Search

It has been suggested that the well-known health benefits associated with exercise can be explained by a placebo effect, and that greater effort should be given to convince people that their current behaviors have desirable health consequences. The overall purpose of this study was to test the efficacy of this "mind-set matters" hypothesis through the use of an expectancy-based intervention with adolescents. The study involved a four week randomized controlled trial with 348 Grade 9 adolescents (M (age) = 14.07 yrs, SD = 0.30), from four high schools, taking part in either a placebo-intervention condition (n = 188) or a control condition (n = 160). Participants in the placebo-intervention condition were informed that what they are already doing in school physical education lessons fulfills current recommendations for an active lifestyle. Participants in the control condition were not given this information. Four weeks after the intervention, adolescents in the placebo-intervention condition did not demonstrate significant changes in physiological health-related measures (diet, weight, body mass index, percentage body fat, heart rate, mean arterial pressure). The findings question the external validity of the "mind-set matters" hypothesis with adolescents, and suggest that simply encouraging adolescents to believe that they are healthy may not enable them to respond with improved indicators of physical health.

Zinc oxide nanoparticles (ZnO NPs) demonstrate potential positive effects on reproduction. However, their protective role against the reproductive toxicity pollutants has not yet been adequately studied at the molecular level. This study was designed to assess this objective using Benzo[α]pyrene B[a]P as reproductive toxic agent . Forty-eight mature male rats were randomly distributed into six groups: Group1 (negative control); Groups 2 and 3 (positive control I and II, wherein the animals were treated with 10 and 30 mg ZnO NPs/kg BW, respectively); Group 4 (B[a]P group; treated with 150 mg B[a]P/kg BW); and Groups 5 and 6 (subjected to B[a]P treatment co-administered with different concentrations of ZnO NPs). We investigated oxidative stress biomarkers; cholesterol side-chain cleavage enzyme (CYP11A1), steroidogenic acute regulatory protein (StAR), and 3β-hydroxysteroid dehydrogenase (3β-HSD) gene expression; testosterone levels; and histopathology of the liver, kidney, and testicles. The B[a]P-treated group showed significant deterioration in all reproductive parameters and displayed induced oxidative stress. ZnO NPs remarkably reduced oxidative stress, effectively upregulated the mRNA levels of CY11A1, StAR, and 3β-HSD, and improved the histological pictures in the examined organs. At their investigated doses and given their NPs properties, ZnO NPs demonstrated a marked ameliorative effect against the reproductive toxic effects of B[a]P. Further studies are needed to thoroughly investigate the molecular mechanisms of ZnO NPs.

Background: Cerebral palsy (CP) is defined asneurode-velopmental disorder that occurs due to disturbance in the development of the central nervous system. Laser acupuncture is a technique with minimal side effects and safe for chil-drenand can be used as an alternative line of treatment in several chronic pediatric diseases. Aim of Study: To assess the efficacy of using a six weeks course of laser acupuncture on muscle strength in children with diplegic or quadriplegic cerebral palsy. Patients and Methods: A randomized controlled trial; that was conducted in the neuropediatric clinic of the National Research Centre, Egypt and Outpatient Clinic of Ain Shams Children Hospital from 2018 to February 2020. It included 40 children with spastic CP. They were randomly assigned to two equal groups (n=20), laser acupuncture group and control group. Both groups received physiotherapy for successive 6 weeks; while only the study group also received low-level laser 650nm with 50mW powers that was applied at each acupoint for 30s giving an energy density of 1.8J/cm2). Results: Power of hip flexors and knee extensors were significantly increased in laser acupuncture group compared to the control group, p < 0.001. Conclusion: Laser acupuncture has a beneficial effect on hip flexors and knee extensors muscles strength compared with the control group in children with spastic cerebral palsy.

Background Peripheral neuropathy (PN) is the damage and dysfunction of neurons of the peripheral nervous system. The present study was conducted to estimate the effectiveness of low-power laser therapy (LPLT) in the management of PN in a rats’ model. Methods PN was induced by giving dichloroacetate (DCA) (250 mg/kg/day) for up to 12 weeks. Four groups of rats were used: control group, PN group, PN group treated with gabapentin and PN group treated with LPLT. The study was conducted for 8 weeks. The management of PN was estimated by behavioral tests which included hot plate and Morris water maze tests. Blood biochemical analysis were carried out. Results Using of hot plate test indicated thermal hypoalgesia and using Morris water maze test showed cognitive decline in PN rats. Treatment with LPLT or gabapentin improved both the pain sensations and deteriorated memory that occurred in the PN rats. Biochemical analysis showed that LPLT significantly decreased the elevated beta-endorphin level in PN rats, while gabapentin could not reduce it. Treatment PN rats with LPLT or gabapentin shifted the high levels of TNF-α, IL-1β and IL-10 cytokines back to their normal values. Serum nitric oxide and MDA significantly increased in the PN group together with significant reduction in the rGSH level, these values were significantly improved by LPLT application while this was not the case with gabapentin treatment. Furthermore, treatment with gabapentin or LPLT significantly reduced serum ALAT and ASAT activities which are otherwise increased in the PN group. S100B, PGE2, total cholesterol, triglycerides, LDL-cholesterol, HDL-cholesterol, urea and creatinine showed insignificant changes among all groups. Conclusions Our results showed that treatment with LPLT is more efficient than gabapentin in ameliorating the peripheral neuropathy induced by xenobiotics.

Background: The foot is the most complex body’s structure; it is highly susceptible to disorders because of its loading pattern. The complexity of the foot structure geometry implies the use of reverse engineering tools to obtain a model that can accurately mimic the biomechanical behavior of the foot. Objective: The objective of this study is to establish a state-of-the-art ankle-foot finite element (FE) model with anatomically realistic geometry and structure in order to get the model that will suit all cases for future studies on stress injuries and foot insole designs under different loading conditions.Material and Methods: In this analytical study, tomography images were imported in DICOM format, after that, the object was exported in the form of three-dimensional structures in STL file format to define and assemble the structures. After that, the computer simulation on numerical model was done. One-way Analysis of variance (ANOVA) test was performed, and a threshold (pResults: The results showed no significant differences (P>0.05) between the values of the plantar pressure corresponding to neutral standing condition with other foot models in literature. The stresses transferred to the bone structure show that the relatively higher stress was located in the fifth, fourth and third tarsometatarsal, where the maximum von Mises stress in the bone structure was 2155.4 kPa. Conclusion: The state-of-the-art ankle-foot FE model with anatomically realistic geometry and structure will be very helpful for future studies on stress injuries and foot insole designs under different loading conditions.

