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Purpose: To estimate the pattern and frequency of ocular morbidity in the urban and Rural school going children aged 5-15 years. Materials and Methods: It was cross sectional study conducted through various school camps including 1000 students in both rural and urban schools between age group 5-15 years. Results: Ocular morbidity was found in 30.5% among all the students, 31.5% in the rural schools and 29.4% in the urban schools. Ocular morbidity was more in female students (33.5%), as compared to male (27.5%). Refractive errors was found to be the most common ocular morbidity 18.8%, followed by allergic conjunctivitis 2.2% and blepharitis 2.2%. Vitamin A deficiency was more in rural schools compared to urban schools. Conclusion: Our study concluded that ocular morbidity was more in rural areas as compared to urban areas seen more among the females belonging to the age group of 11-15 years where Refractive error was seen as the most common cause.

Choudhary et al. show lipid droplet (LD) biogenesis from discrete ER subdomains in yeast. Fld1 together with Nem1 localize to discrete ER subdomains independent of each other and of LDs, but both are required to recruit triacylglycerol (TAG)-synthases and LD biogenesis factors for localized TAG production and droplet assembly., Lipid droplets (LDs) are fat storage organelles that originate from the endoplasmic reticulum (ER). Relatively little is known about how sites of LD formation are selected and which proteins/lipids are necessary for the process. Here, we show that LDs induced by the yeast triacylglycerol (TAG)-synthases Lro1 and Dga1 are formed at discrete ER subdomains defined by seipin (Fld1), and a regulator of diacylglycerol (DAG) production, Nem1. Fld1 and Nem1 colocalize to ER–LD contact sites. We find that Fld1 and Nem1 localize to ER subdomains independently of each other and of LDs, but both are required for the subdomains to recruit the TAG-synthases and additional LD biogenesis factors: Yft2, Pex30, Pet10, and Erg6. These subdomains become enriched in DAG. We conclude that Fld1 and Nem1 are both necessary to recruit proteins to ER subdomains where LD biogenesis occurs., Graphical Abstract

Most cells store metabolic energy in lipid droplets (LDs). LDs are composed of a hydrophobic core, covered by a phospholipid monolayer, and functionalized by a specific set of proteins. Formation of LDs takes place in the endoplasmic reticulum (ER), where neutral lipid biosynthetic enzymes are located. Recent evidence indicate that this process is confined to specific ER subdomains, where proteins meet to initiate LD assembly. The lipodystrophy protein Seipin, is emerging as a major coordinator of LD biogenesis. Seipin forms a large oligomeric toroidal structure, which traps neutral lipids to promote LD nucleation. Here, we discuss the role of LD biogenesis factors that associate with Seipin to assemble functional LDs.

Background: Aims of this study was assess the ideal time of laparoscopic cholecystectomy after endoscopic retrograde cholangiopancreatography in cases of cholelithiasis with Choledocholthiasis Methods: The present study was carried out on 150 patients admitted in Department of General surgery National Institute of Medical Sciences and Research, Jaipur, diagnosed with cholelithiasis along with choledocholithiasis from 1stJanuary 2019 to 30thJune, 2020. Results: In group 1 out of 75 patients 7 patients developed complications in post operative period compared to 35 patients out of 75 patients developed complications in group 2. The The mean hospital stay (in day) in group 1 patients was 2.26, median 2 and the mean of group 2 was 5.26, median 5. Conclusion: To conclude in our study there has been found significant advantage of early laparoscopic cholecystecomy following ERCP over the late group to minimize complications and promote early recovery of the patients. Keywords: ERCP, Cholecystectomy, Endoscopic retrograde cholangiopancreatography

