Significance of ca15-3 in carcinoma of the breast with Visceral metastases.

Background: The most common malignancy and second most common cause of death is breast cancer among women. About 2.09 million fatalities from breast cancer happened in 2018. The objective was to evaluate the elevated CA15-3 in breast cancer patients with visceral metastases presenting at the tertiary care hospital of Karachi. Methods: It was a cross-sectional study conducted at the Department of Oncology of Jinnah Postgraduate Medical Center from 15th December 2018 to 15th November 2019. Female patients aged 26-80 years diagnosed with visceral metastatic (defined as metastasis to lung, liver, brain and adrenal glands) breast cancer were included in the study. The diagnosis of breast cancer was confirmed on histopathology whereas the metastatic sites were evaluated using physical examination and imaging. The serum CA15-3 concentration was assessed using assay kits. The serum CA15-3 level of 0-32 U/ml was taken as normal range for all the patients whereas CA15-3 level greater than 32 U/L was considered as elevated CA15-3. SPSS version 23 was used to enter and analyze data. Results: A total of 139 females were included in the study. The mean age & BMI of the patients were reported as 46.5 years & 26.69 kg/m2. In the majority of the patients' metastases were detected in the liver (n=54), 92 in the lungs+ parenchymal disease, 20 in adrenal glands, 12 in pleural effusion and 10 in the brain. Out of 139 patients with visceral metastases, 52(37.4%) had normal CA15-3 level whereas 87 (62.6%) had elevated serum CA15-3 levels (>32 U/L). Conclusion: The serum CA15-3 tumour marker is elevated significantly in visceral metastases and can be used as a prognostic marker in metastatic breast cancer patients.

Effect Of Tamoxifen On Plasma Lipid Profile In Patients Of Breast Cancer.

BACKGROUND: To evaluate the effect of Tamoxifen on plasma lipid profile in breast cancer patients presenting at tertiary care hospitals. METHODS: It was a longitudinal study conducted at the Department of Oncology of Jinnah Postgraduate Medical Center from December 2018 to November 2019. Eighty-eight females aged 26-66 years diagnosed with breast cancer were included in the study using a non-probability consecutive sampling technique. Detailed gynaecological and clinical investigations and detailed history were taken. The blood samples of all the patients were collected and the plasma lipid profile was measured before initiation of Tamoxifen treatment and three- and six-months post-treatment at the clinical laboratory. The plasma lipid profile includes the measurement of Total cholesterol (mg/dl), Triglyceride(mg/dl), High-density Lipoprotein (mg/dl) & Low-density Lipoprotein (mg/dl). SPSS version 23 was used to analyse data. RESULTS: After treatment, there was a significant reduction in serum cholesterol & Low-density Lipoprotein level by 20.54 mg/dl & 16.46 mg/dl at 3 months (p<0.05), moreover there was a significant increase in Triglyceride by 22.14 at 3 months (p<0.05). No significant difference was observed in High density lipoprotein level at 3 months after using Tamoxifen. At 6 months there was a significant reduction in serum cholesterol and low-density lipoprotein by 32.29mg/dl and 24.11 mg/dl at 6 months (p<0.05), moreover there was a significant increase in Triglyceride level by 42.19 mg/dl at 6 months (p<0.05). No significant difference was observed in High-density lipoprotein level at 6 months after using Tamoxifen. CONCLUSIONS: Total cholesterol and Low-density Lipoprotein levels showed significant reduction over the period of six months from the baseline with the use of Tamoxifen. Hence Tamoxifen should be considered to have an added advantage on lipid metabolism and therefore, can reduce the risk of cardiovascular events.

Prevalence of Hypertension and its Determinants among Policemen in a City of Haryana, India.

