Serum-free generation and quantification of functionally active Leukemia-derived DC is possible from malignant blasts in acute myeloid leukemia and myelodysplastic syndromes

Functional dendritic cells (DC) are professional antigen presenting cells (APC) and can be generated in vitro from leukemic cells from acute myeloid leukemia AML patients, giving rise to APC of leukemic origin presenting leukemic antigens (DC_leu). We have already shown that DC can be successfully generated from AML and myeloplastic syndromes (MDS) cells in serum-free standard medium (X-vivo + GM-CSF + IL-4 +TNF + FL) in 10–14 days. In this study, we present that DC counts generated from mononuclear cells (MNC) varied between 20% (from 55 MDS samples), 34% (from 100 AML samples) and 25% (from 38 healthy MNC samples) medium. Between 53% and 58% of DC are mature CD83⁺ DC. DC harvests were highest in monocytoid FAB types (AML-M4/M5, MDS-CMML) and independent from cytogenetic risk groups, demonstrating that DC-based strategies can be applied for patients with all cytogenetic risk groups. Proof of the clonal derivation of DC generated was obtained in five AML and four MDS cases with a combined FISH/immunophenotype analysis (FISH-IPA): The clonal numerical chromosome aberrations of the diseases were regularly codetectable with DC markers; however, not with all clonal cells being convertible to leukemia-derived DC_leu (on average, 53% of blasts in AML or MDS). To the contrary, not all DC generated carried the clonal aberration (on average, 51% of DC). In 41 AML and 13 MDS cases with a suitable antigen expression, we could confirm FISH-IPA data by Flow cytometry: although DC_leu are regularly detectable, on average only 57% of blasts in AML and 64% of blasts in MDS were converted to DC_leu. After coculture with DC in mixed lymphocyte reactions (MLR), autologous T cells from AML and MDS patients proliferate and upregulate costimulatory receptors. The specific lysis of leukemic cells by autologous T cells could be demonstrated in three cases with AML in a Fluorolysis assay. In six cases with only few DC_leu or few vital T cells available after the DC/MLR procedure, no lysis of allogeneic or autologous leukemic cells was seen, pointing to the crucial role of both partners in the lysis process. We conclude: (1) the generation of DC is regularly possible in AML and also in MDS under serum-free conditions. (2) Clonal/leukemia-derived DC_leu can be regularly generated from MDS and AML-MNC; however, not with all blasts being converted to DC_leu and not all DC generated carrying leukemic markers. We recommend to select DC_leu for vaccinations or ex vivo T-cell activations to avoid contaminations with non-converted blasts and non-leukemia-derived DC and to improve the harvest of specific, anti-leukemic T cells. DC and DC-primed T cells could provide a practical strategy for the immunotherapy of AML and MDS.     

Prognostic value of GLUT-1 expression in ovarian surface epithelial tumors: a morphometric study

OBJECTIVE: To investigate the reported increase in the expression of the glucose transporter GLUT-1 in borderline and malignant ovarian epithelial tumors and its relationship to prognosis. STUDY DESIGN: In this study, areas in which immunohistochemical membranous staining with GLUT-1 were most evident were selected, and the proportions of GLUT-1 expression in 46 benign, 11 borderline and 42 malignant cases of ovarian epithelial tumors were determined quantitatively with a computer and Zeiss Vision KS 400 3.0 (G?ttingen, Germany) for Windows (Microsoft, Redmond, Washington, U.S.A.) image analysis. RESULTS: GLUT-1 expression was determined in all borderline tumors (11 of 11) and in 97.6% of malignant tumors (41 of 42). No GLUT-1 expression was observed in benign tumors. The intensity of GLUT-1 staining was lower in borderline tumors than in malignant cases. This was statistically significant (p = 0.005). As differentiation in malignant tumors increased, proportions of GLUT-1 expression showed a relative increase, but this difference was not statistically significant (p = 0.68). CONCLUSION: When GLUT-1 expression in borderline and malignant ovarian epithelial tumors was analyzed against prognosis, no statistically significant difference was identified. Assessment of GLUT-1 expression using the image analysis program was more reliable, with higher reproducibility than in previous studies.     

