Ex vivo culture of human CD34+ cord blood cells with thrombopoietin (TPO) accelerates platelet engraftment in a NOD/SCID mouse model

OBJECTIVES: Hematopoietic recovery, in particular platelet reconstitution, can be severely delayed after transplantation with cord blood (CB) stem cells (SC). Expansion of CB SC may be one way to improve the recovery, but there is concern that ex vivo expansion compromises the repopulating ability of SC. METHODS: We used a short-term expansion protocol with TPO as single growth factor. The expanded cells were tested in the NOD/SCID mouse model and both platelet recovery and repopulation capacity were examined and compared with unexpanded CD34+ CB cells of the same CB donor. RESULTS: Platelet recovery started 1 week earlier in mice transplanted with TPO-expanded CD34+ cells and at days 5 and 8 after transplantation, 6.2 +/- 2.6 and 13.9 +/- 6.7 plt/microL were observed, respectively. At similar time intervals 0.0 and 1.5 +/- 0.2 plt/microL respectively were detected in mice receiving the unmanipulated CD34+ grafts. This was accompanied by a higher number of CFU-Mk in the bone marrow (BM) 7 days after transplantation. Moreover, the BM engraftment and the lineage differentiation of human cells at 6 weeks after transplantation was similar, suggesting that long-term engraftment was not compromised by the expansion procedure. CONCLUSION: Ex vivo expansion with TPO as single growth factor results in an accelerated platelet recovery in NOD/SCID mice and appears not to affect the long-term repopulation capacity.     

Production of a novel influenza vaccine using insect cells: protection against drifted strains

A recombinant trivalent hemagglutinin (HA) vaccine produced in cell culture using the baculovirus expression system provides an attractive alternative to the current egg‐based influenza vaccine (Trivalent Inactivated Influenza Vaccine [TIV]) manufacturing process. The HA genes from the annual World Health Organization‐recommended strains are cloned, expressed, and purified using a general purification process. Here, we provide an overview of the expression technology used to make the annual adjustment of the vaccine and the clinical studies completed to date with recombinant HA. The highly purified protein vaccine, administered at three times higher antigen content than TIV, results in stronger immunogenicity, a long‐lasting immune response and provides cross‐protection against drift variant influenza viruses. Furthermore, the vaccine does not contain egg proteins, adjuvants, preservatives, endotoxins, or antibiotics and can therefore be used in a broader population.     

Unconjugated bile salts shuttle through hepatocyte peroxisomes for taurine conjugation

Bile acid-CoA:amino acid N-acyltransferase (BAAT) conjugates bile salts to glycine or taurine, which is the final step in bile salt biosynthesis. In addition, BAAT is required for reconjugation of bile salts in the enterohepatic circulation. Recently, we showed that BAAT is a peroxisomal protein, implying shuttling of bile salts through peroxisomes for reconjugation. However, the subcellular location of BAAT remains a topic of debate. The aim of this study was to obtain direct proof for reconjugation of bile salts in peroxisomes. Primary rat hepatocytes were incubated with deuterium-labeled cholic acid (D(4)CA). Over time, media and cells were collected and the levels of D(4)CA, D(4)-tauro-CA (D(4)TCA), and D(4)-glyco-CA (D(4)GCA) were quantified by liquid chromatography-tandem mass spectrometry (LC/MS/MS). Subcellular accumulation of D(4)-labeled bile salts was analyzed by digitonin permeabilization assays and subcellular fractionation experiments. Within 24 hours, cultured rat hepatocytes efficiently (>90%) converted and secreted 100 μM D(4)CA to D(4)TCA and D(4)GCA. The relative amounts of D(4)TCA and D(4)GCA produced were dependent on the presence of glycine or taurine in the medium. Treatment of D(4)CA-exposed hepatocytes with 30-150 μg/mL digitonin led to the complete release of D(4)CA, D(4)GCA, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (cytosolic marker). Full release of D(4)TCA, catalase, and BAAT was only observed at 500 μg/mL digitonin, indicating the presence of D(4)TCA in membrane-enclosed organelles. D(4)TCA was detected in fractions of purified peroxisomes, which did not contain D(4)CA and D(4)GCA. Conclusion: We established a novel assay to study conjugation and intra- and transcellular transport of bile salts. Using this assay, we show that cholic acid shuttles through peroxisomes for taurine-conjugation.     

