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Objective: To compare antiplatelet drugs as compared to antiplatelet drugs plus antithrombotic drugs in patients of antiphospholipid syndrome. Study Design: Randomized Control Trial. Setting: Bahawal Victoria Hospital Bahwalpur. Period: October 2018 to October 2019. Material & Methods: Two hundred ten were included in the study.105 patients were treated with loprin and 105 patients were treated with loprin and low molecular weight heparin (clexane). Pregnancy outcome were studied in both groups in terms of live fetal birth. Results: The mean age in group-A was 26.24. In group-B the mean age of patients was 26.50 years. 145 patients delivered full term babies while 65 patients delivered premature babies or presented with abortion.63 patients in group-A and 82 patients in group-B delivered full term babies while 42 patients in group-A and 23 patients in group-B presented with abortion or delivered premature babies with significant p values of 0.005. Conclusion: Use of loprin and clexane 40 mg subcutaneously daily in patient with recurrent pregnancy loss due to antiphospholipid syndrome resulted in high live birth rates compared to patients who took only loprin.

Background: Defining hypertension in pregnancy, it is taken as systolic blood pressure of 140 or more and diastolic blood pressure of 90mg or more. Presence of proteinurea differentiates chronic hypertension and gestational hypertension from pre eclampsia .Prevalence and incidence of hypertension in pregnancy along with its complications like end-organ damage, preeclampsia, eclampsia, and postpartum eclampsia are increasing. Gestational hypertension and its sequels are major causes of fetal and maternal morbidity and mortality and affect almost 8% of pregnancies. Labetalol and methyldopa were effective in reducing systolic and diastolic BP significantly but labetalol was found to control BP more rapidly and efficiently. Safety profile of both drugs was similar. The rationale of this study is Labetalol is more effective in comparison with Methyldopa in pregnancy induced hypertensive females in terms of control of blood pressure within 48hours of drug intake, in pregnant patients between POG of 28 to 32 weeks. Design: Randomized control trial. Place and Duration of Study: Department of Gynaecology Unit 2 Bahawal Victoria Hospital, Bahawalpur from 1st January 2022 to 31st December 2022. Methodology: One hundred 100 patients having gestational hypertension between periods of gestation from 28to 32 weeks were enrolled. They were divided into 2 groups; group A treated with labetalol and B treated with methyldopa respectively. Results: Statistical no significant difference in systolic and diastolic blood pressure was observed before starting treatment in both groups but a significant fall in diastolic blood pressure was observed in group A after 48 hrs of initiation of treatment i.e., with labetalol (P = 0.007) Conclusion: Both labetalol and methyldopa reduce blood pressure in women with gestational hypertension but labetalol is more effective than methyldopa as proved by our study. Keywords: Gestational hypertension, Diastolic blood pressure, Labetalol, Methyldopa

Aim: The aim of this study was to compare the efficacy of nasolabial flap, split thickness skin graft and collagen sheet in increasing postoperative mouth opening as a reconstructive material in surgical management of Oral Submucous Fibrosis (OSMF). Materials and Methods: 15 patients were included in this prospective cohort study and were randomly divided into 3 groups of 5 patients, based on type of graft material used for reconstruction. Group 1 patients received nasolabial flap, Group 2 patients received split thickness skin grafts and Group 3 patients received collagen sheet after resection of fibrous bands. Histologically proven cases of OSMF with mouth opening less than 25 mm were included in the study. Patients were evaluated at regular intervals and mouth opening was documented preoperatively, intraoperatively, and at 1 and 3 months of follow-up. Results: In all three groups, mouth opening differed substantially at all periods of follow-up from preoperative values. At 1 month follow up mean mouth opening increased to 28.4 mm in Group 1, 26.2 mm in Group 2 and 25.0 mm in Group 3. At the end of 3 months of follow up mean increase in mouth opening was greater in Group 1 (29.6 mm) in comparison to Group 2 (20.2 mm) and Group 3 (19.2 mm). Conclusion: No statistically significant difference in mouth opening was observed in three groups. Nasolabial flaps, split thickness skin grafts and collagen sheet are good options for reconstruction of defects created by resection of fibrous bands in patients with OSMF.

Objective: The objective of this study was to assess average improvement in maximal mouth opening, pain at rest, and active mouth opening after platelet-rich plasma (PRP) injections using ultrasonography (USG)-guided platelets rich plasma injection in the management of internal derangement of the joint. Materials and Methods: Ten patients with internal derangement of the temporomandibular joint (TMJ) underwent PRP injections guided by the ultrasonography. The extent of maximal mouth opening, pain, and tenderness of the TMJ and the masticatory muscles at rest, motion, and mastication were thoroughly assessed at the beginning of the study and scheduled for next follow-up at 2, 4, 12, and 24 weeks. Results: USG-guided platelet-rich plasma injection was significantly effective in improvements of the extent of maximal mouth opening, pain at rest, and on movement. Statistics result demonstrated a significant reduction in the visual analog scale values of pain at rest and on movement. Conclusion: USG-guided PRP injection to the upper TMJ space provided improvement in signs and symptoms of patient with internal derangement of the TMJ.

A pilot study was conducted to analyze and compare the stability of dental implants using 2 different graft materials (autologous dentin graft and an alloplastic hydroxyapatite [HA] crystal) in the same patient. As far as we know, this study is first of its kind where socket preservation has been done using 2 different grafts in the same patient. Ten patients undergoing extraction of at least 2 teeth were selected. Atraumatic extractions were performed. Out of the 2 extraction sockets, 1 was grafted with autogenous tooth graft and other with HA crystals. Computed tomography scan was performed 3 months after grafting to evaluate bone mineral density. Primary implant stability was measured using resonance frequency analysis. It was observed that among both sites, autogenous tooth grafted sites showed better results.

