Effect of a high-calcium energy-reduced diet on abdominal obesity and cardiometabolic risk factors in obese Brazilian subjects.

BACKGROUND: Clinical trials designed to examine the effects of calcium supplementation on abdominal obesity have had ambiguous results. AIMS: This study aimed to evaluate, during energy restriction, the effects of a high-calcium diet (HCD) on measures of abdominal obesity and cardiometabolic risk factors in Brazilian obese subjects of multiethnic origin. METHODS: We conducted a randomised clinical trial. Fifty obese subjects of both sexes, aged 22-55 years, with stable body weight and a low calcium intake were randomised into the following outpatient dietary regimens: (i) a low-calcium diet (LCD; < 500 mg/day) or (ii) a HCD [1200-1300 mg/day, supplemented with non-fat powdered milk (60 g/day)]. Both groups followed an energy-restricted diet (-800 kcal/day) throughout the study (16 weeks). RESULTS: Thirty-nine participants completed the study. After 16 weeks of energy restriction, a significant reduction was observed in all anthropometric parameters, metabolic variables (except for high-density lipoprotein cholesterol) and blood pressure levels in both the groups. Insulin was significantly reduced only in the HCD group. Subjects on the HCD compared with those on the LCD exhibited a greater reduction in waist circumference (p = 0.002), waist-to-hip ratio (p = 0.0001), diastolic blood pressure (p = 0.04) and mean blood pressure (p = 0.03). CONCLUSIONS: Our study suggests that increased calcium intake may enhance the beneficial effects of energy restriction on abdominal obesity and blood pressure.

Weight gain post-renal transplantation and its association with glomerular filtration rate.

OBJECTIVE: Obesity may be a risk factor for renal graft loss. The purpose of this study was to assess weight gain and its association with glomerular filtration rate after kidney transplant. METHODS: This retrospective analysis of 152 adult renal transplant outpatients (77 males, 75 females; mean age, 45.6 +/- 10.1 years) with at least 6 months posttransplantation (post-tx) included body weight and creatinine clearance (CrCl) measured pretransplantation (pre-tx) as well as at 6 months (post-tx1), 1 year (post-tx2), 5 years (post-tx3), 10 years (post-tx4), 15 years (post-tx5), and 20 years (post-tx6) post-tx. Weight gain was determined by subtracting patient preoperative weight from the post-tx weights. RESULTS: The weight gain post-tx versus pre-tx was progressive and significant (P < .001) post-tx1, 8.7 +/- 10.9% (n = 152); post-tx2, 12.4 +/- 11.5% (n = 150); post-tx3, 16.6 +/- 14.8% (n = 102); post-tx4, 20.1 +/- 18.1% (n = 47); post-tx5, 20.8 +/- 19.8% (n = 17); and post-tx6, 21.0 +/- 32.5% (n = 11). There was a positive and significant correlation between weight gain and CrCl during most times post-tx: post-tx2 (r = 0.32; P < .000); post-tx3 (r = 0.54; P < .0001); post-tx4 (r = 0.47; P = .01); and post-tx6 (r = 0.92; P < .001). CONCLUSIONS: There was a great increase in body weight after kidney transplantation. The excessive body weight gain was associated with a better glomerular filtration rate.

Prevalence of risk factors for cardiovascular disease in Brazilian renal transplant recipients.