Micronutrients play an important role in enhancing the immune system, therefore, proper nutritional support of micronutrients could have a positive impact on COVID-19 outcome.

Diabetic neuropathy (DN) is the highly occurred complication of diabetes mellitus; it has been defined as an event of peripheral nerve dysfunction characterized by pain, allodynia, hyperalgesia, and paraesthesia. The current study was conducted to evaluate the efficacy of low-level laser therapy (LLLT) in the management of neuropathy in diabetic rats. The used animals were divided into the following groups: negative control, streptozotocin-induced diabetic rats, and diabetic rats with peripheral neuropathy (DNP) and DNP treated with gabapentin or with LLLT. Behavioral tests were carried out through hotplate test for the determination of pain sensations and the Morris water maze test for spatial reference memory evaluation. Blood samples were collected at the end of treatment for biochemical determinations. In the current study, the latency of hind-paw lick decreased significantly when DNP are treated with gabapentin or LLLT. The Morris water maze test showed that LLLT treatment improved memory that deteriorated in DNP more than gabapentin do. The results of the biochemical study revealed that LLLT could not affect the level of beta-endorphin that decreased in DNP but significantly decreased S100B that rose in DNP. PGE2 and cytokines IL-1β, IL-10, and TNF-α showed significant increase in DNP compared with control group. The gabapentin administration or LLLT application significantly reversed the levels of the mentioned markers towards the normal values of the controls. Levels of serum MDA and nitric oxide increased significantly in the DNP but rGSH showed significant decrease. These markers were improved significantly when the DNP were treated with gabapentin or LLLT. The treatment with gabapentin or LLLT significantly decreased the raised level in total cholesterol in DNP but could not decrease the elevated level of triglycerides, while LDL cholesterol decreased significantly in DNP treated with gabapentin but not affected by LLLT. Values of serum alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), urea, and creatinine increased significantly in the DPN and diabetic rats without peripheral neuropathy (PN) compared with control group. The treatment of DNP with gabapentin induced significant increases in ALAT and ASAT activities but LLLT treatment induced significant decreases in ALAT and ASAT activities as compared with DNP group. Neither gabapentin nor LLLT could improve the elevated levels of serum urea and creatinine in the DNP. It could be concluded that LLLT is more safe and effective than gabapentin in the management of neuropathy in diabetic rats.

Background: Awareness of various aspects of Diabetes Mellitus (DM) is essential for the prevention, management and control of the disease. Saudi Arabia is one of the most countries have the highest incidence of diabetes. To our knowledge, this was the first research evaluating the level of awareness in a diabetic population of Taif. In this study, we assessed the awareness of DM among diabetic patients attending in the hospitals and diabetes clinics, in Taif city, KSA. Methods: A cross-sectional study conducted in Taif city from December 2017 to February 2018. An attached questionnaire was filled after a personal interview, having 11 items. The correct answers were marked as aware and incorrect as unaware and made conversions to a percentage. Total 210 diabetic patients were assessed. The data were analyzed using SPSS version 22 software. Results: The 210 participants interviewed included 114 females and 96 males, 88 with types 1 DM while 122 with type 2. Approximately 84% of participants have previously counseled (Diabetes education), 77.1% of them have a family history of diabetes. Overall sufficient awareness regarding diabetes mellitus was reported among the majority of the participants with a significant effect of gender and type in some specific items. Conclusion: This is the first study examining the knowledge and awareness of diabetes among the only diabetic population in KSA. Our data conducted that the diabetic population at Taif city have enough general awareness of DM regarding complication, sugar monitoring, safe fasting, life style modifications, and gestational diabetes which reflect the very well health care services provided at Taif.

Nanotechnology plays a unique and novel role to develop new methods for adjuvant preparation, whichplay an important role in the efficacy of vaccines. In this study we have studied the effects of Calciumphosphate nanoparticles (CaP) magnitude and type of immunity elicited in response to inactivated FMD trivalent vaccine. Acomprehensive sero-immunological study was conduced to reveal the adjuvant's effect of Calciumphosphate nanoparticles on the immune response to oil adjuvanted trivalent Foot and Mouth Disease(FMD) in vaccinated calves. This study was conducted in three calve groups; group (A) vaccinatedsubcutaneously with trivalent oil FMD vaccine, group (B) vaccinated subcutaneously with trivalentFMD vaccine adjuvanted with Calcium Phosphate nonaoparticles (10 mg/dose).While group (C)vaccinated subcutaneously with trivalent FMD vaccine adjuvanted with both oil and CaP nanoparticles.The humeral and cellular immunoresponses were monitored in different tested groups. Results indicatedthat the incorporation of Calcium phosphate nanoparticles into inactivated FMD vaccine induces anincrease of the specific protective immune response. Higher and longer period of immuneresponses were found in calves vaccinated with both oil and Calcium phosphate nanoparticlesadjuvanted vaccine up to 40 week, while those vaccinated with Calcium Phosphate nanoparticles andwith oil vaccine showed protected immunity up to 36 and 32 weeks respectively

Objective: To learn more about reproductive physiology of adult female Arabian horses over a period of 24 months and to examine the effect of breed’s difference between Arabian and European female horses over a period of 36 months on the circulatory levels of both metabolic hormones (IGF-1 and leptin). Methods: Thirty female Arabian mares and 22 European non pregnant brood mares exported from Swedish and Germany of ages from 3 to 7 years belonging to Mubarak Police Academy (Abaseia horse farm) was used. Rectal ultrasonography was conducted in Arabian horses to monitor ovarian activity which classified into cyclic ovarian activity, no ovarian activity, and ovarian tumor, pregnancy and postpartum mares were also included in this study. Blood samples from these mares were collected and analyzed for progesterone and Leptin, IGF-1 and Nitric oxide (NO). In the same time, blood samples were collected from Arabian and foreign breeds for IGF-1 and leptin analysis Results: There are significant increase in the IGF-1 (778.1±15.7 ng/mL and NO concentrations (39.83±9.15 μM/mL) in case of ovarian tumor. Significant decrease in leptin concentration was recorded (0.61±0.31 ng/mL) in case of postpartum cases. Inactive ovaries mare and pregnant one recorded significant increase in progesterone levels (10.1±1.46 and 22.6±2.0 ng/mL, respectively). On other hand Leptin recorded significant decrease in Arabian horses than European horses (0.86± 0.14 vs. 1.73±1.34), while IGF-1 have no significant change between two breeds. Conclusion: The knowledge of the normal and abnormal metabolic and sex hormones concentrations will help us to understand the role of these hormones in reproductive physiological and additionally, potential diagnostic and prognostic uses in both human and veterinary medicine, and will provide information for further research on this equine breeds as well as in human diseases.