INTRODUCTION: The treatment of temporomandibular joint is mainly surgical and aims at restoring the function of joint and correcting the aesthetics deformity caused by ankylosis. Studies are available in the literature which showed that there is osteogenic potential in condylar stump remaining after gap arthroplasty, especially in children. Our objective was to assess the growth of ramus operated after surgery using orthopantomogram as the tool of assessment. MATERIALS AND METHOD: Fifteen patients of prospective study design were selected for study. This study was undertaken to evaluate the mandibular ramal growth in TMJ ankylosis patients treated without growth center transplantation using orthopantomogram as a tool of assessment. RESULTS: Mean ramal length of the operated right side was found to be 42.02 mm in immediate postoperative, 43.18 mm in sixth month postoperative and 44.57 mm in 1 year postoperative follow-up. Mean ramal length of the operated left side was found to be 40.15 mm in immediate postoperative, 42.10 mm in sixth month postoperative and 44.80 mm in 1 year postoperative follow-up. CONCLUSION: Our investigations showed that mandibular growth rate increases after successful treatment of ankylosis in children. One drawback of this treatment protocol that we observed was that the total growth deficit manifested in all our patients could not be made up.

In the conventional method of fixation of symphyseal fractures, 2 miniplates are fixed in the interforamina region of the mandible at the lateral cortex. However, with the better understanding of tension and compression areas defined according to the place of energy insertion, it is now possible to fix miniplates at the lower border of the mandible. In this study we assumed that the application of perpendicular miniplates, in contrast to the placement of the plates on the lateral surface of the mandibular cortex, will constrict the motion in horizontal as well as vertical planes leading to better functional outcomes, which can be evaluated by measuring bite forces. The results obtained were affirmative with the hypothesis.

Summary Background Dietary salts sodium (Na+), potassium (K+), magnesium (Mg2+), and calcium (Ca2+) are important in metabolic diseases. Yet, we do not have sufficient understanding on the salts global molecular network in these diseases. In this systematic review we have pooled information to identify the general effect of salts on obesity, insulin resistance and hypertension. Aims To assess the roles of salts in metabolic disorders by focusing on their individual effect and the network effect among these salts. Methods We searched articles in PubMed, EMBASE and Google Scholar. We selected original laboratory research, systematic reviews, clinical trials, observational studies and epidemiological data that focused on dietary salts and followed the preferred reporting items for systematic review in designing the present systematic review. Results From the initial search of 2898 studies we selected a total of 199 articles that met our inclusion criteria and data extraction. Alterations in metabolic pathways associated with the sensitivity of sodium, potassium, magnesium and calcium may lead to obesity, hypertension, and insulin resistance. We found that the results of most laboratory research, animal studies and clinical trials are coherent but some research outcome are either inconsistent or inconclusive. Conclusion Important of salts in metabolic disorder is evident. In order to assess the effects of dietary salts in metablic diseases, environmental factors, dietary habits, physical activity, and the microbiome, should be considered in any study. Although interest in this area of research continues to grow, the challenge is to integrate the action of these salts in metabolic syndrom.

ADSD (Agile Distributed Software Development) is a growing trend for software development organizations to develop quality software with limited cost and time. However, it gives rise to additional situational challenges. Situational variations result in unstable agile architecture which gets highly affected. Although the present literature focuses on agile architecture but effect of situational variation on the agile architecture still needs consideration. One possible solution is development of conceptual model and incorporation of multiple situational factors. This research study aims to identify the most relevant situations and propose situational ADSD approach for the development of situation-based software architecture framework for agile distributed projects. The study focuses on agile methods, rendering identified situational variations. The approach is developed through analyzing data from literature and their associated work documents. Comprehensive survey helps to bridge the gaps from conceptual to architecture model. The experimental results are acquired through both practical and statistical analysis. The results support Conway’s law, e.g. correlation that maps architecture with the communication and coordination needs. Correlation results show that architecture has major contribution in ADSD and validate the relationship among conceptual model and architecture model. The results also suggest that stable architecture in ADSD can positively affect the product.

Management of temporomandibular joint (TMJ) ankylosis is mainly through surgical intervention. Interpositional materials are a necessity when it comes to prevention of TMJ re-ankylosis after arthroplasty. Early aggressive postoperative physiotherapy is essential for the prevention or treatment of TMJ hypomobility or ankyloses. Recently, it has been shown that abdominal dermis fat helps promote smooth, pain-free joint function and it is stable after interposition and less prone to fragmentation. The purpose of this study was to assess that whether dermal fat is a good choice of interpositional material when it comes to decreased pain perception during aggressive physiotherapy after release of ankyloses thus ensuring good compliance by the patient. We also assessed the fate of the graft material on computed tomography to evaluate any volume changes if occurred after interposition.