OBJECTIVE: Cardiovascular disease (CVD) is the major cause of death in renal transplant recipients. The aim of this study was to assess the prevalence of CVD risk factors among Brazilian adult renal transplant recipients. METHODS: This cross-sectional study included 192 cases in which the evaluated CVD risk factors were hypertension, diabetes mellitus, impaired fasting glucose, obesity, overweight, abdominal obesity, metabolic syndrome, and dyslipidemia. RESULTS: Hypertension, abdominal obesity, and hypercholesterolemia were the highest prevalent risk factors among the total population with prevalences of approximately 60%. The prevalence of obesity was significantly higher (P < .001) in recipients with normal graft function (28%) than in those with impaired graft function (7%). Abdominal obesity was also higher (P = .02) in the normal graft function group (77%) than in recipients with creatinine clearance (CrCl) values <60 mL/min (61%). There were positive, significant correlations between CrCl and body mass index (BMI) (r = 0.47; P < .001) and between CrCl and waist circumference (WC) (r = 0.44; P < .001). BMI (r = 0.31; P < .001) and WC (r = 0.27; P < .001) were also positively associated with triglyceride levels. There were negative associations of high-density lipoprotein (HDL)-cholesterol (HDL-c) with BMI (r = -0.28; P < .01) and WC (r = -0.32; P < .01). CONCLUSIONS: The high prevalence of CVD risk factors among renal transplant recipients emphasizes the importance of taking appropriate therapeutic measures to reduce modifiable risk factors, reducing CVD and its consequences.

Association between -250G/A polymorphism of the hepatic lipase gene promoter and coronary artery disease and HDL-C levels in a Southern Brazilian population.

Hepatic lipase (HL) is a glycoprotein that plays a major role in remodeling high-density lipoprotein (HDL). The effect of the -250G/A promoter polymorphism on coronary artery disease (CAD) and lipid levels was studied in 231 male CAD patients and in a population-based sample of men and women (n = 514). A sample of 140 men was chosen among those included in the population-based sample as controls for the CAD sample. In the total group of CAD patients, the frequency of the -250A allele was somewhat lower (25% in CAD patients and 32% in controls; p = 0.06), but when the control samples were compared only with the CAD(+) sample (more than 60% of luminal stenosis in at least one coronary artery or major branch segment) the -250A allele was significantly less frequent (23% in the patients vs 32% in controls; p = 0.02). A multiple logistic regression analysis showed that this association was independent of classical CAD risk factors [odds ratio (OR) = 1.79, p = 0.025]. Using multiple linear regression analyses, it has been shown that this polymorphism was a significant factor affecting HDL-C levels in men from the population-based sample (p = 0.001), an interaction between -250G/A variant and wine consumption was also detected (p = 0.001). Thus, our results show that the -250G/A polymorphism in the HL gene is associated with significant variations in HDL-C levels and CAD risk in males.

Interaction between SREBP-1a and APOB polymorphisms influences total and low-density lipoprotein cholesterol levels in patients with coronary artery disease.

In this study, we examined the insertion/deletion (Ins/Del) and XbaI polymorphisms of the apolipoprotein B (APOB) gene and the -36delG polymorphism in the sterol regulatory element binding protein-1a (SREBP-1a) gene in 298 patients with non-diabetic angiographically assessed coronary artery disease (CAD), and 188 healthy controls, from a Brazilian population of European descent. Del/X+ haplotype carriers had higher levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in patients (TC, p = 0.05; LDL-C, p = 0.049) and controls (TC, p = 0.004; LDL-C, p = 0.013). No association was detected between the SREBP-1a-36delG polymorphism and lipid levels, but a significant interaction effect between APOB and SREBP-1a polymorphisms was observed in the patient sample on TC (p = 0.005) and on LDL-C (p = 0.019) levels. Carriers of the APOB Del/X+ haplotype and SREBP-1a G-G- genotype showed the highest levels of these lipid parameters. This effect of interaction was not observed in the control sample. Despite the associations with lipids, these polymorphisms were not associated with CAD risk or severity in this sample.

SEM study of water activity and temperature effects on the initial growth of Aspergillus ochraceus, Alternaria alternata and Fusarium verticillioides on maize grain. Scanning electron microscopy.