INTRODUCTION: Noncommunicable diseases (NCDs) kill 40 million people each year, equivalent to 70% of all deaths globally. Cardiovascular diseases account for most NCD deaths or 17.7 million people annually. Police work has been regarded by some researchers as one of the most stressful occupations in the world, and coronary heart disease has been identified as a major cause of mortality in this population. MATERIALS AND METHODS: A cross-sectional study was carried out during July 2016-June 2017 among 450 policemen posted in Rohtak city of Haryana selected randomly. The investigator made two measurements of blood pressure. Data were collected using predesigned, pretested, semi-structured interview schedule, and analyzed using the Statistical Package for the Social Sciences version 20.0. RESULTS: Of 450 participants, 164 (36.4%) participants were found to be hypertensive. Age of study participants, duration of service, rank, and education are significantly associated with the prevalence of hypertension (HTN) among policemen. CONCLUSION: HTN in policemen has emerged as an important public health problem. Knowledge of risk factors for HTN may give tracks for prevention in this population. Therefore, it is the need of hour to devise a sound screening strategy to diagnose HTN among policemen and devise a comprehensive strategy for the management of HTN.

An Awake Throughout Approach for Awake Craniotomy: A Perspective from a Resource-Limited Country.

BACKGROUND: An awake throughout (AT) approach for awake craniotomy is mostly under utilized. The purpose of this study was to review the efficacy of the technique at our tertiary care center. The primary objective was to identify the incidence of perioperative complications. The secondary objective was to review the patients' satisfaction, satisfaction of surgical team, length of stay (LOS) in special care unit (SCU), and overall LOS in the hospital. METHODS: The study was a retrospective review of patients data. All patients were treated with the AT technique. This included preoperative assessment, psychologic preparation, and institution of scalp block. The incidence of perioperative complications, including satisfaction of surgical team was noted. The patients' satisfaction and the LOS in SCU and in the hospital was also recorded. RESULTS: In total, the data from 55 patients were reviewed. Their mean age was 41 years, and 63% were reported to have seizures at presentation. The AT approach was successful in 100% of cases. The incidence of intraoperative seizures was 7.4%, of vomiting was 5.4%, and of conversion to general anesthesia was 0%. The surgical team was able to perform gross total resection in 53% of patients and rated a satisfaction score of 8 out of 10. Postoperative seizures occurred in 5.4% of patients and vomiting in 3.6%. The mean LOS in SCU was 1.2 days, and the overall hospital LOS was 4 days. The patients remained fully satisfied, as evidenced by a mean satisfaction score of 8.6. CONCLUSION: An AT approach might be very useful in resource-limited setups because of the low incidence of complications, the use of resources, and significant surgeon and patient satisfaction.

Anaesthetic Management of Supratentorial Tumor Craniotomy Using Awake-Throughout Approach.

The authors are reporting an anaesthetic management of patient presenting with left parietal lobe space occupying lesion and scheduled for Awake-craniotomy. Awake-throughout approach using scalp block was planned. Among techniques reported for keeping patient awake during the surgery, this one is really underutilized. The successful conduct requires thorough preoperative assessment and psychological preparation. We used powerpoint presentation as a preoperative teaching tool. The anatomical landmark technique was used to institute scalp block, where individual nerves were targeted bilaterally. Patient remained stable throughout and participated actively in intraoperative neurological monitoring. Postoperative period showed remarkable recovery, better pain control, and shorter length of stay in hospital.

Renal Biopsy Findings in Patients with Hypothyroidism: Report of 16 cases.

INTRODUCTION: Hypothyroidism is prevalent in India. Its association with renal diseases though not very common but have been described in many studies. Here we are reporting renal biopsy findings in 16 cases, all of whom were already diagnosed cases of hypothyroidism. AIM: To study renal parenchymal diseases associated in patients with hypothyroidism. MATERIALS AND METHODS: Formalin fixed paraffin embedded sections of renal biopsy were examined after staining with H&E, PAS and Acid Fuschin Orange G (AFOG) stain. Serum urea/creatinine measurements done by semi-autoanalysers and urine analysis were done by using urine strips and light microscopy. RESULTS: In 16 cases, M:F ratio was 9:7. Duration of disease varied from 6 months to 14 years. Blood urea and serum creatinine were raised in 10 cases (62.5%) and nephrotic range proteinuria was present in 13 cases (81.25%). Two of the patients had co existing systemic lupus erythaematous. Renal pathology revealed membranous glomerulonephritis (GN) in both cases. In renal biopsy seven cases (43.75%) had pure Membranous Glomerulonephritis (MGN), 4 cases (25%) had mixture of Mesan-gial cell proliferation and membranous Glomerulonephritis(GN) also called MembranoProliferative GN (MPGN). Another four cases (25%) had Focal Segmental Glomerulosclerosis (FSGS) with chronic interstitial nephritis and one case was having minimal change disease. CONCLUSION: Thus present study concludes that hypothyroidism can cause renal parenchymal disease like membranous GN, mesangiocapillary GN which is also called as membranoproliferative GN and FSGS.

Frequency of Hyperthermia in Acute Ischemic Stroke Patients Visiting a Tertiary Care Hospital.

OBJECTIVE: To determine the frequency of hyperthermia in acute ischemic stroke patients visiting a tertiary care hospital in a developing country. STUDY DESIGN: Cross-sectional, observational study. PLACE AND DURATION OF STUDY: Medical Wards of Civil Hospital, Karachi, from January to June 2013. METHODOLOGY: Patients aged ≥18 years of either gender with acute ischemic stroke presenting within 24 hours of onset of symptoms were included. Written informed consent was obtained from all participants as well as approval of ethical review committee of the institute. Axillary temperature by mercury thermometer was monitored at the time of admission and after every 6 hours for 3 days. The data was analyzed using SPSS version 17.0 (SPSS Inc., IL, Chicago, USA). RESULTS: Atotal of 106 patients of ischemic stroke were included. The mean age of enrolled participants was 60.1 ±9.5 years. Among these, 61 (57.5%) were males and 45 (42.5%) females. Among all patients, 51.9% presented with loss of consciousness, 30.2% with slurred speech, 77.4% with limb weakness, and 9.4% with decrease vision. Atotal of 17 (16%) patients with ischemic stroke developed hyperthermia. When the prevalence of hyperthermia was stratified according to age, among patients of < 60 years of age, 26% developed hyperthermia compared to 7.1% in patients of ≥60 years of age (p=0.008). On gender stratification, among male patients, 14.8% developed hyperthermia compared to 17.8% in female patients (p=0.43). CONCLUSION: It is concluded from this study that the frequency of hyperthermia in ischemic stroke was 16% and it should be looked for as it has significant impact on the outcome. The hyperthermia was significantly more common in younger adults as compared to older adults. However, gender had no influence on the prevalence rate of hyperthermia.

Diabetes education program for people with type 2 diabetes: An international perspective.

