The Immune Reactivity Role of HCV-Induced Liver Infiltrating Lymphocytes in Hepatocellular Damage

Liver infiltrating lymphocytes (LIL) were isolated from HCV-positive (+) and HCV-negative (–) end-stage livers. Phenotypic analysis and functional studies using proliferative and lymphocytotoxic assays were performed with the isolated LIL. Two CD3+ lymphocyte populations were found in LIL using FITC anti-CD3 monoclonal antibodies (mAb). One was a bright fluorescence intensity population (as in PBL), and the other dim. We calculated the number of FITC-anti-CD3 mAbs bound per lymphocyte on PBL and LIL and found 80,040 ± 4628 and 39,615 ± 3932, respectively. Therefore, HCV+ and HCV– patient PBL contained approximately twice the number of CD3 molecules per cell than patient CD3+ LIL. LIL also contained approximately a threefold higher concentration of TCR+, CD4–CD8–, and CD56,16 (NK) cells than the patient PBL. Thus, a major subset of LIL is phenotypically similar to mouse NK1.1+ intermediate T cells. LIL freshly isolated from HCV+ livers exhibited weak CTL activity against EBV- or Con A-transformed lymphoblast targets infected with vaccinia–HCV recombinant virus (rHCV) or primary hepatocyte cultured cells. However, after in vitro coculture of LIL with rHCV, these cells developed a strong cytotoxicity for the above targets. In contrast, LIL from HCV– livers were not cytotoxic against the same targets. Histochemical studies (in situ) demonstrated that these hepatocytes express CD95, and stains demonstrated apoptosis. The HCV+ hepatocytes also express class I MHC molecules and ICAM-1. The addition of mAb specific for these adhesion molecules inhibited CML activity. Short-term cultured hepatocytes (targets) from HCV+ and HCV– patients produced low levels of cytokines IL-1, IL-2, IL-6, TNF, and IFN- but a high level of IL-8. It is speculated that LIL expressing reduced numbers of CD3 molecules may even function as immune regulators as proposed for intermediate T cells in mice.     

Diverse Effects of Combinations of Maternal-Neonatal VDR Polymorphisms and 25-Hydroxyvitamin D Concentrations on Neonatal Birth Anthropometry: Functional Phenocopy Variability Dependence,Highlights the Need for Targeted Maternal 25-Hydroxyvitamin D Cut-Offs during Pregnancy

Vitamin D receptor (VDR) polymorphisms have been associated with a plethora of adverse pregnancy and offspring outcomes. The aim of this study was to evaluate the combined effect of maternal and neonatal VDR polymorphisms (ApaI, TaqI, BsmI, FokI, Tru9I) and different maternal and neonatal 25(OH)D cut-offs on neonatal birth anthropometry. This cross-sectional study included data and samples from a cohort of mother–child pairs at birth. A detailed neonatal anthropometry analysis at birth was also conducted. Different 25(OH)D cut-offs for neonates and mothers were included, according to their vitamin D status at birth: for neonates, cut-offs of [25(OH)D ≤ 25 and > 25 nmol/L] and [25(OH)D ≤ 50 nmol/L] were adopted, whereas for mothers, a 25(OH)D cut-off of [25(OH)D ≤ 50 and > 50 nmol/L)] was investigated. Following this classification, maternal and neonatal VDR polymorphisms were evaluated to investigate the potential different effects of different neonatal and maternal 25(OH)D cut-offs on neonatal birth anthropometry. A total of 69 maternal-neonatal dyads were included in final analysis. Weight, neck rump length, chest circumference, abdominal circumference, abdominal circumference (iliac), high thigh circumference, middle thigh circumference, lower arm radial circumference, and lower leg calf circumference of neonates who had the TAQl SNP TT genotype and maternal 25(OH)D < 50 nmol/L were significantly higher than that of neonates who had the Tt or tt genotypes (p = 0.001, Hg = 1.341, p = 0.036, Hg = 0.976, p = 0.004, Hg = 1.381, p = 0.001, Hg = 1.554, p = 0.001, Hg = 1.351, p = 0.028, Hg = 0.918, p = 0.008, Hg = 1.090, p = 0.002, Hg = 1.217, and p = 0.020, Hg = 1.263, respectively). Skin fold high anterior was significantly lower in neonates who had the BSMI SNP BB genotype compared to that of neonates with Bb or bb genotypes (p = 0.041, Hg = 0.950), whereas neck rump length was significantly higher in neonates who had the FOKI SNP FF genotype compared to that of neonates who had Ff or ff genotypes (p = 0.042, Hg = 1.228). Regarding neonatal VDR polymorphisms and cut-offs, the abdominal circumference (cm) of neonates who had the TAQI SNP TT genotype and 25(OH)D < 25 nmol/L were significantly higher than that of neonates who had the Tt or tt genotypes (p = 0.038, Hg = 1.138). In conclusion, these results indicate that the maternal TAQI VDR polymorphism significantly affected neonatal birth anthropometry when maternal 25(OH) concentrations were <50 nmol/L, but not for a higher cut-off of >50 nmol/L, whereas this effect is minimally evident in the presence of neonatal TAQI polymorphism with neonatal 25(OH)D values <25 nmol/L. The implication of these findings could be incorporated in daily clinical practice by targeting a maternal 25(OH)D cut-off >50 nmol/L, which could be protective against any effect of genetic VDR variance polymorphism on birth anthropometry.     

