Prevalence of ESBL Encoding Genes in Acinetobacter baumannii Strains Isolated from Various Samples of a Tertiary Care Hospital in Mymensingh.

The aim of this study was to find the prevalence of ESBL genes among A. baumannii isolates. In this cross sectional study, 49 Acinetobacter spp. were isolated from various clinical samples from March 2019 to February 2020 conducted in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Clinical samples including endotracheal aspirates, wound swab/pus, urine and blood. A total of 380 samples were analyzed. Growth was obtained in 34.21% of the samples yielding 130 organisms. Out of 130 organisms, 49(37.69%) were Acinetobacter spp. Among 49 Acinetobacter spp, 39(79.59%) were Acinetobacter baumannii which was identified by PCR targeting OXA-51 like gene. Amplification of the ESBL encoding genes, namely CTX-M, TEM, SHV done by molecular technique PCR. The most antibacterial resistance was against ceftriaxone (79.48%) and lower resistance only showed in colistin (12.82%). All the isolates were sensitive to tigecycline. The distribution of ESBLs genes such as TEM 20(51.28%), CTX-M 16(41.02%) and SHV 0(0%). The high resistance to most of the antibiotics among the studied strains and also a high prevalence of TEM gene in A. baumannii strains found in our study gives alarming sign towards the treatment complexity of these strains.

Risk Factors of UTI in Children with Nephrotic Syndrome.

Urinary tract infection (UTI) is a common complication in nephrotic children and leads to most morbidity and mortality in developing countries like Bangladesh. This case control study was conducted in the Department of Pediatrics and Pediatric Nephrology ward of Dhaka Medical College Hospital, Dhaka from July 2016 to June 2018 to identify the risk factors of UTI in children with nephrotic syndrome. Total 90 patient of nephrotic children aged 2-12 years, who were fulfilling the inclusion and exclusion criteria were selected as Group I (case) and Group II (control) according to urine culture report. Group I was UTI positive and Group II was UTI negative. The mean age of Group I was 5.26±3.18 years and Group II was 6.03±2.85 years. There was male predominance in both groups. No significant difference has been found regarding age and sex (p>0.05). No significant difference was also observed regarding economic status and educational level of mother among both groups (p>0.05). Fever, dysuria, abdominal pain, anasarca, vomiting and pallor were found as common presentations in Group I. Children with dysuria and abdominal pain were significantly higher in Group I than Group II (p value <0.001). UTI was found more in relapsed cases than initial attack. E. coli was the most common etiologic agent (37.8%). Mean Hb (gm/dl), serum total protein, serum albumin and serum IgG level were found significantly lower and spot urine protein creatinine ratio was significantly higher in Group I, which implies that those biochemical factors were associated with development of UTI in nephrotic children. Younger age group (<6 years), Constipation and uncircumcised male were found as risk factors and has association with UTI in nephrotic children.

Antibody responses to Lucilia cuprina in sheep selected for resistance or susceptibility to L. cuprina.

Sheep bred for resistance (R) or susceptibility (S) to fleece rot and myiasis (blowfly strike) have been shown to differ in inflammatory response to intradermal administration of blowfly (Lucilia cuprina) antigens and artificial challenge. The current paper describes analysis of antibody responses to L. cuprina antigens in the R and S animals. Serum antibody titres and specificities to larval antigens were examined and the specificity of wound exudate antibodies was also investigated in animals artificially challenged with L. cuprina. Titres of L. cuprina specific serum IgA, IgM, IgG2 and IgG1 were measured by ELISA, while specificities were examined on two-dimensional immunoblots of larval homogenates. Exposure to L. cuprina stimulated the production of specific antibody in both R and S animals, however antibody titres did not differ between the R and S animals. There was large variation in antibody specificity between individual animals and some L. cuprina proteins appear to be more frequently recognized by sera from either resistant or susceptible animals, however the recognition of a specific protein could not be solely attributed to the resistance status of the animal. It appears that resistance in these animals may be independent of serum antibody and is likely to be an innate response. Despite high levels of IgG in wound exudates, this antibody recognized few antigens in comparison with serum from the same animal, suggesting that exudate contains little functional antibody in comparison to serum.

A comparison of inflammatory exudates released from myiasis wounds on sheep bred for resistance or susceptibility to Lucilia cuprina.