Aims Hypoxia-induced adipokine release has been attributed mainly to HIF-1α. Here we investigate the role of intracellular calcium and NF-kB in the hypoxia-dependent release of leptin, VEGF, IL-6 and the hypoxia-induced inhibition of adiponectin release in human adipocytes. Main methods We used intracellular calcium imaging to compare calcium status in preadipocytes and in adipocytes. We subjected both cell types to hypoxic conditions and measured the release of adipokines induced by hypoxia in the presence and absence of HIF-1α inhibitor YC-1, NF-κB inhibitor SN50 and intracellular calcium chelator BAPTA-AM. Key findings We demonstrate reduced intracellular calcium oscillations and increased oxidative stress as the cells transitioned from preadipocytes to adipocytes. We show that differentiation of preadipocytes to adipocytes is associated with distinct morphological changes in the mitochondria. We also show that hypoxia-induced secretion of leptin, VEGF, IL-6 and hypoxia-induced inhibition of adiponectin secretion are independent of HIF-1α expression. The hypoxia-induced leptin, VEGF and IL-6 release are [Ca++]i dependent whereas adiponectin is NF-kB dependent. Significance Our work suggests a major role for [Ca++]i in preadipocyte differentiation to adipocytes and that changes in mitochondrial morphology in the adipocytes might underlie the reduced calcium oscillations observed in the adipocytes. It also demonstrates that multiple signaling pathways are associated with the hypoxia-induced adipokine secretion.

Collaborative effort is required in DSD (Distributed Software Development) in order to develop software in manageable components. Immense effort is taking place due to the cultural, geographical and temporal distances among project teams in Pakistani Software Industry to this end. The purpose of this study is to propose a solution during DSD that will help the distributed teams in coordinating their activities. Our solution enhances the management associated activities and assists in optimal usage of team coordination in dispersed settings. The method that we used is single case study type, which involved distributed software business to assess the usefulness and efficiency related to Pakistan’s DSD industries, and has shown strong tendency of participants towards the effectiveness of proposed team coordination framework. Results indicate that the predictable solution will look up the group management, which is considered as a vigorous issue of Pakistani DSD industry. Furthermore, it will assist in resolving the problems of team coordination in distributed situation. One of the most significant current discussion about the proposed solution suggested that it helps in human communication methods team coordination in a distributed environment.

PURPOSE: The purpose of this study was to suggest a protocol for the management of odontogenic space infections in patients with hemophilia which is often incompletely addressed by clinicians because of the fear of excessive bleeding leading to prolonged morbidity. METHODS AND RESULTS: Six diagnosed cases of hemophilia (four cases of hemophilia A and two cases of hemophilia B) with odontogenic fascial space infection were included in the study. Apart from routine investigations, assessments were also done to identify inhibitors to factor VIII/IX. Factor VIII/IX was replaced along with infusion of factor eight inhibitor bypass activity (FEIBA) one hour before incision and drainage. All the cases were treated by incision and drainage along with removal of foci under local anesthesia. The postoperative outcome was uneventful in all cases except one in which postoperative bleeding was noted from the extraction site on the third day which was again managed following the designed protocol. CONCLUSION: Maintaining good oral hygiene and seeking early dental care are of prime importance in patients with hemophilia to avoid invasive procedures. However, if odontogenic infection develops in these patients, performing timely incision and drainage using our protocol will minimize the complications and give successful outcome.

We isolated and characterized pistachio hull polysaccharides (PHP). The PHP was a heteropolysaccharide mainly contained 75.50% (w/w) total sugar and 9.51% (w/w) uronic acid. As determined by GPC analysis, the polysaccharide with a molecular weight of 3.71×106 D (83.2%) was the most dominant fraction. Moreover, HPLC analysis indicated that PHP was predominantly composed of xylose, glucose, arabinose, and fructose with a molar ratio of 1.00:2.50:19.67:28.81. FT-IR and NMR analysis also confirmed the results obtained by HPLC and characterized preliminary structure features of the PHP. Functional properties of the PHP including water holding capacity (WHC: 2.44±0.05g water/g DM), and oil holding capacity (OHC: 11.53±0.04g oil/g DM) were significant compared to inulin used as reference prebiotic (p

We describe a case of maxillofacial trauma in a paediatric patient in which the left mandibular condyle was superolaterally displaced into the temporal fossa, medial to the zygomatic arch which was intact and the right condyle was also displaced similarly but the zygomatic arch was also fractured on this side. Also, there was an associated mandibular symphyseal fracture. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12663-017-1042-7) contains supplementary material, which is available to authorized users.

Cesarean section is the most commonly performed abdominal operations on women worldwide. Its rate has increased markedly in recent years in most developed countries. Due to insufficient available data regarding the best suture materials for cesarean skin closure, we conducted a study “to compare the efficacy of 2 suture materials non absorbable prolene 2-0 and absorbable suture vicryl (2-0), brand Ethicon as subcuticular skin stitches in women undergoing elective cesarean section. Study Design: Prospective, observational study. Setting: Department of Obstetrics and Gynecology, Bahawal Victoria Hospital Bahawalpur. Period: 6 months. Materials and Methods: A total of 200 women fulfilling inclusion and exclusion criteria as above were enrolled for the present study. They were divided into 2 groups of 100 each by systematic sampling technique. Results: There is statistically significant difference related to post-operative wound complication parameters between non absorbable vs absorbable suture material on day 8, 15, 30 and 45. More number of patients having absorbable suture had pain, tenderness swelling and indurations. Maximum number of participants, that is, 86 (86%) and 93 (93%) from Group 1 showed excellent wound healing at day 30 and day 45 of caesarean section as compared to 63 (63%) and 66 (68%) from Group 2. Conclusion: The non-absorbable suture (prolene) was better in terms of wound healing and cosmesis as compared to absorbable suture used in our study (vicryl).

AIM AND OBJECTIVE: To perform the comparative finite element analysis of conventional and perpendicular plating in mandibular symphyseal fracture. MATERIAL AND METHOD: Two FE model of mandible were developed and symphyseal fracture was created in both of them. Each fractured model was fixed by conventional method 2.0 mm system and perpendicular method 2.00 mm system. Stresses which developed in plates and mandible after application of load were observed in the model. RESULTS: Results of the study indicated that perpendicular method was more resistant to displacing forces and rest of the parameters was within the permitted limits. CONCLUSION: Perpendicular method was reasonably effective as conventional method of fixation for mandibular symphyseal fracture.