Postoperative pulmonary function after laparoscopic and open cholecystectomy

In this prospective, randomized study, we compared 42 patients undergoing laparoscopic cholecystectomy and 40 undergoing open cholecystectomy to determine if laparoscopic cholecystectomy results in less respiratory impairment and fewer respiratory complications. Pulmonary function tests, arterial blood-gas analysis and chest radiographs were obtained in both groups before operation and on the second day after operation. Postoperative pain scores and analgesic requirements were also recorded. After operation, a significant reduction in total lung capacity, functional residual capacity (FRC), forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and mid-expiratory flow (FEF25-75%) occurred after both laparoscopic and open cholecystectomy. The reductions in FRC, FEV1, FVC and FEF25-75% were smaller after laparoscopic (7%, 22%, 19% and 23%, respectively) than after open (21%, 38%, 32% and 34%, respectively) cholecystectomy. Laparoscopic cholecystectomy was also associated with a significantly lower incidence (28.6% vs 62.5%) and less severe atelectasis, better oxygenation and reduced postoperative pain and analgesia use compared with open cholecystectomy. We conclude that postoperative pulmonary function was impaired less after laparoscopic than after open cholecystectomy.      

Physical training modulates proinflammatory cytokines and the soluble Fas/soluble Fas ligand system in patients with chronic heart failure

OBJECTIVES: We sought to investigate the effects of physical training on circulating proinflammatory cytokines and the soluble apoptosis mediators Fas (sFas) and Fas ligand (sFasL) in patients with chronic heart failure (CHF). BACKGROUND: Recent investigations have shown an overexpression of circulating proinflammatory cytokines and soluble apoptosis mediators in patients with CHF, which may be related to their exercise intolerance and clinical deterioration. METHODS: Plasma levels of tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptors I and II (sTNF-RI and sTNF-RII, respectively), interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), sFas and sFasL were measured in 24 patients with stable CHF (New York Heart Association functional class II/III; left ventricular ejection fraction 23.2 +/- 1.3%) and in 20 normal control subjects before and after a 12-week program of physical training in a randomized, crossover design. Functional status of patients with CHF was evaluated by using a cardiorespiratory exercise test to measure peak oxygen consumption (VO2max). RESULTS: Physical training produced a significant reduction in plasma levels of TNF-alpha (7.5 +/- 1.0 pg/ml vs. 4.6 +/- 0.7 pg/ml, p < 0.001), sTNF-RI (3.3 +/- 0.2 ng/ml vs. 2.7 +/- 0.2 ng/ml, p < 0.005), sTNF-RII (2.6 +/- 0.2 ng/ml vs. 2.3 +/- 0.2 ng/ml, p = 0.06), IL-6 (8.3 +/- 1.2 pg/ml vs. 5.9 +/- 0.8 pg/ml, p < 0.005), sIL-6R (34.0 +/- 3.0 ng/ml vs. 29.2 +/- 3.0 ng/ml, p < 0.01), sFas (5.5 +/- 0.7 ng/ml vs. 4.5 +/- 0.8 ng/ml, p = 0.05) and sFasL (34.9 +/- 5.0 pg/ml vs. 25.2 +/- 4.0 pg/ml, p < 0.05), as well as a significant increase in VO2max (16.3 +/- 0.7 ml/kg per min vs. 18.7 +/- 0.8 ml/kg per min, p < 0.001). Good correlations were found between a training-induced increase in VO2max and a training-induced reduction in levels of the proinflammatory cytokine TNF-alpha (r = -0.54, p < 0.01) and the apoptosis inducer sFasL (r = -0.57, p < 0.005) in patients with CHF. In contrast, no significant difference in circulating cytokines and apoptotic markers was found with physical training in normal subjects. CONCLUSIONS: Physical training reduces plasma levels of proinflammatory cytokines and the sFas/sFasL system in patients with CHF. These immunomodulatory effects may be related to the training-induced improvement in functional status of patients with CHF.     

