Purification of Proteins: Between Meaning and Different Methods

Purification of protein attracts the scientists’ attention toward science in 1926, as Somnar started purification and crystallization of urease from yeast. As years go forward, protein purification strategies updated from using dextrose passing through DEAE-cellulose and ended by affinity chromatography. In this chapter we will describe some of the concepts and the differences between traditional and novel purification methods, in order to point out how a researcher can start the protein purification techniques. Different fundamental purification steps from plant sample will be discussed in this chapter including isolation, concentration, ion exchanger, and affinity chromatography, as well as the important additives that a researcher should add in order to gain a high purification fold

Cooperation and Underlay Mode Selection in Cognitive Radio Network

In this research, we proposes a new method for cooperation and underlay mode selection in cognitive radio networks. We characterize the maximum achievable throughput of our proposed method of hybrid spectrum sharing. Hybrid spectrum sharing is assumed where the Secondary User (SU) can access the Primary User (PU) channel in two modes, underlay mode or cooperative mode with admission control. In addition to access the channel in the overlay mode, secondary user is allowed to occupy the channel currently occupied by the primary user but with small transmission power. Adding the underlay access modes attains more opportunities to the secondary user to transmit data. It is proposed that the secondary user can only exploits the underlay access when the channel of the primary user direct link is good or predicted to be in non-outage state. Therefore, the secondary user could switch between underlay spectrum sharing and cooperation with the primary user. Hybrid access is regulated through monitoring the state of the primary link. By observing the simulation results, the proposed model attains noticeable improvement in the system performance in terms of maximum secondary user throughput than the conventional cooperation and non-cooperation schemes

Vitamin D deficiency and low hemoglobin level as risk factors for severity of acute lower respiratory tract infections in Egyptian children: A case-control study

AbstractObjectiveAcute lower respiratory tract infection (ALRTI) is an important cause of morbidity in the developed world, and both morbidity and mortality in the developing world. Vitamin D has a major role in both acquired and innate immunity. Anemic children have less oxygen carrying capacity of blood. This study was done to determine the relation between vitamin D deficiency, anemia and the severity of ALRTIs in hospitalized children.MethodsThis study included 96 hospitalized infants with ALRTI, 48 diagnosed with pneumonia and 48 with bronchiolitis. Mean age was 10.67±3.143months. Matched age and sex infants with no respiratory illness were included. Serum 25 hydroxy vitamin D was measured in all cases and controls by Radio-immune assay. Hemoglobin level was measured by Coulter.ResultsVitamin D deficiency and low hemoglobin level were positively correlated with the severity of ALRTIs (r=0.798 and P=0.001) and (r=0.708, P=0.028), respectively. Low vitamin D level was significantly correlated with low hemoglobin level (r=0.708, P=0.028).ConclusionVitamin D deficiency was associated with severity of ALRTIs. Low hemoglobin level was more prevalent in those children. Improving the nutritional status in children by preventing vitamin D deficiency and low hemoglobin might influence the outcome of children with ALRTI

LIPID METABOLISM IN DIABETIC RATS AS AFFECTED BY CANOLA AND MUSTARD SEED SPROUTS

Canola (Brassica juncea L.) and mustard (Sinapis alba L.) seed sprout effects on diabetic rats  have no available information and to clarify their effects, both sprouts were investigated in streptozotocin (STZ) induced diabetic and normal rats. Rats were fed on a semi-modified diet containing 10% of canola or mustard sprouted using tap or saline water for sprouting ad-libitum for 6 weeks. STZ showed increases in blood sugar, low density lipoprotein cholesterol (LDL-c), vary low density lipoprotein cholesterol (VLDL-c) and triglycerides (TG). The addition of canola and mustard with or without salinty at 10% to diabetic rats diet as semimodified diet resulted a significant decrease in blood glucose, TG and VLDL-c and data was more pronounced using mustard sprouted or saline water without changes in the HDL-c   parameter. These results showed that canola and mustard especially mustard sprouted in saline water had a hypoglycemic activity in diabetic rats and partly improved lipid metabolism in the experimental rats, with non-toxic to rats in doses given  over 6 weeks period in this study