Objectives: To Compare metformin and regular insulin in blood sugar control and neonatal outcome in patients with gestational diabetes mellitus. Material and Method:… Study Design: Randomized control trial. Place of the Study: Department of Gynecology and Obstetrics, Bahawal Victoria hospital Bahawalpur. Duration of the study.1 year 1st January 2017- 31st December 2017. Sample Size: N=200, one half (group A) receiving Metformin and other half (group B) receiving regular insulin. Results: Blood sugar control depicted by fasting, 2 hr post parandial levels and HbA1c % was similar in both group.coexisting hypertensive complications were seen more in insulin group. Weight gain was also significantly more in insulin group 17 kg on an average as compared to metformin group where average weight gain was 13 kg.Convenience and satisfaction regarding their treatment,more number of patients were satisfied in metformin group. There were significant increase in the mean birth weight of the newborns, need for admission in NICU and Neonatal hypoglycemia in insulin group as compared to metformin group. Conclusion: Metformin when compared to insulin has equal efficacy in controlling maternal blood sugar levels but better neonatal outcome, patient satisfaction and convenience in treatment of gestational Diabetes Mellitus.

Hypoxia-induced adipokine release has been attributed mainly to HIF-1α. Here we investigate the role of intracellular calcium and NF-kB in the hypoxia-dependent release of leptin, VEGF, IL-6 and the hypoxia-induced inhibition of adiponectin release in human adipocytes.We used intracellular calcium imaging to compare calcium status in preadipocytes and in adipocytes. We subjected both cell types to hypoxic conditions and measured the release of adipokines induced by hypoxia in the presence and absence of HIF-1α inhibitor YC-1, NF-We demonstrate reduced intracellular calcium oscillations and increased oxidative stress as the cells transitioned from preadipocytes to adipocytes. We show that differentiation of preadipocytes to adipocytes is associated with distinct morphological changes in the mitochondria. We also show that hypoxia-induced secretion of leptin, VEGF, IL-6 and hypoxia-induced inhibition of adiponectin secretion are independent of HIF-1α expression. The hypoxia-induced leptin, VEGF and IL-6 release are [CaOur work suggests a major role for [Ca

Background Lipid metabolism dysfunction leading to excess fat deposits (obesity) may cause tumor (cancer) development. Both obesity and cancer are the epicenter of important medical issues. Lipid metabolism and cell death/proliferation are controlled by biochemical and molecular pathways involving many proteins, and organelles; alteration in these pathways leads to fat accumulation or tumor growth. Mammalian Kruppel-like factors, KLFs play key roles in both lipid metabolism and tumor development. Scope of review Substantial epidemiological and clinical studies have established strong association of obesity with a number of human cancers. However, we need more experimental verification to determine the exact role of this metabolic alteration in the context of tumor development. A clear understanding of molecules, pathways and the mechanisms involved in lipid metabolism and cell death/proliferation will have important implications in pathogenesis, and prevention of these diseases. Major conclusion The regulatory role of KLFs, in both cell death/proliferation and lipid metabolism suggests a common regulation of both processes. This provides an excellent model for delivering a precise understanding of the mechanisms linking altered expression of KLFs to obesity and tumor development. General significance Currently, mouse and rats are the models of choice for investigating disease mechanisms and pharmacological therapies but a genetic model is needed for a thorough examination of KLF function in vivo during the development of an organism. The worm Caenorhabditis elegans is an ideal model to study the connectivity between lipid metabolism and cell death/proliferation.

A retrospective review of maxillofacial trauma cases reported over a period of two years among urban Indian population with age

Background: Stromal response to cancer is usually characterized by intense lymphoplasmacytic infiltrate. However, recently, the attention has shifted to tumor-associated tissue eosinophils (TATE). Tumor-associated blood eosinophils (TABE) are rare in solid cancers; however, carcinoma of the head and neck shows its prevalence. Aim: The aim of the study was to investigate the prevalence and relationship of tissue and blood eosinophils in various grades of oral cancer. The purpose of the article is to emphasize the possible clinical and biological significance of eosinophils in patients of oral squamous cell carcinoma (OSCC) so that appropriate therapeutic strategies can be devised accordingly. Study Design: Thirty histologically confirmed cases of oral squamous cell carcinoma were divided into well, moderate and poorly differentiated carcinoma. Eosinophilic infiltration in the tissue was graded as low, moderate and massive TATE. The number of eosinophils per 100 WBCs was taken as the differential eosinophil count. Blood eosinophilia (BE) >6% was considered to be TABE. Materials and Methods: Hematoxylin and eosin-stained tissue sections at 5 μ were evaluated. Prolonged staining in dilute 0.05% aqueous eosin demonstrated eosinophils selectively. Blood smears were stained by Leishman stain. Statistical Analysis: Student's t-test, Chi-square test, ANOVA, Newman–Keuls Multiple Comparison Test and Karl Pearson correlation coefficient® method were used. Results: The mean TATE value was highest in poorly differentiated carcinoma. TABE was seen only in a few cases and was associated mostly with poorly differentiated OSCC. Conclusion: There was a statistically significant correlation between TATE and histological grades of OSCC. Eosinophilia of the peripheral blood is an adverse sign in patients with carcinoma.

