Anticholinergic drug atropine diminishes newly formed fear memory in male rats

Post-traumatic stress disorder [PTSD] is a condition which is triggered shortly after experiencing traumatic events. PTSD is complicated by the fact that people with PTSD often develop additional disorders such as phobias, addiction, depression, panic disorder and obsessive-compulsive disorder. Beta-adrenergic and cholinergic system both are involved in memory formation as well as in emotional response associated with memory. It is reported that the administration of beta-adrenergic and cholinergic antagonist results in the impairment in memory formation. Here, we examined the potential of beta-adrenergic antagonist propranolol and muscarinic cholinergic antagonist atropine for impairing the recently formed fear memory associated with PTSD. Reconsolidation is the memory process during which labile memory converts into permanent memory. In this study it is hypothesized that if recently formed fear memory is disturbed during reconsolidation phase by pharmacological intervention then it could be possible to impair wellconsolidated fear memory. Atropine and propranolol were injected in separate set of rats [n=6] just after the reactivation of fear memory. Short term memory and long term memory were monitored after 2 h and 24 h of reactivation respectively. Results of current study demonstrated that only atropine showed significant impairment of reconsolidation of newly formed fear memory whereas propranolol did not show fear memory disrupting effects. The results emphasize the significance of pharmacological intervention to impair reconsolidation of newly formed fear memory

In vitro inhibitory analysis of consensus siRNAs against NS3 gene of hepatitis C virus 1a genotype

OBJECTIVE@#To explore inhibitory effects of genome-specific, chemically synthesized siRNAs (small interference RNA) against NS3 gene of hepatitis C virus (HCV) 1a genotype in stable Huh-7 (human hepatoma) cells as well as against viral replication in serum-inoculated Huh-7 cells.@*METHODS@#Stable Huh-7 cells persistently expressing NS3 gene were produced under antibiotic gentamycin (G418) selection. The cell clones resistant to 1000 μg antibiotic concentration (G418) were picked as stable cell clones. The NS3 gene expression in stable cell clone was confirmed by RT-PCR and Western blotting. siRNA cell cytotoxicity was determined by MTT cell proliferation assay. Stable cell lines were transfected with sequence specific siRNAs and their inhibitory effects were determined by RT-PCR, real-time PCR and Western blotting. The viral replication inhibition by siRNAs in serum inoculated Huh-7 cells was determined by real-time PCR.@*RESULTS@#RT-PCR and Western blot analysis confirmed NS3 gene and protein expression in stable cell lines on day 10, 20 and 30 post transfection. MTT cell proliferation assay revealed that at most concentrated dose tested (50 nmol/L), siRNA had no cytotoxic effects on Huh-7 cells and cell proliferation remained unaffected. As demonstrated by the siRNA time-dependent inhibitory analysis, siRNA NS3-is44 showed maximum inhibition of NS3 gene in stable Huh-7 cell clones at 24 (80%, P = 0.013) and 48 h (75%, P = 0.002) post transfection. The impact of siRNAs on virus replication in serum inoculated Huh-7 cells also demonstrated significant decrease in viral copy number, where siRNA NS3-is44 exhibited 70% (P < 0.05) viral RNA reduction as compared to NS3-is33, which showed a 64% (P < 0.05) decrease in viral copy number. siRNA synergism (NS3-is33 + NS3-is44) decreased viral load by 84% (P < 0.05) as compared to individual inhibition by each siRNA (i.e., 64%-70% (P < 0.05)) in serum-inoculated cells. Synthetic siRNAs mixture (NS5B-is88 + NS3-is33) targeting different region of HCV genome (NS5B and NS3) also decreased HCV viral load by 85% (P < 0.05) as compared to siRNA inhibitory effects alone (70% and 64% respectively, P < 0.05).@*CONCLUSIONS@#siRNAs directed against NS3 gene significantly decreased mRNA and protein expression in stable cell clones. Viral replication was also vividly decreased in serum infected Huh-7 cells. Stable Huh-7 cells expressing NS3 gene is helpful to develop anti-hepatitis C drug screening assays. siRNA therapeutic potential along with other anti-HCV agents can be considered against hepatitis C.

