Immunological role of CD4<Superscript>+</Superscript>CD28<Superscript>null</Superscript> T lymphocytes,natural killer cells,and interferon-gamma in pediatric patients with sickle cell disease: relation to disease severity and response to therapy

Sickle cell disease (SCD) is associated with alterations in immune phenotypes. CD4⁺CD28^null T lymphocytes have pro-inflammatory functions and are linked to vascular diseases. To assess the percentage of CD4⁺CD28^null T lymphocytes, natural killer cells (NK), and IFN-gamma levels, we compared 40 children and adolescents with SCD with 40 healthy controls and evaluated their relation to disease severity and response to therapy. Patients with SCD steady state were studied, focusing on history of frequent vaso-occlusive crisis, hydroxyurea therapy, and IFN-gamma levels. Analysis of CD4⁺CD28^null T lymphocytes and NK cells was done by flow cytometry. Liver and cardiac iron overload were assessed. CD4⁺CD28^null T lymphocytes, NK cells, and IFN-gamma levels were significantly higher in patients than controls. Patients with history of frequent vaso-occlusive crisis and those with vascular complications had higher percentage of CD4⁺CD28^null T lymphocytes and IFN-gamma while levels were significantly lower among hydroxyurea-treated patients. CD4⁺CD28^null T lymphocytes were positively correlated to transfusional iron input while these cells and IFN-gamma were negatively correlated to cardiac T2* and duration of hydroxyurea therapy. NK cells were correlated to HbS and indirect bilirubin. Increased expression of CD4⁺CD28^null T lymphocytes highlights their role in immune dysfunction and pathophysiology of SCD complications.     

Vildagliptin Recruits Regulatory T Cells in Patients Undergoing Primary Percutaneous Coronary Intervention

Regulatory T cells (Treg) has been documented to be protective against myocardial ischemia–reperfusion injury (MIRI). The administration of drugs which recruit Treg cells may participate in the cardioprotection of MIRI. The purpose of the present study was to investigate whether the add-on vildagliptin (vild) to standard treatment of MIRI prior to reperfusion could increase Treg recruitment, anti-inflammatory, and antioxidant effects of the standard treatment or not. Sixty diabetic patients with ST-segment elevation myocardial infarction were randomly divided into two equal groups: control group was given the standard medical treatment and vild group was given the standard medical treatment plus vild. There were no statistical differences between the mean of percentage of changes in nitric oxide, ischemia modified albumin, highly sensitive C reactive protein, and interferon-gamma levels in the studied groups. While, the percentages of changes of myeloperoxidase level, CD4+CD25+ Treg cells count, and transforming growth factor-beta1 level were significantly higher in vild group compared with control group. We concluded that addition of vild to standard medical treatment of MIRI could increase its effectiveness through recruitment of CD4+CD25+ Treg cells.     

Follow-up of the patients after bone marrow transplantation for red blood cell chimerism using flow cytometry

The importance of chimerism is that it is useful to demonstrate engraftment. It allows for early intervention which might facilitate treatment of emerging relapse in leukemia, and it may help to prevent graft rejection. The aim of this study is to evaluate mixed red cell population and red blood cell chimerism after hematopoietic stem cell transplantation to predict the final outcome of mixed chimerism which may help in deciding interventions and preventing graft rejection. This study was conducted on 22 bone marrow-transplanted patients admitted to Bone Marrow Transplantation (BMT) Unit, Nasser institute, using simple agglutination method and flow cytometery (FCM) technique. In our study, three recipients had complete chimerism after 6 month which was detected by agglutination method, and one recipient had complete chimerism after 1 year which was detected by both simple agglutination method and FCM. Two recipients had mixed-field agglutination after 30 days which was detected by agglutination method and FCM. Evaluating erythrocyte repopulation by the agglutination method is feasible, easy, and cost-effective. FCM analysis is a simple, sensitive test and an objective method, which could be used as a protocol for follow-up to detect chimerism after allogeneic BMT. However, due to the difficulty found in retrieving the patients for follow-up, we suggest the use of alternative methods for detection of chimerism as variable number of tandem repeats and XY fluorescence in situ hybridization probe.     

Antifungal and Antihepatotoxic Effects of Sepia Ink Extract Against Oxidative Stress as a Risk Factor of Invasive Pulmonary Aspergillosis in Neutropenic Mice

Background

There is a great need for novel strategies to overcome the high mortality associated with invasive pulmonary aspergillosis (IPA) in immunocompromised patients. To evaluate the antifungal and antihepatotoxic potentials of Sepia ink extract, its effect on liver oxidative stress levels was analyzed against IPA in neutropenic mice using amphotercin B as a reference drug.

