Generation and characterization of human cardiac resident and non-resident mesenchymal stem cell

Despite the surgical and other insertional interventions, the complete recuperation of myocardial disorders is still elusive due to the insufficiency of functioning myocardiocytes. Thus, the use of stem cells to regenerate the affected region of heart becomes a prime important. In line with this human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) have gained considerable interest due to their potential use for mesodermal cell based replacement therapy and tissue engineering. Since MSCs are harvested from various organs and anatomical locations of same organism, thus the cardiac regenerative potential of human cardiac-derived MSCs (hC-MSCs) and human umbilical cord Wharton’s Jelly derived MSC (hUC-MSCs) were tested concurrently. At in vitro culture, both hUC-MSCs and hC-MSCs assumed spindle shape morphology with expression of typical MSC markers namely CD105, CD73, CD90 and CD44. Although, hUC-MSCs and hC-MSCs are identical in term of morphology and immunophenotype, yet hUC-MSCs harbored a higher cell growth as compared to the hC-MSCs. The inherent cardiac regenerative potential of both cells were further investigated with mRNA expression of ion channels. The RT-PCR results demonstrated that both MSCs were expressing a notable level of delayed rectifier-like K+ current (I KDR ) ion channel, yet the relative expression level was considerably varied between hUC-MSCs and hC-MSCs that Kv1.1(39 ± 0.6 vs 31 ± 0.8), Kv2.1 (6 ± 0.2 vs 21 ± 0.12), Kv1.5 (7.4 ± 0.1 vs 6.8 ± 0.06) and Kv7.3 (27 ± 0.8 vs 13.8 ± 0.6). Similarly, the Ca2+-activated K+ current (I KCa ) channel encoding gene, transient outward K+ current (I to ) and TTX-sensitive transient inward sodium current (I Na.TTX ) encoding gene (Kv4.2, Kv4.3 and hNE-Na) expressions were detected in both groups as well. Despite the morphological and phenotypical similarity, the present study also confirms the existence of multiple functional ion channel currents IKDR, IKCa, Ito, and INa.TTX in undifferentiated hUC-MSCs as of hC-MSCs. Thus, the hUC-MSCs can be exploited as a potential candidate for future cardiac regeneration

Repeated infections of dengue (serotype DENV-2) in lung cells of BALB/c mice lead to severe histopathological consequences

To determine the effect of DENV (serotype 2) repeated infections on lung cells is the main goal of this study. From the result, lung histology of control BALB/c mice showed normal alveolar morphology, while vehicle control BALB/c mice highlighted a slight thickening of the alveolar septum. Lung histopathology of BALB/c mice infected twice by DENV-2 showed the presence of hemorrhage, plasma leakage and presence of hemosiderin-laden macrophages (HLMs). Notably, in the lung of BALB/c mice infected four times by DENV-2, we observed thickening and disruption of the alveolar septum, inflammatory cell infiltration, plasma leakage and increased cellularity. Megakaryocyte releasing platelets were also found into the lung alveolus. Overall, our findings showed severe histopathological damage in lungs repeatedly infected by DENV-2, allowing us to argue that they can be linked to pulmonary complication. Result also showed that the number of infections with similar total DENV-2 titer led to different histopathological changes

A diagnostic kit for the detection of early acute leptospirosis

The present invention relates to a diagnostic method for the detection of acute leptospirosis. The method of the invention allows for a quick, specific and sensitive detection of the disease caused by pathogenic Leptospira. The described method is based on the immunological detection of the protein LipL21, a membrane protein expressed specifically by pathogenic strains of Leptospira. Furthermore, the present invention relates to the development of a diagnostic device that allows positive diagnosis to be conducted in the early and critical stage of the infection. The preferred diagnostic device of the invention is a lateral flow test, which as a hand-held assay, is easy to use and facilitates diagnosis of leptospirosis in the first 5 days of infection. Further disclosed is the antibody raised against the LipL21 protein as well as its respective uses in the diagnostic methods and devices. Finally, a diagnostic kit is disclosed containing the inventive materials described herein to conduct diagnosis of acute leptospirosis during the critical early stage of infection. The first 5 days of the infection is where the subject or patient could succumb to the disease. Diagnosis of the disease at this critical stage of infection would then advise the affected patient to have immediate systemic antibiotic treatment and thereby, avoid succumbing to the disease

Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis

Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection