Comparison of recovery times by using bispectral index monitoring versus end-tidal agent concentration monitoring in patients undergoing inhalational general anaesthesia.

Background and Aims: End-tidal anaesthetic gas concentration (ETAG) and bispectral index (BIS) are both used to monitor depth of anaesthesia. Maintaining an accurate depth of anaesthesia helps in early post-operative recovery. This study compared the recovery times from sevoflurane-nitrous oxide anaesthesia using ETAG monitoring with BIS monitoring. Methods: Four hundred and two patients undergoing elective surgeries under sevoflurane- nitrous oxide anaesthesia were enroled in this double blinded parallel group prospective randomised trial and allocated into two groups. The depth of anaesthesia was monitored using BIS in BIS group (n = 202) and end-tidal sevoflurane concentration (EtSevo) in ETAG group (n = 200). The time to extubation and recovery were compared between the groups. Parametric, non-parametric and categorical variables were compared using Student's 't' test, Wilcoxon's rank sum test and Chi-square test, respectively. Results: Time to extubation (min) [BIS group - 10, 5; ETAG group - 10, 5 (median, inter-quartile range, IQR), P = 0.32] and time to recovery (min) [BIS group - 14, 6; ETAG group - 13.5, 7 (median, IQR), P = 0.34] did not differ significantly between the two groups. The EtSevo concentration (vol%) was significantly higher in the BIS group at 5 min [BIS group - 1.2, 0.4; ETAG group - 1.0, 0.4 (median, IQR), P < 0.001], 30 min [BIS group - 1.1, 0.4; ETAG group - 1.0, 0.3 (median, IQR), P = 0.002] and 120 min [BIS group - 1.11 ± 0.28; ETAG group - 0.96 ± 0.27 (mean ± standard deviation), P = 0.014] after induction of anaesthesia. Conclusions: BIS and ETAG monitoring are associated with comparable recovery profiles. ETAG monitoring is associated with significantly less sevoflurane consumption.

RalA and PLD1 promote lipid droplet growth in response to nutrient withdrawal.

Lipid droplets (LDs) are dynamic organelles that undergo dynamic changes in response to changing cellular conditions. During nutrient depletion, LD numbers increase to protect cells against toxic fatty acids generated through autophagy and provide fuel for beta-oxidation. However, the precise mechanisms through which these changes are regulated have remained unclear. Here, we show that the small GTPase RalA acts downstream of autophagy to directly facilitate LD growth during nutrient depletion. Mechanistically, RalA performs this function through phospholipase D1 (PLD1), an enzyme that converts phosphatidylcholine (PC) to phosphatidic acid (PA) and that is recruited to lysosomes during nutrient stress in a RalA-dependent fashion. RalA inhibition prevents recruitment of the LD-associated protein perilipin 3, which is required for LD growth. Our data support a model in which RalA recruits PLD1 to lysosomes during nutrient deprivation to promote the localized production of PA and the recruitment of perilipin 3 to expanding LDs.

One vector-based method to verify predicted plant miRNAs, target sequences, and function modes.

Many microRNAs (miRNAs) have been predicted from small RNA sequencing data, but little was experimentally verified due to the lack of effective methods. Here, we developed a simple and reliable dual gene expression cassette vector-based method to verify predicted plant miRNAs. We cloned osa-miR528 as a known miRNA, hvu-miRX as a predicted miRNA and TaDREB3 open reading frame as a non-miRNA into the first gene expression cassette and fused their complementary or noncomplementary sequences as predicted target or nontarget sequences with the 3' untranslated region of green fluorescent protein (GFP) in the second one. When these constructs were bombarded into plant cells, only the construct containing both osa-miR528 or hvu-miRX and its complementary sequence did not generate green fluorescence. Stem-loop reverse-transcription polymerase chain reaction detected mature osa-miR528 or mature hvu-miRX in the cells, while northern analysis showed that GFP messenger RNA from the construct containing both osa-miR528 or hvu-miRX and its complementary sequence was degraded. Taken together, the results indicate that hvu-miRX is an authentic miRNA like osa-miR528, miRNA's complementary sequence is its target sequence, and both osa-miR528 and hvu-miRX silenced the GFP expression via a cleavage mode. Our method thus facilitates the verification of predicted plant miRNAs, target sequences, and function modes.

