Cellular Mechanism of Exocrine Pancreatic Insufficiency in Diabetes Mellitus

Diabetes mellitus (DM) is associated with the compromised digestion of carbohydrates. This complication is described as exocrine pancreatic insufficiency. Whilst this causes malnutrition in patients and contributes to diabetic morbidity, the physiology and molecular biology leading to this state is not well defined. This disease-induced inability to digest foodstuffs could have many levels of regulation. Obvious candidates are ligand proteins involved in stimulating secretion, receptor defects, intracellular ion levels and post-receptor signal transduction encompassing transcription and translation. To address these points, the current studies aimed to characterise the effects of experimental type I DM upon both physiological and molecular events. The first study investigated the effects of cholecystokinin-octapeptide (CCK-8) and exogenous insulin on exocrine pancreatic amylase secretion in streptozotocin (STZ)- induced diabetic rats compared to healthy age-matched controls in vivo and in vitro. Seven - eight weeks after the induction of diabetes, animals were either anaesthetised for the study of in vivo exocrine secretion or humanely killed and pancreatic acinar cells isolated for the measurement of intracellular free calcium and magnesium concentrations ([Ca 2+ ]1 and [Mg2+ ]1), total protein, and amylase output employing the Phadebas method. For rats in both in vivo and in vitro studies, fasting blood glucose in control and diabetic rats was 73.3 ± 3.4 mg dl-1 (n = 44) and 380.0 ± 25.9 mg dl-1 (n = 27), respectively. Basal pancreatic juice flow rate in STZ-diabetic rats was significantly increased (P<0.001) whereas protein and amylase outputs were significantly decreased (P<0.001) compared to control rats. CCK-8 infusion (150 pmol kg -1 h-1 for 100 mm) resulted in marked elevations in flow rate as well as in protein and amylase secretion in control animals (P<0.05 compared with the corresponding basals). In contrast, in diabetic rats, CCK-8 evoked a small increase in flow rate, which was not significant when compared to basal. In these animals, CCK-8 stimulated the secretion of amylase and protein output, but the secretory rates were dramatically lower compared with those in control rats. Administration of insulin (1 U, I.P.) in healthy rats significantly increased pancreatic flow rate, amylase secretion, protein output and blood glucose levels in vivo compared to basal (P<0.05). Infusion of CCK-8 together with insulin (1 U) in control rats markedly potentiated pancreatic juice flow and amylase secretion. Pretreatment with atropine (0.2 mg kg-1, I.P.) abolished the effects of insulin on secretory parameters despite a similar reduction in glycaemia. In diabetic rats, insulin (4 U, I.P.) did not modify exocrine pancreatic secretion either alone or in combination with CCK-8. In vitro experiments revealed that either (ACh (10-8 – 10-4 M) or CCK-8 (10" - 10-8 M)) can evoke total amylase release which was elevated in healthy control pancreatic acinar cells compared to diabetic acinar cells. In contrast, 10-6 M insulin produced a significant increase (Pc0.05) in the amount of total amylase output in control acinar cells compared to diabetic acinar cells. Combining insulin (10-8 – 10-6 M) with either ACh or CCK-8 had little or no effect on total amylase release in both control and diabetic acinar cells. There were no significant differences among the groups in unstimulated [Ca2+]j and [Mg2+]i. However, the peak [Ca 2+ ]i induced by 10-8 M CCK-8 was depressed (P<0.05) in diabetic cells (275.3 ± 11.5 nM n = 8) compared to (359.7 ± 27.5 nM, n = 6) control cells. Similarly, CCK-8 significantly decreased (P<0.05) [Mg2 ']1 in diabetic acinar cells compared to control. On a molecular level, the gene encoding amylase was under transcriptional dysregulation. Healthy control animals had a significantly lower (P>0.05) crossing point value (8.54 ± 0.131. n = 8) compared to STZ-induced diabetic animals (17.96 ± 0.272, n = 7), respectively. On a protein level, those mediators controlling translation such as p70 S6K and 4E-BPI were present at significantly lower (P>0.05) relative concentrations, suggesting an impaired capacity for protein synthesis. Interestingly, the actual activity of these proteins as measured by phosphorylation was slightly increased. It is suggested that this is a cellular mechanism to counteract loss in transcription and/or translation of mRNA encoding these proteins. Protein ubiquitination was also elevated suggesting increased protein breakdown which could be responsible for pancreatic atrophy and net protein loss. The NFkβ protein widely implicated in tissue atrophy was actually lower in STZ-induced DM, and therefore probably does not contribute to pancreatic wasting. To conclude, the results indicate that DM-induced exocrine pancreatic insufficiency is associated with decreased levels of total protein output and amylase secretion and these changes may be in part be associated with derangements in cellular Ca2+ and Mg2+ homeostasis. Furthermore, transcription of the α-amylase gene is reduced suggesting a reduced protein level and thus capacity for stimulus-secretion coupling. Finally, there appears impaired protein translation and elevated ATP-dependent protreasome mediated protein breakdown in STZ-induced DM