In vitro inhibitory analysis of consensus siRNAs against NS3 gene of hepatitis C virus 1a genotype

Objective To explore inhibitory effects of genome-specific, chemically synthesized siRNAs (small interference RNA) against NS3 gene of hepatitis C virus (HCV) 1a genotype in stable Huh-7 (human hepatoma) cells as well as against viral replication in serum-inoculated Huh-7 cells. Methods Stable Huh-7 cells persistently expressing NS3 gene were produced under antibiotic gentamycin (G418) selection. The cell clones resistant to 1 000 μg antibiotic concentration (G418) were picked as stable cell clones. The NS3 gene expression in stable cell clone was confirmed by RT-PCR and Western blotting. siRNA cell cytotoxicity was determined by MTT cell proliferation assay. Stable cell lines were transfected with sequence specific siRNAs and their inhibitory effects were determined by RT-PCR, real-time PCR and Western blotting. The viral replication inhibition by siRNAs in serum inoculated Huh-7 cells was determined by real-time PCR. Results RT-PCR and Western blot analysis confirmed NS3 gene and protein expression in stable cell lines on day 10, 20 and 30 post transfection. MTT cell proliferation assay revealed that at most concentrated dose tested (50 nmol/L), siRNA had no cytotoxic effects on Huh-7 cells and cell proliferation remained unaffected. As demonstrated by the siRNA time-dependent inhibitory analysis, siRNA NS3-is44 showed maximum inhibition of NS3 gene in stable Huh-7 cell clones at 24 (80%, P = 0.013) and 48 h (75%, P = 0.002) post transfection. The impact of siRNAs on virus replication in serum inoculated Huh-7 cells also demonstrated significant decrease in viral copy number, where siRNA NS3-is44 exhibited 70% (P < 0.05) viral RNA reduction as compared to NS3-is33, which showed a 64% (P < 0.05) decrease in viral copy number. siRNA synergism (NS3-is33 + NS3-is44) decreased viral load by 84% (P < 0.05) as compared to individual inhibition by each siRNA (i.e., 64%–70% (P < 0.05)) in serum-inoculated cells. Synthetic siRNAs mixture (NS5B-is88 + NS3-is33) targeting different region of HCV genome (NS5B and NS3) also decreased HCV viral load by 85% (P < 0.05) as compared to siRNA inhibitory effects alone (70% and 64% respectively, P < 0.05). Conclusions siRNAs directed against NS3 gene significantly decreased mRNA and protein expression in stable cell clones. Viral replication was also vividly decreased in serum infected Huh-7 cells. Stable Huh-7 cells expressing NS3 gene is helpful to develop anti-hepatitis C drug screening assays. siRNA therapeutic potential along with other anti-HCV agents can be considered against hepatitis C.

Diagnostic accuracy of CT scan in diagnosing paranasal fungal infection

Objective: To assess the diagnostic accuracy of CT scan in detecting paranasal sinus fungal infections

Study Design: Cross-sectional, observational study

Place and Duration of Study: Department of Radiology, Ziauddin University Hospital, Karachi, from April to September 2014

Methodology: Patients with clinical suspicion of fungal sinusitis were included in this study. Patients with already diagnosed or history of recurrent fungal infections were excluded. All these patients underwent CT scan examination of paranasal sinuses. Findings were recorded. Final diagnosis was based on smear analysis for fungal culture. Accuracy analysis was conducted for CT diagnosis using smear analysis as the gold standard. Kappa analysis was conducted to determine agreement

Results: Out of the 120 patients, 71 [59%] were male. The sensitivity, specificity, positive predictive value and negative predictive value of CT were 96.19%, 93.33%, 99.01%, 77.77%, respectively. The diagnostic accuracy was 95.83%. Kappa statistics showed 82% agreement beyond chance

Conclusion: CT scan is highly accurate in diagnosing and characterizing fungal infection of paranasal sinuses. It also determines the extent of disease which eventually aids in deciding the surgical approach to be used

Hyperlipidemias, diabetes and depression

Background: Factors causing increased cholesterol levels in the body may include inactivity, obesity, genetic factors and an unhealthy diet. The high colesterol levels or hyperlipidemias may contribute to high concentrations of its precursor triglycerides and low density lipoproteins in plasma of the individuals. High triglyceride levels signal insulin resistance. This study was designed to determine an association of hyperlipidemia and diabetes mellitus with depression

Method: This case-control study involved 30 patients diagnosed with type 2 Diabetes Mellitus [DM] and hyperlipidemia [HL], and 30 non-diabetic healthy individuals having normal glucose tolerance test and no other co-morbidity. All subjects were of 30-50 years age. Blood samples from all participants were collected for determination of the HbA1C and lipid profiles. PQ9 score questionnaire for depression was asked from all subjects

Results: Patients suffering with hyperlipidemias and diabetes mellitus had higher incidence of depression compared to healthy subjects [p<0.05]

Conclusion: Depression was more prevalent in hyperlipidemic and diabetic patients