The role of oxidative stress in ovarian toxicity induced by haloperidol and clozapine—a histological and biochemical study in albino rats

Oxidative stress has been implicated in reproductive toxicity induced by antipsychotics (APs). This study aims to further investigate the role of AP-induced oxidative stress in reproductive dysfunction. Thirty adult female albino rats were divided into three groups including a control group (n = 10) receiving distilled water, HAL group (n = 10) receiving haloperidol (HAL) (2 mg/kg/day), and CLZ group (n = 10) receiving clozapine (CLZ) (20 mg/kg/day). After 28 days, the rats were anesthetized, blood was withdrawn from their hearts, and ovaries were removed before they were sacrificed. Serum prolactin concentrations were measured. For each rat, one ovary was used for biochemical studies including mitochondrial complexes I and III activities and oxidative stress markers (lipid peroxidation, super oxide dismutase [SOD], catalase [CAT], and reduced glutathione [GSH]). The other ovary was used for histopathological examination and immunohistochemistry staining for p53 and Ki-67. HAL-treated rats showed significantly (p ≤0.001) higher serum prolactin concentrations compared with other groups. HAL significantly inhibited complexes I (p ≤ 0.001) and III activities (p ≤ 0.05), while CLZ inhibited only complex I (p ≤ 0.001). Lipid peroxidation was increased by HAL (p ≤ 0.001) and CLZ (p ≤ 0.01). HAL caused significant (p ≤ 0.001) reductions in SOD, CAT, and GSH. CLZ caused a significant decrease in SOD (p ≤ 0.001) and GSH (p ≤ 0.01) with no effect on CAT. Histopathological studies of CLZ- and HAL-treated ovaries showed features suggestive of hyperprolactinemia and oxidative stress. Ki-67- and P53-immunostained sections were suggestive of disruption of cellular proliferation. These findings support the hypothesis that HAL and CLZ induce reproductive dysfunction through mechanisms involving ovarian mitochondrial dysfunction and oxidative stress

Viral etiology, seasonality and severity of hospitalized patients with severe acute respiratory infections in the Eastern Mediterranean Region, 2007-2014.

INTRODUCTION: Little is known about the role of viral respiratory pathogens in the etiology, seasonality or severity of severe acute respiratory infections (SARI) in the Eastern Mediterranean Region. METHODS: Sentinel surveillance for SARI was conducted from December 2007 through February 2014 at 20 hospitals in Egypt, Jordan, Oman, Qatar and Yemen. Nasopharyngeal and oropharyngeal swabs were collected from hospitalized patients meeting SARI case definitions and were analyzed for infection with influenza, respiratory syncytial virus (RSV), adenovirus (AdV), human metapneumovirus (hMPV) and human parainfluenza virus types 1-3 (hPIV1-3). We analyzed surveillance data to calculate positivity rates for viral respiratory pathogens, describe the seasonality of those pathogens and determine which pathogens were responsible for more severe outcomes requiring ventilation and/or intensive care and/or resulting in death. RESULTS: At least one viral respiratory pathogen was detected in 8,753/28,508 (30.7%) samples tested for at least one pathogen and 3,497/9,315 (37.5%) of samples tested for all pathogens-influenza in 3,345/28,438 (11.8%), RSV in 3,942/24,503 (16.1%), AdV in 923/9,402 (9.8%), hMPV in 617/9,384 (6.6%), hPIV1 in 159/9,402 (1.7%), hPIV2 in 85/9,402 (0.9%) and hPIV3 in 365/9,402 (3.9%). Multiple pathogens were identified in 501/9,316 (5.4%) participants tested for all pathogens. Monthly variation, indicating seasonal differences in levels of infection, was observed for all pathogens. Participants with hMPV infections and participants less than five years of age were significantly less likely than participants not infected with hMPV and those older than five years of age, respectively, to experience a severe outcome, while participants with a pre-existing chronic disease were at increased risk of a severe outcome, compared to those with no reported pre-existing chronic disease. CONCLUSIONS: Viral respiratory pathogens are common among SARI patients in the Eastern Mediterranean Region. Ongoing surveillance is important to monitor changes in the etiology, seasonality and severity of pathogens of interest