Drp1 Promotes KRas-Driven Metabolic Changes to Drive Pancreatic Tumor Growth.

Mitochondria undergo fission and fusion to maintain homeostasis, and tumors exhibit the dysregulation of mitochondrial dynamics. We recently demonstrated that ectopic HRasG12V promotes mitochondrial fragmentation and tumor growth through Erk phosphorylation of the mitochondrial fission GTPase Dynamin-related protein 1 (Drp1). However, the role of Drp1 in the setting of endogenous oncogenic KRas remains unknown. Here, we show that Drp1 is required for KRas-driven anchorage-independent growth in fibroblasts and patient-derived pancreatic cancer cell lines, and it promotes glycolytic flux, in part through the regulation of hexokinase 2 (HK2). Furthermore, Drp1 deletion imparts a significant survival advantage in a model of KRas-driven pancreatic cancer, and tumors exhibit a strong selective pressure against complete Drp1 deletion. Rare tumors that arise in the absence of Drp1 have restored glycolysis but exhibit defective mitochondrial metabolism. This work demonstrates that Drp1 plays dual roles in KRas-driven tumor growth: supporting both glycolysis and mitochondrial function through independent mechanisms.

Integration of mental health into primary healthcare: perceptions of stakeholders in Pakistan.

BACKGROUND: In Pakistan, there is high prevalence of mental health disorders, but mental health services to address these are not well developed. To provide effective mental health services, the World Health Organization emphasizes the integration of mental health into primary health care (PHC). OBJECTIVES: This study aimed to assess the views of key stakeholders about integration of mental health into PHC in Karachi, Pakistan. METHODS: A qualitative, exploratory study was conducted between June and September 2013 among 15 decision-making (from the Department of Health) and implementation-level stakeholders (mental health and public health professionals and primary care staff) from both the public and private sectors. Face-to-face, in-depth interviews were conducted using a semi-structured interview guide. Data were collected until theoretical saturation was achieved and conventional content analysis was carried out. RESULTS: Although there was general support among all the stakeholders for integration of mental health services within PHC, there were also a number of reservations. First was the perceived lack of support within the system in terms of resource allocation and acceptance from the community. Second was the lack of human resources in the field of mental health. In addition, resistance at the PHC level is likely as staff are already burdened with other preventive care services. CONCLUSIONS: The study suggests that strong political commitment, adequate human and financial resources, and strong advocacy are needed for the integration of mental health into PHC in Pakistan.

Identification and characterisation of a previously unknown drought tolerance-associated microRNA in barley.

Drought is the most serious abiotic stress, and causes crop losses on a worldwide scale. The present study identified a previously unknown microRNA (designated as hvu-miRX) of 21 nucleotides (nt) in length in barley. Its precursor (designated pre-miRX) and primary transcript (designated pri-miRX) were also identified, with lengths of 73 and 559 nt, respectively. The identified upstream sequence of pri-miRX contained both the TATA box and the CAAT box, which are both required for initiation of transcription. Transient promoter activation assays showed that the core promoter region of pri-miRX ranged 500 nt from the transcription start site. In transgenic barley overexpression of the wheat DREB3 transcription factor (TaDREB3) caused hvu-miRX to be highly expressed as compared with the same miRNA in non-transgenic barley. However, the high expression was not directly associated with TaDREB3. Genomic analysis revealed that the hvu-miRX gene was a single copy located on the short arm of chromosome 2 and appeared to be only conserved in Triticeae, but not in other plant species. Notably, transgenic barley that overexpressed hvu-miRX showed drought tolerance. Degradome library analysis and other tests showed that hvu-miRX targeted various genes including transcription factors via the cleavage mode. Our data provides an excellent opportunity to develop drought stress tolerant cereals using hvu-miRX.