Aim: To determine the minimal effective dose of Foot and Mouth disease (FMD) serotypes (A, O, SAT2) according toantigenic content (146S) in order to produce a potent trivalent FMD vaccine.Materials and Methods: Monovalent ISA 206 vaccines were prepared with 3 final concentration of 146S (1.6, 2.2, 2.8μg/dose). The vaccine potency was evaluated by the determination of guinea pig protective dose 50 (GPPD )for each 50concentration of 146S for each type of FMD monovalent vaccine where a fourfold dilution of the vaccines was constructedand each dilution was inoculated as 0.5 ml S/C in each of 5 guinea pigs.Results: The obtained results revealed that by using 1.6 μg of 146S for type O Pan Asia-2, AIran O5 and SAT/EGY/2012, theGPPD was 40.4, 19.75 and 31.6 respectively, while the use of 2.2 μg of 146S resulted in GPPD 78.6, 78.6 and 105.8 for the 50 50three types respectively, and by using 2.8 μg of 146S resulted in GPPD 161.7, 105.8 and 161.7 for the three subtypes (A, O, 50SAT2) respectively. So it is clear that the lowest 146S dilution inducing good protection (more than 72 GPPD ) was 2.2 μg for 50each serotype of used FMD monovalent vaccines. Depending on this result, the trivalent vaccine was formulated as 2.2 μg of146S payload from each virus type/dose with equal volume of montanide ISA206 oil as adjuvant. For more confirmation theprepared trivalent vaccine potency was evaluated by Guinea pig protective dose 50 which was found to be 88 GPPD .Also 50mean SNT antibody titer was detected in serum of the test Guinea pigs 1.56, 1.68 and 1.68 log /ml against FMDVserotype O 10Pan Asia-2, AIran O5 and SAT/EGY/2012 respectively in a higher level than the recommended protective titer (PT=1.2). Alsofor further confirmation the formulated trivalent vaccine which contain 2.2 μg/serotype/dose were evaluated in cattle tomeasure the antibody titer against the three serotypes and the antibody against the three serotypes were found to be higher thanthe recommended titer (1.5) which extended for 32 WPVand these results came in parallel manner to the GPPD and antibody 50titer of the Guinea pigs of the prepared trivalent vaccine with 146S (2.2 μg/serotype/dose).Conclusion: It could be concluded that the minimum content of antigenic 146S of FMDVserotype O pan Asia, AIran O5 andSAT2/EGY/2012 should not be less than 2.2 μg/dose/ from each serotype in the trivalent vaccine aiming to induce thepermissible protection in vaccinated livestock.

Type 2 diabetes is a polygenic disease characterized by interaction of environmental and genetic factors. The paraoxonase 1 gene (PON1) 55 and 192 polymorphisms have been reported to be associated with type 2 diabetes and its complications. Our aim is to study the PON1 55, 192 gene polymorphisms and enzyme activity in type 2 diabetic Egyptian population with complications. 100 type 2 diabetic patients with complications (34 with cardiac and 66 with microvascular complications (neuropathy, retinopathy and/or nephropathy)). This was in addition to 100 healthy control subjects of matched age and sex were taken. PON1 55 L⁄M and 192 Q⁄R gene polymorphisms and PON1 enzyme activity serum levels were detected. The LL genotype of PON1 55 polymorphism and QR and QQ genotypes of PON1 192 polymorphism were more frequent among the patients with diabetic complications. The PON1 enzyme activity levels were lower among the diabetic patients than in control subjects. PON1 55 and 192 polymorphisms and enzyme activity seems to be related to diabetic complications in this population.

Type 2 diabetes mellitus is the most common type of diabetes worldwide with serious macro- and microvascular complications. It is a polygenic disease characterized by interaction of environmental and genetic factors. The paraoxonase 1 gene (PON1) 55 and 192 polymorphisms have been reported to be associated with type 2 diabetes and its complications. Our aim is to study the PON1 55 , 192 gene polymorphisms and enzyme activity in type 2 diabetic Egyptian population with complications. 100 type 2 diabetic patients with complications (34 with cardiac and 66 with microvascular complications (neuropathy, retinopathy and/or nephropathy). This was in addition to 100 healthy control subjects of matched age and sex were taken. PON1 55 L⁄M and 192 Q⁄R gene polymorphisms and PON1 enzyme activity serum levels were detected. The LL genotype of PON1 55 polymorphism and QR and QQ genotypes of PON1 192 polymorphism were more frequent among the patients with diabetic complications. The PON1 enzyme activity levels were lower among the diabetic patients than in control subjects. PON1 55 and 192 polymorphisms and enzyme activity seems to be related to diabetic complications in an Egyptian type 2 diabetic patients.

Aim: The aim of this study was to investigate whether subcutaneous (S/C) or intramuscular (I/M) is better in vaccination against foot-and mouth disease (FMD), in addition, to determination of the most protective dose concerning the economic point of view. Forty animals were divided into ten groups ,each group eight of sheep were vaccinated with different doses of the bivalent oil FMD vaccine (with 1 ml, 0.5 ml, 0.25 ml and 0.125 ml doses) either S/C or I/M. The experiment included challenged non-vaccinated control group and non-challenged-non-vaccinated control groups. Four weeks post vaccination all animal groups were challenged by inoculation intradermolingual (IDL) with 10.000 ID50 (infective dose fifty) sheep adapted challenge FMD (O & A types) virus and subjected to clinical observation for 8 days. The results showed that the vaccinated animal with a full, ½, ¼ dose injected either S/C or I/M were able to withstand the virus infection while animals vaccinated with 1/8 dose didn’t

Avascular necrosis (AVN) is the final common pathway resulting from insufficient blood supply to bone, commonly the femoral head. There are many postulated etiologies of non-traumatic AVN, including corticosteroids, bisphosphonates, and radiotherapy (RT). However, it is unclear whether there is a dose threshold for the development of RT-induced AVN. In this case report, we describe a patient with prostate cancer metastatic to bone diagnosed with AVN after receiving single-fraction palliative RT to the left femoral head. Potential contributing factors are discussed, along with a review of other reported cases. At present, the RT dose threshold below which there is no risk for AVN is unknown, and therefore detrimental impact from the RT cannot be excluded. Given the possibility that RT-induced AVN is a stochastic effect, it is important to be aware of the possibility of this diagnosis in any patient with a painful hip who has received RT to the femoral head.