A 10-year aerobiological study (1994-2003) in the Mediterranean island of Crete, Greece: trees, aerobiologic data, and botanical and clinical correlations.

Pollen grains from the plant cover of a given area participate largely in the composition of aeroflora (pollen and molds) of this area. Association of allergic respiratory disorders with concentration of allergenic particles in the atmosphere is well documented, and aerobiologic studies are of great relevance. A 10-year volumetric aerobiologic study was conducted in the city of Heraklion, located in the center of the north-shore of the island of Crete, Greece. Main allergenic families and genera encountered were, in descending order of frequency Oleaceae, Quercus, Platanaceae, Cupressaceae, Pinaceae, Populus, Moraceae, and Corylaceae. Concentrations noted for most of these aeroallergens were much lower than those reported from other European regions. In parallel, an atopic population of 576 individuals, exhibiting allergic symptoms mainly of the respiratory tract were subjected to a battery of skin-prick tests. A fair degree of agreement between total pollen counts and positive skin-prick test frequencies for the families of Oleaceae, Platanaceae, and Cupressaceae was noted. On the contrary a poor degree of concordance was noted for the rest of the families and genera.     

Does Pregnancy-Associated Breast Cancer Imply a Worse Prognosis? A Matched Case-Case Study

Zusammenfassung

Hintergrund: Die Auswirkungen einer Schwangerschaft auf die Prognose des Mammakarzinoms werden in der Literatur kontrovers diskutiert. Wir haben eine gepaarte Fall-Fall-Studie konzipiert, in der schwangerschaftsasso-ziierte Mammakarzinom (SAM)-Fälle entsprechend ihres Stadiums, Alters und Jahr der Diagnosestellung mit Mammakarzinom-Patientinnen gepaart wurden. Patientinnen und Methoden: 39 aufeinanderfolgende SAM-Fälle wurden mit 39 prämenopausalen Fällen von Brust-krebs gepaart. Univariate und multivariate Überlebens-analysen mit Anpassung an Stadium, Grad, Östrogen-rezeptorstatus und Alter zum Zeitpunkt der Diagnose wurden durchgeführt. Ergebnisse: Hinsichtlich des Gesamtüberlebens deutete die univariate Analyse auf ein längeres Gesamtüberleben für nicht-SAM-Fälle vs. SAM-Fälle hin. Gleichzeitig war ein fortgeschritteneres Stadium ein Prädiktor für ein kürzeres Überleben. Die multivariate Analyse bestätigte die unabhängige ver-schlechternde Auswirkung einer Schwangerschaft. Inter-essanterweise ergab eine genestete Post-hoc-Analyse der SAM-Fälle Hinweise auf ein kürzeres Gesamtüberleben für das dritte Trimester. Die oben erwähnten Ergebnisse für das Gesamtüberleben konnten desweiteren bei der Untersuchung des rezidivfreien Überlebens reproduziert werden. Schlussfolgerung: Mit ihrem gepaarten Fall-Fall-Design deutet die vorliegende Studie darauf hin, dass Schwangerschaft ein schlechter Prognosefaktor beim Mammakarzinom ist.     

Sexual dysfunction in testicular cancer patients subjected to post‐chemotherapy retroperitoneal lymph node dissection: a focus beyond ejaculation disorders

Post‐chemotherapy retroperitoneal lymph node dissection (PC‐RPLND) represents an integral part of multidisciplinary treatment of advanced germ cell cancer; however, it is associated with a high complications rate. The present study aimed to describe sexual disorders in 53 patients with testicular cancer who underwent full bilateral, non‐nerve‐sparing PC‐RPLND in our institution, focusing beyond ejaculatory dysfunction. The International Index for Erectile Function (IIEF) questionnaire was used as diagnostic tool of male sexual functioning pre‐operatively and three months after RPLND, while post‐operatively patients were asked to describe and evaluate changes in selected sexual parameters. Study findings demonstrate mixed pattern of changes in sexual functioning, with no difference in erectile functioning before and after operation. However, orgasmic function and intercourse and overall sexual satisfaction were found significantly impaired post‐operatively. Sexual desire and frequency of attempted sexual intercourses were found significantly increased post‐operatively, in comparison with pre‐operative levels. With regard to patients' subjective perception on sexual functioning alterations after PC‐RPLND, a significant number of patients reported higher levels of sexual desire, no difference in erectile function and worse orgasmic function and satisfaction post‐operatively. Thus, patients subjected to PC‐RPLND should be closely and routinely evaluated due to close relationship of sexual dissatisfaction with secondary psychological disorders.     