BACKGROUND: In the Palestinian community, lifestyle changes, rapid urbanization and socioeconomic development, stress, smoking, and changes in food habits has increased the risk of non-communicable diseases especially diabetes mellitus. Diabetes complications can be prevented if the glycemic status of patients with diabetes is maintained within a nearly normal range. Therefore, patient education is critical in controlling blood glucose levels within the normal range. OBJECTIVE: This study aimed at measuring the effect of diabetes educational intervention program for patients suffering from type 2 diabetes attending the Diabetes Clinic in Tulkarim Directorate of Health. METHODS: A short duration observational study involving pre- and post-test educational intervention program was carried out on a relatively small number of type 2 diabetes patients at the Diabetes Clinic in Tulkarim Directorate of Health. In total, 215 patients attended a group-based 4h educational intervention session about diabetes. The program included explaining diabetes mellitus-symptoms, risk factors, types, treatment and complications and main aspects of self-care of the disease (foot care, eye care, and blood glucose monitoring), main aspects of dietary management, weight reduction, blood pressure, smoking cessation, periodic investigations, home monitoring and importance of physical activity. Knowledge evaluation questionnaire were evaluated pre- and post-study. Anthropometric measurements such as body weight (WT), body mass index (BMI) and laboratory tests such as fasting blood glucose (FBG), hemoglobin A1C (HbA1c), cholesterol (Chol), and triglycerides (TG) were measured both at the beginning and at the end of the study. Significance of the results was assessed by paired t-test at 95% confidence interval. RESULTS: The participant's mean age was 51.07 that ranged between 31 and 70 years. For a total of 215 participants, 41.4% were males and 58.6% were females. The mean weight before educational intervention was 80.81±14.95kg (82.6kg for males and 79.5kg for females) that decreased to 78.9±14.33kg (81.1kg for males and 77.3kg for females) after educational intervention program. The BMI also decreased significantly after educational intervention. The mean fasting blood sugar was 188.65±71.45mg/dL before educational intervention that decreased to 177.7±66.11mg/dL after the educational intervention (p=0.049). The mean glycosylated hemoglobin was 8.57±1.21 before educational intervention that decreased to 7.95±1.42 after educational intervention. The mean value of cholesterol before educational intervention was 183.27±37.74mg/dL that decreased to 169.57±34.23mg/dL after educational intervention. The mean triglycerides value decreased after educational intervention from 209.85±171.04mg/dL to 183.28±152.4mg/dL (p=0.025). The mean score of knowledge questionnaire before educational intervention was 60.6±20.65 that increased to 78.1±13.4 after conducting educational intervention. CONCLUSIONS: Diabetes education was found to be effective on BMI, FBG, HbA1c, Chol, TG, and knowledge. RECOMMENDATIONS: Diabetes education is a cornerstone in the management and care of diabetes and should be an integral part of health planning involving patient's family, diabetes care team, community, and decision makers in the education process.

The Presence of Human Chorionic Gonadotropin/Luteinizing Hormone Receptors in Pancreatic ß-Cells.

We investigated the possible presence of functional human chorionic gonadotropin (hCG)/luteinizing hormone (LH) receptors in ß-cells of pancreas, using a combination of techniques on hCG/LH receptor knockout mice, immortalized rat insulinoma cells, and human pancreatic islets. The results showed the presence of receptors and their activation resulted in a dose-dependent increase in glucose-induced release of insulin. These findings place hCG and LH among the regulators of insulin release with potential implications for insulin-level changes during the periods of altered hCG and LH secretion.

Validation of Grace Risk Score in predicting in-hospital mortality in patients with non ST-elevation myocardial infarction and unstable angina.

OBJECTIVE: To validate the global registry of acute coronary event (grace) risk score in a Pakistani population at Tabba Heart Institute Karachi in patients with non ST-Elevation Myocardial Infarction (NSTEMI) and Unstable Angina (UA). METHODS: In this prospective Observational registry study, 530 adults hospitalized patients with a diagnosis of Non-ST-Elevation Myocardial Infarction and unstable angina were enrolled between March 2012 and August 2012 at the Tabba Heart Institute, Karachi, Pakistan. For each patient, the grace risk score was calculated and its discrimination evaluated and correlated with in-hospital mortality using the Kendall's tau-b bivariate correlation test. Each patient was grouped either into high, intermediate or low risk groups according to their GRS. RESULTS: A total of 530 patients with NSTEMI and UA were included; the overall mean grace risk score in our population was 131.87 +/- 41.56. The GRACE Risk Score showed good discrimination, with Area under the ROC curve of 0.803 (95% CI 0.705-0.902, P < 0.001). During the in-hospital stay, total of 19 (3.6%) patients died, and out of those 15 (8.4%) patients belonged to high risk group. CONCLUSION: GRACE RS strongly validates the in-hospital mortality among our patient population presenting with a wide spectrum of complications. However, more multicentre registries on a larger population with long-term follow up are required to study detailed trends in our population.

Differential proteomics of Picrorhiza kurrooa Royle ex Benth. in response to dark stress.