Materials and Methods

Eighty neutropenic infected mice were randomly assigned into four main groups. The 1^st group was treated with saline, neutropenic infected (NI), the 2^nd group was treated with ink extract (200 mg/kg) (IE) and the 3^rd group was treated with amphotericin B (150 mg/kg) (AMB) and 4^th group was treated with IE plus AMB. Treatment was started at 24 h after fungal inoculation (1×10⁹ conidia/ml).

Results

The present study revealed good in vitro and in vivo antifungal activity of IE against A. fumigatus. IE significantly reduced hepatic fungal burden and returns liver function and histology to normal levels. Compared with the untreated infected group, mice in the IE, AMB, and IE+ AMB groups had increased glutathione reduced (GSH) and superoxide dismutase (SOD) and significantly reduced malondialdehyde (MDA) levels at 24 and 72 h after inoculation with A. fumigatus conidia.

Conclusion

It is then concluded that in combination with antifungal therapy (AMB), IE treatment can reduce hepatic fungal burden, alleviate hepatic granulomatous lesions and oxidative stress associated with IPA in neutropenic mice     

Effect of Potassium Bromate on the Liver of Adult Male Albino Rat and A Possible Protective Role of Vitamin C: Histological,Immunohistochemical, and Biochemical Study

Potassium bromate (KBrO₃) is a food additive which is used primarily as a maturing agent for flour. It is proved as a toxic agent with significant reduction in the activities of antioxidant capacity. The therapeutic efficacy of vitamin C as antioxidant may provide a possible solution to KBrO₃ mediated oxidative damage. Twenty four adult male albino rats were used to evaluate the protective role of vitamin C against KBrO₃ induced hepatotoxicity and divided into four groups; Group 1 (control), Group 2: received 30 mg/Kg/day vitamin C orally for 4 weeks, Group 3: received 20 mg/Kg/dose KBrO₃ orally twice weekly for 4 weeks and Group 4: received both KBrO₃ and vitamin C. Liver specimens were processed for histological study by light and electron microscopes and stained immunohistochemically to detect glial fibriller acidic protein (GFAP). Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were estimated as well as the levels of malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) activities in all dissected tissues were determined. KBrO₃ induced histological alterations in the form of degeneration, cellular infiltration and significant increase in collagen deposition in portal tracts with a significant increase in immunoexpression of GFAP. Significant rise in serum levels of AST, ALT, and MDA in liver tissues were recorded. However, levels of GSH and SOD were significantly decreased. Most of these changes were improved by vitamin C treatment. In conclusion, vitamin C ameliorates the histological and biochemical alterations of the liver induced by KBrO₃. Anat Rec, 299:1256–1269, 2016. © 2016 Wiley Periodicals, Inc.     

Novel 5-flucytosine-resistant clade of Candida dubliniensis from Saudi Arabia and Egypt identified by Cd25 fingerprinting

DNA fingerprinting of Candida dubliniensis isolates using the species-specific probe Cd25 previously showed that this species consists of two distinct groups, termed Cd25 group I and Cd25 group II. The present study investigated the population structure of 30 C. dubliniensis oral isolates from Saudi Arabia and Egypt using Cd25 fingerprinting and rRNA gene internal transcribed spacer region-based genotyping. Cd25 fingerprinting analysis of these isolates revealed two distinct populations, the first of which consisted of 10 closely related genotype 1 isolates (average similarity coefficient [S(AB)] value, 0.86). The second population of 20 isolates was much more heterogeneous (average S(AB) value, 0.35) and consisted of two distinct subpopulations, one of which consisted of genotype 3 isolates (n = 13) and the other of genotype 4 isolates (n = 7). A mixed dendrogram generated from the fingerprint data from the 30 Saudi Arabian and Egyptian isolates, 5 Israeli isolates, and 51 previously characterized international isolates (32 of Cd25 group I and 19 of Cd25 group II) revealed the presence of three distinct main clades. The first corresponded to the previously described Cd25 group I and contained all the Saudi Arabian, Egyptian, and Israeli genotype 1 isolates mixed with international isolates. The second clade corresponded to the previously described Cd25 group II and contained three Israeli isolates, one genotype 2 isolate, one genotype 3 isolate, and a genotype 4 variant isolate, which were mixed with international isolates. The third clade has not been described before and consisted solely of the 20 Saudi Arabian and Egyptian genotype 3 and 4 isolates identified in this study and a previously described genotype 4 Israeli isolate. All 20 Cd25 group III isolates exhibited high-level resistance to 5-flucytosine (MIC > or = 128 microg/ml), whereas all Cd25 group I and Cd25 group II isolates tested (10 Saudi Arabian and Egyptian, 16 Israeli, and 24 international) were susceptible to 5-flucytosine (MIC < or = 0.125 microg/ml). The results of this study show for the first time the presence of a novel 5-flucytosine-resistant clade of C. dubliniensis (Cd25 group III) that is predominant among isolates from Saudi Arabia and Egypt and absent from a previously characterized international collection of 98 isolates from 15 countries.     