Integration of mental health into primary healthcare: perceptions of stakeholders in Pakistan

Background: In Pakistan, there is high prevalence of mental health disorders, but mental health services to address these are not well developed. To provide effective mental health services, the World Health Organization emphasizes the integration of mental health into primary health care [PHC]. Objectives: This study aimed to assess the views of key stakeholders about integration of mental health into PHC in Karachi, Pakistan. Methods: A qualitative, exploratory study was conducted between June and September 2013 among 15 decision-making [from the Department of Health] and implementation-level stakeholders [mental health and public health professionals and primary care staff] from both the public and private sectors. Face-to-face, in-depth interviews were conducted using a semi-structured interview guide. Data were collected until theoretical saturation was achieved and conventional content analysis was carried out. Results: Although there was general support among all the stakeholders for integration of mental health services within PHC, there were also a number of reservations. First was the perceived lack of support within the system in terms of resource allocation and acceptance from the community. Second was the lack of human resources in the field of mental health. In addition, resistance at the PHC level is likely as staff are already burdened with other preventive care services. Conclusions: The study suggests that strong political commitment, adequate human and financial resources, and strong advocacy are needed for the integration of mental health into PHC in Pakistan

Contexte : Au Pakistan, la prévalence des troubles de santé mentale est élevée, mais les services de santé mentale visant à prendre en charge ces troubles ne sont pas bien développés. Afin de fournir des services de santé mentale efficaces, l'Organisation mondiale de la Santé met l'accent sur l'intégration de la santé mentale dans les soins de santé primaires. Objectifs : La présente étude visait à évaluer les points de vue des parties prenantes clés au sujet de l'intégration de la santé mentale dans les soins de santé primaires à Karachi [Pakistan]. Méthodes : Une étude exploratoire qualitative a été conduite entre juin et septembre 2013 parmi 15 parties prenantes chargées de la prise de décisions [relevant du Département de la Santé] et de la mise en œuvre [professionnels de la santé mentale et de la santé publique et équipes de soins primaires] issues des secteurs public et privé. Des entretiens approfondis en face-à-face ont été menés en recourant à un guide d'entretiens semi-structurés. Les données ont été recueillies jusqu'à saturation théorique, et une analyse de contenu conventionnelle a été réalisée. Résultats : Bien que les parties prenantes soutiennent dans leur ensemble l'intégration de la santé mentale aux soins de santé primaires, certaines d'entre elles ont émis des réserves. La première concernait le manque de soutien perçu au sein de ce système en ce qui concerne l'allocation des ressources et l'acceptation par la communauté. Ensuite venait le manque de ressources humaines dans le domaine de la santé mentale. En outre, il est probable de rencontrer une résistance au niveau des soins de santé primaires, les équipes étant déjà surchargées par d'autres services de soins préventifs. Conclusions : Cette étude suggère qu'un engagement politique fort, des ressources humaines et financières suffisantes et un plaidoyer énergique sont requis en vue de l'intégration de la santé mentale aux soins de santé primaires au Pakistan