Picrorhiza (Picrorhiza kurrooa Royle ex Benth.) an important medicinal herb of western Himalayan region has been used to treat various diseases and disorders. Over-harvesting and lack of cultivation has led to its entry in Red Data Book as an endangered species. Further, its very restrictive habitat and lesser biomass production are major limitations for bringing it under commercial cultivation. All these issues necessitate deeper insights into mechanisms governing its growth and interaction with the environmental cues. Light may be one of the important factors to be studied for its role in regulating growth and adaptation of Picrorhiza as in natural habitat it prefers shady niches. Keeping this in view, proteome of Picrorhiza kept under light vis-à-vis under dark was analysed and compared. Leaf as well as root proteome of Picrorhiza was studied. Denaturing two dimensional gel electrophoresis and mass spectrometry techniques were used to detect and identify differentially expressed proteins, respectively. Twenty two proteins from leaf and 25 proteins from root showed differential expression levels under dark and light conditions. Among the differentially expressed proteins, majority were those involved in metabolism, protein synthesis, and stress and defense response. Other differentially expressed proteins were those involved in photosynthetic process, photorespiration and few proteins were with unknown function indicating that many different processes work together to establish a new cellular homeostasis in response to dark and light conditions. Proteins found to be differentially expressed under light vis-à-vis dark conditions suggested a range of biochemical pathways and processes being associated with response of plant to dark conditions. The identified proteins may be utilized for developing strategies for improving the biomass production/performance of Picrorhiza under varied light/dark habitats.

Translation initiation factor 5A in Picrorhiza is up-regulated during leaf senescence and in response to abscisic acid.

Translation initiation, the first step of protein synthesis process is the principal regulatory step controlling translation and involves a pool of translation initiation factors. In plants, from recent studies it is becoming evident that these translation initiation factors impact various aspects of plant growth and development in addition to their role in protein synthesis. Eukaryotic translation initiation factor eIF5A is one such factor which functions in start site selection for the eIF2-GTP-tRNAi ternary complex within the ribosomal-bound preinitiation complex and also stabilizes the binding of GDP to eIF2. In the present study we have cloned and analysed a gene (eIF5a) encoding eIF5A from Picrorhiza (Picrorhiza kurrooa Royle ex Benth.) a medicinal plant of the western Himalayan region. The full length eIF5a cDNA consisted of 838 bp with an open reading frame of 480 bp, 88 bp 5' untranslated region and 270 bp 3' untranslated region. The deduced eIF5A protein contained 159 amino acids with a molecular weight of 17.359 kDa and an isoelectric point of 5.59. Secondary structure analysis revealed eIF5A having 24.53% α-helices, 8.81% ß-turns, 23.27% extended strands and 43.40% random coils. pk-eIF5a transcript was found to be expressing during the active growth phase as well as during leaf senescence stage, however, highest expression was observed during leaf senescence stage. Further, its expression was up-regulated in response to exogenous application of abscisic acid. Both high intensity as well as low intensity light decreased the expression of pk-eIF5a. The findings suggest eIF5a to be an important candidate to develop genetic engineering based strategies for delaying leaf senescence.

The impact of computerized physician order entry on medication errors and adverse drug events.

OBJECTIVES: Medication errors and adverse drug events (ADEs) are common, costly, and clinically important problems. This research was conducted to determine whether computerized physician order entry (CPOE) improves the quality of care by increasing patient safety and decreasing medication errors at the King Fahad Medical City Hospital (KFMCH) of the Kingdom of Saudi Arabia (KSA). METHODS: The study utilized a cross-sectional research design. Questionnaires were distributed to physicians in various departments who used the system for more than six months. The study was conducted in Riyadh at KFMCH, which is the largest medical complex hospital in the Middle East, in the outpatient setting. KEY FINDINGS: Ninety-three physicians participated in the study; the response rate was 31 percent. Only descriptive analyses were conducted. Results showed that 88 percent of the physicians agreed that the use of CPOE improved their performance and 76 percent reported that the use of CPOE increased their productivity. In addition, 56 percent of the participants agreed that CPOE was a simple system and 64 percent reported that it was easy to use. However, 44 percent of the physicians agreed that CPOE lacked a user guide during medication ordering and 55 percent reported that it created new types of errors. Results showed that 234 physicians always changed their order, 179 physicians changed their order often, 175 physicians rarely changed their order, and 74 physicians never changed their order. Furthermore, 72 percent of the physicians agreed that CPOE helped them to decrease ADEs. Finally, 91 percent of the physicians agreed that CPOE reduced errors related to hand-written prescriptions.