Background: Intestinal perforation is a surgical emergency often, encountered. Usually, coelotomy is performed to manage intestinal perforation. It has been the gold standard approach to deal with intestinal perforation for the past decades in conjunction with various procedures. Post laparotomy wound related complications have been the biggest challenge for general surgeons till date, so much as to adding up to morbidity, as severe as burst abdomen. Laparoscopic management of perforation is a recent technique. Aim: The present study is aimed to assess the efficacy of laparoscopic management of intestinal perforation. Material and Method: Prospective study of 75 patient admitted and operated in National Institute of Medical Sciences & Research. Intestinal perforation (traumatic and not traumatic) will be managed through laparoscopic procedure. The perforated intestine will be – Identified and exteriorized with or without repair of perforation. Adequate peritoneal toilet would be performed. Sub diaphragmatic and pelvic spaces will be drained. The outcome of the patients will be assessed in regards to- Total hospital stay, Any associated complications – local or distant. Cause of mortality. Results: In present study the perforation peritonitis was found to be in 70.6% with 58.6% ileal perforation 17.1% jejunal perforation. In our study 52% patients managed laparoscopically 33.3% laparoscopic with exteriorization and 14.6% were converted to open surgery. Various cause for open surgery includes 27.2% multiple perforation,9.2% cardiac disease and 63.6v % site not identified laparoscopically. Patients managed laparoscopically had less complications as compared to open. Conclusion: dealing with intestinal perforation or associated peritonitis using minimal access technique is feasible and in turn helpful in minimizing the most dreaded morbidity i.e. the surgical site infections which can be reduced with improving skills and thorough lavage. Other factors associated with post-operative morbidity i.e. ambulation can be started as early as post operative day one. Prolonged hospital stay can be also be reduced and patients can be sent home early decreasing risk for nosocomial Keywords: Laparoscopic, intestinal, perforation, perforation peritonitis

Lipid droplets (LDs) are intracellular organelles dedicated for fat storage and play critical roles in cellular homeostasis. Recently, LD biology has moved to the forefront of biomedical research due to their involvement in a variety of diseases that are affected by lipid imbalance, such as obesity, type 2 diabetes, fatty liver, cardiovascular diseases, Alzheimer’s disease, and cancer. Growing evidence suggests that majority of intracellular pathogens, be they viral, bacterial, or protozoan, rely on host LDs for completing some steps of their life cycle, thus emphasizing the importance of LDs in host-pathogen interactions. Host and pathogen lipids play vital role in the ability of the pathogen to evade host immune system. Therefore, droplet homeostasis and pathogen replication are intricately linked, the mechanisms of which are largely unknown. This chapter summarizes our current understanding of how unique aspect of LD biology is exploited by pathogens for their replication and propagation in the host. Advancement in the field of lipidomics for performing lipid-profiling of host-pathogen interactions will shed light on many novel and unanticipated findings in disease pathogenesis aimed at discovery of novel biomarkers and identification of therapeutic interventions.

Fat storage-inducing transmembrane (FIT or FITM) proteins have been implicated in the partitioning of triacylglycerol to lipid droplets and the budding of lipid droplets from the ER. At the molecular level, the sole relevant interaction is that FITMs directly bind to triacyglycerol and diacylglycerol, but how they function at the molecular level is not known. Saccharomyces cerevisiae has two FITM homologues: Scs3p and Yft2p. Scs3p was initially identified because deletion leads to inositol auxotrophy, with an unusual sensitivity to addition of choline. This strongly suggests a role for Scs3p in phospholipid biosynthesis. Looking at the FITM family as widely as possible, we found that FITMs are widespread throughout eukaryotes, indicating presence in the last eukaryotic common ancestor. Protein alignments also showed that FITM sequences contain the active site of lipid phosphatase/phosphotransferase (LPT) enzymes. This large family transfers phosphate-containing headgroups either between lipids or in exchange for water. We confirmed the prediction that FITMs are related to LPTs by showing that single amino-acid substitutions in the presumptive catalytic site prevented their ability to rescue growth of the mutants on low inositol/high choline media when over-expressed. The substitutions also prevented rescue of other phenotypes associated with loss of FITM in yeast, including mistargeting of Opi1p, defective ER morphology, and aberrant lipid droplet budding. These results suggest that Scs3p, Yft2p and FITMs in general are LPT enzymes involved in an as yet unknown critical step in phospholipid metabolism.

