Reoperation following Pancreaticoduodenectomy.

Introduction. The literature on reoperation following pancreaticoduodenectomy is sparse and does not address all concerns. Aim. To analyze the incidence, causes, and outcome of patients undergoing reoperations following pancreaticoduodenectomy. Methods. Retrospective analysis of 520 consecutive patients undergoing pancreaticoduodenectomy from May 1989 to September 2010. Results. 96 patients (18.5%) were reoperated; 72 were early, 18 were late, and 6 underwent both early and late reoperations. Indications for early reoperation were post pancreatectomy hemorrhage in 53 (68%), pancreatico-enteric anastomotic leak in 10 (13%), hepaticojejunostomy leak in 3 (3.8%), duodenojejunostomy leak in 4 (5%), intestinal obstruction in 1 (1.2%) and miscellaneous causes in 7 (9%). Patients reoperated early did not fare poorly on long-term follow up. Indications for late reoperations were complications of index surgery (n = 12), recurrence of the primary disease (n = 8), complications of adjuvant radiotherapy (n = 3), and gastrointestinal bleed (n = 1). The median survival of 16 patients reoperated late without recurrent disease was 49 months. Conclusion. Early reoperations following pancreaticoduodenectomy, commonly for post pancreatectomy hemorrhage, carries a high mortality due to associated sepsis, but has no impact on long-term survival. Long-term complications related to pancreaticoduodenectomy and adjuvant radiotherapy can be managed successfully with good results.

Rice bran lysolecithin as a source of energy in broiler chicken diet.

1. Rice bran lysolecithin (RBL) was evaluated in broiler chicken diets. In the first experiment, RBL was included in diet at 0, 0·5, 2, 8 and 32 g/kg and fed to 250 broiler chickens from 0 to 42 d of age. In the second experiment, RBL was fed at 0, 25 and 50 g/kg diet to 405 day-old broiler chickens until 21 d of age, while during the finisher phase (22-35 d of age) chickens receiving each concentration of RBL were given all three concentrations of RBL in a 3 × 3 factorial manner. The diets were isocaloric. 2. Body weight, food consumption and food conversion efficiency were unaffected by feeding RBL, while the weight of pancreas increased at ≥2 g/kg of RBL in diet (experiment 1). In experiment 2, body weight was greater in the chickens receiving RBL at either 25 or 50 g/kg (21 d) and 50 g/kg (35 d of age). At 21 d of age, food consumption was greater at 25 or 50 g RBL/kg diet, while food conversion efficiency improved with 50 g RBL/kg diet. 3. Fat digestibility increased with RBL at 32 g/kg (experiment 1) and ≤25 g/kg (experiment 2). Rice bran lysolecithin increased ready to cook weight at 50 g/kg during starter phase and decreased abdominal fat at 25 and 50 g/kg during finisher phase (experiment 2). Liver and meat fat content were not affected. 4. It is concluded that lysolecithin from rice bran oil could be used as energy supplement in broiler chicken diet.

Lipase-mediated conversion of vegetable oils into biodiesel using ethyl acetate as acyl acceptor.

Ethyl acetate was explored as an acyl acceptor for immobilized lipase-catalyzed preparation of biodiesel from the crude oils of Jatropha curcas (jatropha), Pongamia pinnata (karanj) and Helianthus annuus (sunflower). The optimum reaction conditions for interesterification of the oils with ethyl acetate were 10% of Novozym-435 (immobilized Candida antarctica lipase B) based on oil weight, ethyl acetate to oil molar ratio of 11:1 and the reaction period of 12h at 50 degrees C. The maximum yield of ethyl esters was 91.3%, 90% and 92.7% with crude jatropha, karanj and sunflower oils, respectively under the above optimum conditions. Reusability of the lipase over repeated cycles in interesterification and ethanolysis was also investigated under standard reaction conditions. The relative activity of lipase could be well maintained over twelve repeated cycles with ethyl acetate while it reached to zero by 6th cycle when ethanol was used as an acyl acceptor.

Lipase-mediated transformation of vegetable oils into biodiesel using propan-2-ol as acyl acceptor.