Turkish Healthcare Providers’ Level Of Knowledge, Attitude And Practice Toward Diagnosis Related Group System – A Cross Sectional Study

Diagnosis-related group (DRG) system is patient classification system designed to produce limited number of classes which are relatively similar in terms of resource consumption and clinical characteristics. The aim of this study was to assess the level of knowledge, attitude and practice (KAP) of Turkish health care providers toward DRG system implemented in Turkey.A total of 238 healthcare providers were randomly selected from two urbanand one rural hospital in Turkey.A questionnaire was used for data collection; contacting 32 items (10 items about knowledge, 12 items about attitude and 10 items about the practice) and its validity and reliability were confirmed. Data analysis was performed using chi-square and multivariate logistic regression.In this study,only one third of healthcare providers showed good knowledge (35.7%) and good practice (37.4%) about DRG system,compared to 54.2% of them showed good attitude.There was significant difference between age, gender, occupation groups and whether the respondents have attended a workshop for DRG system in terms of KAP (p > 0.05).These results indicated the need for further actions to implement DRG system in terms of creation of suitable environment and increasing awareness among healthcare providers, especially male, medical doctors, nurses, elderly, and those who have never attended a workshop, in addition to regular review to ensure the program would reach its targets.

Diagnostic accuracy of Ultrasound B scan using 10 MHz linear probe in ocular trauma; Results from a high burden country

Objective: To study the diagnostic accuracy of Ultrasound B scan using 10 MHz linear probe in ocular trauma

Methods: A total of 61 patients with 63 ocular injuries were assessed during July 2013 to January 2014. All patients were referred to the department of Radiology from Emergency Room since adequate clinical assessment of the fundus was impossible because of the presence of opaque ocular media. Based on radiological diagnosis, the patients were provided treatment [surgical or medical]. Clinical diagnosis was confirmed during surgical procedures or clinical follow-up

Results: A total of 63 ocular injuries were examined in 61 patients. The overall sensitivity was 91.5%, Specificity was 98.87%, Positive predictive value was 87.62 and Negative predictive value was 99%

Conclusion: Ultrasound B-scan is a sensitive, non invasive and rapid way of assessing intraocular damage caused by blunt or penetrating eye injuries

Application of Bromocresol Green and Bromothymol Blue for the Extractive Spectrophotometric Determination of Anti-hypertensive Drugs.

Simple, selective and highly sensitive spectrophotometric methods are proposed for the rapid and accurate determination of anti-hypertensive drugs namely telmisartan (TEL), propranolol (PRO), bisoprolol (BIS) and carvedilol (CRV) in tablets and biological fluids using bromocressol green (BCG) and bromothymol blue (BTB). The developed methods involve formation of stable yellow colored dichloromethane extractable ion-pair complexes of the amino derivative of four antihypertensive drugs such as TEL, PRO, BIS and CRV with two sulphonphthalein acid dyes, namely; BCG and BTB in acidic buffer. The effect of optimum conditions via pH on the ion-pair formation, reagent concentration, time and temperature and solvent was studied. The composition of the ion-pairs was found 1: 1 by Job’s method. The established methods having high sensitivity and good selectivity could be applied to the determination of the studied drugs in pharmaceutical, urine and blood serum samples with satisfactory results. The results obtained are good agreement with experimental data. The reaction mechanism was also discussed.

2, 3, 7, 8-tetrachloro-dibenzo-p-dioxin induced testicular toxicity in rats and the protective effect of quercetin: Biochemical, histopathological and immunohistochemical studies.

Humans and animals are most sensitive to toxicant exposure during development. Dioxin, as an endocrine disruptor, is known to impair testicular functions and fertility. The present study was carried out to investigate the effects of quercetin on TCDD-induced toxicity in the testicular tissue of rats. Forty male albino rats were randomly divided into four groups (n = 10/group). Group I represent the control group; Group II administrated TCDD (27.5 μg/kg) via gavage for four week; Group III received quercetin (20 mg/kg bw.) Via gavage before TCDD administration; Group IV received quercetin alone (20 mg/kg bw). Biochemical markers included levels of testicular malondialdehyde formation and reduced glutathione as well as monitoring the activities of testicular superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase were studied. Also, serum hormonal profiles of luteinizing hormone and testosterone were reported. Our results show that administration of TCDD induces testicular damage concerning oxidative stress parameters, serum hormone level and sperm parameters. In addition, the microscopic structures of the testis, including histological and immunohistochemical studies were evaluated. Exposure to TCDD induces histopathological changes in rats testis including degeneration of seminiferous tubules, tubular necrosis, intratubular vacuolization, widened lumen and deshaped germ cells. Marked increase of apoptotic activity was observed. Also, our results clearly demonstrate the ameliorative potential of quercetin in dioxin induced testicular damage.