Liu et al. show that ER–Golgi tethering increases during ER stress in yeast. The protein Nvj2p is required for this tethering, which promotes nonvesicular ceramide transport from the ER to the Golgi to alleviate ceramide toxicity., Ceramides are key intermediates in sphingolipid biosynthesis and potent signaling molecules. However, excess ceramide is toxic, causing growth arrest and apoptosis. In this study, we identify a novel mechanism by which cells prevent the toxic accumulation of ceramides; they facilitate nonvesicular ceramide transfer from the endoplasmic reticulum (ER) to the Golgi complex, where ceramides are converted to complex sphingolipids. We find that the yeast protein Nvj2p promotes the nonvesicular transfer of ceramides from the ER to the Golgi complex. The protein is a tether that generates close contacts between these compartments and may directly transport ceramide. Nvj2p normally resides at contacts between the ER and other organelles, but during ER stress, it relocalizes to and increases ER–Golgi contacts. ER–Golgi contacts fail to form during ER stress in cells lacking Nvj2p. Our findings demonstrate that cells regulate ER–Golgi contacts in response to stress and reveal that nonvesicular ceramide transfer out of the ER prevents the buildup of toxic amounts of ceramides.

Joshi et al. show that Pex30p and Pex31p contain reticulon-like ER tubulating domains. Like reticulons, they localize to the edges of ER sheets and tubules but are only present in subdomains. These subdomains are devoid of reticulons and are the sites of pre-peroxisome vesicle biogenesis., Saccharomyces cerevisiae contains three conserved reticulon and reticulon-like proteins that help maintain ER structure by stabilizing high membrane curvature in ER tubules and the edges of ER sheets. A mutant lacking all three proteins has dramatically altered ER morphology. We found that ER shape is restored in this mutant when Pex30p or its homologue Pex31p is overexpressed. Pex30p can tubulate membranes both in cells and when reconstituted into proteoliposomes, indicating that Pex30p is a novel ER-shaping protein. In contrast to the reticulons, Pex30p is low abundance, and we found that it localizes to subdomains in the ER. We show that these ER subdomains are the sites where most preperoxisomal vesicles (PPVs) are generated. In addition, overproduction or deletion of Pex30p or Pex31p alters the size, shape, and number of PPVs. Our findings suggest that Pex30p and Pex31p help shape and generate regions of the ER where PPV biogenesis occurs.

Nascent lipid droplet (LD) formation occurs in the endoplasmic reticulum (ER) membrane but it is not known how sites of biogenesis are determined. We previously identified ER domains in S. cerevisiae containing the reticulon homology domain (RHD) protein Pex30 that are regions where preperoxisomal vesicles (PPVs) form. Here, we show that Pex30 domains are also sites where most nascent LDs form. Mature LDs usually remain associated with Pex30 subdomains, and the same Pex30 subdomain can simultaneously associate with a LD and a PPV or peroxisome. We find that in higher eukaryotes multiple C2 domain containing transmembrane protein (MCTP2) is similar to Pex30: it contains an RHD and resides in ER domains where most nascent LD biogenesis occurs and that often associate with peroxisomes. Together, these findings indicate that most LDs and PPVs form and remain associated with conserved ER subdomains, and suggest a link between LD and peroxisome biogenesis., Lipid droplets (LDs) and peroxisomes are both generated by budding off the endoplasmic reticulum (ER). Here, the authors show that the yeast protein Pex30 marks ER subdomains where both LD and peroxisome biogenesis occurs, and identify MCTP2 as the putative mammalian Pex30 ortholog.

