and borax. Periodate oxidized alginate rapidly cross-links proteins such as gelatin in the presence of borax to give in situ forming hydrogels that are both non-toxic and biodegradable. The composite matrix has the haemostatic effect of gelatin, the wound healing-promoting feature of alginate and the antiseptic property of borax to make it a potential wound dressing material. The hydrogel was found to have a fluid uptake of 90% of its weight which would prevent the wound bed from accumulation of exudates. The water vapour transmission rate (WVTR) of the hydrogel was found to be 2686+/-124 g/m2/day indicating that the hydrogel can maintain a moist environment over wound bed in moderate to heavily exuding wound which would enhance epithelial cell migration during the healing process. The wound healing efficacy of hydrogel was evaluated in experimental full thickness wounds using a rat model which demonstrated that within 2 weeks, the wound covered with gel was completely filled with new epithelium without any significant adverse reactions. These in situ forming hydrogels fulfil many critical elements desirable in a wound dressing material.

Evaluation of soft tissue response to a poly(urethane urea).

Variations in the performance of vascular prostheses constructed of polyurethanes, and some evidence which suggested that these variations could be due not to the properties of the polymer itself, but to differences in the cellular response to the various microstructures of porous polyurethanes require investigation. Experiments were performed to evaluate quantitatively the extent of the cell behaviour adjacent to a series of polyurethane samples. It was shown that, with Biomer, a polyurethane urea, the profile of cell behaviour as a function of distance from the implant surface and of time following implantation, the response of cells in general and macrophages in particular, varied considerably with different internal microstructure. This supports the suggestion that the cellular response to different structures and susceptibility to degradation are related.

Studies on the effect of degree of hydrophilicity on tissue response of polyurethane interpenetrating polymer networks.

Interpenetrating polymer networks of polyurethane and vinyl monomers such as polyacrylamide, polyvinyl pyrrolidone, poly(hydroxyethyl methacrylate) and poly(methyl methacrylate) were implanted intramuscularly in rabbits. Attempts were made to correlate the morphological aspects of the interpenetrating polymer networks to their histological response. A relatively increased hydrophilicity of hydrophobicity of the interpenetrating polymer networks as in the case of polyurethane-polyvinyl pyrrolidone and polyurethane-poly(methyl methacrylate) interpenetrating polymer networks, respectively, could elicit an inert response whilst degradation of materials promoted reactivity.

Use of glutaraldehyde-gentamicin-treated bovine pericardium as a wound dressing.

Glutaraldehyde (GA)-pretreated gentamicin post-fixed bovine pericardium has been evaluated as a wound dressing in this study. Two excisions approximately 7 x 4 cm, each of full thickness skin, from the upper and lower parts down to, but not including, the panniculus carnosus were made from the back of the guinea pig. The skin excised from the upper part was placed on the wound bed of the lower part as an autograft, whereas the upper wound was closed using 5% sodium chloride-trypsin-0.1% GA-0.048% gentamicin-treated bovine pericardium and sutured for comparative study. The wounds were inspected every 3-6 d for infection and exudation. Histopathological studies were performed at weekly intervals in the post-operative period. At the fifth week, a very thin linear scar on the epidermal aspect without remarkable contracture was observed and histopathology showed the completion of epithelization across the wounds in all cases. This study demonstrates that GA-pretreated, gentamicin-post-fixed bovine pericardium may be used as an alternative biological dressing in the case of large wounds.

Structural studies on bovine bioprosthetic tissues and their in vivo calcification: prevention via drug delivery.