Impact of <Emphasis Type="Italic">ERCC2</Emphasis> gene polymorphism on HIV-1 disease progression to AIDS among North Indian HIV patients

HIV/AIDS remains to be one of the killing diseases of mankind. Host genetic response is one of the factor which determine susceptibility to HIV and disease progression to AIDS. The aim of the present study was to evaluate the impact of ERCC2 Lyc ⁷⁵¹ Gln (excision repair cross complementing rodent repair deficiency, complementation group 2) polymorphism on HIV-1 susceptibility and disease progression to AIDS, as this gene has been reported to intervene in degrading retroviral cDNA before it integrates with the host DNA. This case control study included 300 HIV seropositive cases and an equal number of HIV seronegative controls. DNA was isolated from the blood samples of study subjects and genotyping of ERCC2 was conducted by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) method. The Gln/Gln genotype showed a significant variation between cases and controls (P = 0.047, OR 1.71, 95% CI 1.00–2.93), indicating a possible role of susceptibility in reference to controls and disease progression when compared within cases.     

Screening by coral green fluorescent protein (GFP)-like chromoproteins supports a role in photoprotection of zooxanthellae

Green fluorescent protein (GFP)-like pigments are responsible for the vivid colouration of many reef-building corals and have been proposed to act as photoprotectants. Their role remains controversial because the functional mechanism has not been elucidated. We provide direct evidence to support a photoprotective role of the non-fluorescent chromoproteins (CPs) that form a biochemically and photophysically distinct group of GFP-like proteins. Based on observations of Acropora nobilis from the Great Barrier Reef, we explored the photoprotective role of CPs by analysing five coral species under controlled conditions. In vitro and in hospite analyses of chlorophyll excitation demonstrate that screening by CPs leads to a reduction in chlorophyll excitation corresponding to the spectral properties of the specific CPs present in the coral tissues. Between 562 and 586 nm, the CPs maximal absorption range, there was an up to 50 % reduction of chlorophyll excitation. The screening was consistent for established and regenerating tissue and amongst symbiont clades A, C and D. Moreover, among two differently pigmented morphs of Acropora valida grown under identical light conditions and hosting subclade type C3 symbionts, high CP expression correlated with reduced photodamage under acute light stress.     

Determinants of Visceral Leishmaniasis: A Case-Control Study in Gedaref State,Sudan

Background

Improving knowledge on local determinants of visceral leishmaniasis (VL) is crucial to guide the development of relevant control strategies. This study aimed to identify individual and household level determinants of primary VL in 24 highly endemic villages of Tabarak Allah hospital’s catchment area, Gedaref State, Sudan.

Methods

From September 2012 to July 2013, in an unmatched case-control design, 198 patients with primary VL were compared to 801 controls free of VL symptoms and with a negative VL rapid test. Using random spatial sampling, controls were selected with a distribution of age, sex and village of residence proportionate to the distribution of the target population. Data were collected using a structured questionnaire.

Results

Children and men were at higher risk of VL. Reporting VL patient(s) in the household in the previous year was the strongest VL risk factor. In a multivariate analysis, VL risk increased with household size, sleep location (outside the yard, not in the farm), evening outdoor activities in the rainy season (playing, watching TV, radio listening), use of ground nut oil as animal repellent and of smoke of Acacia seyal as indoor repellent, presence of dogs in the yard at night, Acacia nilotica in the yard’s immediate surroundings and of a forest at eye range. VL risk appeared to decrease with the use of drinking water sources other than the village water tank, a buffer distance from the adjacent house yard, and with the presence of animals other than dogs in the yard at night. In contrast with previous studies, housing factors, mosquito-net use, black cotton soil, ethnicity, socioeconomic index, presence of Balanites aegyptica and Azadirachta indica in the yard were not independent VL determinants.

Discussion and conclusion

Although these results do not provide evidence of causality, they provide useful suggestions for guiding further intervention studies on VL preventive measures.     