الخلفية: تنتشر اضطرابات الصحة النفسية على نطاق واسع في باكستان، إلا أن خدمات الصحة النفسية المقدمة للتصدي لتلك الاضطرابات ليستمتطورة بشكل كاف. ولتوفير خدمات صحة نفسية فعالة، تشدد منظمة الصحة العالمية على دمج الصحة النفسية في الرعاية الصحية الأولية.الأهداف: هدفت هذه الدراسة إلى تقييم آراء الأطراف المعنية الرئيسية بشأن دمج الصحة النفسية في الرعاية الصحية الأولية في كراتشي بباكستان.طرق البحث: أُجريت دراسة كيفية استكشافية في الفترة ما بين يونيو/حزيران إلى سبتمبر/أيلول 2013 شملت 15 طرفاً من الأطراف المعنية علىمستوى صنع القرار )من إدارة الصحة( والتنفيذ )المختصين في مجال الصحة النفسية والصحة العمومية وموظفي الرعاية الأولية( من القطاعين العاموالخاص على حدٍ سواء. وأُجريت مقابلات مباشرة ومعمقة باستخدام دليل شبه منظم لإجراء المقابلات. وجُعت بيانات إلى أن تم بلوغ مستوى التشبع النظري وأجري تحليل تقليدي للمحتوى.النتائج: برغم إبداء جميع الأطراف المعنية بوجه عام دعمها لدمج خدمات الصحة النفسية في الرعاية الصحية الأولية، فقد أُبدي عدد من التحفظات.وتمثل التحفظ الأول في نقص الدعم داخل المنظومة من حيث تخصيص الموارد والقبول المجتمعي لذلك. أما التحفظ الثاني فتمثل في نقص المواردالبشرية في مجال الصحة النفسية. وفضلاً على ذلك، من المرجح حدوث مقاومة على مستوى الرعاية الصحية الأولية حيث يعاني العاملون فيها منثقل العبء الملقى على عاتقهم بالفعل من أثر تقديم خدمات الرعاية الوقائية الأخرى.الاستنتاجات: تشير الدراسة إلى الحاجة إلى وجود التزام سياسي قوي وموارد بشرية ومالية كافية وأنشطة مناصرة قوية لدمج الصحة النفسية في الرعاية الصحية الأولية في باكستان

Deletion of the Rab GAP Tbc1d1 modifies glucose, lipid, and energy homeostasis in mice.

Tbc1d1 is a Rab GTPase-activating protein (GAP) implicated in regulating intracellular retention and cell surface localization of the glucose transporter GLUT4 and thus glucose uptake in a phosphorylation-dependent manner. Tbc1d1 is most abundant in skeletal muscle but is expressed at varying levels among different skeletal muscles. Previous studies with male Tbc1d1-deficient (Tbc1d1(-/-)) mice on standard and high-fat diets established a role for Tbc1d1 in glucose, lipid, and energy homeostasis. Here we describe similar, but also additional abnormalities in male and female Tbc1d1(-/-) mice. We corroborate that Tbc1d1 loss leads to skeletal muscle-specific and skeletal muscle type-dependent abnormalities in GLUT4 expression and glucose uptake in female and male mice. Using subcellular fractionation, we show that Tbc1d1 controls basal intracellular GLUT4 retention in large skeletal muscles. However, cell surface labeling of extensor digitorum longus muscle indicates that Tbc1d1 does not regulate basal GLUT4 cell surface exposure as previously suggested. Consistent with earlier observations, female and male Tbc1d1(-/-) mice demonstrate increased energy expenditure and skeletal muscle fatty acid oxidation. Interestingly, we observe sex-dependent differences in in vivo phenotypes. Female, but not male, Tbc1d1(-/-) mice have decreased body weight and impaired glucose and insulin tolerance, but only male Tbc1d1(-/-) mice show increased lipid clearance after oil gavage. We surmise that similar changes at the tissue level cause differences in whole-body metabolism between male and female Tbc1d1(-/-) mice and between male Tbc1d1(-/-) mice in different studies due to variations in body composition and nutrient handling.

Role of microRNAs in plant drought tolerance.

Drought is a normal and recurring climate feature in most parts of the world and plays a major role in limiting crop productivity. However, plants have their own defence systems to cope with adverse climatic conditions. One of these defence mechanisms is the reprogramming of gene expression by microRNAs (miRNAs). miRNAs are small noncoding RNAs of approximately 22 nucleotides length, which have emerged as important regulators of genes at post-transcriptional levels in a range of organisms. Some miRNAs are functionally conserved across plant species and are regulated by drought stress. These properties suggest that miRNA-based genetic modifications have the potential to enhance drought tolerance in cereal crops. This review summarizes the current understanding of the regulatory mechanisms of plant miRNAs, involvement of plant miRNAs in drought stress responses in barley (Hordeum vulgare L.), wheat (Triticum spp.) and other plant species, and the involvement of miRNAs in plant-adaptive mechanisms under drought stress. Potential strategies and directions for future miRNA research and the utilization of miRNAs in the improvement of cereal crops for drought tolerance are also discussed.