Background: Liver abscess is a constant source of mortality and morbidity in general and tropical countries. India has second highest incidence of Liver abscess. This study were planned to identify the clinical profile of liver abscess in terms of demographic profile, clinical presentation, etiology, evaluation of different modalities of investigations, effectiveness of various treatment modalities and to study influence of alcohol and diabetes. Methods: This study was carried out in a tertiary care centre in Northern India between the period of January 2013 to June 2014. An attempt was made in this study to define the various symptoms complex, treatment options and complications occurring in cases of liver abscess. Results: In our study highest incidence occurred in 31-60 years age group (71%). Males were more commonly affected (92%). Abdominal pain (98%) was the most common symptom. Alcohol was the single most common etiological factor in 84% of cases of Liver abscess. Laboratory investigations were leucocytosis (72%), anaemia (23%), Raised B. Urea (21%), RBS >200mgm% (29%), raised alkaline phosphatase (83%), hypoalbuminemia (84%), PT >20sec (29%), raised SGOT (52%), SGPT (45%). Aspirated pus culture from liver abscess was sterile in 59.45% cases suggestive of amoebic etiology. Ultrasonography revaled solitary abscess in 78%. Isolated Right Lobe abscess was seen in 74%.72% of patients were subjected to percutaneous aspiration in abscess >100 ml or >3 cm; Recurrences were noted in 8% of cases in conservative management group. Conclusions: Amoebic abscess is much more common than pyogenic liver abscess. Lack of proper sanitary condition, DM and addiction to alcohol are the most important predisposing condition for liver abscess. Ultrasound is simple, inexpensive and quick to perform, with a diagnostic accuracy of 90%.

Recent studies have reported intrinsic metabolic reprogramming in Pkd1 knock-out cells, implicating dysregulated cellular metabolism in the pathogenesis of polycystic kidney disease. However, the exact nature of the metabolic changes and their underlying cause remains controversial. We show herein that Pkd1 k o /ko renal epithelial cells have impaired fatty acid utilization, abnormal mitochondrial morphology and function, and that mitochondria in kidneys of ADPKD patients have morphological alterations. We further show that a C-terminal cleavage product of polycystin-1 (CTT) translocates to the mitochondria matrix and that expression of CTT in Pkd1 ko/ko cells rescues some of the mitochondrial phenotypes. Using Drosophila to model in vivo effects, we find that transgenic expression of mouse CTT results in decreased viability and exercise endurance but increased CO2 production, consistent with altered mitochondrial function. Our results suggest that PC1 may play a direct role in regulating mitochondrial function and cellular metabolism and provide a framework to understand how impaired mitochondrial function could be linked to the regulation of tubular diameter in both physiological and pathological conditions.

Visualization of nascent lipid droplets reveals that they form lens-like structures inside the ER membrane bilayer and that FIT proteins are necessary for lipid droplet protrusion toward the cytoplasm., Lipid droplets (LDs) are found in all cells and play critical roles in lipid metabolism. De novo LD biogenesis occurs in the endoplasmic reticulum (ER) but is not well understood. We imaged early stages of LD biogenesis using electron microscopy and found that nascent LDs form lens-like structures that are in the ER membrane, raising the question of how these nascent LDs bud from the ER as they grow. We found that a conserved family of proteins, fat storage-inducing transmembrane (FIT) proteins, is required for proper budding of LDs from the ER. Elimination or reduction of FIT proteins in yeast and higher eukaryotes causes LDs to remain in the ER membrane. Deletion of the single FIT protein in Caenorhabditis elegans is lethal, suggesting that LD budding is an essential process in this organism. Our findings indicated that FIT proteins are necessary to promote budding of nascent LDs from the ER.

Nascent lipid droplet (LD) formation occurs in the ER membrane1-4. It is not known whether LD biogenesis occurs stochastically in the ER or at subdomains with unique protein and lipid composition. We previously identified ER subdomains in S. cerevisiae that contain Pex30, a reticulon-like ER-resident membrane protein5. There are ~25 Pex30-containing puncta in the ER per cell. These sites are regions where preperoxisomal vesicles (PPVs) are generated5. Here we show that Pex30 subdomains are also the location where most nascent LDs form. Mature LDs remain associated with Pex30 subdomains and the same Pex30 subdomain can simultaneously associate with a LD and a PPV. Pex30 subdomains become highly enriched in diacylglycerol (DAG) during LD biogenesis, indicating they have a unique lipid composition. We find that in higher eukaryotes multiple C2 domain containing transmembrane protein (MCTP2) is the functional homologue of Pex30; MCTP2 resides in ER subdomains where most nascent LD biogenesis occurs and that are often associated with peroxisomes. Together, these findings indicate that most LDs and PPVs form and remain associated with conserved ER subdomains and suggest a link between LD and peroxisome biogenesis.