Cardiovascular calcification, the formation of calcium phosphate deposits in cardiovascular tissue, is a common end-stage phenomenon affecting a wide variety of bioprostheses. To study the process of calcification in tissue prosthetics, glutaraldehyde-treated bovine pericardium, dura mater and fascialata were implanted subcutaneously in rats and retrieved 21 days later and thereby morphological findings were correlated with biochemically determined levels of calcium. Transmission electron microscopy showed that calcification primarily involved the surface of collagen fibrils and the interfibrillar spaces. The deposition of calcium was higher with dura and fascia prostheses compared to pericardium. However, the release of Fe3+ ions from chitosan matrix had substantially inhibited the deposits of calcium in all implanted tissues. It seems that the structural and anatomical features of the tissue is one of the important factors for tissue-associated calcification. It is also confirmed that glutaraldehyde-preserved pericardium is the most suitable material for the development of cardiac prosthesis, with an appropriate drug combination therapy for prevention of pathological calcification.

Long term tissue response to titanium coated with diamond like carbon.

Diamond like carbon (DLC) coatings were deposited on to Ti substrates by plasma enhanced chemical vapor deposition technique. Ti and DLC/Ti samples were implanted in skeletal muscle of rabbits. The samples were explanted after 1, 3, 6 and 12 months and the tissue-cell interaction was studied. Our data indicate both DLC/Ti and bare Ti to be compatible with skeletal muscle.

Casein as a carrier matrix for 5-fluorouracil: drug release from microspheres, drug-protein conjugates and in-vivo degradation of microspheres in rat muscle.

Glutaraldehyde cross-linked casein microspheres were loaded with 5-fluorouracil (5-FU) from concentrated aqueous solutions of the drug after the microspheres were synthesized and cleaned. In-vitro release of the drug was examined in phosphate buffer in the absence and in the presence of protease at 37 degrees C. Drug release data showed that only about 20% of the drug is released in the absence of protease even after 5 days, while digestion of the matrix with protease released the entrapped drug completely in about 24 h. A protein-drug conjugate was synthesized via carbamoyl linkage using 6-(5-FU-1-yl)hexyl isocyanate and the drug release was examined in phosphate buffer at 37 degrees C. Release from the protein-5-FU conjugate was slower compared with the release from microspheres in the presence of protease. Implantation of placebo microspheres of different cross-linking densities in the gluteal muscle of rats showed no adverse tissue reactions over a one-year period. Histopathological examination of the tissues containing injected microspheres suggested that the biological life of casein microspheres in muscle is about 6 months, which is three times that of cross-linked albumin microspheres.

Effects of double cross-linking technique on the enzymatic degradation and calcification of bovine pericardia.

The strength, resorption rates, and biocompatibility of collagenous biomaterials are profoundly influenced by the method of cross-linking. The in vitro and in vivo calcification and enzymatic degradation of bovine pericardia (BP) after a series of surface modifications were studied as a function of exposure time. Collagenase degradations of modified BP were monitored by scanning electron microscopy and tensile strength measurements. Bovine pericardium was modified by a combination of different tissue fixatives such as glutaraldehyde (GA), carbodiimide (EDC), diisocyanate (HMDIC), and polyethylene glycol (PEG). GA-PEG-EDC-PEG and GA-PEG-HMDIC-PEG combination treated BP retained maximum stability in collagenase digestion compared to GATBP. In vitro calcification studies and in vivo rat subcutaneous implantations of modified pericardium have shown substantial reduction in the calcification of double cross-linked BP with PEG modification. Further, the biocompatibility aspects of pericardial tissues were established by platelet adhesion and octane contact angle. It seems that cross-links involving amino and carboxyl residues may provide new ways of controlling biodegradation and calcification.

The influence of photodynamic therapy on the wound healing process in rats.

