Improvement of antibody affinity by introduction of basic amino acid residues into the framework region

Antibodies are widely used not only as therapeutic agents but also as research tools and diagnostic agents, and extensive efforts have been made to generate antibodies that have higher affinity. It was recently reported that introduction of charged residues into the framework region of an antibody improved its affinity; however, the underlying molecular mechanism has not been elucidated. In this study, we used kinetic and thermodynamic analyses of the antibody–antigen interaction to investigate the molecular mechanism by which an antibody with introduced charged residues recognizes its antigen with higher affinity. The introduction of basic amino acid residues resulted in improvement of the affinity whereas the introduction of acidic residues weakened the interaction. For two mutant antigen-binding fragments (Fabs) with improved affinity (named K5- and R5-mutants), the balance between the association rate constant k_on and the dissociation rate constant k_off was distinct despite each mutant having the same number of charged residues. Moreover, thermodynamic analysis of the interactions in the transition state revealed a difference between the K5- and R5-mutants in terms of enthalpic energy change following formation of the encounter complex with the antigen. These results suggest that the affinity of the K5- and R5-mutants is improved by distinct mechanisms. Although the mutations destabilize the Fab and necessitate further studies, our strategy is expected to become a versatile and simple means to improve the affinity of antibodies to their antigens.     

The preparation and kinetic properties of multiple forms of chicken brain acetylcholinesterase

A method for preparing various forms of acetylcholinesterase (A ChE) from chicken brain has been developed and they have been characterized in terms of kinetic parameters such as K_m, rate constant (k), turnover number (k_p), specificity constant (k_sp), V_max and half-life (t_1/2). The solubility experiments show that, there are two major forms of A ChE i.e. water-soluble and membrane-bound A ChE (MBA ChE). The MBA ChE shows several subforms, and on the basis of percentage activity only three MBA ChE forms have been selected for complete characterization by various kinetic parameters. It was found that these three forms of MBA ChE demonstrate significant differences in their kinetic properties.     

The comparative gastrointestinal morphology of five species of muroid rodents found in Saudi Arabia

Meriones rex (King jird), Meriones libycus (Libyan jird), Acomys dimidiatus (Eastern spiny mouse), Acomys cahirinus (Egyptian spiny mouse), and Dipodillus dasyurus (Wagner's dipodil) are five species of small rodents of the superfamily Muroidea with distributions in Eastern Africa, Egypt, and the desert regions of the Arabian Peninsula. Water is scarce in these regions and may result in relatively low‐digestible food. The aim of the present study is to describe and compare the gastrointestinal tract morphology and morphometry of these five species in order to elucidate whether morphology is influenced by phylogeny or dietary preference. Each segment of the gastrointestinal tract of each species was macroscopically examined and the length and basal surface area of each segment was measured. Standard histologic procedures were performed to determine a surface enlargement factor to determine the mucosal luminal surface area. A unilocular‐hemiglandular stomach was observed in all the species examined. The caeca of all the species were long and arranged into a loose spiral toward the caecal tip with the ileocaecal and caeco‐colic openings positioned close together. Two rows of oblique folds could be observed in the proximal colon of all species except in D. dasyurus which had longitudinal folds. Morphometric analysis showed the largest stomach in A. cahirinus and the largest caecum and colon in M. libycus. All the species can be grouped in the family Muridae in two subfamilies and similarities were observed including the hemiglandular stomach and relatively large caecum. It could be concluded that phylogeny plays an important role in determining gastrointestinal morphology while diet plays a subordinate role in the desert rodents in the present study. J. Morphol. 275:980–990, 2014. © 2014 Wiley Periodicals, Inc.     

Are patients affected by chronic non-communicable diseases aware of their own clinical and laboratory parameters? A cross-sectional study from the south of Saudi Arabia

BackgroundPatient’s awareness of their clinical and laboratory parameters is an indicator of the degree of involvement in achieving their management goals. This investigation aimed to identify awareness of patients affected by chronic non-communicable diseases of their clinical and laboratory parameters and factors associated with the awareness.MethodsThis study was a cross-sectional investigation conducted in the Jazan region, between January and August 2020. Data was collected during phone interviews utilizing a semi-structured questionnaire. Odds ratios (ORs) were calculated to estimate the likelihood of awareness of each clinical and laboratory parameter according to the measured demographic variables.ResultsThe total number of recruited patients was 675. The mean age of participants was 53.7 years and the 28.7% of patients were illiterate. About 17% of the patient do not attend follow-up visits to any healthcare provider. When patients were asked about their parameters, 87% of them were able to report their body weight and 74% were able to report their height. However, less than half of patients were aware of their glycated hemoglobin level (HbA1c) (271/675 patients) and systolic blood pressure (BP) level (329/ 675 patients), and a minority were aware of their total cholesterol level (71/675 patients). Being female, resident in a rural area, illiterate, and older than 53 was strongly associated with high odds of limited awareness about their own clinical and laboratory parameters (P values < 0.05).ConclusionAwareness of patients affected by chronic non-communicable diseases of their own clinical and laboratory parameters in the Jazan region is sub-optimal where this limited awareness is likely to be associated with the lower engagement of patients with achieving their desired management targets.     

Effect of neurogenic acupoint cupping on high sensitive C-reactive protein and pain perception in female chronic pelvic pain: A randomized controlled trial

Objectives:To determine the effect of neurogenic acupoint dry cupping therapy on high sensitive C-reactive protein (hs-CRP) level, pain perception & intensity, and life impact of pelvic pain in women with chronic pelvic pain (CPP), with regard to the biological and neurophysiological impacts of dry cupping on acupoint.Methods:Thirty women with CPP were randomly divided into two equal groups; the study group received dry cupping on neurogenic acupoints plus lifestyle modifications for 8 weeks (n=15), while the control group received only lifestyle modifications for 8 weeks (n=15). Women were assessed pre- and post-rehabilitation program with the hs-CRP blood test, the short-form McGill Pain Questionnaire (SF-MPQ), and the pelvic pain impact questionnaire (PPIQ).Results:Comparing both groups post-treatment revealed that there were significant reductions in levels of hs-CRP, and scores of SF-MPQ & PPIQ (p<0.05) in the study group compared with the control group. Also, there were significant positive correlations between hs-CRP and both SF-MPQ “Visual Analogue Scale (VAS), Present Pain Intensity (PPI) index & Pain Rating Index (PRI)” and PPIQ (p<0.05).Conclusion:Neurogenic acupoint cupping therapy had significantly improving effects on the degree of inflammation, pain perception & intensity, and life impact of pelvic pain in women with CPP.