Lipid droplets (LDs) store fats and play critical roles in lipid and energy homeostasis. They form between the leaflets of the endoplasmic reticulum (ER) membrane and consist of a neutral lipid core wrapped in a phospholipid monolayer with proteins. Two types of ER-LD architecture are thought to exist and be essential for LD functioning. Maturing LDs either emerge from the ER into the cytoplasm, remaining attached to the ER by a narrow membrane neck, or stay embedded in the ER and are surrounded by ER membrane. Here, we identify a lipid-based mechanism that controls which of these two architectures is favored. Theoretical modeling indicated that the intrinsic molecular curvatures of ER phospholipids can determine whether LDs remain embedded in or emerge from the ER; lipids with negative intrinsic curvature such as diacylglycerol (DAG) and phosphatidylethanolamine favor LD embedding, while those with positive intrinsic curvature, like lysolipids, support LD emergence. This prediction was verified by altering the lipid composition of the ER in S. cerevisiae using mutants and the addition of exogenous lipids. We found that fat-storage-inducing transmembrane protein 2 (FIT2) homologs become enriched at sites of LD generation when biogenesis is induced. DAG accumulates at sites of LD biogenesis, and FIT2 proteins may promote LD emergence from the ER by reducing DAG levels at these sites. Altogether, our findings suggest that cells regulate LD integration in the ER by modulating ER lipid composition, particularly at sites of LD biogenesis and that FIT2 proteins may play a central role in this process.

Summary Pathogenesis-related proteins played a pioneering role fifty years ago in the discovery of plant innate immunity as a set of proteins that accumulated upon pathogen challenge. The most abundant of these proteins, PATHOGENESIS-RELATED 1 (PR-1) encodes a small antimicrobial protein that has become, as a marker of plant immune signaling, one of the most referred to plant proteins. However, the biochemical activity and mode of action of PR-1 proteins has remained elusive. Here, we provide genetic and biochemical evidence for the capacity of PR-1 proteins to bind sterols and demonstrate that the inhibitory effect on pathogen growth is caused by sterol sequestration from pathogens. In support of our findings, sterol-auxotroph pathogens such as the oomycete Phytophthora are particularly sensitive to PR-1 whereas sterol-prototroph fungal pathogens become highly sensitive only when sterol-biosynthesis is compromised. Our results are in line with previous findings showing that plants with enhanced PR-1 expression are particularly well protected against oomycete pathogens. This article is protected by copyright. All rights reserved.

Proteins belonging to the CAP superfamily are present in all kingdoms of life and have been implicated in different physiological processes. Their molecular mode of action, however, is poorly understood. Saccharomyces cerevisiae expresses three members of this superfamily, pathogen-related yeast (Pry)1, -2, and -3. We have recently shown that Pry function is required for the secretion of cholesteryl acetate and that Pry proteins bind cholesterol and cholesteryl acetate, suggesting that CAP superfamily members may generally act to bind sterols or related small hydrophobic compounds. Here, we analyzed the mode of sterol binding by Pry1. Computational modeling indicates that ligand binding could occur through displacement of a relatively poorly conserved flexible loop, which in some CAP family members displays homology to the caveolin-binding motif. Point mutations within this motif abrogated export of cholesteryl acetate but did not affect binding of cholesterol. Mutations of residues located outside the caveolin-binding motif, or mutations in highly conserved putative catalytic residues had no effect on export of cholesteryl acetate or on lipid binding. These results indicate that the caveolin-binding motif of Pry1, and possibly of other CAP family members, is crucial for selective lipid binding and that lipid binding may occur through displacement of the loop containing this motif.