A scanning electron microscopy (SEM) study of the effect of water activity (a(w)), temperature and fungal interactions on the very early phases of Aspergillus ochraceus, Alternaria alternata and Fusarium verticillioides development on maize grains was carried out. Germination and growth of individual fungal strains were assayed at 0.92, 0.95 and 0.98 a(w), and 20 and 30 degrees C. Hyphal lengths were measured on micrographs taken by SEM at different periods of incubation. A. alternata had the highest linear growth at 0.98 a(w), and was the only species able to grow under all conditions tested, whereas A. ochraceus was not able to germinate at 0.92 a(w) at any temperature assayed. F. verticillioides demonstrated a different behaviour depending on growth temperature. Fungal interactions were studied at 0.95a(w) and 30 degrees C. A. ochraceus germination was inhibited when it was co-inoculated with one or two of the other species. A. alternata showed an increased growth rate when growing together with the other fungi, whereas growth of F. verticillioides was significantly inhibited when paired with A. ochraceus.

HSV and glycoprotein J inhibit caspase activation and apoptosis induced by granzyme B or Fas.

HSV-1 inhibits apoptosis of infected cells, presumably to ensure that the infected cell survives long enough to allow completion of viral replication. Because cytotoxic lymphocytes kill their targets via the induction of apoptosis, protection from apoptosis could constitute a mechanism of immune evasion for HSV. Several HSV genes are involved in the inhibition of apoptosis, including Us5, which encodes glycoprotein J (gJ). Viruses deleted for Us5 showed defects in inhibition of caspase activation after Fas ligation or UV irradiation. Transfected cells expressing the Us5 gene product gJ were protected from Fas- or UV-induced apoptosis, as measured by morphology, caspase activation, membrane permeability changes, or mitochondrial transmembrane potential. In contrast, caspase 3 activation in mitochondria-free cell lysates by granzyme (gr)B was inhibited equivalently by Us5 deletion and rescue viruses, suggesting that gJ is not required for HSV to inhibition this process. However, mitochondria-free lysates from transfected cells expressing Us5/gJ were protected from grB-induced caspase activation, suggesting that Us5/gJ is sufficient to inhibit this process. Transfected cells expressing Us5/gJ were also protected from death induced by incubation with purified grB and perforin. These findings suggest that HSV has a comprehensive set of immune evasion functions that antagonize both Fas ligand- and grB-mediated pathways of CTL-induced apoptosis. The understanding of HSV effects on killing by CTL effector mechanisms may shed light on the incomplete control of HSV infections by the immune system and may allow more rational approaches to the development of immune modulatory treatments for HSV infection.

Magnetic properties of diruthenium(II,III) carboxylate compounds. Crystal structures of Ru2Cl(mu-O2CCH=CHCH=CHMe)4 and Ru2Cl(mu-O2CCH2OMe)4.

The reaction of Ru2Cl(mu-O2CMe)4 with 2,4-hexadienoic and 2-methoxyacetic acids affords the compounds Ru2Cl(mu-O2CR)4 [R = CH=CHCH=CHCH3 (1), CH2OMe (2)]. The structures of both complexes have been determined by X-ray crystallography. 1 crystallizes in the triclinic space group P-1 with a = 9.264(1) A, b = 12.661(8) A, c = 12.839(5) A, alpha = 106.09(3) degrees, beta = 77.89(2) degrees, gamma = 97.73(3) degrees, and Z = 2. 2 crystallizes in the nonstandard monoclinic space group P2(1)/c with a = 12.132(4) A, b = 11.570(2) A, c = 13.674(2) A, beta = 91.18(2) degrees, and Z = 4. Complexes 1 and 2 show [Ru2(mu-O2CR)4]+ units linked by chloride ions, giving zigzag chains with Ru-Cl-Ru angles of 119.43(4) degrees and 110.11(7) degrees, respectively. The Ru-Ru bond distances are 2.2857(9) A (1) and 2.290(1) A (2). A magnetic study, in the 2-300 K temperature range, of the new compounds and the previously described Ru2Cl(mu-O2CR)4 [R = CHMe2 (3), CMe3 (4), C4H4N (5)] is described. The polymeric complexes 1 and 2 and the nonpolymeric 3-5 show a large zero-field splitting which varies from 53.9 to 68.1 cm-1. These complexes also show a weak, but not negligible, through-space intermolecular antiferromagnetic coupling not observed in the previous magnetic studies carried out on these types of compounds.

