Extraction and Characterization of Lipopolysaccharide from Salmonella typhi

There is no standard process available for high purity LPS isolation, and it may be appropriate to mix two purification steps. For Salmonella typhi LPS that were subjected to normal microbiological & biochemical screening protocols, updated phenol-water extraction protocol & non-phenolic extraction methods were also used in present research. Pellet of crude LPS obtained with wet weight was 2.0 gm. yield of LPS was more by hot phenol method 1.94 mg/ml as compared to non-phenolic method 0.40 mg/ml. relative purity of LPS obtained by hot phenol method was more as compared to non-phenolic method, as protein content was 23.60 mg/ml in LPS extracted from hot phenol method & 26.62 mg/ ml in LPS extracted from non-phenolic method. However, Nucleic acid contain was comparable in LPS extracted from both methods. Qualitative analysis showed ladder like bands of LPS extracted by hot phenol method as compared to single band obtained for LPS extracted by non-phenolic method. Findings of silver staining clearly revealed ladder pattern of multi-rung bands that are characteristics of smooth form of gram negative bacteria due to difference in length of carbohydrate chain of O-antigen component. In order to clarify diseased conditions & better work on LPS profiling, this research may be crucial to lead to more successful diagnosis and care.

Extending import detection algorithms for concept import from two to three biomedical terminologies

Background While enrichment of terminologies can be achieved in different ways, filling gaps in the IS-A hierarchy backbone of a terminology appears especially promising. To avoid difficult manual inspection, we started a research program in 2014, investigating terminology densities, where the comparison of terminologies leads to the algorithmic discovery of potentially missing concepts in a target terminology. While candidate concepts have to be approved for import by an expert, the human effort is greatly reduced by algorithmic generation of candidates. In previous studies, a single source terminology was used with one target terminology. Methods In this paper, we are extending the algorithmic detection of “candidate concepts for import” from one source terminology to two source terminologies used in tandem. We show that the combination of two source terminologies relative to one target terminology leads to the discovery of candidate concepts for import that could not be found with the same “reliability” when comparing one source terminology alone to the target terminology. We investigate which triples of UMLS terminologies can be gainfully used for the described purpose and how many candidate concepts can be found for each individual triple of terminologies. Results The analysis revealed a specific configuration of concepts, overlapping two source and one target terminology, for which we coined the name “fire ladder” pattern. The three terminologies in this pattern are tied together by a kind of “transitivity.” We provide a quantitative analysis of the discovered fire ladder patterns and we report on the inter-rater agreement concerning the decision of importing candidate concepts from source terminologies into the target terminology. We algorithmically identified 55 instances of the fire ladder pattern and two domain experts agreed on import for 39 instances. In total, 48 concepts were approved by at least one expert. In addition, 105 import candidate concepts from a single source terminology into the target terminology were also detected, as a “beneficial side-effect” of this method, increasing the cardinality of the result. Conclusion We showed that pairs of biomedical source terminologies can be transitively chained to suggest possible imports of concepts into a target terminology.

The pattern of peri-hilar and hilar arterial branching in kidney allografts of living donors

Introduction: Kidney transplantation gives us the opportunity to study kidney vascular anatomical variations and their probable effect on allograft survival. Objectives: The aim of this study was to evaluate the renal arteries’ branching patterns of the engrafted kidneys and their impact on transplant outcomes. Patients and Methods: Three hundred patients who received kidney transplantation between 2014 -2017 were included. Peri-hilar and hilar branching patterns of the engrafted kidney were studied by reviewing the archived CT angiographies of donors and then they categorized based on the existing knowledge in this field. Clinical data were also gathered from medical records and recipients’ latest clinical and laboratory evaluations. Results: Based on peri-hilar and their corresponding hilar branching patterns, our morphology findings were classified into 17 groups. From different peri-hilar branching patterns, the fork pattern was more common which is detected in 95% (242) while the ladder pattern was observed in 5% (13) of kidney grafts. In a later branching sub-categorization, among the fork pattern, 65.2% (158) were duplicated and 34.7% (84) had triplicated hilar branching patterns. There was not a statistically significant correlation between each of those patterns and allograft function (P > 0.05). Conclusion: Peri-hilar and hilar branching patterns of the kidney allografts’ renal artery were different, but they follow certain patterns. Although it may not influence the allograft survival, it provides us precise knowledge about renal vasculature patterns and outcome of probable vascular events meanwhile; it could be useful in the field of transplantation.

Adult intestinal malrotation: a case report

Midgut malrotation is an incomplete rotation of the intestine which occurs during foetal development and usually presents in the neonatal period. The bowel is not fixed adequately and is thus held by a precariously narrow-based mesentery. The incidence of malrotation has been estimated at 1 in 6000 live births, it is rare for malrotation to present in adulthood. Author report a 40-year-old man presented to emergency department with generalized abdominal pain. His symptoms began 2 days before admission. The patient really weak and his mental state was somnolence. His abdomen was slightly extended, and intestinal peristalsis was obscure. Abdominal X-rays revealed dilatation of gastric, coiled spring appearance without free air and step ladder pattern. The patient then underwent laparotomy exploration. Upon entering the abdomen, ileum was noted to completely mobilized and founded volvulus segment, a clockwise twisting three times, about 30 cm proximal from ICJ, and soon author released it. The ligament of Treitz misplaced, there was in the right lower quadrant, close by ICJ. Then about 110 cm segment of ileum necrotic, the rest of the small bowel was normal, author found internal hernia in the right lower quadrant and there was Ladd's band. Clinicians should be aware of this serious cause of abdominal pain. The diagnosis of malrotation in adulthood is often delayed. Complete resolution of acute obstruction or chronic abdominal pain is the result of a high index of suspicion for malrotation, appropriate diagnostic studies, and aggressive treatment.

Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm

Background and Objectives: Burkholderia mallei is the leading cause of glanders, a highly transmittable and an OIE-no- tifiable disease of equidae. Despite the importance of B. mallei, little is known about serodiagnosis of glanders. The present study aimed to develop an immunoblotting assay based on whole-cell proteome of B. mallei to enable accurate serodiagnosis of glanders. Materials and Methods: Three farm horses were subcutaneously immunized with a crude suspension (106 cfu/ml) of heat-inactivated B. mallei formulated with incomplete Freund's adjuvant (IFA) to achieve a hyperimmune sera panel. The immunization was done for 1, 14 and 28 days with 1 dose of 1 ml antigen containing 106 cfu/ml. The hyperimmunity of sera was confirmed by CFT. B. mallei whole-cell proteome was prepared through sonication and the protein content was visu- alized by SDS-PAGE and quantified by Western blot using HRP-conjugated rabbit anti-horse IgG. A comprehensive set of positive and negative horse sera validated the test. Results: A ladder pattern of the B. mallei immunoreactive antigens was seen within the region of 20-90 kDa clearly and the immunoblot was scored positive, while no reaction was seen for the negative sera. The Western blot assay indicated a noticeably higher diagnostic specificity for positive or negative sera of glanders. Conclusion: The whole-cell proteome-based immunoblot proved reliable and straightforward in our study. The prepared an- tigen was adaptable for application in immunoblotting. We assumed this improved immunoblotting system provides appro- priate sensitivity and also specificity expected in serodiagnosis of glanders in endemic areas and typically in less-developed countries.

Mechanisms of chromium hexavalent-induced apoptosis in rat testes

Hexavalent chromium (CrVI)-containing compounds, present in industrial settings and in the environment, are known as carcinogens and mutagens. The present study is designed to test the hypothesis that oxidative stress mediates CrVI-induced apoptosis in testis. Male Wistar rats received an intraperitoneal injection of potassium dichromate at doses of 1 and 2 mg kg−1. Superoxide anion production was assessed by the determination of the reduction of cytochrome c and iodonitrotetrazolium, lipid peroxidation (LPO), metallothioneins (MTs), and catalase (CAT) activity. Apoptosis was evaluated by DNA fragmentation detected by agarose gel electrophoresis. Germinal cells apoptosis was detected by toluidine blue staining. The expression of Bax and Bcl-2 proteins (Pts) was also investigated. After 15 days of treatment, an increase of LPO and MT levels occurred, while CAT activity was decreased. Testicular tissues of treated rats showed pronounced degradation of the DNA into oligonucleotides as seen in the typical electrophoretic DNA ladder pattern. Intense apoptosis was observed in germinal cells of Cr-exposed rats. Bax Pt expression was induced in spermatogonia and spermatocytes cells of CrVI-treated rats. In contrast, Bcl-2 Pt was occasionally observed in germ cells of CrVI-exposed rats. These results clearly suggest that CrVI subacute treatment causes oxidative stress in rat testis leading to apoptosis.

POTENCY OF BRASSICACEAE EXTRACTS AS AN APOPTOSIS ENCHANCER IN BLOOD CELLS MICE EXPOSED TO LEAD ACETATE

<p>Lead (Pb) is an environmental pollutant that is widely used in various industries. It has genotoxic effect that can trigger cancer formation (carcinogenic). The member of Brassicaceae family (white headed cabbage, pak choi, and Chinese cabbage) is known to have glucosinolates, isothiocyanates, phenethilisothiocyanates, and alyl isothiocyanates which are anticarcinogenic. The aim of this study was to examine the potential of Brassicaceae extracts to enhance apoptosis in mouse blood cell exposed to lead acetate. The research method was descriptive with five treatments and four replications. Mice were pretreated with white headed cabbage, pak choi, and Chinese cabbage extracts by cumulative dose 156 mg/kg bw for 13 days prior to exposure of lead acetate by cumulative dose 150 mg/kg bw. All treatment was given orally. Blood samples were taken on the thirteenth day from the tail venous vessel. Furthermore, the DNA isolation and electrophoresis were done to examine DNA fragmentation. The result showed that the white headed cabbage extract enhanced the formation of a 200 bp DNA ladder pattern that is characteristic of apoptosis in the mice blood cells due to lead acetate exposures.</p><p><br /><strong>Keywords</strong>: apoptosis, lead acetate, white headed cabbage, pak choi, Chinese cabbage</p>

DEVELOPMENT OF REVERSE-TRANSCRIPTION LOOP-MEDIATED ISOTHERMAL AMPLIFICATION FOR AVIAN LEUKOSIS VIRUS SUBGROUPS A AND J

Neoplasic diseases in chickens are frequently caused by avian leukosis virus (ALV), including subgroup A (ALV-A) and subgroup J (ALV-J) mostly. The objective of the present study was to develop a one-step, single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) test for detecting ALV-A and ALV-J separately. Primers were designed from the conserved region of the pol and env genes of ALV-A and ALV-J for RT-LAMP tests. Other avian viruses were used to determine the specificity of these tests. The results showed that a ladder pattern on gel electrophoresis was observed for ALV-A and ALV-J specifically and no cross-reaction was observed with other avian viruses. Serial dilutions of ALV-A and ALV-J were used to evaluate the detection limits of these tests. The present RT-LAMP tests showed 10–100 times more sensitivity than the conventional reverse transcription polymerase chain reaction. Thus, these tests are rapid and accurate for the detection of ALV-A and ALV-J.