Cellular mechanism of exocrine pancreatic insufficiency in diabetes mellitus

Diabetes mellitus (DM) is associated with the compromised digestion of carbohydrates. This complication is described as exocrine pancreatic insufficiency. Whilst this causes malnutrition in patients and contributes to diabetic morbidity, the physiology and molecular biology leading to this state is not well defined. This disease-induced inability to digest foodstuffs could have many levels of regulation. Obvious candidates are ligand proteins involved in stimulating secretion, receptor defects, intracellular ion levels and post-receptor signal transduction encompassing transcription and translation. To address these points, the current studies aimed to characterise the effects of experimental type I DM upon both physiological and molecular events. The first study investigated the effects of cholecystokinin-octapeptide (CCK-8) and exogenous insulin on exocrine pancreatic amylase secretion in streptozotocin (STZ)- induced diabetic rats compared to healthy age-matched controls in vivo and in vitro. Seven - eight weeks after the induction of diabetes, animals were either anaesthetised for the study of in vivo exocrine secretion or humanely killed and pancreatic acinar cells isolated for the measurement of intracellular free calcium and magnesium concentrations ([Ca 2+ ]1 and [Mg2+ ]1), total protein, and amylase output employing the Phadebas method. For rats in both in vivo and in vitro studies, fasting blood glucose in control and diabetic rats was 73.3 ± 3.4 mg dl-1 (n = 44) and 380.0 ± 25.9 mg dl-1 (n = 27), respectively. Basal pancreatic juice flow rate in STZ-diabetic rats was significantly increased (P0.05) crossing point value (8.54 ± 0.131. n = 8) compared to STZ-induced diabetic animals (17.96 ± 0.272, n = 7), respectively. On a protein level, those mediators controlling translation such as p70 S6K and 4E-BPI were present at significantly lower (P>0.05) relative concentrations, suggesting an impaired capacity for protein synthesis. Interestingly, the actual activity of these proteins as measured by phosphorylation was slightly increased. It is suggested that this is a cellular mechanism to counteract loss in transcription and/or translation of mRNA encoding these proteins. Protein ubiquitination was also elevated suggesting increased protein breakdown which could be responsible for pancreatic atrophy and net protein loss. The NFkβ protein widely implicated in tissue atrophy was actually lower in STZ-induced DM, and therefore probably does not contribute to pancreatic wasting. To conclude, the results indicate that DM-induced exocrine pancreatic insufficiency is associated with decreased levels of total protein output and amylase secretion and these changes may be in part be associated with derangements in cellular Ca2+ and Mg2+ homeostasis. Furthermore, transcription of the α-amylase gene is reduced suggesting a reduced protein level and thus capacity for stimulus-secretion coupling. Finally, there appears impaired protein translation and elevated ATP-dependent protreasome mediated protein breakdown in STZ-induced DM.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Smart economics: evaluation of Australian aid support for women’s economic empowerment

This report evaluates how effective Australia has been in achieving gender equality outcomes in economic development programs. Executive summary Promoting women’s economic empowerment is ‘smart economics’. When women are fully involved in economic development, countries become more equitable and prosperous. Worldwide, there has been steady progress for women and girls in the key sectors of health and education. However, Australia—like most donors—has had limited success in achieving gender equality outcomes in economic development programs. Last financial year, approximately 25 per cent of the Australian aid budget was invested in the economic sectors, ranging from primary industry through to the production of goods and provision of services. Australian aid support for economic development is mainly concentrated on agriculture, rural development and transport. Smaller, but still significant, amounts of aid focus on energy, trade, and business and banking. Less than a third of this economic sector investment can demonstrate an explicit focus on gender equality. This is a concern, especially given Australia’s longstanding policy of ‘mainstreaming’ gender equality in its aid program

Anterior cervico-vaginal tear along with posterior bladder wall rupture: a rare case report