Background The underlying pathogenesis of isolated coronary artery ectasia (CAE) is still unknown. The aim of this study was to shed light on the potential mechanisms underlying the development of isolated CAE and its relation to carotid intima media thickness (IMT) and certain inflammatory markers especially adhesion molecules and uric acid. Methods The study included 16 patients with isolated CAE, 16 patients with obstructive coronary artery disease (CAD) without CAE, and 10 gender and age matched subjects with normal coronary arteries as control group. All patients underwent diagnostic coronary angiography, B-mode ultrasonography to measure carotid IMT, and serum levels of soluble intercellular adhesion molecule (ICAM-1), E-selectin and uric acid. Results Serum ICAM-1 levels were found to be significantly higher in patients with isolated CAE compared to CAD and control subjects ( p = 0.0001). E-selectin levels showed no difference between the three groups, while serum uric acid was significantly higher in patients with isolated CAE and patients with obstructive CAD compared to control group ( p = 0.004). There were no difference in carotid IMT between isolated CAE and CAD. Univariate analysis showed that the carotid IMT, serum levels of ICAM-1, E-selectin, and uric acid were related with CAE. ICAM-1 was the independent variable most strongly associated with CAE by multiple linear regression analysis ( p = 0.0001). Conclusion Isolated CAE reflects atherosclerosis associated with high grade vascular inflammation out of proportion to, atherosclerotic involvement. Serum levels of ICAM-1 were the most independent predictor of vascular inflammation.

The use of short-term feed restriction (R) without or with subsequent refeeding (F) as biostimulant of rabbit fertility was examined in this study. A total of 40 mature, non-pregnant, non-lactating New Zealand white female rabbits were allocated to five treatments. The rabbits were individually caged and fed a complete pelleted diet (16.7% CP; 13.1 CF; 2490kcalDE/kg). Rabbits on the control (C) treatment received 150g/d of the diet. Two groups of 8 rabbits received 70% of the control daily feed intake (105g/d; moderate restriction; M) and the other two groups received 50% of the control feed intake (75g/d; severe restriction; S) for 21d. At the end of this period, one group each of M and S fed rabbits were slaughtered for oocyte recovery. Rabbits in the remaining three groups (C, MF and SF) were retained for a further 8d before slaughter and fed the control level of the diet during this period. The effects on body weight, oocyte number and quality, GDF-9 gene expression in oocytes, and changes in serum levels of leptin and IGF-1 were recorded. Initial mean body weights were not significantly different ranging from 2.50±0.33kg (S) to 2.58±0.24kg (C). After 3wk on treatment the C rabbits were significantly heavier (2.65±0.32kg; P0.05) than rabbits on the M (2.26±0.33kg) or S (2.10±0.33kg) treatments. Following 8d of refeeding, the remaining group of S treated rabbits (SF) were still significantly lighter (2.40±0.21kg; P0.05) than C (2.71±0.31kg) with MF rabbits having an intermediate weight (2.50±0.20kg). The number of mature grade A oocytes recovered per ovary was significantly lower for control (3.3±0.35) than the refed treatments (MF 4.0±0.30; SF 4.5±0.39; P0.05). Semi-quantitative PCR analysis of GDF-9 expression showed that control mature grade A oocytes had significantly lower levels of expression (1.27±0.20; P0.05) than those of refed rabbits (MF 1.60±0.10; SF 1.39±0.01). Leptin and IGF-1 values for refed rabbits were significantly higher (P0.05) than at the end of feed restriction and the start point. It was concluded that this biostimulant method has the potential to improve the fertility of rabbits.

BackgroundAtrial fibrillation (AF) is the most common type of arrhythmia and recognized as a risk factor for thromboembolism. Endothelial damage or dysfunction may contribute to increase the risk of thromboembolism via the mediation of a prothrombotic or hypercoagulable state.ObjectivesThe aim of the current study is to investigate endothelial dysfunction (represented by brachial flow-mediated dilatation “FMD”) and inflammation (represented by hs-CRP) in patients with paroxysmal atrial fibrillation.Subjects and methodsForty-two patients with AF taken from the Cardiology Department and Outpatients Clinic, Specialized Medical Hospital, Mansoura University, in the period between February 2011 and May 2011 were enrolled in our study, the patients were then subsequently divided according to the clinical type of AF into Group I: comprised 20 patients with paroxysmal AF (PAF) with mean age 57.35y. Group II: comprised 22 patients with chronic AF (CAF) with mean age 57.68y. Twenty control subjects without AF were enrolled in this study (Group III). Patients and control groups were subjected to clinical evaluation, electrocardiography (ECG), echocardiography and brachial FMD (using external brachial ultrasonography. Serum level of hs-CRP was assessed in all subjects. The diameter change induced by FMD was expressed as the percent change relative to that at the initial scan (FMD%) according to the following equation:FMD%=Maximum diameter-baseline diameterBaseline diameter×100.ResultsLeft atrial diameter was significantly increased when compared either GI or GII with control group (3.96±0.27; 4.7±0.48 vs 3.05±0.35cm) (P

Context.—Epidermal growth factor receptor (EGFR) is overexpressed in up to 80% of colorectal and endometrial carcinomas. Deletions of the polyA tract in the 3′ untranslated region (3′ UTR) have been reported in microsatellite instability–high (MSI-H) colonic carcinomas, but their impacts on EGFR expression and downstream pathways are unclear. This phenomenon has not been reported in other MSI-H tumors. Objective.—To assess the 3′ UTR polyA tract of EGFR in both endometrial and colorectal carcinomas and the mutational status of EGFR downstream pathways. Design.—Ninety-eight colorectal carcinomas and 47 endometrial carcinomas were included. EGFR 3′ UTR polyA status was detected by capillary electrophoresis and Sanger sequencing. EGFR gene expression, EGFR copy numbers, and KRAS and BRAF mutation status were analyzed accordingly. Results.—The 3′ UTR polyA tract was deleted in 18 of 23 (78%) MSI-H versus 0 of 24 microsatellite-stable endometrial carcinomas (P < .001). Similar observations were seen in colorectal carcinomas, in which 29 of 36 (81%) MSI-H, 1 of 62 (1.6%) microsatellite instability–low, and none of the microsatellite-stable tumors harbored the deletion (P < .001). A moderate increase in EGFR mRNA level was observed in endometrial carcinomas with 3′ UTR polyA deletions versus those with wild-type polyA tract. Amplification of the EGFR gene was not observed. Deletions in polyA tract do not seem to affect the frequency of KRAS and BRAF mutations. Conclusions.—Deletions of EGFR 3′ UTR polyA are frequent in endometrial and colorectal carcinomas, are confined almost exclusively to MSI-H tumors, and do not affect KRAS and BRAF mutations.