Prolonged treatment of Salmonella enterica serovar Typhimurium with commercial disinfectants selects for multiple antibiotic resistance, increased efflux and reduced invasiveness

OBJECTIVES: To study how disinfectants affect antimicrobial susceptibility and phenotype of Salmonella enterica serovar Typhimurium SL1,344. METHODS: Wild-type strain SL1,344 and its isogenic gyrA mutant were passaged daily for 7 days in subinhibitory concentrations, and separately for 16 days in gradually increasing concentrations of a quaternary ammonium disinfectant containing formaldehyde and glutaraldehyde (QACFG), an oxidizing compound blend (OXC), a phenolic tar acids-based disinfectant (TOP) and triclosan. The MICs of antimicrobials and antibiotics for populations and representative isolates and the proportion of cells resistant to the MICs for the wild-type were determined. Expression of acrB gene, growth at 37 degrees C and invasiveness of populations in Caco-2 intestinal epithelial cells were assessed. RESULTS: QACFG and triclosan showed the highest selectivity for variants with reduced susceptibility to chloramphenicol, tetracycline, ampicillin, acriflavine and triclosan. Populations treated with the above biocides had reduced invasiveness in Caco-2 cells, and altered growth kinetics. Resistance to disinfectants was observed only after exposure to gradually increasing concentrations of triclosan, accompanied with a 2000-fold increase in its MIC. Growth in OXC and TOP did not affect the MICs of antibiotics, but resulted in the appearance of a proportion of cells resistant to the MIC of acriflavine and triclosan for the wild-type. Randomly selected stable variants from all populations, except the one treated with TOP, over-expressed acrB. CONCLUSIONS: In vitro exposure to QACFG and triclosan selects for Salmonella Typhimurium cells with reduced susceptibility to several antibiotics. This is associated with overexpression of AcrAB efflux pump, but accompanied with reduced invasiveness.     

Islet Preparation Purity Is Overestimated,and Less Pure Fractions Have Lower Post-Culture Viability Before Clinical Allotransplantation

Background

Replacement of β-cells with the use of isolated islet allotransplantation (IT) is an emerging therapy for type 1 diabetics with hypoglycemia unawareness. The current standard protocol calls for a 36–72-hour culture period before IT. We examined 13 clinical islet preparations with ≥2 purity fractions to determine the effect of culture on viability.

Methods

After standard islet isolation and purification, pure islet fractions were placed at 37°C with 5% CO₂ for 12–24 hours and subsequently moved to 22°C, whereas less pure fractions were cultured at 22°C for the entire duration. Culture density was targeted at a range of 100–200 islet equivalents (IEQ)/cm² adjusted for purity. Islets were assessed for purity (dithizone staining), quantity (pellet volume and DNA), and viability (oxygen consumption rate normalized to DNA content [OCR/DNA] and membrane integrity).

Results

Results indicated that purity was overestimated, especially in less pure fractions. This was evidenced by significantly larger observed pellet sizes than expected and tissue amount as quantified with the use of a dsDNA assay when available. Less pure fractions showed significantly lower OCR/DNA and membrane integrity compared with pure. The difference in viability between the 2 purity fractions may be due to a variety of reasons, including hypoxia, nutrient deficiency, toxic metabolite accumulation, and/or proteolytic enzymes released by acinar tissue impurities that are not neutralized by human serum albumin in the culture media.

Conclusions

Current clinical islet culture protocols should be examined further, especially for less pure fractions, to ensure the maintenance of viability before transplantation. Even though relatively small, the difference in viability is important because the amount of dead or dying tissue introduced into recipients may be dramatically increased, especially with less pure preparations.     

Treatment of small cell lung cancer

Small cell lung cancer (SCLC) is a very chemo- and radiosensitive systemic disease. Combination chemotherapy produces a survival advantage resulting in median survival of 9 to 12 months in extensive disease. Platinum and etoposide in combination with concurrent, early, hyperfractionated chest radiotherapy, in patients with limited disease, produces median survival of 20 months. Prophylactic cranial radiotherapy, in patients with complete response following induction chemotherapy, reduces the incidence of brain metastases and improves survival. Triplet combinations, dose intensification, and maintenance therapy have not demonstrated meaningful survival improvements. Recurrent disease can be treated with the same chemotherapy, as in the first line treatment if the progression-free interval exceeds 3 months; otherwise, monotherapy with a novel compound is suggested. Camptothesins (topotecan, irinotecan) appear the most promising new compounds and may become first-line agents for SCLC in the near future. Molecular advances have provided many new targets for SCLC therapy. Many studies, ongoing or planned, evaluate the effectiveness of new agents developed to attack these targets.