Propan-2-ol was used as an acyl acceptor for immobilized lipase-catalyzed preparation of biodiesel. The optimum conditions for transesterification of crude jatropha (Jatropha curcas), karanj (Pongamia pinnata) and sunflower (Helianthus annuus) oils were 10% Novozym-435 (immobilized Candida antarctica lipase B) based on oil weight, alcohol to oil molar ratio of 4:1 at 50 degrees C for 8 h. The maximum conversions achieved using propan-2-ol were 92.8, 91.7 and 93.4% from crude jatropha, karanj and sunflower oils, respectively. Reusability of the lipase was maintained over 12 repeated cycles with propan-2-ol while it reached to zero by 7(th) cycle when methanol was used as an acyl acceptor, under standard reaction conditions.

Phospholipase-mediated preparation of 1-ricinoleoyl-2-acyl-sn-glycero-3-phosphocholine from soya and egg phosphatidylcholine.

1-Ricinoleoyl-2-acyl-sn-glycero-3-phosphocholine was prepared by incorporating ricinoleic acid completely in the sn-1 position of egg and soya phosphatidylcholine (PC) using immobilized phospholipase A(1) as the catalyst. The optimum reaction conditions for maximum incorporation of ricinoleic acid into PC through transesterification were 10% (w/w) immobilized enzyme (116 mg), a 1:5 mol ratio of PC (soya, 387 mg; egg, 384 mg) to methyl ricinoleate (780 mg) at 50 degrees C for 24 h in hexane.

An immunochromatographic serological assay for the diagnosis of Mycobacterium tuberculosis.

A rapid serological test for tuberculosis (TB) infection was designed using antigens specific to Mycobacterium tuberculosis. Tuberculosis infection, TB vaccination and exposure to environmental Mycobacteria cannot be distinguished using skin tests based on tuberculin protein derivatives. The standard diagnostic techniques such as skin tests, X-rays and DNA techniques are time consuming, expensive, and not practical for screening large populations. We used the 38, 63, 64, 14, 59-kDa antigens of M. tuberculosis to develop a rapid immunochromatographic test kit. This study evaluates the diagnostic potential of the rapid test kit using TB positive and TB negative serum samples from various hospitals in India. The samples were obtained from patients infected with or exposed to bacteria and viral pathogens. The results demonstrated that the combination of antigens improved the diagnostic specificity and sensitivity. The specificity of the test was 99.42% with sensitivity of 98.52% (n = 241). In case of multiple infections, the specificity was 93.15% with a low sensitivity of 73.52% n = 141). The test kit may offer an improved alternative to purified protein derivative (PPD). This rapid TB test kit may be a useful tool for first-line testing of suspected cases, epidemiological studies and in designing a quality health system to reduce health hazards in resource-poor countries.

Semiliki forest virus vector carrying the bovine viral diarrhea virus NS3 (p80) cDNA induced immune responses in mice and expressed BVDV protein in mammalian cells.

Bovine viral diarrhea virus (BVDV) is a primary pathogen responsible for bovine enteric, respiratory and reproductive failure. A genetic region is encoding the p80 (NS3) of BVDV as the most conserved protein among Pestiviruses. BVDV infection in cattle induces NS3 specific lymphocyte proliferation and humoral responses. To generate a DNA vaccine against BVDV, the gene for BVDV-NADL NS3 was cloned into an eukaryotic expression vector of Semiliki Forest virus (pSFV-1). Quadriceps muscles of BALB/c mice were injected with recombinant DNA generated statistically significant cytotoxic T-lymphocyte activity (CTL) and cell mediated immune (CMI) responses against cytopathic and noncytopathic BVDV. Whereas, the BVDV-NS3 did not generate neutralizing antibodies against BVDVin mice. pSFV-1-NS3 DNA was subjected to in vitro transcription into mRNA. The mRNA was transfected into baby hamster kidney cells (BHK-21) and Madin-Darby bovine kidney cells (MDBK). The recombinant cells were used in the detection of DNA antigen responses by immunological assays. This report establishes the ability of BVDV-NS3 DNA inoculation to induce a strong cellular immune responses in mice.

The effect of different bovine viral diarrhea virus genotypes and biotypes on the metabolic activity and activation status of bovine peripheral blood mononuclear cells.

