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Afzal, M. A.

Publications and source records attributed to Afzal, M. A..

2 recordsLinked to original sources

Intracellular lipopolysaccharide regulates ER remodeling upon bacterial infection

Selective autophagy of the endoplasmic reticulum (ER), termed ERphagy or reticulophagy, plays a key role in organelle remodeling and cellular homeostasis. However, whether and how ERphagy is regulated during Gram-negative bacteria infection to influence host responses remains unclear. Here, we show that Salmonella enterica serovar Typhimurium releases lipopolysaccharide (LPS) that colocalizes with RETREG1/FAM134B, a reticulon-like ER-resident receptor for ERphagy. Cytosolic delivery of LPS, either during infection or via transfection, markedly increases RETREG1- and LC3B-decorated ER fragments. Mechanistically, affinity-isolation assays demonstrate that LPS directly binds RETREG1 through interactions between lipid A and positively charged residues within its amphipathic helices and C-terminal region. This interaction promotes RETREG1 oligomerization and drives ER membrane fragmentation, a process further amplified by the O-antigen moiety of LPS. The resulting ER fragments accumulate around LC3-positive Salmonella-containing vacuoles, facilitating bacterial clearance. Importantly, both intracellular and extracellular Salmonella exploit outer membrane vesicles (OMVs) to deliver LPS into the host cytosol, triggering RETREG1 activation and ER remodeling. Collectively, our findings reveal a previously unrecognized host response by which LPS of Gram-negative bacteria are sensed by the host ERphagy machinery to promote xenophagy and enhance antibacterial defense.

cell biology↗

Diagnostic efficacy of hand-held digital refractometer for determining total serum protein in indigenous sheep of Pakistan

The study was designed to ascertain the diagnostic efficacy of hand-held digital refractometer in determining total protein. The Sipli sheep (n=141) were grouped as per gender (females=99, males=29) and age (G1=up till 1 year, G2=from 1 to 2 years, G3=above 2 years). The results regarding the overall mean ({+/-}SE) values and RIs for the TPs attained through serum chemistry analyzer (TP1) and hand-held digital refractometer (TP2) were non-significantly (P[≤]0.05) different (59.2{+/-}1.6g/L and 59.8{+/-}0.5g/L, respectively). However, the RIs were quite different between the two TPs being 45.1-95.7g/L and 57.0-67.0g/L for TP1 and TP2, respectively. Similar results were seen for gender-wise and group-wise results. On the contrary, the results regarding correlation coefficient and logilinear regression showed a negative correlation between the two TPs (r=-0.0244) with an adjusted r-square of 0.059 (5.9% probability). Furthermore, the results for Cronbach alpha and intraclass correlation coefficient between TP1 and TP2 showed that the values for single measure and average values were lower between TP1 and TP2 being - 0.135 and -0.313. Bland and Altman test between TP1 and TP2 also showed a weak level of agreement between the two methods of detecting TP. A proportional bias on the distribution of data around the mean difference line was noticed between TP1 and TP2 (Mean= 0.5; 95% CI= 39.8 to -40.9) with a standard deviation of biasness being 20.58. In a nutshell, the hand-held digital refractometer cannot be used as an on-farm POCT device for determining serum TP in sheep. However, certain other models of refractometers with higher sensitivity and specificity may be utilized in future studies to establish these conclusions for other species of livestock.

zoology↗