Background Assessment of right ventricular (RV) function remains difficult because of the RV complex shape. Data regarding RV performance in patients with diabetes are incomplete The aim of this study was to assess the feasibility of pulsed wave tissue Doppler imaging and myocardial performance index (MPI) for the assessment of right ventricular function in diabetic patients without coronary artery disease. Methods The study included 20 diabetic patients, 20 diabetic hypertensive and 20 gender and age matched healthy subjects underwent standard echocardiography with tissue Doppler imaging (TDI) to assess RV function. Patients with myocardial ischemia, impaired left ventricular systolic function, valvular heart disease or other diseases which could alter the right ventricular performance were excluded. Results Myocardial performance index was significantly higher in diabetes compared to control group (0.41 ± 0.05 versus 0.27 ± 0.04, p = 0.001). Peak myocardial systolic velocity (Sa), early diastolic myocardial velocity (Ea), and late diastolic myocardial velocity (Aa) were significantly lower in patients with diabetes mellitus (DM) compared to the control group (p = 0.0001). Isovolumetric relaxation time (IVRT) was significantly higher in DM group compared to control group (p = 0.003). MPI was significantly higher in diabetic hypertensive group versus DM alone group (0.46 ± 0.050 versus 0.41 ± 0.05, p = 0.01). There was no correlation between MPI and blood glucose level and duration of diabetes. Conclusion Myocardial performance index is a useful noninvasive tool for the detection of early right ventricular systolic and diastolic dysfunction in diabetic patients, regardless of coexisting hypertension.

Background and aim Appropriate assessment and prevention of cardiovascular (CV) disease is one of the most important medical tasks worldwide. Carotid artery intima-media thickness (CIMT) is a marker of atherosclerosis, which has been associated with CV events. We examined the associations of a panel of different clinical, laboratory, and ultrasound variables simultaneously and individually with CIMT; to reveal the presence of additional surrogate markers of atherosclerosis to CIMT. Subjects and methods 407 Consecutive non-diabetic individuals (220 men) who underwent comprehensive CV evaluation were included. The maximum IMT of the common and internal carotid artery on the right and left side of the neck were recorded by ultrasonography, and CIMT was calculated as the average of the four measurements. Ten-year Reynolds risk score (RRS) for CV events was calculated online for men and women. CAVI was measured using a VaSera vascular screening system, and the averages of the right and left CAVI were used for analysis after being adjusted for age. Results Univariate linear regression models were constructed to test the association of each of the independent variables with the log-transformed CIMT values [Ln (average CIMT) + 2] separately for men and women. Only the variables with non-adjusted p ⩽ 0.1 were included in the final multivariately-adjusted stepwise linear regression model. After multivariate adjustment, only two variables were significantly and independently associated with log-transformed CIMT values in males (the log-transformed Reynolds risk score and the average age-matched CAVI) and females (the average age-matched CAVI and systolic blood pressure). Conclusion RRS could be a candidate marker of atherosclerosis in men but not in women, while age-adjusted CAVI could be considered a marker of atherosclerosis in both genders.

Background: Myocardial infarction (MI) results in impairment of left ventricular (LV) systolic and diastolic functions to various degrees. The tissue Doppler image derived Tei index (TDI-Tei index) has recently been used to assess LV function, and like the conventional Tei index, it was also found to be useful for global function. There are a small number of studies which apply TDI-Tei index to assess regional function and whether it varies according to the degree of changes in wall motion in various LV segments. Objectives: This study was conducted to assess the changes of segmental myocardial Tei index by TDI according to the degree of changes of wall motion in the patient with acute ST-segment elevation myocardial infarction (STEMI) and its correlation with Tei index derived by conventional Doppler. Patients and methods: This study was carried out on thirty patients with acute STEMI selected from Coronary Care Unit, Cardiology Department, Mansoura Specialized Hospital, Mansoura University, Egypt. The study group was subjected to clinical assessment, electrocardiography (ECG), routine laboratory profile and Doppler echocardiography (conventional and pulsed wave tissue Doppler imaging “PW-TDI”). Tei index was calculated from the sum of isovolumetric contraction time (IVCT) and isovolumetric relaxation time (IVRT), divided by ejection time (ET) which were measured from pulsed wave Doppler imaging of the trans-mitral inflow and LV outflow tracts. Myocardial velocities and TDI-Tei index were measured at basal and mid-segments of LV walts from apical 4, 2 and 5-chamber views using 16-segment model. Average values of myocardial velocities and TDI-Tei indices were obtained from normal, hypokinetic and akinetic segments then compared. Results: The mean values of left ventricular ejection fraction (LVEF), mitral E/A ratio and conventional Tei index were 51.83 ± 5.15%, 1.18 ± 0.15 and 0.63 ± 0.07 ms, respectively. Segmental myocardial TDI-Tei index was correlated positively with conventional Tei index (r = 0.648, P

Background: To investigate the rate of all cause and cardiovascular mortality in patients with symptomatic or asymptomatic peripheral arterial disease (PAD) compared to those without PAD. Methods and results: All the subjects were inpatients at high risk of atherosclerosis and enrolled from February to November, 2006. A total of 320 were followed up until an end-point (death) was reached or until February 2010. The mean follow-up time was 37.7 ± 1.5 months. Compared with non-PAD, PAD patients had significantly higher rates of hypertension, diabetes mellitus, and smoking (P

Deposits of iron and hemosiderosis in the kidney have been observed in diseases with intravascular hemolysis, including paroxysmal nocturnal hemoglobinuria, and valvular heart diseases and prosthetic heart valve implants. However, the decrease in kidney function associated with hemolysis caused by cardiac valvular disease or prostheses is less well recognized. We present a case of intravascular hemolysis after repair and banding of the mitral valve that resulted in massive renal tubular deposition of hemosiderin with decreased kidney function. We discuss the pathophysiologic process of both acute and chronic tubular injury from heme and heme proteins, including injury to organelles resulting in autophagic vacuoles containing damaged organelles, such as mitochondria. We conclude that tubular injury resulting from heme proteins should be considered as a cause of decreased kidney function in all patients with a cardiac valvular disease or prosthesis.