Occurrence of fumonisins in Spanish beers analyzed by an enzyme-linked immunosorbent assay method.

This study describes the occurrence of fumonisin in a survey of 32 Spanish beers, including some non-alcoholic beers. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method and, in the range assayed (0-100 ng fumonisin B1/ml spiked-beer), recovery rates were about 81%, with variation coefficients that did not exceed 4.2%. The detection limit with this method was 3 ng fumonisins/ml beer. Results for the fourteen positive samples (43.8% of total samples) showed that the lowest amount of fumonisins found in a positive sample was 4.76 ng/ml and the highest was 85.53 ng/ml.

Identification and cloning of a mobile transposon from Aspergillus niger var. awamori.

Aspergillus niger var. awamori contains multiple copies of a transposable element, Vader. This element was detected as a 437-bp insertion in four independently isolated spontaneous mutants of the niaD (nitrate reductase) gene. The Vader element is present in approximately 15 copies in both A. niger var. awamori and A. niger. A single copy of Vader was detected from only one of the two laboratory strains of A. nidulans which were also examined. Insertion of the Vader element into the niaD gene of A. niger var. awamori caused a 2-bp duplication (TA) of the target sequence. The Vader element is flanked by a 44-bp inverted repeat. The genetic stabilities of the inserted Vader elements at niaD were examined by studying reversion frequencies resulting in colonies able to grow on nitrate as a sole nitrogen source. Mutants niaD392 and niaD436 reverted at a frequency of 9x10(-3) and 4x10(-2), respectively. Two of the mutants, niaD587 and niaD410, reverted at a lower frequency of 6x10(-4).

Intragastric infection of conventional and germfree mice with Giardia lamblia.

The effects of experimental infection with Giardia lamblia were studied in 30-day old conventional and germfree CFW mice (7 animals in each group) of both sexes. Cysts were observed in the feces of both groups 6 to 7 days after intragastric infection of each animal with about 2.5 x 10(5) G. lamblia trophozoites. Fecal cyst level was statistically higher in germfree mice (about 10(5) cysts/g feces) when compared with the conventional group (about 10(4) cysts/g feces). The peak of infection in the conventional group apparently occurred on the 10th day after infection as indicated by an increase of fecal weight and by histopathological examination. Intense infiltration of the lamina propria and high reactional hyperplasia of the lymphoid component were observed in the conventional group. There was no infiltration or hyperplasia in germfree infected mice and fecal weight was relatively constant throughout the experiment. These results suggest that, as is the case for other intestinal pathogenic protozoa, the intestinal microflora is indispensable for the expression of the pathogenicity but not for the multiplication of G. lamblia.

Intragastric infection of conventional and germfree mice with Giardia lamblia

The effects of experimental infection with Giardia lamblia were studied in 30-day old conventional and germfree CFW mice (7 animals in each group) of both sexes. Cysts were observed in the feces of both groups 6 to 7 days after intragastric infection of each animal with about 2.5 x 10 5 G. lamblia trophozoites. Fecal cyst level was statisticaly higher in germfree mice (abouth 10 5 cysts/g feces) when compared with the conventional group (about 10 4 cysts/g feces). The peak of infection in the conventional group apparently occurred on the 10th day after infection as indicated by an increase of fecal weight and by histopathological examination. Intense infiltration of the lamina propria and high reactional hyperplasia of the lymphoiud component were observed in the conventional group. There was no infiltration or hyperplasia in germfree infected mice and fecal weight was relatively constant throughout the experiment. These results suggest that, as is the case for other intestinal pathogenic protozoa, the intestinal microflora is indispensable for the expression of the pathogenicity but not for the multiplication of G. lamblia

Extracellular release of protease III (ptr) by Escherichia coli K12.

Escherichia coli strains carrying the protease III structural gene (ptr) on a plasmid secreted the protein into the growth medium. Plasmid-encoded beta-lactamase and chloramphenicol acetyl transferase, which served as periplasmic and cytoplasmic markers during cell fractionation, were not released into the growth medium. There appeared to be some strain dependence on the proficiency of the secretion system. Protease III was not detectably processed upon export through the outer cell membrane.