Sterols and related membrane-perturbing agents are subject to a quality control cycle. Compounds that fail to pass this control are acetylated and secreted into the culture media, whereas lipids that pass the cycle are deacetylated and retained within the cell. Here we describe the identification of a family of conserved proteins, the Pathogen-Related Yeast (PRY) proteins, as a class of sterol-binding proteins. Saccharomyces cerevisiae has three members of this family, two of which, Pry1 and Pry2, are secreted, whereas Pry3 is a cell wall–associated protein. Cells lacking both PRY1 and PRY2 have a complete block in secretion of the acetylated lipid and Pry1 and Pry2 proteins bind free cholesterol and cholesteryl acetate in vitro. PRY proteins belong to a large protein superfamily of unknown mode of action, the CAP protein superfamily [i.e. cysteine-rich secretory proteins (CRISP), antigen 5, and pathogenesis related 1 proteins]. The conserved CAP domain of Pry1 is necessary and sufficient for lipid export and sterol binding. Expression of a human CAP superfamily member, the cysteine-rich secretory protein 2 (CRISP2), rescues the phenotype of yeast mutants lacking Pry function and purified CRISP2 binds cholesterol in vitro, indicating that lipid binding is a conserved function of the CAP superfamily proteins.

Eukaryotes store metabolic energy in form of neutral lipids, which are deposited within a dedicated organelle, termed lipid droplet (LD). While neutral lipids are synthesized by ER localized integral membrane proteins, the fate of these lipids after their synthesis and the mechanism resulting in their accumulation in LDs are not well understood. We have recently shown that LDs are functionally connected to the ER membrane allowing for a bidirectional and energy-independent transport of integral membrane proteins and possibly lipids between the two compartments during lipogenesis or lipolysis. To further characterize the nature of this connection, we investigated the topology of triacylglycerol (TAG) formation. Here we show that the active site residues of the TAG biosynthetic enzyme in yeast, Lro1, a homolog of the lecithin cholesterol acyltransferase (LCAT)-related proteins, are located within the ER luminal domain of the enzyme, suggesting that TAG formed by Lro1 is initially present in the ER luminal leaflets of the ER membrane. The topology of TAG formed by Lro1 thus contrasts that of the second TAG biosynthetic enzyme, Dga1, which has a cytosolic acyl-CoA binding domain and thus is likely to catalyze TAG formation in the cytosolic leaflet of the ER membrane. Since TAG formed by either Dga1 or Lro1 can be efficiently packed into LDs we conclude that neutral lipids from both the cytosolic as well as the luminal leaflets of the ER membrane can be concentrated and packed into LDs.

Cells store metabolic energy in the form of neutral lipids that are deposited within lipid droplets (LDs). In this study, we examine the biogenesis of LDs and the transport of integral membrane proteins from the endoplasmic reticulum (ER) to newly formed LDs. In cells that lack LDs, otherwise LD-localized membrane proteins are homogenously distributed in the ER membrane. Under these conditions, transcriptional induction of a diacylglycerol acyltransferase that catalyzes the formation of the storage lipid triacylglycerol (TAG), Lro1, is sufficient to drive LD formation. Newly formed LDs originate from the ER membrane where they become decorated by marker proteins. Induction of LDs by expression of the second TAG-synthesizing integral membrane protein, Dga1, reveals that Dga1 itself moves from the ER membrane to concentrate on LDs. Photobleaching experiments (FRAP) indicate that relocation of membrane proteins from the ER to LDs is independent of temperature and energy, and thus not mediated by classical vesicular transport routes. LD-localized membrane proteins are homogenously distributed at the perimeter of LDs, they are free to move over the LD surface and can even relocate back into the ER, indicating that they are not restricted to specialized sites on LDs. These observations indicate that LDs are functionally connected to the ER membrane and that this connection allows the efficient partitioning of membrane proteins between the two compartments.