The differential diagnosis for groin pain is quite long, and among the many etiologies can include such diagnoses as chronic appendicitis, diverticulitis, urologic diseases, and gynecological processes. One must not forget, however, the importance of the inguinal hernia in the workup for groin pain, as it is the cause of 4.7 million visits each year. Many of these hernias are evident on clinical exam, though an occult hernia may be suspected in absence of a palpable bulge.

Background : Left ventricular hy­pertrophy (LVH) is an independent risk factor for cardiovascular morbidi­ty and mortality in hypertensive pa­tients. The identification of risk factors for the initiation of LVH in patients with hypertension (HTN) is important including microalbuminunuria (MAU) and hyperaldosteronism. Objectives : Evaluation of the rela­tionship of MAU and plasma aldoste-rone to blood pressure (BP) and LVH in patients with essential HTN. Subjects and Methods 467 Thirty male patients with essential HTN and 15 healthy subjects as a control group were subjected to thorough clinical examination, transthoracic echocardiography, lipid profile, serum potassium, serum aldosterone esti­mation. MAU was evaluated with dip­stick Micral-ll Test of fasting mid­stream morning urine on two successive days. Left ventricular mass index (LVMI) was calculated and values >134gm/m2 were consid­ered as LVH

The low serum level of testosterone in the elderly subjects may be a contributing factor for coronary artery disease (CAD) through many pathways.The aim of this work is to study serum levels of free testosterone in elderly men with CAD (both those with acute coronary syndrome (ACS) and those with stable CAD). This study was conducted on 100 elderly males with CAD ,one half of them (50 patients) was presented with ACS (with mean age 69.6± 2.44 year), and the other half (50 patients) was presented with stable CAD (with mean age 69.42± 2.14year), in addition to 50 apparently healthy elderly males (with mean age 69.06 ± 1.64year) as a control group. We detected the levels of serum free testosterone, cortisol, fibrinogen, plasminogen activator inhibitor-1(PAI-1), high sensitive C-reactive protein(hsCRP), interleukin-6(IL-6). Cases with CAD had significant lower values of free testosterone and HDL-c, but they had significant higher values of cortisol, fibrinogen, PAI-1, IL-6, hsCRP, in comparison to control group. Cases with ACS had significant higher values of cortisol, hsCRP, IL-6, fibrinogen, PAI-1, total cholesterol and BMI more than those with stable CAD. The free testosterone had significant negative correlation with fibrinogen, PAI-1, , hsCRP and IL-6 in both groups of patients. The lower value of serum free testosterone in elderly male subjects may contribute to CAD in them via multiple mechanisms including increase blood pressure, dyslipidemia, obesity, increased blood coagulation and increased inflammatory activity., Yaşlı bireylerde düşük testosteron düzeyi, birçok mekanizma ile koroner arter hastalığı (KAH) gelişimine katkıda bulunabilir. Bu çalışmada KAH olan (akut koroner sendromu(AKS) veyastabil KAH olan) yaşlı erkeklerde serbest testosteron düzeylerinin incelenmesi amaçlanmıştır. Çalışmaya 50AKS’lu (ortalama yaş 69.6± 2.44), 50 stabil KAH( ortalama yaş 69.42± 2.14) olan toplam 100 hasta ve kontrol grubu olarak 50 sağlıklı yaşlı (ortalama yaş 69.06± 1.64) alındı. Çalışmaya alınan kişilerin serumserbest testosteron düzeyleri,kortizol, fibrinojen, plazminojenetkinleştirici inhibitör- 1 (PAI-1),yüksek duyarlı C- reaktif protein (hsCRP), interlökin-6 (IL-6) düzeylerine bakıldı.Kontrol grubu ile karşılaştırıldığında, KAH olan olgularda serbest testesteron ve HDL-c düzeyleri anlamlı olarak düşük bulunurken, kortizol, fibrinojen, PAI-1, hsCRP,IL- 6 düzeyleri yüksek bulundu. AKS’lu hastalarda Kortizol, fibrinojen, PAI-1, hsCRP,IL- 6, total kolesterol ve BMI düzeyleri stabil KAH olan hastalardan önemli derecede yüksekti. Her iki grupta da serbest testesteron ile fibrinojen, PAI-1, hsCRP,IL-6 düzeyleri arasında negatif korelasyon vardı. Yaşlı erkek hastalarda serum serbest testosteron düzeyleri, tansiyon artışı, dislipidemi, aşırı şişmanlık, kogülasyon ve enflamatuar aktivite artışı gibi çoklu mekanizmalar yoluyla KAH’na katkıda bulunabilir

A conclusive study was conducted for preparation and evaluation of combined inactivated entero-4 vaccine containing bovine rotavirus (BRV), bovine coronavirus (BCV), E. coli K99 and toxoid of C. perfringens type "C". Laboratory and field evaluations were conducted on laboratory animals, calves and late pregnant cows with monitoring the active and passive antibodies in vaccinated cows and their offspring respectively. Laboratory evaluation proved purity, safety and high efficacy of the vaccine without interference between different vaccine ingredients. Field evaluation gave satisfactory results when pregnant cows vaccinated at late stage of pregnancy with high neutralizing antibody titers against BRV, BCV and C. perfringens as well as high E. coli agglutinating titers. Maternal immunity passively protected offspring during the critical period of age and remained protected till the end of sampling time (30th day post parturition).