Components of antioxidant system of Picrorhiza kurrooa exhibit different spatio-temporal behavior.

Antioxidant system is one of the important factors in regulating plant growth, development and adaptation. Thus, in order to have better insights into molecular mechanisms of growth and adaptation of a plant it is prerequisite to have known the status of various components of the antioxidant system of the plant. Here we studied the status of enzymatic and non-enzymatic components of the antioxidant system of picrorhiza (Picrorhiza kurrooa). Picrorhiza is an important medicinal herb of western Himalayan region and has been listed in the Red Data Book as an endangered species. Spatio-temporal analysis of ascorbic acid and glutathione in leaf, root and rhizome during different stages of development revealed differential status of these antioxidant molecules. Of the three tissues, ascorbic acid was found to be highest in leaves and lowest in roots. Interestingly, just opposite to that, glutathione was highest in roots and lowest in leaves. Using degenerate primers based approach followed by rapid amplification of complementary DNA (cDNA) ends method, full length cDNAs of three important genes namely Picrorhiza kurrooa ascorbate peroxidase (pkapx), Picrorhiza kurrooa monodehydroascorbate reductase (pkmdhar) and Picrorhiza kurrooa glutathione reductase (pkgr) of antioxidant system were cloned from picrorhiza. Complementary DNAs of pkapx, pkmdhar and pkgr contained 1,049, 2,016 and 1,664 bp, respectively. Expression analysis showed differential spatio-temporal expression of these genes. Expressions of all the three genes were found higher in roots as compared to rhizome and leaves. Temporal expression analysis of pkapx, pkmdhar and pkgr revealed differential transcript levels. Expression of pkapx exhibited negative correlation with the light intensity. Just opposite to the pkapx, expression pattern of pkgr revealed its positive correlation with light intensity. Expression pattern of pkmdhar revealed its light independent expression behavior. The findings may be useful to assess the role of cloned genes in picrorhiza growth, adaptation and can further be utilized for transgenic development for desired trait(s).

A connectivity model for assessment of HIV transmission risk in injection drug users (IDUs).

The purpose of this study was to produce models composed of mapping of connectivity networks of HIV transmission risk in injection drug users (IDUs). This methodology provided a novel approach and diagnostic tool for understanding HIV infection transmission risk and drug use in the typical niche of IDUs, i.e., a "shooting gallery" (a gathering site for injection drug activity). Furthermore, component IDUs may have memberships in multiple "shooting galleries" revealing subsequent interconnectivities. Charting of IDU connectivity diagrams illustrated the relationships of peripheral sites to the critical central core of high HIV transmission risk. Members of this highly interlinked and infectious central core of IDUs had high HIV transmission risk and severe drug use-producing high morbidity and mortality that resulted in great public health concern. In addition, connectivity diagrams reveal very high HIV transmission risk in component IDUs in "dual memberships", i.e., membership in more than one central core (with the highest number of partners). Therefore, IDUs with "dual memberships" were the most infectious members of the "shooting gallery". In summation, network mapping of HIV transmission risk in IDUs allows for subsequent socio-behavioral analysis and the development of focused individual and programmatic interventions.

Reactive oxygen species via redox signaling to PI3K/AKT pathway contribute to the malignant growth of 4-hydroxy estradiol-transformed mammary epithelial cells.