Cells of Saccharomyces cerevisiae lacking Apq12, a nuclear envelope (NE)-endoplasmic reticulum (ER) integral membrane protein, are defective in assembly of nuclear pore complexes (NPCs), possibly because of defects in regulating membrane fluidity. We identified BRR6, which encodes an essential integral membrane protein of the NE-ER, as a dosage suppressor of apq12δ. Cells carrying the temperature-sensitive brr6-1 allele have been shown to have defects in nucleoporin localization, mRNA metabolism and nuclear transport. Electron microscopy revealed that brr6-1 cells have gross NE abnormalities and proliferation of the ER. brr6-1 cells were hypersensitive to compounds that affect membrane biophysical properties and to inhibitors of lipid biosynthetic pathways, and displayed strong genetic interactions with genes encoding non-essential lipid biosynthetic enzymes. Strikingly, brr6-1 cells accumulated, in or near the NE, elevated levels of the two classes of neutral lipids, steryl esters and triacylglycerols, and over-accumulated sterols when they were provided exogenously. Although neutral lipid synthesis is dispensable in wild-type cells, viability of brr6-1 cells was fully dependent on neutral lipid production. These data indicate that Brr6 has an essential function in regulating lipid homeostasis in the NE-ER, thereby impacting NPC formation and nucleocytoplasmic transport.

This paper proposes an ingenious photonic device which measures non-invasively hemoglobin and glucose concentration of human blood. The proposed device is cheap, portable, infection and pain free and provides real time measurement.

Most cells store neutral lipids in a dedicated compartment, the lipid droplet (LD). These LDs are structurally and functionally conserved across species. In higher eukaryotes, LDs are covered by abundant scaffolding proteins, such as the oleosins in plants and perilipins (PLINs) in animal cells. S. cerevisiae, however, has no homologues of these scaffolding proteins. To analyze a possible function of these proteins in the biogenesis of LDs, oleosin and perilipin family members (PLIN1, ADRP/PLIN2, and TIP47/PLIN3) were expressed in yeast cells and their targeting to LDs, membrane association and function in neutral lipid homeostasis and LD biogenesis were analyzed. When expressed in wild-type cells, these proteins were properly targeted to LDs. However, when expressed in cells lacking LDs, oleosin was localized to the ER bilayer and was rapidly degraded. PLINs, on the other hand, did not localize to the ER membrane in the absence of LDs and lost their membrane association. Photobleaching experiments revealed that PLIN2 and PLIN3 rapidly exchanged their LD association but PLINs did not move over the LD surface as quickly as did an integral membrane protein, such as oleosin. Interestingly, expression of these scaffolding LD proteins in mutant cells containing elevated levels of neutral lipids within the ER bilayer resulted in the formation of LDs. These results suggest that these LD scaffolding proteins promote the sequestration of neutral lipids from the ER bilayer and thereby induce LD formation. Consistent with this proposition, addition of a cell permeable diacylglycerol (DAG) was sufficient to promote LD formation in cells expressing the LD scaffolding proteins but lacking the capacity to synthesize storage lipids.

Sterols are essential lipid components of eukaryotic membranes. They are synthesized in the endoplasmatic reticulum (ER) from where they are efficiently transported to the plasma membrane, which harbors ~90% of the free sterol pool of the cell. The molecular mechanisms that govern this lipid transport, however, are not well characterized and are challenging to analyze. Saccharomyces cerevisiae offers the opportunity to circumvent some of the technical limitations associated with studying this forward transport of sterols from the ER to the plasma membrane, because the organism can also take up sterols from the environment, incorporate them into the plasma membrane and transport them back to the ER, where the free sterol is converted to steryl esters. This reverse sterol transport, however, occurs only under anaerobic conditions, where the cells become sterol auxotroph, or in mutant cells that cannot synthesize heme. The reverse sterol transport pathway, however, is more amenable to experimental studies, because arrival of the sterol in the ER membrane can be monitored unambiguously by following the formation of steryl esters. Apart from sterol acylation, we have recently described a reversible sterol acetylation cycle that is operating in the lumen of the ER. Acetylation occurs on both cholesterol and pregnenolone, a steroid precursor, and serves as a signal for export of the acetylated sterols into the culture media. The time-dependent appearance of acetylated sterols in the culture supernatant thus provides a new means to monitor the forward transport of chemically modified sterols out of the ER.