Lipid metabolism is coordinately regulated through signaling networks that integrate biochemical pathways of fat assimilation, mobilization and utilization. Excessive diversion of fat for storage is a key risk factor for many fat-related human diseases. Dietary lipids are absorbed from the intestines and transported to various organs and tissues to provide energy and maintain lipid homeostasis. In humans, disparity between triglycerides (TG) synthesis and removal, via mitochondrial β-oxidation and VLDL (very low density lipoprotein) secretion, causes excessive TG accumulation in the liver. The mutation in Caenorhabditis elegans KLF-3 leads to high TG accumulation in the worm's intestine. Our previous data suggested that klf-3 regulates lipid metabolism by promoting fatty acid β-oxidation. Depletion of cholesterol in the diet has no effect on fat deposition in klf-3 (ok1975) mutants. Addition of vitamin D in the diet, however, increases fat levels in klf-3 worms. This suggests that excess vitamin D may be lowering the rate of fatty acid β-oxidation, with the eventual increase in fat accumulation. We also demonstrate that mutation in klf-3 reduces expression of C. elegans dsc-4 and/or vit genes, the orthologs of mammalian microsomal triglyceride transfer protein and apolipoprotein B, respectively. Both microsomal triglyceride transfer protein and apolipoprotein B are essential for mammalian lipoprotein assembly and transport, and mutation in both dsc-4 (qm182) and vit-5 (ok3239) results in high fat accumulation in worm intestine. Genetic interactions between klf-3 and dsc-4, as well as vit-5 genes, suggest that klf-3 may have an important role in regulating lipid assembly and secretion.

Background Dietary salts are important factors in metabolic disorders. They are vital components of enzymes, vitamins, hormones, and signal transduction that act synergistically to regulate lipid metabolism. Our previous studies have identified that Krüppel-like factor −3 (KLF-3) is an essential regulator of lipid metabolism. However, it is not known if KLF-2 also regulates lipid metabolism and whether KLF-2 and −3 mediate the effects of dietary salts on lipid metabolism. Methods In this study, we used klf mutants [homozygous klf-2 (ok1043) V and klf-3 (ok1975) II mutants] to investigate the role of dietary salts in lipid metabolism. All gene expression was quantified by qRT-PCR. Localization of KLF-2 was analyzed by the expression of klf-2::gfp (in pPD95.75 vector) using a fluorescent microscope. Fat storage was measured by Oil Red O staining. Results Klf-2 was identified to express in the intestine during all stages of Caenorhabditis elegans development with peak expression at L3 stage. Mutation of klf-2 increased fat accumulation. Under regular growth media free of Ca2+, the expression of both klf-2 and −3 was inhibited slightly; further their expression reduced significantly in WT worms fed on 10X Ca2+ diet. When klf-3 was mutated, the expression of klf-2 increased under 10X Ca2+ diet; but when klf-2 was mutated, the expression of klf-3 was not altered under 10X Ca2+ diet. Overall, Mg2+ and K+ were less effective on the gene expression of klfs. KLF target gene Ce-C/EBP-2 showed elevated expression in WT and klf-3 (ok1975) worms with changed Ca2+ concentrations but not in klf-2 (ok1043) worms. However, high Ca+2 diet exhibited inhibitory effect on Ce-SREBP expression in WT worms. Conclusion Dietary Ca2+ is most effective on fat storage and klf-2 expression, wherein high Ca2+ diet decreased klf-2 expression and reduced fat buildup. Mechanistic study identified Ce-C/EBP (C48E7.3; lpd-2) and Ce-SREBP (Y47D3B.7; lpd-1) as the target genes of klf-2 and/or klf-3 to mediate lipid metabolism. This study identifies a new function of klf-2 in inhibiting fat buildup and reveals the interplay between dietary salts and klf-2 and klf-3 in lipid metabolism.

Coordinated regulation of fat storage and utilization is essential for energy homeostasis, and its disruption is associated with metabolic syndrome and atherosclerosis in humans. Across species, Krüppel-like transcription factors (KLFs) have been identified as key components of adipogenesis. In humans, KLF14 acts as a master transregulator of adipose gene expression in type 2 diabetes and cis-acting expression quantitative trait locus associated with high-density lipoprotein cholesterol. Herein we report that, in Caenorhabditis elegans, mutants in klf-3 accumulate large fat droplets rich in neutral lipids in the intestine; this lipid accumulation is associated with an increase in triglyceride levels. The klf-3 mutants show normal pharyngeal pumping; however, they are sterile or semisterile. We explored important genetic interactions of klf-3 with the genes encoding enzymes involved in fatty acid (FA) β-oxidation in mitochondria or peroxisomes and FA synthesis in the cytosol, namely acyl-CoA synthetase (acs-1 and acs-2), acyl-CoA oxidase (F08A8.1 and F08A8.2), and stearoyl-CoA desaturase (fat-7). We show that mutations or RNA interference in these genes increases fat deposits in the intestine of acs-1, acs-2, F08A8.1, and F08A8 animals. We further show that acs-1 and F08A8.1 influence larval development and fertility, respectively. Thus, KLF3 may regulate FA utilization in the intestine and reproductive tissue. We demonstrate that depletion of F08A8.1 activity, but not of acs-1, acs-2, F08A8.2, or fat-7 activity, enhances the fat phenotype of the klf-3 mutant. Taken together, these results suggest that klf-3 regulates lipid metabolism, along with acs-1, acs-2, F08A8.1, and F08A8.2, by promoting FA β-oxidation and, in parallel, may contribute to normal reproductive behavior and fecundity in C. elegans.

In vertebrates, adipose tissue stores energy in the form of fat. Fat storage is tightly controlled by and dynamically balanced with energy expenditure under physiological settings; the perturbation of fat in either excess (obese) or deficit (lipodystrophy) has devastating pathologic consequences in the fueling of homeostasis and organismal fitness. The process by which fat storage is coordinated through positive and negative feedback signals is still poorly understood. To address potential mechanisms underlying fat storage we study a Caenorhabditis elegans Krüppel-like transcription factor, Ce-klf-3 and demonstrate that klf-3 is a hitherto unrecognized key regulator of fat metabolism in C. elegans. The Ce-klf-3 is highly expressed during larval development and predominantly present in intestine: the site of fat digestion, absorption, storage, and utilization. We found a strong positive correlation between klf-3 expression and fat deposition in a worm's intestine. Significantly, a klf-3 (ok1975) loss-of-function mutation, characterized by the deletion of a 1658-bp sequence spanning the 3' end of exon 2 through to the 5' end of exon 3 of klf-3, enhanced fat deposition in the intestine and caused severe defects in worm reproduction. Although klf-3 mutants seemed very similar to wild type worms in appearance and life span, 70% of mutants became semi-sterile, each producing 40-50 viable progenies, and the remaining 30% were rendered completely sterile toward adulthood. Notably, both mutant types displayed extensive deposition of fat in the intestine. Our study also demonstrates that klf-3 is critical for maintaining normal fatty acid composition by regulating genes involved in a fatty acid desaturation pathway. Strikingly, klf-3 mutant animals with impaired fatty acid beta-oxidation pathway genes resulted in fat accumulation in the mutant worm. We present the first clear in vivo evidence supporting essential regulatory roles of KLF-3 in fat storage in C. elegans and shed light on the human equivalent in disease-gene association.

In mammals, adipose tissue stores energy in the form of fat. The ability to regulate fat storage is essential for the growth, development and reproduction of most animals, thus any abnormalities caused by excess fat accumulation can result in pathological conditions which are linked to several interrelated diseases, such as cardiovascular diseases, dia- betes, and obesity. In recent years significant effort has been applied to understand basic mechanism of fat accumulation in mammalian system. Work in mouse has shown that the family of Kruppel-like factors (KLFs), a conserved and important class of transcrip- tion factors, regulates adipocyte differentiation in mammals. However, how fat storage is coordinated in response to positive and negative feedback signals is still poorly under- stood. To address mechanisms underlying fat storage we have studied two Caenorhabdi- tis elegans KLFs and demonstrate that both worm klfs are key regulators of fat metabo- lism in C. elegans. These results provide the first in vivo evidence supporting essential regulatory roles for KLFs in fat metabolism in C. elegans and shed light on the human counterpart in disease-gene association. This finding allows us to pursue a more com- prehensive approach to understand fat biology and provides an opportunity to learn about the cascade of events that regulate KLF activation, repression and interaction with other factors in exerting its biological function at an organismal level. In this review, we provide an overview of the most current information on the key regulatory components in fat biology, synthesize the diverse literature, pose new questions, and propose a new model organism for understanding fat biology using KLFs as the central theme.

We describe what is to our knowledge the first report of a patient who presented with a superolateral dislocation of the mandible, in which the medial portion of the left condyle was fractured and the lateral fragment was hooked on the zygomatic arch without any other associated fracture.

We describe the successful use of RNA interference (RNAi) to investigate gene function in the human filarial parasite Onchocerca volvulus third-stage larvae (L3). We targeted two specific gene products, the O. volvulus cathepsin L ( Ov -CPL) and cathepsin Z-like ( Ov -CPZ) cysteine proteases, which were proposed to function during O . volvulus L3 molting. We show that fluorescent-labeled Cy™3-dsRNA corresponding to cpl or cpz regions encoding the mature enzymes can enter the larvae. The molting rate of larvae treated overnight with 0.5 mg ml −1 cpl was reduced by 92% and 86% in comparison to normal control worms. It appeared that although the larvae started the molting process the last stage of molting, ecdysis was inhibited. The effect was gene specific, as larvae that did not molt in the presence of cpl or cpz dsRNA expressed the other cysteine protease, CPZ and CPL, respectively. This was confirmed by immunoelectron microscopy using antibodies directed against each enzyme. Our present study validate conclusively that both enzymes are essential for the molting of O. volvulus L3 to fourth-stage larvae. We also confirmed that the activity of the enzymes is specific to the changes that occur during the molting process on days 1–3, when the separation between the cuticles is in progress. The development of RNAi in O. volvulus L3 could further help study many of the abundant L3 and molting L3 genes identified through the filarial genome project, many of which, although have no attributed function, were identified as vaccine candidates or potential drug targets.

We have analyzed the expression and function of Cecpz-1, a Caenorhabditis elegans cathepsin Z-like cysteine protease gene, during development of the worm. The cpz-1 gene is expressed in various hypodermal cells of all developmental stages and is specifically expressed in the gonads and the pharynx of adult worms. Disruption of cpz-1 function by RNA interference or cpz-1(ok497) deletion mutant suggests that cpz-1 has a role in the molting pathways. The presence of the native CPZ-1 protein in the hypodermis/cuticle of larval and adult stages and along the length of the pharynx of adult worms, as well as the cyclic expression of the transcript during larval development, supports such function. We hypothesize that the CPZ-1 enzyme functions directly as a proteolytic enzyme degrading cuticular proteins before ecdysis and/or indirectly by processing other proteins such as proenzymes and/or other proteins that have an essential role during molting. Notably, an impressive level of the CPZ-1 native protein is present in both the new and the old cuticles during larval molting, in particular in the regions that are degraded prior to shedding and ecdysis. The similar localization of the related Onchocerca volvulus cathepsin Z protein suggests that the function of CPZ-1 during molting might be conserved in other nematodes. Based on the cpz-1 RNA interference and cpz-1 (ok497) deletion mutant phenotypes, it appears that cpz-1 have additional roles during morphogenesis. Deletion of cpz-1 coding sequence or inhibition of cpz-1 function by RNA interference also caused morphological defects in the head or tail region of larvae, improperly developed gonad in adult worms and embryonic lethality. The CPZ-1 native protein in these affected regions may have a role in the cuticular and the basement membrane extracellular matrix assembly process. The present findings have defined a critical role for cathepsin Z in nematode biology.