The effects of cytopathic (cp) and non-cytopathic (ncp) bovine viral diarrhea virus (BVDV) on the cellular metabolic activity and activation status of bovine peripheral blood mononuclear cells (PBMC) were investigated. Cellular DNA and protein synthesis was determined by [3H]thymidine and [3H]valine incorporation, respectively, in phytohemagglutinin (PHA)-stimulated PBMC. All cp strains and most ncp BVDV strains significantly inhibited DNA synthesis in PHA-stimulated PBMC; however, only cp BVDV strains inhibited protein synthesis. A plaque assay and immunofluorescence test confirmed productive BVDV infection of PBMC. In addition, viral RNA synthesis was demonstrated in BVDV-infected PBMC by RT-PCR. The interleukin-2 receptor (IL-2R) was used as a marker for the activation status of BVDV-infected PBMC. The expression of IL-2R was preserved in virus-infected cells, even though DNA and protein synthesis was suppressed. These findings suggest a novel mechanism of virus-induced immune suppression in which BVDV inhibits basic metabolic activities of bovine PBMC. The activation signals, however, are maintained.

Application of recombinant bovine viral diarrhea virus proteins in the diagnosis of bovine viral diarrhea infection in cattle.

The National Animal Disease Laboratory (NADL) vaccine strain of bovine viral diarrhea virus (BVDV) genes for gp48 and p80 were expressed in Escherichia coli. The BVDV-NADL gene for gp62 was integrated into a baculovirus genome for expression in Spodoptera frugiperda (Sf-9) insect ovarian cells. The antigenicity of baculovirus expressed BVDV protein was detected by anti-BVDV specific antibodies in an enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescent assay (IFA) and radio-immunoprecipitation (RIP). The recombinant proteins isolated from bacteria showed antigenic properties when analyzed by ELISA and immunoblotting using BVDV antibodies. The recombinant proteins were then used in ELISA or IFA to detect BVDV infection by testing 54 independent bovine serum samples. The baculovirus-expressed BVDV protein was used as an ELISA and IFA antigen, and the bacteria-expressed proteins were used as ELISA antigens. BVDV-NADL-infected Madin-Darby bovine kidney (MDBK) cell monolayers served as a control antigen. Statistical analysis showed a high degree of correlation between the reactivity of recombinants and natural antigens in ELISA using bovine sera. The results of ELISA or IFA proved there is a high degree of correlation with the virus neutralization. In the comparative ELISA assays, the insect-cell-mediated expression revealed greater specificity and sensitivity than the bacterial expression or the natural BVDV antigens produced by cell cultures.

A 50 kDa membrane protein from bovine kidney cells is a putative receptor for bovine viral diarrhea virus (BVDV).

A 50 kDa cell surface protein from MDBK cells has been identified as a putative receptor for bovine viral diarrhea virus (BVDV) by using a BVDV specific anti-idiotypic antibody (Anti-D89). This study delineates further characterization of the receptor protein. Protease treatment of cultured MDBK cells adversely affected the receptor thus abolishing the binding of anti-D89 to the cells. However, pretreatment of the cells with either phospholipases or glycosidases did not signficantly alter the extent of anti-D89 binding. Additionally, pretreatment of cell monolayers with proteases decreased BVDV attachment and replication in the cells. These results suggested that the receptor for BVDV is a protein in nature, and glycosylation and phosphorylation of the receptor protein may not play a direct role in BVDV attachment to cells. The BVDV receptor protein gradually regenerated on cells when they were maintained in culture following protease treatment. The purified 50 kDa receptor protein also significantly inhibited BVDV infection in a plaque reduction assay.

Antigenic differences between a field isolate and vaccine strains of bovine viral diarrhea virus.

Antigenic diversity among the bovine viral diarrhea virus (BVDV) cytopathic strains 87-2552 (field isolate) and NADL and Singer (prototype strains) was demonstrated with monoclonal antibodies (MAbs) in enzyme-linked immunosorbent, immunofluorescence, virus neutralization, and immunoprecipitation assays. Two MAbs against BVDV strain 87-2552, designated D11 and B7, strongly neutralized this field strain and were specific for the 48-kDa glycoprotein of the virus. These two MAbs have different subisotypes, immunoglobulin G1 for D11 and immunoglobulin G3 for B7. MAbs against BVDV strains 87-2552 and NADL were specific for their respective strains in virus neutralization assays. The results indicated significant antigenic differences between BVDV strain 87-2552 and the NADL and Singer strains.