Factors Influencing Pap Smear Practice Among Primary School Teachers In Diyala City, Iraq

Cervical cancer is the seventh most common cancer among women in Iraq. This study aimed to assess the factors influencing Pap smear practice among married primary school teachers in Diyala City, Iraq. A total of 222 teachers (aged 20 years to 63 years) residing in Diyala City were enrolled in a cross sectional study from 1st August till 30th September 2012. The sampling method was a self-administered survey using a multistage sampling technique. The prevalence of Pap smear practice was very low (12.6%). Relatives and friends were the main source of information about the Pap smear test for more than one third (41.9%) of the respondents. However, the medical doctor’s advice was the most important reason for undergoing a Pap smear test in more than two third (60.7%) of respondents. Only 32.4% of the teachers have good knowledge about cervical cancer and Pap smear test, but 76.1% of them showed a positive attitude toward learning about cervical cancer and Pap smear. Based on multiple logistic regression analysis results only the husband’s encouragement (p= 0.003, prevalence odds ratio [POR] = 0.256, 95%CI 0.10-0.63) was significantly associated with Pap smear practice. A more effective educational program is needed to educate Iraqi women about cervical cancer and screening tests, including Pap smear.

Central retinal artery occlusion in a healthy pregnant woman: a suspected case of susac syndrome

An eighteen-year-old pregnant woman presented with history of acute loss of vision in the right eye for 4 days. Clinical examination revealed central retinal artery occlusion in the right eye. All investigations were normal except MRI brain. MRI report suggested the possibility of acute vasculitis or Susac syndrome; it should be considered in all cases of acute retinal artery occlusions occurring in apparently healthy young females without any precipitating factors. The syndrome may not manifest 'with full clinical triad

Association between the polymorphisms of angiotensin converting enzyme [Peptidyl-Dipeptidase A] INDEL mutation [I/D] and Angiotensin II type I receptor [A1166C] and breast cancer among post menopausal Egyptian females

Renin-angiotensin system [RAS] is involved in the regulation of cardioascular homeostasis. Angiotesin [Ang II] is converted from angiotensin converting enzyme [ACE]. Ang II exerts it effects by binding to two types of receptors; AT[1]R and AT[2]R. Ang II effect on AT[1]R promotes proliferation angiogenesis and metastasis in breast tissues. ACE [I/D] polymorphism is an insertion/deletion of a 287 bp DNA fragment within intron 16 of ACE gene. A1166C is a single nucleotide polymorphism [SNP] in the 3-UTR of AT[1]R gene. Both [D] and [C] alleles were found to be related to RAS overactivation. One hundred and twenty postmenopausal Egyptian females were included in the present study and were divided into control group [fifty apparently healthy women] and patients group [seventy breast cancer patients]. Detailed history taking was done with stress on age, family history, menstrual, obstetric, medical and drug history. Physical examination including body mass index calculation was done. Histopathological examination was done for tumor grading and staging. Detection of ACE gene [I/D] polymorphism by PCR and AT[1]R AT[1]66C SNP using PCR/RFLP were done

Health System Reform From The People’s Point Of View: Development Of Reliable And Valid Questionnaire

Health system reform has been a major concern for different countries. The aim of this research was to develop a reliable and valid questionnaire suitable to assess the consequences of health reform process from people’s perspective. An extensive literature review used to extract a set of statements as possible indicators for health system reform. Expert panel used to determine the content validity rate (CVR) and the content validity index (CVI). The first version produced in Turkish language and pre-piloted with 20 heads of household. Qualified committee used to translate the Turkish version to English version. Group of eighteen academics and graduate students recruited to tests both versions for parallel test validity. The construct validity of the questionnaire was determined using principal components analysis with Varimax rotation method (PCA). Internal consistency and questionnaire’s reliability were calculated by Cronbach’s alpha and the test–retest reliability test. A 17- items questionnaire was developed through the qualitative phase. The Bartlett’s test was significant (p < 0.001), and the KMO value (0.842) showed that using principal component analysis (PCA) was suitable. Eigenvalues equal or higher than 1 were considered significant and chosen for interpretation. By PCA, 4 factors were extracted (accessibility, attitude and preference, quality of care and availability of resources) that jointly accounted for 85.2% of observed variance. The Cronbach’s alpha coefficient showed excellent internal consistency (alpha=0.97), and test-retest of the scale with 2-weeks intervals indicated an appropriate stability for the scale (Intra-class coefficient = 0.96). The findings showed that the designed questionnaire was valid and reliable and can be used easily to assess the consequences of health reform process by comparing the situation before and after the reform from people’s perspective.