In photodynamic therapy (PDT), photosensitisers (PS) are used along with lasers for the treatment of tumors. The combined effect of photosensitisers and lasers on the wound healing process is studied using delta-aminolevulinic acid (ALA) (5 mg/kg) and hematoporphyrin derivative (HPD) (5 mg/kg) as photosensitisers in the open excision wounds of rats. The lasers used were He-Ne laser (3 J/cm2) and Nd:YAG laser (30 J/cm2). This study is important for understanding the healing process involved after PDT. Open excision wounds treated with He-Ne lasers in animals that received ALA as photosensitiser showed complete wound closure at the earliest by 13 +/- 1 days, and with results obtained for HPD and the combination of lasers with complete closing by 14 +/- 1 days. However, the control group of animals that received ALS or HPD with no laser treatment showed wound healing on the twentieth and eighteenth days with a deviation of one day and two days, respectively. ALA with the combination of Nd:YAG and He-Ne lasers and HPD with He-Ne laser alone does not show quicker wound healing effects. Histopathological results also gave similar results. Tensile strength measurements do not vary significantly from control group to the test group. ALA along with He-Ne laser of HPD along with the combination of He-Ne and low power Nd-YAG lasers are found to be ideal methods for quickening the wound healing process in rat.

Cellular basis for failure of joint prosthesis.

In recent times, loosening of joint prosthesis resulting in failure, is of grave concern to orthopedicians. It is estimated that 50% of total hip replacements become loose after 15 years, and most of them require either revision surgery or resection orthoplasty. Neither, newer operative techniques, change in design, use of novel materials, nor surface modifications have helped to circumvent the problem. It is in this context, that attention has been focussed on the role of tissue surrounding the implant, in the loosening of the prosthesis. Tissue response around prosthesis results in either formation of a fibrous layer around the implant, ingrowth into fenestrations on implant or direct bone apposition on prosthesis. Long-term implantation results in implant debris being released into surrounding tissue. These particles initiate a chronic granulonatous inflammation with a significant number of activated macrophages and foreign body type of giant cells, all engaged in attempts to get rid of the debris. These features have been found to be invariably associated with peri-prosthetic lysis of bone. Since such bone resorption is also observed around non-cemented prosthesis, possibly causes other than cement are responsible for the osteolysis. Retrospective studies on failed implants suggest that peri-prosthetic osteolysis is mediated by activated macrophages. Cytokines are capable of stimulating bone resorbing cells, the osteoclasts. Bone resorption results in further loosening of the prosthesis, changes in stress, frictional wear, release of more wear debris and recruitment of more macrophages. Bone death and proliferation of macrophages, thus appear to be the cause for pain and loosening of prosthesis.

Pulmonary vasospasm in rabbits infused with stroma free haemoglobin solution.

Effect of infusion of stroma free haemoglobin solution into mice and rabbits was studied. Though no abnormalities were noted in mice, pulmonary vasospasm and hepatocytic necrosis were seen in all rabbits. In addition to reports that low adenosine deaminase levels in the solution could be a cause of vasoconstriction, it is proposed on the basis of present results that either denaturation of haemoglobin molecule itself or release of super oxide anion during oxygenation of haemoglobin, could cause cellular damage leading to toxicity.

Efficacy of Chitra granule and powder (hydroxyapatite) in alveolar bone regeneration in rabbits. A histological evaluation.

The reconstruction or restoration of osseous defects caused by inflammatory periodontal disease is a continuing challenge in periodontal therapy. Great strides are being made to this effect using alloplasts such as hydroxyapatite. The present study was designed in Newzealand dwarf rabbits to observe the biologic response of periodontal tissues to synthetically prepared hydroxyapatite in both powder and granule forms by Srichitra Tirunal Institute for Medical Sciences & Technology, Trivandrum and to compare it with that of a commercially available, pure resorbable hydroxyapatite, OsteoGen (HA Resorb). The test materials were implanted in the artificially created bonydefects in the mandible via an intraoral approach. The wounds were allowed to heal upto 26 weeks postimplantation. The clinical evaluation at 12 weeks and 26 weeks postimplantation revealed neither any evidence of inflammation, infection or abscess formation nor any exposure or exfoliation of test materials. The histological examination of the implant sites at 12 and 26 weeks postimplantation revealed varying extent of formation of new osseous tissue and periodontal fibers. A comparison between test materials and control suggested that Chitra granules exhibit a relatively greater potential for newbone and periodontal fibre formation.