Sheep bred for resistance (R) or susceptibility (S) to fleece rot and myiasis (blowfly strike) were experimentally infected with L. cuprina larvae. Exudates released from the wound site were collected during the infection at 6, 12, 18 and 24 h. The exudates were separated using two-dimensional gel electrophoresis, and proteins were silver stained and identified by immunoblotting with specific antibody and by their isoelectric points and molecular weights. Comparisons of exudate composition were made over time and between R and S sheep. Between 6 and 12 h post larval implantation the exudate was rich in IgG and fibrinogen, which is before extensive tissue damage and suggests that the exudate is not simply tissue haemorrhage but the result of an inflammatory response by the sheep to L. cuprina. The exudate grew in complexity between 12 and 18 h and contained a maximum of 74 distinct peptide spots by 24 h. Exudate from wounds on resistant sheep contained many more peptides in the first 12 h of infection, suggesting a more rapid inflammatory response. The source of proteins from the exudate remains speculative; it appears to be composed of many acute-phase proteins, large amounts of immunoglobulin G and proportionally low levels of serum albumin. Exudate composition is likely to be influenced by the local synthesis of acute-phase proteins and perhaps immunoglobulins, selective transport to the infection site and also enzymic degradation by L. cuprina larval enzymes. The more rapid exudation of acute-phase and serum proteins at infection sites on R sheep may allow the inhibition of the establishment of fleece rot bacteria or L. cuprina larvae under natural challenge.

Crystallization and preliminary X-ray diffraction studies of the soluble 14 kDa beta-galactoside-binding lectin from bovine heart.

The soluble 14 kDa beta-galactoside-binding lectin from bovine heart, a member of the S-type lectin family, has been crystallized in a form suitable for X-ray diffraction analysis. The crystals, in the absence of a saccharide ligand, diffract beyond 2.5 A resolution. They are obtained from polyethylene glycol 6000 at pH 6.0. Crystals grow as monoclinic plates, space group P2(1), with cell dimensions: a = 35.47 A, b = 64.33 A, c = 58.78 A and beta = 91.7 degrees. The asymmetric unit contains two molecules related by a 2-fold non-crystallographic axis. Two lectin monomers in the asymmetric unit give a Vm of 2.4 A3/Da, i.e. a solvent content of approximately 50%. The complex of lectin with the saccharide ligand, N-acetyllactosamine, crystallizes in the space group P2(1)2(1)2 with cell dimensions: a = 63.55 A, b = 82.13 A and c = 62.39 A. Crystals of this complex diffract beyond 2.0 A resolution. Two complexes in the asymmetric unit lead to a Vm value of 2.8 A3/Da (57% solvent).

Crystallization of pancreatic procolipase and of its complex with pancreatic lipase.

The human pancreatic lipase-porcine procolipase complex has been crystallized in space group P3(2)21 (a = b = 80.3 A and c = 251 A) from a solution containing polyethylene glycol, NaCl and beta-octyl glucoside. The crystals diffract to 2.6 A on a synchrotron beam. The complex in the presence of bile salts and phospholipids crystallizes in a tetragonal space group P4(2)2(1)2 (a = b = 133.4 A, c = 92.6 A). Crystals of procolipase alone were obtained under slightly different experimental conditions (space group I432, a = b = c = 164.3 A).

Crystallization and preliminary X-ray diffraction study of Lathyrus ochrus isolectin II complexed to the human lactotransferrin N2 fragment.

Isolectin II (LOL II) isolated from the seeds of Lathyrus ochrus has been crystallized in the presence of the N2 fragment (18,500 Da) isolated from human lactotransferrin, which contains an N-acetyllactosamine type biantennary glycan linked to Asn137. This is the first example of a legume lectin crystallized with an N-glycosylprotein. Crystals of the LOL II-N2 complex belong to the tetragonal space group (P4(1)2(1)2 or the enantiomorph) with cell dimensions: a = b = 63.5 A, c = 251.9 A. They diffract well up to at least 3.5 A resolution and more weakly up to 2.8 A resolution. Assuming one functional half-entity in the asymmetric unit, an alpha, beta monomer complexed to one N2 fragment (24,500 Da + 18,500 Da) would give a Vm of 2.95 A3/Da and a solvent content of approximately 58%. SDS/polyacrylamide gels of the dissolved crystals show the presence of both the LOL II and N2 fragment.

Variation in skin inflammatory responses between sheep bred for resistance or susceptibility to fleece rot and blowfly strike.

Sheep which have been bred for resistance (R) or susceptibility (S) to fleece rot and blowfly strike, were tested for intradermal inflammatory responses to excretory and secretory products of Lucilia cuprina larvae. R rams and lambs gave significantly larger skin weals than S animals. In addition, R and S rams were infected with L cuprina first instar larvae and wound exudates were collected. In the first 12 hours of infection R rams released significantly more exudate protein at the wound site than S rams. Correlations suggested that exudate production was stimulated by both larval burden and inflammatory responses, however, in the R group the inflammatory correlation was positive while in the S group it was negative. The results imply that inflammatory responses may play a role in innate resistance to L cuprina. The difference in inflammatory responses suggests genetic differences between the flocks and therefore could show some potential as a trait for indirect selection for resistance to fleece rot and body strike.