Peoples Expectations From Healthcare Providers – A Turkish Perspective

There is high expectation from the population on part of the healthcare providers. These include; skilful and timely medication administration; and knowledge, honesty, listening skills, availability and professional attitude. The aim of this paper is to evaluate the expectation of population with regards to the healthcare providers in Turkey. A cross- sectional study was conducted in Turkey, including both rural and urban population, carried out from October 2011 till January 2012. A total of 540 household heads were selected using multistage random sampling technique. Data was collected using modified self-administered 16-items QUOTE (Quality of Care Through the Patients’ Eyes) questionnaire. The questionnaire measures communication/ accessibility, organizational skills and professional skills. The response rate was (77.1%) and data was analyzed by using SPSS version 16.0. All the aspects measured using QUOTE questionnaire were found to be important by the majority of respondents, but with varying degrees of priority. The quality aspects related to the professional skills of physicians was ranked first followed by communication/ accessibility and last but not the least is the organizational skills of health care providers. This study explored the Turkish people priorities and expectations regarding healthcare providers. The public priorities and expectation were different across population. This may reflect the need to understand people’s expectations before providing the services to avoid complaints that may occur after the services have been rendered.

Abdominal cocoon [sclerosing encapsulating peritonitis]: a rare cause of intestinal obstruction

A 24 years old lady presented with classical history of acute intestinal obstruction. There was a background history of chronic abdomen for 9 years. There was asymmetrical abdominal distension. On laparotomy, the entire small intestine was cocooned and enclosed in a yellowish white thick fibrotic membrane resulting in obstruction of the small intestine. When the membrane was carefully peeled off the small intestine, the underlying small gut was found to be absolutely healthy. The histopathology report was consistent with non-specific dense fibrosis. Based on these findings, a diagnosis of abdominal cocoon or sclerosing encapsulating peritonitis was made which is an extremely rare cause of small bowel obstruction

Apache-II score correlation with mortality and length of stay in an intensive care unit

To correlate the APACHE-II score system with mortality and length of stay in ICU. Cohort study. The Intensive Care Unit [ICU] of the Aga Khan University Hospital, Karachi, from May 2005 to May 2006. All adult patients who were admitted in the ICU were included. APACHE-II score was calculated at the second and seventh days of admission in the ICU. Patients who were discharged alive from the ICU or died after first APACHE-II Score [at 2nd day] were noted as the primary outcome measurement. Second APACHE-II score [at 7th day] was used to predict the length of stay in the ICU. Pearson's correlation coefficient [r] was determined with significance at p < 0.05. In the lowest score category 3-10, 27 out of 30 patients [90%] were discharged and only 3 [10%] died. Out of those 39 patients whose APACHE-II score was found in high category 31 - 40, 33 [84.6%] deaths were observed. This revealed that there might be more chances of death in case of high APACHE-II score [p=0.001]. Insignificant but an inverse correlation [r = -0.084, p < 0.183] was observed between APACHE-II score and length of ICU stay. The APACHE-II scoring system was found useful for classifying patients according to their disease severity. There was an inverse relationship between the high score and the length of stay as well higher chances of mortality

Efficiency of Pa catheters in the ICU of a tertiary care hospital

Existing randomized controlled trials on Pulmonary Artery Catheter [PAC]-guided strategies reveal a modest risk reduction that does reach statistical significance. An observational, prospective, controlled study was carried out in the ICU of a tertiary care hospital. Incidence, indications, complication rate and outcome of Pulmonary Artery [PA] catheter over a period of 3 months was looked at, comparing cases to matched controls. Despite being a limited study, it is obvious that the cost effectiveness and outcome of patients with the PA catheter seems ambiguous. In a developing country where resources are limited, thought must be given to the risk and benefit ratio of placing this invasive monitor and use of the information provided properly justified

Apache II score as a predictor of length of stay and outcome in our ICUS

The APACHE II [acute physiology and chronic health evaluation] is used widely for predicting probability of hospital mortality and length of stay in the ICU. APACHE II forms were available to all ICU residents within 24 hours of admission, and a score was assigned to them. Based on our results the APACHE II score has reliably predicted an outcome of the least amount of length of stay [LOS] in the ICU as well as a 100% probability of being shifted out of the ICU for a score of <10 [according to international benchmarks]. This reliable scoring system can be used for predicting mortality and length of stay and therefore, resource allocation, antibiotic use and ethical decisions regarding counseling families about end of life decisions - all within 24 hours of admissions