Antimicrobial resistance genes in pathogenic Escherichia coli isolated from diseased broiler chickens in Egypt and their relationship with the phenotypic resistance characteristics Mohamed M. Amer, Hoda M. Mekky, Aziza M. Amer and Hanaa S. Fedawy

Aim: The aim of this study was to determine the relationship between phenotypic resistance and genotypic resistance of isolated serotyped pathogenic Escherichia coli isolates from the clinically diseased broiler. Materials and Methods: A total of 160 samples (heart, liver, kidney, and lung) were collected from 18 to 34 days old clinically diseased broiler from 40 broiler farms (3-5 birds/farm) reared in Giza and Kaluobaia Governorates for the isolation of pathogenic E. coli. Various E. coli isolates were tested for the pathogenicity based on Congo red (CR) dye binding assay. The obtained CR-positive E. coli isolates were subjected to serological identification using slide agglutination test. Disc diffusion test was used to study the sensitivity pattern of E. coli isolates to available 12 antibiotics. Polymerase chain reaction was performed for the detection of antimicrobial resistance genes in the studied pathogenic E. coli isolates. Results: The results revealed that 56 samples (35 %) were positive for E. coli. The results of the CR assay indicates that 20 isolates of 56 (35.7%) were positive and 36 isolates (64.3%) were negative. Identified E. coli serotypes of CR-positive isolates were 1 (O24), 2 (O44), 2 (O55), 5 (O78), 2 (O86), 1 (124), 3 (O127), 1 (O158), and 3 untyped. Resistance rate in disc diffusion test was 85% to oxytetracycline and kanamycin; 80% to ampicillin (AMP), clindamycin, and streptomycin (S); 75% to enrofloxacin; 65% to chloramphenicol; 55% to cefotaxime and gentamicin (CN); 45% to trimethoprim+sulfamethoxazole; 35% to erythromycin (ERI); and 30% to oxacillin. All strains are multidrug-resistant (MDR). Antibacterial resistance genes CITM, ere, aac (3)-(IV), tet(A), tet(B), dfr(A1), and aad(A1) were detected in 14 (70%), 12 (60%), 12 (60%), 8 (40%), 11 (55%), 8 (40%), and 9 (45%) of tested 20 isolates, respectively. Multidrug resistance was detected in the form of resistance to 42%-83.3% of tested 12 antibiotics. Three isolates (15%) of 20 tested isolates showed a relationship between phenotype and genotype and 17 (85%) showed irregular relation. Strains are sensitive and show resistant gene (P-G+) presented in three isolates for AMP (beta-lactam), one for ERI (Macrolide), as well as five isolates for trimethoprim (pyrimidine inhibitor). E. coli isolates had resistance and lacked gene (P+ G-) reported meanly in one isolate for CN (aminoglycoside), two isolates for tetracycline, four isolates for ERI, seven isolates for trimethoprim, and eight isolates for S (aminoglycoside). Conclusions: The study demonstrates that E. coli is still a major pathogen responsible for disease conditions in broiler. E. coli isolates are pathogenic and MDR. Responsible gene was detected for six antibiotics in most of the isolates, but some do not show gene expression, this may be due to few numbers of resistance genes tested or other resistance factors not included in this study

Molecular identification of Mycoplasma synoviae from breeder chicken flock showing arthritis in Egypt