The purpose of this study was to investigate the effects of 17-ß-estradiol (E2)-induced reactive oxygen species (ROS) on the induction of mammary tumorigenesis. We found that ROS-induced by repeated exposures to 4-hydroxy-estradiol (4-OH-E2), a predominant catechol metabolite of E2, caused transformation of normal human mammary epithelial MCF-10A cells with malignant growth in nude mice. This was evident from inhibition of estrogen-induced breast tumor formation in the xenograft model by both overexpression of catalase as well as by co-treatment with Ebselen. To understand how 4-OH-E2 induces this malignant phenotype through ROS, we investigated the effects of 4-OH-E2 on redox-sensitive signal transduction pathways. During the malignant transformation process we observed that 4-OH-E2 treatment increased AKT phosphorylation through PI3K activation. The PI3K-mediated phosphorylation of AKT in 4-OH-E2-treated cells was inhibited by ROS modifiers as well as by silencing of AKT expression. RNA interference of AKT markedly inhibited 4-OH-E2-induced in vitro tumor formation. The expression of cell cycle genes, cdc2, PRC1 and PCNA and one of transcription factors that control the expression of these genes - nuclear respiratory factor-1 (NRF-1) was significantly up-regulated during the 4-OH-E2-mediated malignant transformation process. The increased expression of these genes was inhibited by ROS modifiers as well as by silencing of AKT expression. These results indicate that 4-OH-E2-induced cell transformation may be mediated, in part, through redox-sensitive AKT signal transduction pathways by up-regulating the expression of cell cycle genes cdc2, PRC1 and PCNA, and the transcription factor - NRF-1. In summary, our study has demonstrated that: (i) 4-OH-E2 is one of the main estrogen metabolites that induce mammary tumorigenesis and (ii) ROS-mediated signaling leading to the activation of PI3K/AKT pathway plays an important role in the generation of 4-OH-E2-induced malignant phenotype of breast epithelial cells. In conclusion, ROS are important signaling molecules in the development of estrogen-induced malignant breast lesions.

Calcium oscillations and waves in cells.

From beginning of the life to final moment of the life, Ca(2+) functions as an important signaling messenger. The intracellular Ca(2+) concentration, [Ca(2+)](i), in resting cells is normally maintained at around 100 nM with a very steep ∼20,000 times concentration gradient of Ca(2+) between extracellular and intracellular compartments. Ca(2+) signals in the form of time-dependent changes in [Ca(2+)](i) appear as brief spikes that are organized into regenerative Ca(2+) waves. The release of Ca(2+) from internal stores plays a key role in regulating such Ca(2+) signals. Since global Ca(2+) oscillations arise from Ca(2+) waves initiated locally, it results in generation of stochastic Ca(2+) oscillations. In addition, the hierarchical organization of signaling structures translate the molecular fluctuations of single channels to the whole cell leading to formation of stochastic media. Several recent observations indicate that [Ca(2+)](i) changes are fluctuation driven as opposed to a typical deterministic intracellular reaction-diffusion system model. Elucidation of this signaling mechanism can provide detailed knowledge of relationship between cell signaling and cell physiology of living systems.

Strict glycemic control and mortality risk among US adults with type 2 diabetes.

OBJECTIVE: To assess the relationships between strict HbA1c levels and mortality risk among adults with type 2 diabetes by age, insulin therapy, and hypertension comorbidity. METHODS: Data of adult participants with type 2 diabetes from the third National Health and Nutrition Examination Survey (1988-1994) and its linked mortality file (with follow-up death up to 2000) were used. RESULTS: Having strict glycemic control (i.e., HbA1c ≤6.5%) was associated with a lower risk of mortality (hazards ratio=0.69; 95% confidence interval=0.48-0.98). However, among those with strict glycemic control levels, statistically significant results were not found. CONCLUSION: Reaching strict glycemic control levels in the general US population with type 2 diabetes appears to be associated with lower mortality. Further research is needed as to how strict glycemic control affects certain diabetic groups.

Tumor necrosis factor-α induces transcriptional activation of nuclear factor-κB in insulin-producing ß-cells.