Khat Chewing Habits in the Population of the Jazan Region,Saudi Arabia: Prevalence and Associated Factors

The use of khat (Catha edulis) is a major public health and social problem that is believed to be growing globally. The khat chewing habit is prevalent in all areas of the Jazan region, Kingdom of Saudi Arabia (KSA). However, few studies have been conducted at the community level to investigate the khat chewing habits in this area. This study was conducted with the aim of assessing the prevalence and predictors of khat chewing among the Jazan community population. A cross-sectional study was conducted with a sample (n = 4,500) of the Jizani population who attended primary heath care centers in Jazan region. The participants were selected using a two-stage cluster random sampling. A structured questionnaire was used for data collection. The overall lifetime prevalence of khat chewing was 33.2% (95% CI 31.8–34.7) and was significantly higher for males 42.2% (95% CI 40.4–43.9) than for females 11.3% (95% CI 9.6–13.1) (P < 0.001). Current khat chewers accounted for 28.7% (95% CI 27.4–30.1) of the population sampled; 36.9% (95% CI 35.2–38.6) of whom were males, which is a significantly higher percentage than the 8.7% (95% CI 7.3–10.4) of current khat chewers who were females (P < 0.001). The multivariate logistic regression analysis suggests that the most important independent predictors of khat chewing were having a friend who chewed khat (OR = 20.1, P < 0.001), participant''s smoking status (OR) = 3.9, P < 0.001), friend''s smoking status (OR = 2.2, P < 0.001), gender (OR = 2.2, P < 0.001) and educational level (OR = 1.5, P < 0.05). A large proportion of the Jizani populations chew khat. Government and non-governmental organizations NGOs should design and strengthen community prevention programs to curb the high prevalence of khat use.     

Comparison of Two Methods for Purification of Enterocin B,a Bacteriocin Produced by Enterococcus faecium W3

This study aimed to compare two different approaches for the purification of enterocin B from Enterococcus faecium strain W3 based on the observation that the bacteriocin was found both in cell associated form and in culture supernatant. The first approach employed ammonium sulfate precipitation, cation-exchange chromatography, and sequential reverse-phase high-performance liquid chromatography. The latter approach exploited a pH-mediated cell adsorption–desorption method to extract cell-bound bacteriocin, and one run of reverse-phase chromatography. The first method resulted in purification of enterocin B with a recovery of 4% of the initial bacteriocin activity found in culture supernatant. MALDI-TOF MS analysis and de novo peptide sequencing of the purified bacteriocin confirmed that the active peptide was enterocin B. The second method achieved the purification of enterocin B with a higher recovery (16%) and enabled us to achieve pure bacteriocin within a shorter period of time by avoiding time consuming purification protocols. The purity and identity of the active peptide were confirmed again by matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry (MS) analysis. Although both approaches were satisfactory to obtain a sufficient amount of enterocin B for use in MS and amino acid sequence analysis, the latter was proved to be applicable in large-scale and rapid purification of enterocin B.     

Spatial analysis of rangeland's vegetation intensity as related to selected physical soil variables over ABQAIQ municipality of Saudi Arabia

Through most of the natural rangelands of Saudi Arabia, overgrazing, sand drifting, and off-road vehicles driving are almost the major contributing factors that lead to vegetation damage and land degradation in natural landscapes. The current study aimed at examining the dynamic nature of Abqaiq rangelands' vegetation, generating a vegetation intensity map, and investigating possible impacts caused by the spatial variability of obtained soil texture, sand content (%), and the derived available water capacity (DAW capacity (%)) on the intensity distribution of vegetation cover. Ordinal-nominal correlation type and spatial autocorrelation processes were adopted throughout the study to analyze the spatial correspondences as well as the interrelated patterns of the given variables. The resultant correlation between vegetation and soil texture (ordinal versus nominal variables) has yielded a significant (p-value < 0.001) relevancies, where, silt loam texture class, in particular, has proven to have the most correlated values to the most intensive vegetation habitats, where 5%, 35%, and 28% of the silt loam class were occupied by 80%, 60%, and 40% of vegetation intensities, respectively. Whereas, for the continuous variables, correlation outcomes have achieved a substantial negative spatial autocorrelation concerning vegetation intensity and sand content percentage, revealing a total absence of green biomass over the sandy soils. Additionally, vegetation intensity versus DAW capacity percentage significantly yielded a positive autocorrelation, revealing a high clustering of green biomass cover that associates with high clustering of high-water capacity soils. The autocorrelation strength identifier (Morn's I), produced an approximate value of 0.3 with a pseudo p-value of 0.001, for both relationships. The findings of this study would help researchers and relative authorities to grasp the reasons, consequences and behaviors of rangeland flora, considering Abqaiq's area as an example.