Looking at time dependent differentiation of mesenchymal stem cells by culture media using MALDI-TOF-MS.

Mesenchymal stem cells (MSCs) are multipotent cells which are popular in human regenerative medicine. These cells can renew themselves and differentiate into several specialized cell types including osteoblasts, adipocytes, and chondrocytes under physiological and experimental conditions. MSCs can secret a lot of components including proteins and metabolites. These components have significant effects on their surrounding cells and also can be used to characterize them. This characterization of multipotent MSCs plays a critical role in their therapeutic potential. The metabolic profile of culture media verified by applying matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF-MS) techniques. Also, the differentiation and development of MSCs have monitored through culture media metabolome or secretome (secreted metabolites). Totally, 24 potential metabolites were identified. Between them 12 metabolites are unique to BM-MSCs and 5 metabolites are unique to AD-MSCs. Trilineage differentiation including chondrocytes, osteoblasts, and adipocytes, as well as metabolites that are being differentiated, have been shown in different weeks. In the present study, the therapeutic effects of MSCs analyzed by decoding the metabolome for MSCs secretome via metabolic profiling using MALDI-TOF-MS techniques.

Probing heat and oxidation induced conformational changes of molecular chaperone artemin by excitation-emission fluorescence spectroscopy.

Artemin is a potent molecular chaperone, which protects Artemia embryos undergoing encystment against extreme environmental stresses. In the present work, we have examined the structural changes of artemin from A. urmiana upon exposure to oxidant and heat, by using CD measurements as well as excitation-emission fluorescence spectroscopy as a powerful tool for monitoring the conformational transitions and molecular interactions in proteins. We have also provided here the first document on reporting the three dimensional fluorescence spectra of a protein using ANS. Totally, the fluorescence results indicated that the microenvironments of tyrosine and tryptophan residues and the hydrophobic pockets as well as the polypeptide backbone or secondary structure of the chaperone were influenced in responses to heat and H2O2 in different degrees. Moreover, the native state of artemin did not induce a considerable exposure of the internal non-polar groups to the solvent. Besides, the excitation-emission spectra of heated artemin by ANS revealed new emission peaks at 430-450 nm when it was excited at 330 nm, which suggests probable exposure of new binding sites for hydrophobic or electrostatic interactions of the protein with ANS. The protein also showed a greater conformational sensitivity to the temperature fluctuations compared to oxidation. Here, we presented some evidence in support of the relation between artemin and its stress dependent activation in vitro and in vivo. This study can expect that the EEM fluorescence spectroscopy could provide a promising tool to study conformational transitions of proteins.

Frequency of Hepatic Hydrothorax and its Association with Child Pugh Class in Liver Cirrhosis Patients.

OBJECTIVE: To determine the frequency of hepatic hydrothorax and its association with Child Pugh Class in patients with liver cirrhosis. STUDY DESIGN: Descriptive, analytical study. PLACE AND DURATION OF STUDY: Jinnah Postgraduate Medical Centre, Karachi, Medical Unit-III, (Ward- 7), from June 2012 to May 2013. METHODOLOGY: All patients with established diagnosis of decompensated chronic liver disease were included. Detailed history, thorough physical examination, routine laboratory investigations, chest X-ray and abdominal ultrasound were carried out in all patients to find out the presence of pleural effusion and ascites, respectively. Fifty milliliters of pleural fluid was aspirated in all patients with pleural effusion using the transthoracic approach, taking ultrasound guidance, wherever required. Fluid was sent for microscopic, biochemical, and microbial analysis. SBEM defined if pleural fluid with polymorphonuclear (PMN) cell count > 500 cells/mm3 or positive culture with PMN cell count > 250 cells/mm3 with exclusion of a parapneumonic effusion. RESULTS: Two hundred and six patients met the inclusion criteria, with mean age of 41.25 ±13.59 years. Among them, 149 (72.3%) were males and 57 (27.7%) females. Twenty-three (11.2%) had hydrothorax; right sided involvement was in 18 (78.3%) subjects, 3 (13%) had left sided while bilateral pleural effusion was found in 2 (8.7%) cases. SBEM was found in 07 (30.43%) cases. Mean serum albumin 3.125 ±0.71 gram/dl. There was association between serum albumin levels and hydrothorax. Asignificant association of hydrothorax with Child Pugh scoring system (p=0.018), but not with serum albumin (p=0.15). CONCLUSION: The frequency of hepatic hydrothorax has a significant association with hepatic function as assessed by Child Pugh scoring system, but not with serum albumin.

An unusual side effect of interferon alfa 2A: digital clubbing.

Interferon Alfa has been widely used to treat chronic hepatitis C virus infection. In this report, we present a case series of two patients referred to Sarwar Zuberi Liver Centre, Civil Hospital Karachi, who suffered from chronic hepatitis C. After getting detailed clinical examination and baseline work up prior to starting treatment, these patients were offered therapy with usual recommended dose of 3 million units of alpha-interferon subcutaneously thrice weekly. Both these patients developed clubbing of fingers during the course of treatment, one developing it during the 2nd month while the other during the 4th month. It was of grade II in one patient and of grade III in another and was bilateral in both the cases. Clubbing was not presented prior to start of treatment and no other secondary cause of clubbing was found in any of the case. These patients were not on any other drug that is known to interfere with interferon or can be associated with clubbing. No national or international data regarding such unusual side effect is available. Whether this effect is idiosyncratic or dose related and whether it is reversible or not after completion of treatment is yet to be established.