Cysteine proteases play critical biological roles in both intracellular and extracellular processes. We characterized Ce-cpl-1, a Caenorhabditis elegans cathepsin L-like cysteine protease. RNA interference with Ce-cpl-1 activity resulted in embryonic lethality and a transient delayed growth of larvae to egg producing adults, suggesting an essential role for cpl-1 during embryogenesis, and most likely during post-embryonic development. Cpl-1 gene (Ce-cpl-1:lacZ) is widely expressed in the intestine and hypodermal cells of transgenic worms, while the fusion protein (Ce-CPL-1::GFP) was expressed in the hypodermis, pharynx, and gonad. The CPL-1 native protein accumulates in early to late stage embryos and becomes highly concentrated in gut cells during late embryonic development. CPL-1 is also present near the periphery of the eggshell as well as in the cuticle of larval stages suggesting that it may function not only in embryogenesis but also in further development of the worm. Although the precise role of Ce-CPL-1 during embryogenesis is not yet clear it could be involved in the processing of nutrients responsible for synthesis and/or in the degradation of eggshell. Moreover, an increase in the cpl-1 mRNA is seen in the intermolt period approximately 4 h prior to each molt. During this process Ce-CPL-1 may act as a proteolytic enzyme in the processing/degradation of cuticular or other proteins. Similar localization of a related cathepsin L in the filarial nematode Onchocerca volvulus, eggshell and cuticle, suggests that some of the Ce-CPL-1 function during development may be conserved in other parasitic nematodes.

Lipid metabolic disorder is a critical risk factor for metabolic syndrome,triggering diseases like obesity and diabetes.Obesity raises the risk for many chronic illnesses including diabetes,and hea...

The free-living nematode Caenorhabditis elegans is a tractable experimental model system for the study of both vertebrate and invertebrate biology. Its most significant advantages are its simplicity, both in anatomy and in genomic organization, and the elaborate methods that have been developed to attribute function to previously uncharacterized genes. Importantly, >40% of parasitic nematode genes exhibit high levels of homology to genes within the C. elegans genome. Studying such genes using the C. elegans model should yield new insights into key molecules and their possible implications in parasite survival, leading to the discovery of new drug targets and vaccine candidates.

Submerged tooth is the one that is depressed below the occlusal plane. Dental ankylosis is thought to be a major cause of submergence. Submerged deciduous teeth have the potential to cause malocclusion not only by prevention of their exfoliation and subsequent replacement by permanent teeth but also by causing tilting of proximal teeth and extrusion of opposing tooth. The purpose of this report is to present three different cases of submerged deciduous teeth and their clinical effects.

A strain of the entomopathogenic nematode,Heterorhabditis bacteriophora,was transformed by the addition of a heat-shock protein gene (hsp70A) from the free-living nematodeCaenorhabditis elegans.Laboratory experiments with wild-type and transgenic nematodes were done at 16, 25, 30, and 37°C to investigate infectivity, reproductive capacity, and survival in water. Heat-shocked transgenic nematodes (ones subjected to a heat shock that is lethal to wild-type nematodes) were included in experiments to determine if this treatment caused sublethal damage. We found no significant differences in the ability of wild-type, transgenic, and heat-shocked-transgenic nematodes to infect or to reproduce in last-instarGalleria mellonellalarvae or to survive at these temperatures. We compared the ability of wild-type and transgenic nematodes to infect and kill 11 species of invertebrates, representing groups known to be susceptible and nonsusceptible to wild-type nematodes. Transformation caused no detectable change in virulence of nematodes to any test invertebrates. Transgenic and wild-type nematodes were also injected interperitoneally into mice and transgenic nematodes were fed by injection to mice without mouse mortality or changes in growth rate. We conclude that genetic engineering has provided a precise method to alter the heat shock tolerance ofH. bacteriophorawithout affecting other important life history characteristics and that the transgenic nematode is unlikely to pose any threat to the environment if released on a wide scale.

A heat-shock response is induced when cells are exposed to temperatures slightly higher than their optimal physiological temperature. This response is based on the synthesis of heat-shock proteins encoded by the heat-shock genes. A correlation between the increased thermotolerance and production of 70-kDa heat-shock protein (hsp70) has been observed in many organisms. We tested this hypothesis by transferring a Caenorhabditis elegans heat-inducible hsp70 A-encoding gene into the entomopathogenic nematodes Heterorhabditis bacteriophora Hp88. Suc- cessful transformation of the gene was confirmed by Southern blot hybridization and polymerase chain reaction. Our blot studies showed that the transgenic nematodes contained five to ten copies per genome of the introduced hsp70 A gene. hsp70 mRNA transcripts were detected in both wild- type and transgenic nematodes. Transcripts increased severalfold in transgenic nematodes upon heat shock. Infective juveniles of both transgenic and wild-type nematodes that exposed to a sub- lethal heat treatment (35°C) for 2 h followed by a normally lethal heat treatment (40°C) for 1 h. More than 90% of transgenic nematodes survived heat treatment, compared to 2% to 3% of the wild-type strain. Our observations establish that overexpression of hsp70 A gene resulted an en- hanced thermotolerance in the transgenic nematodes. The transgenic nematodes displayed nor- mal growth and development. J. Exp. Zool. 281:164n170, 1998. © 1998 Wiley-Liss, Inc.