A 25 year old female presented to our emergency as a case of G3P2+0 (2A and H), full term pregnancy with intrauterine foetal demise with obstructed labour with severe anaemia. In view of obstructed labour with severe anaemia suspicion of rupture uterus was raised. Abnormal contour of abdomen also raised suspicion of bladder tumour. Here emergency caesarean section was done, peroperatively she was diagnosed as a case of anterior cervico-vaginal rupture along with posterior bladder wall rupture which is a rare entity. Uterine closure was done along with anterior cervico-vaginal wall with posterior bladder wall repair. This repair was done through trans- bladder route. Unique finding of this case was tear of anterior cervico-vaginal region with associated posterior bladder wall tear without rupture of uterus despite of obstructed labour in multiparous women. Most probable cause behind this type of injury is impacted head in neglected or obstructed labour responsible for ischemia and necrosis

A comparative study of the effect of dexmedetomidine and lignocaine on hemodynamic responses and recovery following tracheal extubation in patients undergoing intracranial surgery

Background: Recovery from general anesthesia and extubation is a period of intense physiological stress for patients. The most feared complications after intracranial surgery are development of an intracranial hematoma and major cerebral edema. Both may result in cerebral hypoperfusion and brain injury. Thus, the anesthetic emergence of a neurosurgical patient should include maintenance of stable respiratory and cardiovascular parameters. Minimal reaction to the endotracheal tube removal prevents sympathetic stimulation and increases in venous pressure. In our study, we compared dexmedetomidine HCl, lignocaine HCl and placebo to blunt stress response and providing a smooth transition from extubation phase.Methods: 75 ASA Grade I and II patients aged 18-60 years scheduled for elective intracranial surgery for intracranial space occupying lesions were randomly divided into three groups of 25 each. Balanced general anesthesia was given. Inhalation anesthetic was discontinued and after return of spontaneous respiration patient in Group D received injection dexmedetomidine 0.5 µg/kg intravenous (IV), Group X received injection lignocaine 1.5 mg/kg IV and Group P received 10 ml normal saline IV over 60 sec. Heart rate (HR), mean arterial pressure (MAP), quality of extubation were measured at 1, 3, 5, 10, 15 mins interval after extubation. Emergence time and extubation time were noted and quality of extubation was evaluated on cough grading.Results: There was a significant decrease in MAPs and HR in Group D as compared to Group L and Group P (p<0.05) at all-time interval after extubation. Extubation quality score of the majority of patients was 1 in Group D, 2 in Group X, and 3 in Group P (p<0.001). The duration of emergence and extubation were comparable in all three groups. Sedation score of the most patient was 3 (44%) in Group D and 2 (56%) in Group X. Six patients in Group D and 1 patient in Group X had bradycardia.Conclusion: Single bolus dose of IV dexmedetomidine HCl 0.5 mg/kg given before tracheal extubation effectively attenuates hemodynamic response to extubation as compared to 1.5 mg/kg lignocaine HCl

Interaction of human tRNA-dihydrouridine synthase-2 with interferon-induced protein kinase PKR

PKR is an interferon (IFN)-induced protein kinase, which is involved in regulation of antiviral innate immunity, stress signaling, cell proliferation and programmed cell death. Although a low amount of PKR is expressed ubiquitously in all cell types in the absence of IFNs, PKR expression is induced at transcriptional level by IFN. PKR's enzymatic activity is activated by its binding to one of its activators. Double-stranded (ds) RNA, protein activator PACT and heparin are the three known activators of PKR. Activation of PKR in cells leads to a general block in protein synthesis due to phosphorylation of eIF2α on serine 51 by PKR. PKR activation is regulated very tightly in mammalian cells and a prolonged activation of PKR leads to apoptosis. Thus, positive and negative regulation of PKR activation is important for cell viability and function. The studies presented here describe human dihydrouridine synthase-2 (hDUS2) as a novel regulator of PKR. We originally identified hDUS2 as a protein interacting with PACT in a yeast two-hybrid screen. Further characterization revealed that hDUS2 also interacts with PKR through its dsRNA binding/dimerization domain and inhibits its kinase activity. Our results suggest that hDUS2 may act as a novel inhibitor of PKR in cells

A study of bacteriological profile and antibiotic sensitivity of culture proven neonatal sepsis