Total peptide YY is a correlate of postprandial energy expenditure but not of appetite or energy intake in healthy women

Peptide YY (PYY) and ghrelin have been associated with the regulation of energy balance. The objectives of this study were to determine whether total ghrelin and PYY after a standardized meal predict appetite scores and ad libitum energy intake (EI) and to examine the relationship between total ghrelin and PYY and postprandial energy expenditure (PEE). Twenty-five premenopausal women (age, 50.4 ± 2.0 years; body mass index, 23.5 ± 2.2 kg/m²) were studied. Total PYY, total ghrelin (enzyme-linked immunosorbent assay), EE (indirect calorimetry), and appetite scores (visual analogue scales) were measured fasting and every 30 minute for 3 hours after the ingestion of a standardized breakfast. Ad libitum EI was measured at lunch with a buffet-type meal. Peptide YY increased (P < .001) and total ghrelin decreased (P < .001) after breakfast. Significant changes in EE (P < .001) and appetite scores (P < .001) were noted postprandially. Appetite scores were consistently associated with ad libitum EI at lunch (r = −0.51 to 0.40, P < .05), whereas no association between EI and prelunch total ghrelin and PYY was observed. Finally, partial correlation analyses revealed that total PYY was a significant independent correlate of PEE at 60, 90, 120, and 150 minutes (r = 0.37-0.51, P ≤ .05). These findings provide evidence that appetite scores are better correlates of EI than are circulating levels of total PYY or ghrelin and that total PYY could be involved in the regulation of PEE.     

Cardiometabolic differences in children born after in vitro fertilization: follow-up study

CONTEXT: Increasing evidence suggests that adverse conditions during early prenatal life are associated with cardiometabolic dysfunction in postnatal life. In vitro fertilization (IVF) conception may be an early prenatal life event with long-term health consequences. OBJECTIVE: Our objective was to investigate several cardiometabolic measures in 8- to 18-yr-old IVF singletons and spontaneously conceived controls born from subfertile parents. DESIGN AND SETTING: This follow-up study was conducted at the VU University Medical Center, Amsterdam, The Netherlands. PARTICIPANTS: Blood pressure was examined in 225 IVF-conceived children and 225 age- and gender-matched spontaneously conceived control children. Several indicators of insulin resistance were studied in a pubertal subpopulation (131 IVF children and 131 controls). MAIN OUTCOME MEASURES: Blood pressure, fasting glucose, and fasting insulin were determined. RESULTS: Systolic and diastolic blood pressure levels were higher in IVF children than controls (109 +/- 11 vs. 105 +/- 10 mm Hg, P < 0.001; and 61 +/- 7 vs. 59 +/- 7 mm Hg, P < 0.001, respectively). Children born after IVF were also more likely to be in the highest systolic and diastolic blood pressure quartiles (odds ratio = 2.1, 95% confidence interval 1.4, 3.3; odds ratio = 1.9, 95% confidence interval 1.2, 3.0, respectively). Furthermore, higher fasting glucose levels were observed in pubertal IVF children (5.0 +/- 0.4 vs. 4.8 +/- 0.4 mmol/liter in controls; P = 0.005). Blood pressure and fasting glucose differences could not be explained by current body size, birth weight, and other early life factors or by parental characteristics, including subfertility cause. CONCLUSIONS: These findings highlight the importance of continued cardiometabolic monitoring of IVF-conceived children and might contribute to current knowledge about periconceptional influences and their consequences in later life.     

Westernization of the Filipino population resident in Rome: obesity, diabetes and hypertension

BACKGROUND: Aims of the present study were to examine the anthropometrical and metabolic characteristics of the Filipino population migrant to the Southern European city of Rome, Italy. METHODS: A cross-sectional study was carried out in the city of Rome. Three hundred thirty-five Filipinos (95 M/240 F, mean age: 44.0+/-9.8 years, mean residence in Italy: 12.9+/-6.3 years) were studied. Data were collected by standardized questionnaires; anthropometrical parameters, arterial pressure, and fasting capillary blood glucose (FCG) were measured. RESULTS: Abdominal obesity was found in 52.5%, and BMI >or= 25 kg/m(2) in 44.5% of subjects. History of type 2 diabetes mellitus (T2DM) and hypertension were reported by 6.0 and 9.0% of subjects, respectively. Impaired fasting glucose was found in 13.1%, and FCG >or= 110 mg/dl in 18.5% of subjects. Altered systolic and/or diastolic blood pressure was found in 34.3% of subjects. About three-fourths of subjects were unaware of being diabetic and/or hypertensive. Years of Italy residence showed a direct significant correlation with the degree of changes in alimentary behaviours (rho=0.18, p=0.001), and with weight gain (rho=0.27, p<0.001). Multivariate analysis showed only age and waist circumference to be associated with both diabetes and hypertension. CONCLUSIONS: In the present study, the first to examine the metabolic disorders in a migrant Filipino population resident in Rome, a high prevalence of obesity, diabetes, and hypertension was found. The alarming results emerging from this study should be seriously considered by public health practitioners and decision makers, and made known to the Filipinos resident in Europe.     