1. 1. All organisms tested respond to a sudden increase of temperature by synthesizing heat-shock proteins, which helps organisms to survive high temperature. A correlation with increased thermotolerance and production of major 70 kDa protein has been observed in many organisms. 2. 2. Many studies have been designed on a large number of animal species to assess their adaptation to different thermal environments. Genetic analysis of the hsp 70 gene in entomopathogenic nematodes inhabiting different environments may provide insight into the physiological roles of hsps in these nematodes and could be useful for ecological studies. 3. 3. To assess variation among species of entomopathogenic nematodes for thermotolerance, we initiated a search for the molecular organization of heat-inducible hsp 70 genes in these nematodes. Five Heterorhabditis species/isolates with different temperature optima for survival and one warm-adapted species of Steinernema were tested 4. 4. PCR and RFLP analyses of hsp 70 in Heterorhabditis species and S. scapterisci demonstrated a putative homology with the Caenorhabditis elegans hsp 70 A gene, thus indicating evolutionary conserved nature among different nematode species. 5. 5. RFLPs with the hsp 70 A gene probe revealed different banding patterns for Heterorhabditis species and isolates. 6. 6. This is the first report on the identification of any hsp 70 gene in entomopathogenic nematodes. 7. 7. Our observation establishes a unique correlation between geographical distribution and polymorphisms for hsp 70 A gene in these nematodes.

Evidence is presented for the presence of a hatching factor or factors in corn seedling leachates that enhances egg hatching in Heterodera zeae. This study was undertaken to gain information on selected characteristics of the hatching factor(s). Fresh corn seedling leachates were heat- and storage-stable. The hatching factor partitioned into diethyl ether but not into hexane or chloroform.

We report the first successful transformation of an entomopathogenic nematode. Foreign genes were introduced in Heterorhabditis bacteriophora HP88 by microinjection using vectors carrying the Caenorhabditis elegans genes coding for the roller phenotype and 16-kDa heat shock protein (hsp16) gene. A translational fusion made by inserting lacZ in frame into hsp16 was expressed in the body musculature, hypodermis, and pharyngeal muscles. The transcription of the hsp16/lacZ transgenes resulted in the rapid synthesis of detectable levels of beta-galactosidase. This research opens new avenues for genetic modification of entomopathogenic nematodes.

The abnormalities caused by excess fat accumulation can result in pathological conditions which are linked to several interrelated diseases, such as cardiovascular disease and obesity. This set of conditions, known as metabolic syndrome, is a global pandemic of enormous medical, economic, and social concern affecting a significant portion of the world’s population. Although genetics, physiology and environmental components play a major role in the onset of disease caused by excessive fat accumulation, little is known about how or to what extent each of these factors contributes to it. The worm, Caenorhabditis elegans offers an opportunity to study disease related to metabolic disorder in a developmental system that provides anatomical and genomic simplicity relative to the vertebrate animals and is an excellent eukaryotic genetic model which enable us to answer the questions concerning fat accumulation which remain unresolved. The stored triglycerides (TG) provide the primary source of energy during periods of food deficiency. In nature, lipid stored as TGs are hydrolyzed into fatty acids which are broken down through β-oxidation to yield acetyl-CoA. Our recent study suggests that a member of C. elegans Kruppel-like factor, klf-3 regulates lipid metabolism by promoting FA β-oxidation and in parallel may contribute in normal reproduction and fecundity. Genetic and epigenetic factors that influence this pathway may have considerable impact on fat related diseases in human. Increasing number of studies suggest the role of mammalian KLFs in adipogenesis. This functional conservation should guide our further effort to explore C. elegans as a legitimate model system for studying the role of KLFs in many pathway components of lipid metabolism.

We report here the identification and characterization of STIP, a multi-domain nuclear protein that contains a G-patch, a coiled-coil, and several short tryptophan-tryptophan repeats highly conserved in metazoan species. To analyze their functional role in vivo, we cloned nematode stip-1 genes and determined the spatiotemporal pattern of Caenorhabditis elegans STIP-1 protein. RNA analyses and Western blots revealed that stip-1 mRNA was produced via trans-splicing and translated as a 95-kDa protein. Using reporter constructs, we found STIP-1 to be expressed at all developmental stages and in many tissue/cell types including worm oocyte nuclei. We found that STIP-1 is targeted to the nucleus and forms large polymers with a rod-like shape when expressed in mammalian cells. Using deletion mutants, we mapped the regions of STIP-1 involved in nuclear import and polymer assembly. We further showed that knockdown of C. elegans stip-1 by RNA interference arrested development and resulted in morphologic abnormalities around the 16-cell stage followed by 100% lethality, suggesting its essential role in worm embryogenesis. Importantly, the embryonic lethal phenotype could be faithfully rescued with Drosophila and human genes via transgenic expression. Our data provide the first direct evidence that STIP have a conserved essential nuclear function across metazoans from worms to humans.

Silicon micromachining has been used to fabricate microprobes for injecting DNA into cells. Arrays of very sharp pyramidal points are etched on a silicon substrate. Pressing these points into a culture of cells, allows biologically active material to cross the cell wall barrier. Using the microprobes, DNA has been injected into plant (tobacco leaves) and animal (nematodes) cells.