Peroxidase labeled immunoglobulins to camelpox virus (CPV) were prepared for use in various techniques of ELISA. Ten rabbits and three goats were inoculated with a mixture of camelpox virusand Freund’s adjuvant. Sera were pooled separately on the 10th day post the last inoculation and immunoglobulins were precipitated using saturated ammonium sulphate. The globulins were 2.8 g/dl and 2.5 g/dl for rabbits and goats respectively and used for peroxidase conjugation. The peroxidase labeled immunoglobulins were titrated and evaluated using direct solid phase ELISA, double antibody sandwich ELISA and dot immunoblot ELISA. The prepared conjugates gave specific and clear positive reactions till the dilution of 2000 and 1500 for rabbits and goats immunoglobulins respectively. The prepared labeled immunoglobulins could be successfully used in detection of camel pox viral antigen of local virulent and standard vaccinal strain of the virus using various ELISA techniques.

In this study we investigate theeffectiveness of intranasal vaccination of calves with live streptomycin-dependent mutant of P.multocida type B vaccine in comparison with inactivated one. Live P.multocida vaccine gave higher mitogenic response of the peripheral blood lymphocytes as measured by lymphocyte blastogenesis assay than inactivated one allover the period of the experiment. The haemagglutinating antibodies as measured by Passive haemagglutination test (HA) and the IgG as measured by ELISA rose from one day to reach the peak at 15 days, and then decline till 12 weeks post vaccination for live vaccine. However for inactivated vaccine it reached the peak at 8 weeks and then decline till 12 week post vaccination. Calves with live P.multocida vaccine gave higher and earlier secretory IgA in nasal secretions than inactivated one. In conclusion, the intranasal administration of live streptomycin-dependent serotype of P.multocida mutant vaccine was efficient and induced local cellular immune response, IgA antibodies and systemic IgG antibodies.

An aggressive dialysis in a grossly azotemic patient, especially one with severe metabolic acidosis, can lead to dialysis disequilibrium syndrome (DDS). Mild forms present as nausea, vomiting, restlessness, and headache. Severe manifestations include seizures, obtundation, coma, and even death. This clinical picture is caused by cerebral edema induced by one or more of the following mechanisms: "Reverse urea effect" - Dialysis removes urea faster from the blood than from the brain; consequently, water enters the brain. "Cerebrospinal fluid acidosis" - Correction of systemic acidosis engenders the condition due to a lowering of brain pH. "Idiogenic osmoles" - As a response to blood hyperosmolar state, osmoles are produced in the brain. As blood osmolality decreases under relatively quick dialysis, idiogenic osmoles tend to induce brain edema. Because the symptoms of DDS can be life-threatening, preventive measures in patients with severe uremia are important. The first strategy relies on raising blood osmolality by introducing solutes (osmoles) into the blood. The second approach, which is the most common, decreases the efficiency of the dialysis treatment by shortening the duration of a dialysis run to 25% - 30% of normal, by lowering dialyzer blood flow or dialysate flow rate, by using a less efficient dialyzer, or by a combination of these maneuvers. Dialysis frequency is increased instead. Anticonvulsant drugs are needed in cases where the preventive measures have not been used or have been unsuccessful.

Favourable specific mechanical properties of polymer matrix composites make them an attractive material for application in many engineering structures for which they offer substantial improvements over metals. The paper deals with fretting behaviour of unidirectional glass epoxy composites/metal contacts. Fretting is a plague for many industries: failures, loss of matter, loss of function can be induced by fretting. It occurs in all quasi-static contacts and appears as a complex wear phenomenon where a lot of parameters have been studied. From the interface tribology concept, the velocity accommodation mechanisms are discussed for different fibre orientations versus the contact surface of the glass fibre reinforced epoxy material. Results were analysed in two steps. From friction logs, Running Conditions Fretting Maps (RCFM) were first plotted in order to give an analysis of contact conditions and determine the associated material responses. The tribological degradations were then analysed. Differences between the different fibre orientations are mainly discussed on the basis of the stiffness of the anisotropic material and the velocity accommodation in the contact.

This prospective cohort study aimed to compare the efficiency of laser versus traditional acupuncture in treating cervical spondylosis (CS) pain. Forty female patients were randomized into two equal groups that received 3 sessions / week for 4 weeks. Group A received needle acupuncture therapy with electrical stimulation for 20 min at standard acupoints, ear points and Ashi point on the average 3 points. Group B received low level laser therapy (LLLT) acupuncture at the same acupoints. The results demonstrated that tenderness disappeared in 65% of patients in group A and 75% of patients in group B with improved percentage of 85.5% and 89.2%. Pain on VAS related to direction of motion at 6 directions was improved in all cases where with improvement percentage 76.45% and 85.88%. Pain on VAS at rest was improved in all patients with improvement percentage of 80.41% and 84.28%. NDIQ score improved in all patients with improvement percentage of 69.78% and 73.77%. Follow up of VAS after 6 months from the last session revealed persistent improvement in 55% of patients of group A vs 80% of patients of group B. Mean serum TNF-α was decreased in 85% of patients of group A vs 95% of patients of group B where serum beta endorphins was increased in all patients. It is concluded that both modes of treatment for CS gave improvement regarding pain intensity, disability and quality of life being more evident in LLLT followed for 6 months supported with improved serum TNFα and beta endorphin.

Background The global epidemic of obesity is one of the major health issues in the 21st century that influences many aspects of public health, including psychosocial and socioeconomic aspects. Hyperlipidemia is one of the health hazards associated with morbid obesity. Patients and methods This was a prospective study conducted in Kasr Al-Aini University Hospital (sleeve gastrectomy group) and the National Nutrition Institute (diet group) during the period from June 2015 till December 2015 with a follow-up of 12 months till December 2016 for both groups. One hundred patients were included in the study and were equally divided into two groups: group A (sleeve gastrectomy group) and group B (diet group). Objective The objective of this study was to compare the effects of laparoscopic sleeve gastrectomy (LSG) and a dietary regimen on hyperlipidemia in morbidly obese patients. Results LSG significantly decreased total cholesterol in 70% of cases and triglycerides in 78% of cases; however, diet caused a decrease of total cholesterol in 30% of cases and triglycerides in 54% of case. Low-density lipoprotein was not significantly changed in both groups. Conclusion LSG is more effective than diet programs in treating obesity-associated hyperlipidemia due to more significant and sustained excess body weight loss.