Sterols are essential lipid components of eukaryotic membranes. They are synthesized in the endoplasmatic reticulum (ER) from where they are efficiently transported to the plasma membrane, which harbors ~90% of the free sterol pool of the cell. The molecular mechanisms that govern this lipid transport, however, are not well characterized and are challenging to analyze. Saccharomyces cerevisiae offers the opportunity to circumvent some of the technical limitations associated with studying this forward transport of sterols from the ER to the plasma membrane, because the organism can also take up sterols from the environment, incorporate them into the plasma membrane and transport them back to the ER, where the free sterol is converted to steryl esters. This reverse sterol transport, however, occurs only under anaerobic conditions, where the cells become sterol auxotroph, or in mutant cells that cannot synthesize heme. The reverse sterol transport pathway, however, is more amenable to experimental studies, because arrival of the sterol in the ER membrane can be monitored unambiguously by following the formation of steryl esters. Apart from sterol acylation, we have recently described a reversible sterol acetylation cycle that is operating in the lumen of the ER. Acetylation occurs on both cholesterol and pregnenolone, a steroid precursor, and serves as a signal for export of the acetylated sterols into the culture media. The time-dependent appearance of acetylated sterols in the culture supernatant thus provides a new means to monitor the forward transport of chemically modified sterols out of the ER.

Background: The incidence of acute pancreatitis (AP) has increased during the past 20 years. Aim of this study was to review various clinical parameters like etiological factors, age, sex, clinical signs & symptoms and various biochemical criteria like serum amylase, lipase, blood sugar, and serum calcium in assessment of acute pancreatitis. Methods: We conducted prospective study on 30 patients admitted in our Hospital, who were diagnosed clinically as acute pancreatitis associated with raised serum amylase levels more than 160 somogyi units/100ml. Results: The incidence was higher in males (66.66%) as compared to females. Most common etiological association was gallstone (50%) followed by alcohol (9%), idiopathic (16.66%). Most common clinical sign was abdominal tenderness (93.33%) followed by tachycardia (66.66%), tachypnoea (53.33%), rigidity (53.33%), jaundice (40%), epigastric lump (13.33%). Most common clinical symptom was abdominal pain (100%), nausea/vomiting (90%), abdominal distension (83.33%), fever (53.33%). Increase of serum lipase may be a better diagnostic tool because it found to be normal in some non-pancreatic conditions also. Blood sugar >200mg%, serum calcium < 8 mg% are important poor prognostic indicators and show increased severity of disease and mortality risk. In our study complications encountered were pleural effusion (10%), renal failure (6.66%), ARDS (3.33%), cardiac arrhythmias (3.33%). Conclusions: Our study emphasizes that early diagnosis and assessment of severity of disease by various clinical signs, symptoms and biochemical criteria supported by radiological investigations is very important to minimize the complications of acute pancreatitis.

All eukaryotes store excess lipids in organelles known as lipid droplets (LDs), which play central roles in lipid metabolism. Understanding LD biogenesis and metabolism is critical for understanding the pathophysiology of lipid metabolic disorders like obesity and atherosclerosis. LDs are composed of a core of neutral lipids surrounded by a monolayer of phospholipids that often contains coat proteins. Nascent LDs bud from the endoplasmic reticulum (ER) but the mechanism is not known. In this commentary we discuss our recent finding that a conserved family of proteins called fat storage-inducing transmembrane (FIT) proteins is necessary for LDs budding from the ER. In cells lacking FIT proteins, LDs remain in the ER membrane. C. elegans has a single FIT protein (FITM-2), which we found is essential; almost all homozygous fitm-2 animals die as larvae and those that survive to adulthood give rise to embryos that die as L1 and L2 larvae. Homozygous fitm-2 animals have a number of abnormalities including a significant decrease in intestinal LDs and dramatic defects in muscle development. Understanding how FIT proteins mediate LD biogenesis and what roles they play in lipid metabolism and development are fascinating challenges for the future.