We previously showed that tumor necrosis factor-α (TNF-α) induces the dysregulation of intracellular calcium [Ca(2+)](i) in ß-cells by decreasing the levels of the cytoplasmic Ca(2+) binding protein calbindin-D(28k). The purpose of the present study was to test the hypothesis that TNF-α-induced dysregulation of [Ca(2+)](i) in insulin-producing ß-cells causes proteolytic degradation of IκBα and consequently leads to the transcriptional activation of nuclear factor-κB (NF-κB). To test this hypothesis, rat insulinoma (RINr 1046-38) cells, which are an insulin-secreting transformed ß-cell line that constitutively expresses calbindin-D(28k), were treated with increasing concentrations of TNF-α. Using the FunctionELISA procedure to measure degradation of the IκBα subunit as Phospho-IκBα, it was found that, while in the control RIN cell lysate there was no Phospho-IκBα present, in the RIN cells exposed to 2, 5, 10, 20 and 30 ng/ml TNF-α, 17.176±2.85, 17.292±4.35, 53.77±5.63, 30.58±4.89 and 12±3.27 ng/ml Phospho-IκBα/mg of total cell protein was observed, respectively (n=3, P<0.05). Upon treatment of RIN cells with 2, 5, 10, 20 and 30 ng/ml TNF-α, the relative increases in the NF-κB transcriptional activities based on the DNA binding activity of NF-κB determined using an ELISA-based kit were 6.86±0.76-, 8.42±1.27-,7.8±2.32-, 10.28±1.96- and 6.3±1.57-fold, respectively (n=3, P<0.05). The nuclear translocation of NF-κB measured by immunofluorescence showed that, while the ratio of fluorescence in nuclei to that in the cytoplasm of untreated RIN cells was 0.2078±0.0778 (n=11), in RIN cells treated with 10 ng/ ml TNF-α, the ratio was 0.6267±0.1186 (n=11), indicating a statistically significant increase (P<0.05) in the nuclear translocation of NF-κB. These observations suggest that, in insulin-producing ß-cells, the TNF-α-induced degradation of IκBα leads to nuclear translocation and the transcriptional activation of NF-κB.

L-histidine sensing by calcium sensing receptor inhibits voltage-dependent calcium channel activity and insulin secretion in ß-cells.

AIMS: Our goal was to test the hypothesis that the histidine-induced activation of calcium sensing receptor (CaR) can regulate calcium channel activity of L-type voltage dependent calcium channel (VDCC) due to increased spatial interaction between CaR and VDCC in ß-cells and thus modulate glucose-induced insulin secretion. MAIN METHODS: Rat insulinoma (RINr1046-38) insulin-producing ß-cells were cultured in RPMI-1640 medium on 25 mm diameter glass coverslips in six-well culture plates in a 5% CO(2) incubator at 37°C. The intracellular calcium concentration, [Ca(2+)](i), was determined by ratio fluorescence microscopy using Fura-2AM. The spatial interactions between CaR and L-type VDCC in ß-cells were measured by immunofluorescence confocal microscopy using a Nikon C1 laser scanning confocal microscope. The insulin release was determined by enzyme-linked immunosorbent assay (ELISA). KEY FINDINGS: The addition of increasing concentrations of L-histidine along with 10 mM glucose resulted in 57% decrease in [Ca(2+)](i). The confocal fluorescence imaging data showed 5.59 to 8.62-fold increase in colocalization correlation coefficient between CaR and VDCC in ß-cells exposed to L-histidine thereby indicating increased membrane delimited spatial interactions between these two membrane proteins. The insulin ELISA data showed 54% decrease in the 1st phase of glucose-induced insulin secretion in ß-cells exposed to increasing concentrations of L-histidine. SIGNIFICANCE: L-histidine-induced increased spatial interaction of CaR with VDCC can inhibit calcium channel activity of VDCC and consequently regulate glucose-induced insulin secretion by ß-cells. The L-type VDCC could therefore be a potential therapeutic target in diabetes.