Background. The low serum level of testosterone in the elderly subjects may contribute to coronary artery disease (CAD). Our aim is to study serum levels of free testosterone in elderly men with CAD.Subjects and Methods. This study was conducted on 100 elderly males with CAD, one half of them was presented with ACS (with mean age year), and the other half was presented with stable CAD (with mean age year), in addition to 50 apparently healthy elderly males (with mean age year) as a control group. We detected the levels of serum free testosterone, cortisol, fibrinogen, plasminogen activator inhibitor-1(PAI-1), high sensitive C-reactive protein(hsCRP), interleukin-6(IL-6).Results. Cases with CAD had significant lower values of free testosterone and HDL-c, but they had significant higher values of cortisol, fibrinogen, PAI-1, IL-6, hsCRP, in comparison to control group. Cases with ACS had significant higher values of cortisol, hsCRP, IL-6, fibrinogen, PAI-1, total cholesterol and BMI more than those with stable CAD. The free testosterone had significant negative correlation with fibrinogen, PAI-1, hsCRP and IL-6 in both groups of patients.Conclusion. The lower value of serum free testosterone in elderly male subjects may contribute to CAD.

1. 1. The effect of perfusion at pH 5.5 of a Heidenhain pouch on acid and pepsin secretion was studied in cats and rabbits. 2. 2. In basal conditions, acid secretion was not modified by the perfusion while pepsin secretion was increased in both species. 3. 3. After a meal, the perfusion did not modify the stimulation of acid secretion in cats nor the absence of secretions in rabbit. Stimulation of pepsin secretion was the same with or without perfusion in both species. 4. 4. In rabbit, in contrast to cat, pepsin and acid secretions were independent during the meal-induced phase of gastric secretion

SummaryWe have investigated the influence of long term oral anticoagulants (OAC) upon the plasma levels of prothrombin fragment 1 + 2 (F1 + 2), of thrombin-antithrombin III complexes (TAT) and of D-Dimer in 20 patients affected by a proximal deep vein thrombosis (DVT) diagnosed by ultrasonic duplex scanning. Patients (63 ± 17 years, mean ± SD) were sampled at the beginning of the OAC treatment (day 1), which was started 1 to 6 days after diagnosis confirmation and full heparinization, and then 8, 35 and 92 days after. The results were compared to those obtained in a blood donor population (39 ± 10 years) and to an age-matched healthy population (63 ± 19 years). The mean INR determined on days 8, 35 and 92 were almost identical (2.8 ± 0.7, 2.9 ± 0.9 and 2.8 ± 0.6 respectively). In contrast, highly significant variations of the three markers were recorded during the observation period. Eight days after the beginning of OAC, increased levels of TAT complexes were associated with subnormal levels of F1 + 2 suggesting persistence of a hypercoagulable state. On the further sampling times, TAT complexes were in the normal range while F1 + 2 were far below the normal range. Between day 1 and day 92, the levels of D-Dimer continuously decreased reflecting a long-term fibrinolytic process.This study clearly indicates that high INR are not systematically associated with very low F1 + 2 levels, particularly in the acute phase of thrombosis. Whether or not it is possible to reduce the intensity of the anticoagulant treatment 1 month after its initiation on the basis of very low levels of F1 + 2 can only be determined by prospective clinical trials.

Background: There is ample evidence that the incidence of operative cholangiography has declined since the introduction of laparoscopic cholecystectomy. It has been suggested that one of the reasons for this decline is the technical difficulty of cystic duct catheterization. A method of cholangiography which does not require such catheterization has been developed by others. This paper examines the ease and effectiveness of operative cholangiography performed by direct puncture of the gall-bladder. Methods: A retrospective study was made of the records of 380 consecutive patients who underwent attempted laparoscopic cholecystectomy. After exclusion of patients in whom laparoscopic cholecystectomy was abandoned in favour of open cholecystectomy and those patients in whom operative cholangiography was not attempted, the records of 325 patients were available to the study. The operation notes of each patient were studied to determine whether, in the operator’s opinion, a successful cholangiogram had been obtained. Results: There were 290 patients in whom cholangiography was attempted using direct gall-bladder puncture (cholecystocholangiography (CCC)). There were 35 patients in whom cholangiography was attempted via cystic duct cannulation (CDC). Cholecystocholangiography was successful in 86% of those cases in which it was attempted. Cystic duct cannulation was successful in 83% of those cases in which it was attempted. Conclusion: Cholecystocholangiography is a valid alternative to cystic duct catheterization in its ability to achieve intra-operative cholangiography in the setting of laparoscopic cholecystectomy.

Background The risk factors for progression of chronic kidney disease (CKD) in type 2 diabetes mellitus (DM) have not been fully elucidated. Although uncontrolled blood pressure (BP) is known to be deleterious, other factors may become more important once BP is treated. Methods All patients seen in the outpatient clinics of our hospital between January 1993 and September 2002 with type 2 DM and clinical evidence of CKD were evaluated. Progression of kidney disease was evaluated by rate of decline of glomerular filtration rate (GFR) as estimated from the simplified MDRD formula. Variables associated with progression in univariate analyses were examined by multivariate analysis to determine the factors independently associated with kidney disease progression. Results 343 patients (mean age 69 years; all male; 77% Caucasian) were studied. Mean BP, glycated hemoglobin, and serum cholesterol during the study period were 138/72 mmHg, 8.1%, and 4.8 mmol/L, respectively. Mean decline of GFR was 4.5 ml min-1 1.73 m2-1 yr-1 (range -14 to +32). Low initial serum albumin (p < 0.001), black race (p < 0.001), and degree of proteinuria (p = 0.002), but not blood pressure, glycated hemoglobin, or serum cholesterol, were independently associated with progression. Conclusion In a cohort of diabetic patients with CKD in whom mean BP was < 140/80 mmHg, the potentially remediable factors hypoalbuminemia and proteinuria but not blood pressure were independently associated with progression of kidney disease. Further understanding of the relationship between these factors and kidney disease progression may lead to beneficial therapies in such patients.

Hemodialysis using a bath with less-than-normal calcium level can cause hypocalcemia. The development of symptomatic hypocalcemia with resultant hypotension after the inadvertent use of a calcium-free dialysate in a maintenance hemodialysis patient is described. It is suggested that the occurrence of similar mishaps in the future can be reduced by close checking of dialysate concentrate labels.

We report on the case of a 54-year-old woman diagnosed as having culture-negative endocarditis (clinical and histopathologic evidence compatible with a recent episode of endocarditis). The responsibility of Chlamydia pneumoniae in this episode of endocarditis was suggested by a serological study and was then confirmed by the positive results of PCR and in situ hybridization tests with aortic and mitral valves tissues. To our knowledge, this is the first case of endocarditis due to C. pneumoniae confirmed by molecular biology-based techniques.