L-Glutamine therapy reduces endothelial adhesion of sickle red blood cells to human umbilical vein endothelial cells

BACKGROUND: We have previously demonstrated that therapy with orally administered L-glutamine improves nicotinamide adenosine dinucleotide (NAD) redox potential of sickle red blood cells (RBC). On further analysis of L-glutamine therapy for sickle cell anemia patients, the effect of L-glutamine on adhesion of sickle RBC to human umbilical vein endothelial cells (HUVEC) was examined. METHODS: The first part of the experiment was conducted with the blood samples of the 5 adult sickle cell anemia patients who had been on L-glutamine therapy for at least 4 weeks on a dosage of 30 grams per day compared to those of patient control group. In the second part of the experiment 6 patients with sickle cell anemia were studied longitudinally. Five of these patients were treated with oral L-glutamine 30 grams daily and one was observed without treatment as the control. t-test and paired t-test were used for determination of statistical significance in cross-sectional and longitudinal studies respectively. RESULTS: In the first study, the mean adhesion to endothelial cells with the autologous plasma incubated cells were 0.97 ± 0.45 for the treated group and 1.91 ± 0.53 for the nontreated group (p < 0.02). Similarly with lipopolysaccharide (LPS) incubated cells the mean adhesion to endothelial cells were 1.39 ± 0.33 for the treated group and 2.80 ± 0.47 for the untreated group (p < 0.001). With the longitudinal experiment, mean decrease in the adhesion to endothelial cells was 1.13 ± 0.21 (p < 0.001) for the 5 treated patients whereas the control patient had slight increase in the adhesion to endothelial cells. CONCLUSION: In these studies, oral L-glutamine administration consistently resulted in improvement of sickle RBC adhesion to HUVEC. These data suggest positive physiological effects of L-glutamine in sickle cell disease

Blood outgrowth endothelial cell migration and trapping in vivo: a window into gene therapy

Human blood outgrowth endothelial cells (hBOEC) may be useful delivery-cells for gene therapy. hBOEC have high expansion capacity and stable phenotype. If incorporated into blood vessels, hBOEC could release therapeutic agents directly into the blood stream. However, little is known about lodging and homing of hBOEC in vivo. We examined the homing patterns of hBOEC in mice, and explored extending cell-based FVIII gene therapy from mice to larger animals. hBOEC were injected into NOD/SCID mice to determine where they localize, how localization changes over time and if there were toxic effects on host organs. The presence of hBOEC in mouse organs was determined by qPCR and immunofluorescence microscopy. hBOEC lodged most notably in mouse lungs at 3 h, but by 24 h there were no differences between 9 organs. hBOEC longevity was assessed up to 7 months in vivo. hBOEC expanded well and then plateaued in vivo. hBOEC from older cultures expanded equally well in vivo as younger. hBOEC caused no noticeable organ toxicity up to 3 days post-injection. When mice were pretreated with antibodies to E-selectin, P-selectin or anti-α4 integrin prior to hBOEC injection, the number of hBOEC in lungs at 3h was inhibited. Preliminary studies infusing hemophilic dogs with autologous canine BOEC over-expressing FVIII (B-domain deleted) showed improvement in whole blood clotting times (WBCT). In conclusion, the survivability, expandability and lack of toxicity of BOEC in vivo indicate that they may be valuable host cells for gene therapy

Hermafroditismo verdadeiro na espécie suína.

Resumo: Os hermafroditas verdadeiros são animais com intersexualidade, nos quais as gônadas masculinas e femininas se desenvolvem simultaneamente no mesmo indivíduo. Este trabalho relata um caso de hermafrodita na espécie suína após descarte de uma fêmea por diagnóstico de anestro. Tratava-se de uma fêmea com 222 dias de idade e 150 kg a qual havia sido selecionada para reprodução e permanecia em anestro. O indivíduo foi descartado e enviado para o abate em frigorífico sob inspeção federal. Após o abate foi observado no sistema genital aspectos anatômicos sugestivos de hermafroditismo. O sistema genital foi coletado, inspecionado macroscopicamente e conservado em formol a 10% para avaliação microscópica. Durante a análise macroscópica foi verificada a presença de vulva e vagina, contendo um pênis rudimentar. O útero, a cérvix, os cornos uterinos e oviduto estavam presentes e com ausência de anormalidade. Havia um ovário esquerdo com aspectos morfológicos de ciclicidade e folículos contendo oócitos de alta qualidade. À direita, entretanto, foi constatada a presença de um testículo, epidídimo e plexo pampiniforme. A avaliação histopatológica comprovou os achados macroscópicos, demonstrando um ovário contendo folículos, corpos lúteos e cistos foliculares; plexo pampiniforme; testículo e epidídimo com aspecto histológico normal, porém, com ausência de células espermatogênicas. Assim, com base nos achados macro e microscópicos descritos nesse relato de caso, o indivíduo foi considerado como hermafrodita verdadeiro. Abstract: True hermaphrodites are intersexual animals, in which both male and female gonads develop simultaneously in the same individual. This paper reports a case of a true hermaphrodite in the swine species after its culling for anestrus. The specimen was a female of 222 days of age and 150 kg of body weight, that was selected for reproduction and was permanently in anestrus. This individual was culled and sent to an abattoir under federal inspection. After slaughtering, it was observed in the reproductive tract anatomical aspects that suggested hermaphroditism. The reproductive tract was collected, evaluated macroscopically, and preserved in formaldehyde at 10 % to be evaluated by light microscopy. During the macroscopic analysis, the presence of a vulva and a vagina with a rudimentary penis was detected. The uterus, the cervix, uterine horns, and the oviduct were present and abnormalities were not detected. The left ovary exhibited morphological signs of cyclicity and contained follicles with high quality oocytes. On the right side, however, it was observed a testicle, epididymis, and a pampiniform plexus. The histopathological assessment corroborated the macroscopic findings, which demonstrated that the ovary had follicles, corpus luteum, and follicular cysts, and pampiniform plexus, testicle and epididymis were histologically normal, though with the absence of spermatogenic cells. Thus, based on the macro and microscopic findings described herein, the individual was considered a true hermaphrodite

Measurement of inclusive D*+- and associated dijet cross sections in photoproduction at HERA

Inclusive photoproduction of D*+- mesons has been measured for photon-proton centre-of-mass energies in the range 130 < W < 280 GeV and a photon virtuality Q^2 < 1 GeV^2. The data sample used corresponds to an integrated luminosity of 37 pb^-1. Total and differential cross sections as functions of the D* transverse momentum and pseudorapidity are presented in restricted kinematical regions and the data are compared with next-to-leading order (NLO) perturbative QCD calculations using the "massive charm" and "massless charm" schemes. The measured cross sections are generally above the NLO calculations, in particular in the forward (proton) direction. The large data sample also allows the study of dijet production associated with charm. A significant resolved as well as a direct photon component contribute to the cross section. Leading order QCD Monte Carlo calculations indicate that the resolved contribution arises from a significant charm component in the photon. A massive charm NLO parton level calculation yields lower cross sections compared to the measured results in a kinematic region where the resolved photon contribution is significant.Comment: 32 pages including 6 figure