Aim: Arthritis is one of the most economic problems facing poultry industry worldwide. The study was done to detect possible causes of arthritis in breeder chicken flock with emphasis on molecular identification of Mycoplasma synoviae (MS). Materials and Methods: This study was carried on chicken from broiler breeder flock of 57 weeks' age in Dakahlia, Egypt, suffered from arthritis with frequently 5-7% decrease in egg production, reduced fertility, and hatchability. Forty blood samples were randomly collected from individual birds in sterile tubes and used for serum separation. Serum samples were tested using serum plate agglutination (SPA) test against colored antigens for Mycoplasma gallisepticum (MG), MS, and Salmonella gallinarum-pullorum (SGP). On the other hand, 24 joint samples were collected. Of those 24 samples, 12 joint samples were subjected to bacteriological examination, while the other 12 were utilized for molecular diagnosis by polymerase chain reaction (PCR) for MS and avian reovirus (ARV). Results: SPA test results revealed the presence of antibodies against MG, MS, and SGP in tested sera in rates of 14/40 (35%), 35/40 (87.5%), and 9/40 (22.5%), respectively. Furthermore, 19 bacterial isolates were recognized from joint samples and identified as five Staphylococcus spp., nine Escherichia coli, three SGP, one Citrobacter, and one Proteus. The identified Staphylococcal isolates were three coagulase-positive staphylococci (two Staphylococcus aureus and one Staphylococcus hyicus) and two coagulase-negative staphylococci (one Staphylococcus epidermidis and one Staphylococcus lentus), while E. coli isolate serotypes were 1 O11, 2 O55, 3 O78, 1 O124, 1 O125, and 1 untyped. PCR proved that 12/12 (100%) samples were positive for MS variable lipoprotein hemagglutinin A (vlhA) gene, while ARV was not diagnosed in any of the examined samples. Four amplified vlhA gene of MS isolates (named MS-2018D1, MS-2018D2, MS-2018D3, and MS-2018D4) was successfully sequenced. Analysis of phylogenetic tree revealed the presence of 100% identity between each two sequenced isolates (isolates MS-2018D1 and MS-2018D4 and also isolates 2018D2 and MS-2018D3). However, the nucleotide similarity between four isolates was 88.6%. On the other hand, our field isolates MS-2018D1, MS-2018D4, MS-2018D2, and MS-2018D3 showed nucleotide identity with vaccine strain MS-H 98.4%, 98.4%, 88.1%, and 88.1%, respectively. Furthermore, the nucleotide similarities with field strains from Argentina ranged between 87.8% and 98.6%. Conclusion: Four field isolates of MS were identified in examined broiler breeder flock. A phylogenetic study of these isolates revealed the variation between isolated MS strains and vaccine strain. Therefore, further studies are required for evaluating the vaccine efficacy against the present field isolates of MS. In addition, application of MS immunization of breeder flocks is necessary for proper control of the disease

LIPID METABOLISM IN DIABETIC RATS AS AFFECTED BY CANOLA AND MUSTARD SEED SPROUTS

Canola (Brassica juncea L.) and mustard (Sinapis alba L.) seed sprout effects on diabetic rats  have no available information and to clarify their effects, both sprouts were investigated in streptozotocin (STZ) induced diabetic and normal rats. Rats were fed on a semi-modified diet containing 10% of canola or mustard sprouted using tap or saline water for sprouting ad-libitum for 6 weeks. STZ showed increases in blood sugar, low density lipoprotein cholesterol (LDL-c), vary low density lipoprotein cholesterol (VLDL-c) and triglycerides (TG). The addition of canola and mustard with or without salinty at 10% to diabetic rats diet as semimodified diet resulted a significant decrease in blood glucose, TG and VLDL-c and data was more pronounced using mustard sprouted or saline water without changes in the HDL-c   parameter. These results showed that canola and mustard especially mustard sprouted in saline water had a hypoglycemic activity in diabetic rats and partly improved lipid metabolism in the experimental rats, with non-toxic to rats in doses given  over 6 weeks period in this study