Analysis of the altered mRNA stability (ams) gene from Escherichia coli. Nucleotide sequence, transcriptional analysis, and homology of its product to MRP3, a mitochondrial ribosomal protein from Neurospora crassa.

The product of the altered mRNA stability (ams) gene of Escherichia coli is involved in decay of mRNA. The complete nucleotide sequence of a 4-kilobase BamHI restriction fragment containing the ams coding sequence was determined. Transcription of the ams gene was analyzed by high resolution S1 mapping. A promoter was found with a homology score of 58% 361 nucleotides upstream from the start codon of ams. The ams structural gene consists of an open reading frame of 2,445 nucleotides. The protein predicted from this open reading frame has a molecular mass of 91,327 Da, which is significantly smaller than that determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Confirmation of the ams coding sequence was obtained by comparing the predicted amino acid sequence with that derived by amino-terminal analysis of gel-purified Ams protein. The predicted protein sequence of the ams gene was screened against translations of the GenBank DNA sequence data base. A homology of 18% over a region of 315 amino acids of the carboxyl terminus of the Ams product was found to MRP3, a mitochondrial ribosomal protein from Neurospora crassa. A smaller region of homology (29% in 86 residues) was found to the human U1 small nuclear ribonucleoparticle 70,000-Da protein.

Transcriptional antitermination in the bgl operon of E. coli is modulated by a specific RNA binding protein.

Regulation of bgl operon expression in E. coli occurs by a mechanism involving antitermination of transcription at two termination sites within the operon. The bglG gene product is absolutely required for this process. Here we provide evidence that BglG is an RNA binding protein that recognizes a specific sequence located just upstream of each of the terminators. The sequence was delimited using a series of specific oligonucleotide probes. Mutational analysis of this sequence indicates that the protein requires a specific RNA secondary structure for recognition. We propose that BglG prevents transcription termination by binding to nascent RNA and blocking formation of the terminator structure.

Cloning of the altered mRNA stability (ams) gene of Escherichia coli K-12.

A temperature-sensitive mutation in the ams gene of Escherichia coli causes an increase in the chemical half-life of pulse-labeled RNA at the nonpermissive temperature. Using lambda clones containing DNA fragments from the 23- to 24-min region on the E. coli chromosome, we have isolated a 5.8-kilobase DNA fragment which, when present in a low-copy-number plasmid, complements the conditional lethality and increased mRNA stability associated with the ams-1 mutation. The approximate initiation site and the direction of transcription of the ams gene were determined from the size of truncated polypeptides produced by Tn1000 insertions and Bal 31 deletions. Overexpression of the ams locus by using a T7 RNA polymerase-promoter system permitted the identification of an ams-encoded polypeptide of 110 kilodaltons.

Chronic calcifying pancreatitis in children: a report of six cases.

1. A study of six patients with Chronic Calcifying Pancreatitis (CCP) occurring over a 10 year period, representing an incidence rate of approximately 1:1,000 of the inpatients in the children's ward of a general hospital in central Brazil, is reported. 2. Major clinical manifestations as well as therapeutic management are described. 3. The possible relationship between CCP and primary protein-calorie malnutrition is discussed and the importance of a CCP diagnosis when dealing with malnourished children who do not respond satisfactorily to common therapy is emphasized.

Chronic calcifying pancreatitis in children: a report of six cases

1. Study of six patients with chronic calcifying pancreatitis (CCP) occurring over a 10 year period, representing an incidence rate of approximately 1:1,000 of the impatients in the children's ward of a general hospital in central Brazil, is reported. 2. Major clinical manifestations as well as therapeutic management are described. 3. The possible relationship between CCP and primary protein-calorie malnutrition is discussed and the importance of a CCP diagnosis when dealing malnourished children who do not respond satisfactorily to common therapy is emphasized