Longitudinal growth of infants in Qatar: Comparison with WHO and CDC growth standards

Longitudinal growth data for infants in Qatar were compared to growth standards published by the CDC and WHO. 300 randomly selected full-term normal infants (150 males, 150 females) in Qatar were followed-up and weight and length were sequentially recorded at 2 months, 4 months, 6 months, 12 months and 18 months age. The mean length for age of girls was higher than those published by the CDC and WHO at 12 and 18 months of age. Using the CDC standard for weight for length detected more wasted infants (9.0% and 6.5%) compared to using WHO standards (6.27% and 6.0%) for males and females, respectively. When WHO and CDC standards are compared, more infants were identified as overweight when the former were used. The WHO standards are preferable because they are based on a leaner breastfed reference and because overweight is likely to be a greater problem in Qatar in the future.     

Carbetocin versus oxytocin for prevention of postpartum hemorrhage in obese nulliparous women undergoing emergency cesarean delivery

Objective: To assess and compare the effectiveness and safety of single IV polus dose of carbetocin, versus IV oxytocin infusion in the prevention of PPH in obese nulliparous women undergoing emergency Cesarean Delivery.

Methods: A double-blinded randomized-controlled trial was conducted on 180 pregnant women with BMI >30. Women were randomized to receive either oxytocin or carbetocin during C.S. The primary outcome measure was major primary PPH >1000?ml within 24?h of delivery as per the definition of PPH by the World Health Organization Secondary outcome measures were hemoglobin and hematocrit changes pre- and post-delivery, use of further ecobolics, uterine tone 2 and 12-h postpartum and adverse effects.

Results: A significant difference in the amount of estimated blood loss or the incidence of primary postpartum haemorrhage (>1000?ml) in both groups. Haemoglobin levels before and 24-h postpartum was similar. None from the carbetocin group versus 71.5% in oxytocin group needed additional utrotonics (p?<?0.01). The uterine contractility was better in the carbetocin group at 2, and 12-h postpartum (p?<?0.05).

Conclusions: A single 100-µg IV carbetocin is more effective than IV oxytocin infusion for maintaining adequate uterine tone and preventing postpartum bleeding in obese nulliparous women undergoing emergency cesarean delivery, both has similar safety profile and minor hemodynamic effect.     

Age related IGF-I changes and IGF-I generation in thalassemia major

We measured serum concentrations of insulin like growth factor-I (IGF-I) in 20 thalassemic males with short stature (height SDS <-2) and/or slow growth velocity (GV <-1 SD) throughout their childhood and adolescence, compared these data with normal reference data validated in our lab, and evaluated their growth hormone secretion in response to clonidine and glucagon stimulation. We also performed IGF-I generation test on 26 patients with beta thalassemia major (BTM) before and after blood transfusion to evaluate the effect of increased hemoglobin (Hb) on IGF-I and its response to GH. We obtained the following results. 1) No statistical difference in age, HSDS, target height SDS or bone age was observed between BTM patients with growth hormone deficiency (GHD) compared to those with normal GH secretion (GHS). 2) The age-related levels in serum total IGF-I in thalassemic males were significantly decreased from early childhood to 18 years of age compared to normal subjects. Thalassemic males with GHD did not show any significant peak of IGF-I levels until 18 years of age, whereas thalassemic males with normal GH response to provocation (GHS) achieved a significant peak level of IGF-I that was attenuated and late compared to normal males. The basal serum IGF-I concentrations at different ages did not differ between the GHD and GHS groups until the age of 12 years. After 12 years of age, IGF-I levels were significantly higher in thalassemic children with GHS. A significant increase in the circulating basal IGF-I concentrations from 53 +/-35 ug/l to 82.6 +/- 39 ug/L was achieved with increasing Hb concentration after blood transfusion. The serum total IGF-I levels increased significantly with the administration of human growth hormone (hGH) for 4 days, both before and after blood transfusion. The peak IGF-I response to GH injections did not differ before compared to after blood transfusion. The percent increment of IGF-I levels generated after GH injections was higher in thalassemic children with GHD as compared to those with GHS both before and after blood transfusion. In conclusion, our results showed that agerelated serum IGF-I concentrations were significantly lower in short thalassemic patients, with and without GHD, during childhood and adolescence, compared to normal standards. Correction of anemia significantly increased serum concentration of IGF-I but does not affect the increase of IGF-I in response to GH stimulation.