Giant chylolymphatic mesenteric cyst and its successful enucleation: A case report.

INTRODUCTION: Cysts of the mesentery are among surgical rarities. The clinical presentation is not characteristic and in addition, the preoperative imaging although suggestive is not diagnostic in this case ultrasound and CTscan was consistent with giant mesenteric cyst. In most cases, the diagnosis is confirmed after surgical exploration. PRESENTATION OF CASE: A 42 yrs old male patient on exploratory laparotomy had a 14cm×10cm×10 cm cysts which was seen arising from the mesentery ofdistal jejunum 80cm from the duodeno jejunal flexure. The cyst was enucleated successfully from themesentery without entailing resection. The cyst contained milky white fluid consistent with a chylolymphatic cyst. The diagnosis was confirmed on histopathology which revealed a cyst wall with lymphoidaggregates. After 3 years of follow-up, the patient is doing well and there is no evidence of recurrence. DISCUSSION: The cysts may be asymptomatic or maymanifest with abdominal pain, distension lump or intestinal obstruction. Our patient was symptomatic with mild and long standing abdominal pain. The definitive diagnosis of these lesions is difficult prior to surgical exploration as there are no pathognomonic symptoms or characteristic imaging findings. CONCLUSION: Cysts of the mesentery are among surgical rarities. In most of the cases the diagnosis is confirmed after surgical exploration and removal of thecyst. We would like to emphasize the importance of successful enucleation of the cyst irrespective of its size due to its independent blood supply as opposed to enterogenous cyst which requires bowel resection and anastomosis.

Ulnar artery ischaemia following corticosteroid injection for carpal tunnel syndrome.

A 36-year-old female with known bilateral carpal tunnel syndrome was admitted to hospital and given a steroid injection on the ulnar side in her right hand. She suffered immediate ischaemia of the 3rd, 4th, and 5th fingers. Imaging showed decreased flow in the 4th and 5th phalageal arteries. Treatment with iloprost infusion commenced 7 days after the injury, with moderate improvement and further managed with a carpal tunnel release. This case report sheds light on an unusual yet very important complication of carpal tunnel management.

Transcriptomes of the desiccation-tolerant resurrection plant Craterostigma plantagineum.

Studies of the resurrection plant Craterostigma plantagineum have revealed some of the mechanisms which these desiccation-tolerant plants use to survive environments with extreme dehydration and restricted seasonal water. Most resurrection plants are polyploid with large genomes, which has hindered efforts to obtain whole genome sequences and perform mutational analysis. However, the application of deep sequencing technologies to transcriptomics now permits large-scale analyses of gene expression patterns despite the lack of a reference genome. Here we use pyro-sequencing to characterize the transcriptomes of C. plantagineum leaves at four stages of dehydration and rehydration. This reveals that genes involved in several pathways, such as those required for vitamin K and thiamin biosynthesis, are tightly regulated at the level of gene expression. Our analysis also provides a comprehensive picture of the array of cellular responses controlled by gene expression that allow resurrection plants to survive desiccation.

Anatomy of the coracoacromial veil.

The coracoacromial (CA) ligament plays an important role in the stability of the shoulder joint by limiting superior translation of the glenohumeral joint. This ligament is further divided into anterolateral and posteromedial bands. Attached to the CA ligament, a supportive structure was noted in some previous studies. The purpose of this study was to learn more about the anatomy of this structure. Twenty-eight shoulders were obtained. Deltoid and trapezius muscles were removed without damaging the rotator cuff and coracoacromial arch. The CA ligament was dissected further to reveal two constituent bands, an anterolateral and a posteromedial band. A connective tissue structure was noted between the posteromedial band, CA ligament, and rotator interval capsule. This structure was oriented as an L-shaped curtain, which the authors termed the "coracoacromial veil." Anatomical position of this veil provides a stabilizing link between the CA ligament and the rotator interval capsule. This structure potentially limits inferior translation of the glenohumeral joint.

Gene expression profiling in women with breast cancer in a Saudi population.

