Effect of Leishmania major Infection on the Expression of TGF Beta in Murine.

Leishmania major is a protozoan parasite that causes cutaneous Leishmaniasis disease in human beings and animals. The disease is prevalent in tropical and semitropical countries and has great health importance. The present study aimed to identify the histological changes in the organs infected with L. major and to provide a sophisticated diagnostic method for infection through detecting TGF-ß cytokine by immunohistochemistry technique(IHC) from October 2020 to January 2021. A total of 40 samples of paraffin blocks were used for different organs including skin, spleen, liver, kidney, and heart of male and female BALB/c mice, aged 6-8 weeks, which were previously infected subcutaneously with L. major promastigotes at a dose of 1×107 promastigotes/moues. The result indicated epidermal hyperplasia with diffuse severe lymphohistiocytic inflammatory cells infiltration in the dermis. Hyperplasia of the lymphoid follicles was observed in infected spleen and scattered polymorphonuclear cells mainly neutrophil masses with a random distribution of microgranulomas foci composed of lymphocytes and macrophages within the liver parenchyma around central veins and portal areas. The infected kidney showed aggregation of perivascular mononuclear cells (lymphocytes and macrophages) in the renal cortex. Mononuclear lymphocytes and macrophages were observed within the heart parenchyma especially around blood vessels. Additionally, evaluation of TGF-ß1 expression was highly strong for skin, spleen, relatively strong for liver, heart, and weak for the kidney. In conclusion, infection was accompanied by clinical and histological changes as well as inflammatory diseases. Furthermore, the determination of TGF-ß expression level depends on the diagnosis of infection. A clear understanding of immune mechanisms is essential for preventing, treating, and controlling strategies of this infection.

The imprinted region on human chromosome 7q32 extends to the carboxypeptidase A gene cluster: an imprinted candidate for Silver-Russell syndrome.

Imprinted gene(s) on human chromosome 7q32-qter have been postulated to be involved in intrauterine growth restriction associated with Silver-Russell syndrome (SRS) as 7-10% of patients have mUPD(7). Three imprinted genes, MEST, MESTIT1, and COPG2IT1 on chromosome 7q32, are unlikely to cause SRS since epigenetic and sequence mutation analyses have not shown any changes. One hundred kilobases proximal to MEST lies a group of four carboxypeptidase A (CPA) genes. Since most imprinted genes are found in clusters, this study focuses on analysing these CPAs for imprinting effects based on their proximity to an established imprinted domain. Firstly, a replication timing study across 7q32 showed that an extensive genomic region including the CPAs, MEST, MESTIT1, and COPG2IT1 replicates asynchronously. Subsequently, SNP analysis by sequencing RT-PCR products of CPA1, CPA2, CPA4, and CPA5 indicated preferential expression of CPA4. Pyrosequencing was used as a quantitative approach, which confirmed predominantly preferential expression of the maternal allele and biallelic expression in brain. CPA5 expression levels were too low to allow reliable evaluation of allelic expression, while CPA1 and CPA2 both showed biallelic expression. CPA4 was the only gene from this family in which an imprinting effect was shown despite the location of this family of genes next to an imprinted cluster. As CPA4 has a potential role in cell proliferation and differentiation, two preferentially expressed copies in mUPD patients with SRS syndrome would result in excess expression and could alter the growth profiles of these subjects and give rise to intrauterine growth restriction.

A Survey of Coliforms and Staphylococcus aureus in Cheese Using Impedimetric and Plate Count Methods 1.

A total of 256 cheese samples were analyzed for coliform plate count using violet red bile agar and for an impedance count using BactometerR Coliform Medium with a correlation coefficient between methods of R=-.91. Fifty-four percent of the samples contained 102 to 107 colony forming units/gram (CFU/g). The highest counts were in cream and fresh cheese products. When 27 Cheddar cheese samples were inoculated with from 102 to 107 CFU of Escherichia coli /g a correlation of R=-.97 was found between methods. Two hundred of the cheese samples were analyzed for Staphylococcus aureus using Baird-Parker medium and impedance count using BactometerR S.aureus Medium. Five samples (2%) contained over 103 CFU/g. The strains isolated were coagulase-positive. When 34 samples of cheese were inoculated with 102 to 107 CFU of staphylococci/g, the correlation between the plate and impedance method was R=0.98.

Detection of Abnormal Milk with Impedance Microbiology Instrumentation.

Mastitic milk was detected by obtaining conductance measurements using an impedance microbiology Bactomatic 120 SC instrument. Conductance readings separated normal and abnormal milks after 30 min at 25°C when readings differed by more than 2 to 3% and exceeded the variance among instrument module wells. Samples blended from four quarters of a cow indicated milk from one quarter was abnormal if the salt level in the abnormal quarter raised the blend conductivity above that of normal samples and variance among the wells. Either solid or liquid substrates that contained stimulants could be used to accelerate bacterial acid production or reduce impedance detection times and did not affect the ability to detect abnormal milk. However, measurements varied with the volume of sample in the well, suggesting that fixed 1-ml liquid volumes of milk be used. Such volumes would allow detection of abnormal milk and bacterial load on the same sample.