Background: Neonatal sepsis is the second most common cause of neonatal mortality in India. Early detection and proper treatment of sepsis are important in reducing neonatal mortality. The emergence of antibiotic resistance among pathogens that infect newborns is of great concern. Hence, this study was done to identify the bacterial agents causing neonatal septicemia along with their antibiotic sensitivity pattern. Methods: This was a prospective study carried out at neonatal intensive care unit of a tertiary care teaching hospital of western Gujarat, India, from October 2018 to August 2020. 2739 neonates were admitted with probable sepsis during the study period. 299 neonates with positive blood cultures were recruited for the study. Antibiotic sensitivity of organisms was noted and compared with other studies. Results: Out of 299 blood culture proven sepsis, most common organism was Klebsiella pneumoniae, isolated in 98 patients (32.7%), followed by coagulase negative staphylococcus aureus in 90 patients (30.2%). Candida was the third most common organism isolated in 45 (15.1%) patients. Other bacteria isolated were Enterococcus in 33 (11.1%), Staphylococcus aureus in 17 (5.6%), Escherichia coli in 7 (2.3%), Acinetobacter spp in 6 (2%) and Pseudomonas in 3 (1%) patients. Gram positive bacteria were isolated in 140 (46.8%) patients, while, gram negative bacteria and fungus were isolated in 114 (38.2%) and 45 (15%) patients respectively. Klebsiella demonstrated maximum sensitivity to Meropenem (95%) and Piperacillin-Tazobactam (73.1%), while it showed high resistance to ampicillin (97.9%) and Cefoperazone (95.9%). Among non-beta lactam antibiotics, Klebsiella showed maximum sensitivity to colistin (100%) and Vancomycin (80%), while showed high resistance to Aminoglycosides and Quinolones. CONS showed maximum sensitivity to Cefoperazone (81.4%) and Cefotaxim (62.4%), but they showed high resistance to Ampicillin (86.5%) and Meropenem (86.9%). Among non-beta lactam antibiotics, CONS showed maximum sensitivity to Linezolid (100%) and Vancomycin (98%), while showed high resistance to Aminoglycosides and Quinolones. Conclusions: The most common organism isolated was Klebsiella and it showed high resistance to 1st, 2nd and 3rd line antibiotics (Ampicillin, Aminoglycosides, Cefoperazone and Quinolones). Due to the emergence of antibiotic resistance in NICU, it is important to know antibiotics sensitivity and resistant pattern of various organisms for neonatal sepsis

Consequence and Prevention of Haemodynamic Stress Response during Laryngoscopy and Endotracheal Intubation with Oral Ivabradine- A Multicentric Randomised Controlled Study

Introduction: Laryngoscopy and intubation cause lots of haemodynamic changes which adversely affects the patient during the perioperative period. Various methods have been applied to reduce stress response in high risk patients. Ivabradine is a unique cardiotonic drug which reduces the heart rate without compromising blood pressure, specially in debilitating and severely ill patients. Aim: To evaluate role of oral ivabradine in attenuating the haemodynamic stress response to laryngoscopy, intubation and extubation in patients undergoing surgical procedure under General Anaesthesia (GA) and to note the side-effects and its complications, if any. Materials and Methods: A randomised controlled multicentric study was conducted in 200 American Society of Anaesthesiologists’ (ASA)-I and II patients undergoing various surgery under general anaesthesia. The patients were randomly divided into two groups: group A (Test group, n=100) received 5 mg oral ivabradine one hour before intubation, group B (Control group, n=100) received placebo. The pulse rate, Systolic Blood Pressure (SBP) and Diastolic Blood Pressure (DBP) and Mean Arterial Pressure (MAP) were recorded at intubation and 10 minutes postintubation along with at extubation and postintubation period till 10 minutes. Patients were monitored for haemodynamic changes as per the protocol. Statistics analysis was done using Statistical Package of Social Science (SPSS) software version 21.0. Results: Demographic findings were comparable in both groups. Heart rate (84.36±4.11 versus 114.19±12.4), SBP (120±10.5 versus 150±17.5), DBP (76.08±4.29 versus 113.2±10.6), MAP (91.3±6.7 versus 124.4±12.8) at 10 minutes postintubation decreased more in test group as compared to control group from baseline (p-value <0.005). Similarly, heart rate (84.13±2.06 versus 110.58±8.92), SBP (123.4±10.06 versus 150.8±13.1), DBP (84.08±2.02 versus 107±10.2), MAP (97.8±6.47 versus 122.06±9.7) at 10 minutes postintubation decreased significantly in test group as compared to control group from baseline (p-value <0.005). Conclusion: Oral ivabradine is a very useful cardiotonic drug which facilitates the fluctuation in heart rate during laryngoscopy and endotracheal intubation