OBJECTIVE: To generate consensus gene expression profiles of invasive breast tumors from a small cohort of Saudi females, and to explore the possibility that they may be broadly conserved between Caucasian and Middle Eastern populations. METHODS: This study was performed at King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia, from January 2005 to January 2007. Gene expression profiles were generated from 38 invasive breast tumors, and 8 tumor adjacent tissues TATs using BD Atlas cDNA expression arrays containing 1176 genes. Results were confirmed by reverse transcriptase polymerase chain reaction, and analyzed by 2-dimensional unsupervised hierarchical clustering. RESULTS: The analysis identified 48 differentially expressed genes in tumors from which 25 are already reported by various western studies. Forty-three of these genes were also differentially expressed in TATs. The same data set has been able to distinguish between tumors and the TATs, interestingly by using only 4 of the differentially expressed genes. Moreover, we were able to group the patients according to prognosis to an extent by hierarchical clustering. CONCLUSION: Our results indicate that expression profiles between Saudi females with breast cancer and the Caucasian population are conserved to some extent, and can be used to classify patients according to prognostic groups. We also suggest 3 differentially-expressed genes IGHG3, CDK6, and RPS9 in tumors may have a novel role in breast cancer. In addition, the role of TATs is much more essential in breast cancer, and needs to be explored thoroughly.

Gene expression profiles of the fibroblasts from breast tumors and normal tissue compared with the tumor expression profiles.

OBJECTIVE: The study was designed to examine whether the gene expression profiles of fibroblast cell lines, established from the tumor and the normal tissue from the same breast, exhibit any similarities with the profiles of the original tissues. METHODS: Fibroblast cell lines were established from invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS) of the breast and the adjacent normal tissues. Isolated total RNA from the cell lines and tissues were used to prepare labeled cDNA which was hybridized to Becton Dickinson Atlas microarrays for obtaining profiles of expressed genes. The profiles of tumors and cell lines were compared. This study was carried out at King Faisal specialist Hospital and Research Center, Riyadh, Kingdom of Saudi Arabia, during 2004 and 2005. RESULTS: Alterations of expression of most of the genes in the tissues were not detectable in the cell lines. The expression of a lower number of genes was altered in DCIS compared with that in IDC tumors. CONCLUSION: Although the fibroblasts discharge important functions, their gene expression profiles do not represent the breast tissue to the extent that any prognostic decisions could be made.

Quick gene expression profiling of promyelocytic cell line HL-60.

OBJECTIVE: A number of techniques have been developed to perform gene expression profiling. We report preliminary results from our exploratory study, using sequential analysis of gene expression (SAGE) technique, to profile the undifferentiated and differentiated HL-60 cells in line with our interest to characterize the cancer phenotype. The aim of the study is to evaluate the technique and to understand the molecular bases of these 2 states of cells. METHODS: HL-60 cells were differentiated after treatment with dimethyl sulfoxide. Tag libraries were prepared from the messenger RNAs of the undifferentiated and differentiated cells according to the SAGE protocol. The search for genes corresponding to the tags was carried out using SAGE software. The tags and the genes from the 2 libraries were compared for their levels of expression. The study was carried out at the King Faisal Specialist Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia during the year 2001. RESULTS: A comparison of tags from the 2 libraries revealed that 151 tags corresponding to 57 genes expressed differentially: 60 tags were elevated and 59 were repressed in the undifferentiated cells. Thirty-two tags were equally expressed in both types of cells. Of the corresponding genes, 25 were expressed at higher, 17 at lower, while 15 were expressed at comparable levels in both cell types. In the profile of undifferentiated cells, the genes involved in mitochondrial function and protein synthesis were prominent, while in the differentiated cells, the genes coding for proteins associated with cell membranes, signal transduction and for cell specific functions were prominent. The genes, expressed equally in both the cell types, were concerned with the maintenance of the living state. CONCLUSION: Sequential analysis of gene expression is a useful technique for gene expression profiling. As previously indicated by others, a dedicated team can generate useful data within reasonable time limits.