Endoscopic treatment of esophagogastric variceal bleeding in patients with noncirrhotic extrahepatic portal vein thrombosis: a long-term follow-up study

BACKGROUND: Esophagogastric variceal bleeding is the most important complication of extrahepatic portal vein thrombosis (EPVT) and is usually treated endoscopically. Little is known about the prognosis of these patients. OBJECTIVES: To investigate the long-term clinical outcome and efficacy of endoscopic treatment in patients with esophagogastric variceal bleeding secondary to EPVT. DESIGN: Retrospective observational study. SETTINGS: Single university center. PATIENTS: Twenty-seven consecutive patients with esophagogastric variceal bleeding, secondary to noncirrhotic, nonmalignant EPVT, who underwent endoscopic treatment between 1982 and 2005. INTERVENTIONS: Endoscopic band ligation and/or endoscopic sclerotherapy. MAIN OUTCOME MEASUREMENTS: The overall rebleeding risk, overall survival, complications of the endoscopic procedures, and predictive values of rebleeding. Analyses were performed by the Kaplan-Meier method and univariate Cox regression. RESULTS: All patients were followed-up after the first endoscopically treated variceal bleeding. A total of 241 endoscopic procedures were performed. In all patients, initial control of bleeding was obtained. The overall rebleeding risk was 23% (95% CI, 0%-24%) at 1 year and 37% (95% CI, 43%-83%) at 5 years. Extension of thrombosis into the splenic vein and the presence of fundal varices were significant predictors of rebleeding, with a nearly 5-fold increased risk for patients with EPVT and fundal varices at the time of the first variceal hemorrhage (hazard ratio 5.07, P = .01). A portosystemic shunt procedure was performed in 5 patients: 4 for variceal bleeding and in one patient for refractory ascites. Seven patients died, none from variceal bleeding. Overall 5-year and 10-year survivals were 100% and 62% (95% CI, 38%-96%), respectively. LIMITATIONS: Retrospective design. CONCLUSIONS: In patients with variceal bleeding secondary to EPVT endoscopic treatment, in particular, band ligation appears safe and effective. EPVT-related mortality is primarily determined by other causes than variceal bleeding.     

Can a self-management education program for patients with chronic obstructive pulmonary disease improve quality of life ?

OBJECTIVE:

To assess the effects of a self-management program on health-related quality of life (HRQoL) and morbidity commonly associated with chronic obstructive pulmonary disease (COPD).

METHODS:

A total of 57 outpatients with stable COPD received four weeks of self-management education, while 45 patients received usual care. Patients were evaluated at baseline, at three months and one year following the educational intervention. The primary outcome variable was HRQoL measured by the St George’s Respiratory Questionnaire (SGRQ). The secondary outcome variables were number of emergency room visits and hospitalizations for exacerbation.

RESULTS:

The intervention group’s HRQoL improved significantly at three months (total score A=−5.0 [P=0.006]) and 12 months (total score A=−6.7 [P<0.001]), as evidenced by decreased scores on the SGRQ. In contrast, the SGRQ scores increased significantly in the control group at three months (total score A=+3.7 [P= 0.022]) and 12 months (total score A=+3.4 [P=0.032]). Global impact appeared to be responsible for the change in the intervention group. Moreover, in the intervention group, the number of hospitalizations dropped from 0.7/person/year to 0.3/person/year (P=0.017), and emergency room visits dropped from 1.1 person/year to 0.2/person/year (P=0.002), while subjects in the control group did not experience any significant decreases in these parameters.

CONCLUSIONS:

A planned education program improved HRQoL while decreasing the number of emergency room visits and hospitalizations in patients with stable COPD; this improvement persisted at 12 months.