Cystic fibrosis in Afro-Brazilians: XK haplotypes analysis supports the European origin of p.F508del mutation.

Cystic fibrosis (CF) is a common autosomal recessive disorder, being the p.F508del the most frequent mutation. Also, a nearby restriction fragment length polymorphism (RFLP) named XK (KM19 and XV2C) is non-randomly associated with specific CF alleles. Our aim was to analyze the occurrence of the p.F508del mutation and XK haplotypes in Afro-Brazilians CF patients and controls, since these data is available for the other two main ethnic groups found in Brazil (Euro-Brazilians and Brazilian Amerindians), contributing for the whole comprehension of these haplotypes in the Brazilian population. A total of 103 patients and 54 controls were studied. PCR and PCR-RFLP methodologies were used to identify the presence of the p.F508del and the XK haplotype in the subjects. The combined data show that 84.2% of p.F508del mutation is associated with haplotype B and only 15.8% with haplotype A; no other haplotypes were found to be associated with this mutation. Our data suggest that the occurrence of p.F508del mutation and haplotype B in Afro-Brazilian patients occurs probably due to admixture with Euro-descendants. Therefore this mutation and haplotype could be used as a admixture marker.

CD43 signals prepare human T cells to receive cytokine differentiation signals.

T cells are increasingly used for passive immunotherapy and bone marrow transplantation. Proper ex-vivo management of the cells is important for the desired therapeutic effects. For differentiation into effector cells of the Th1 and Th2 phenotypes, T-cells require signals from IFNγ and IL-4, respectively. Naïve cells have an extremely low expression of the specific receptors that recognize these cytokines, indicating that in order to differentiate, cells need to perceive other signals that will enable them to sense the cytokine milieu. CD43 has been proposed as one of the molecules that make the initial contacts with antigen presenting cells. We report here that in cord blood, adult naïve and total human T cells, CD43 signals induced the expression of both IFNγ and IL-4 receptors, mediate their capping, increased their signaling and augmented differentiation mediated by these receptors. CD43 signals also stimulated the expression of IFNγ and in neonatal cells that of IL-4 as well. These data demonstrate an important role for CD43 signals in T-cell preparedness for differentiation into effector cells.

Metabolic evaluation of cooled equine spermatozoa.

Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and the metabolism of these samples was accessed by calorimetry. Centrifugation is part of some of these processing protocols and although this procedure has been deleterious for sperm viability and plasma membrane integrity, no decrease in sperm motility was observed. Microcalorimetry was capable of detecting the positive effect of re-suspending the sperm pellet with Kenney extender. Thus, the use of microcalorimetry offered additional information for equine sperm metabolism evaluation and was efficient in detecting important information from sperm cell metabolism.

Comparison of two methods for cleaning breast pump milk collection kits in human milk banks.

BACKGROUND: Appropriate decontamination of breast pump milk collection kits (BPKs) is critical to obtain safe milk for infants and to avoid discarding donor human milk (DHM). AIM: To evaluate two strategies for BPK decontamination by assessing microbial cultures and the proportion of discarded DHM, according to the criteria of the National Institute for Health and Care Excellence for pre-pasteurization cultures. METHODS: Prospective comparative study, allocation ratio 1:1, microbiologist-blind. PARTICIPANTS: 47 new donors in a human milk bank in Madrid. INTERVENTIONS: Study group (N=21): BPKs washed with water and detergent after each use and further steam decontamination within a microwavable bag. Control group (N=26): washing, rinsing and drying only. Five samples: first sample by hand expression and four samples (one per week) collected using the same pump and method. OUTCOMES: Primary: proportion of DHM discarded due to contamination. Secondary: comparison of the microbiota between samples obtained by hand expression and breast pump in both groups. FINDINGS: In total, 217 milk samples were collected: 47 by hand expression and 170 by pump expression (78 from study group). Steam decontamination of BPKs using a microwavable bag after washing resulted in a lower proportion of discarded DHM samples (1.3% vs 18.5%, P<0.001) and samples contaminated with Enterobacteriaceae (1.3% vs 22.8%, P<0.001) and Candida spp. (1.3% vs 14.1%, P<0.05) compared with samples collected with BPKs that were washed but not steam decontaminated. There were no differences in bacterial contamination between samples obtained using steam decontaminated BPKs and those obtained by hand expression. CONCLUSIONS: Steam decontamination of BPKs using a microwavable bag after washing decreases the amount of discarded DHM and the number of samples with potentially pathogenic bacteria.

Bacillus thuringiensis improved isolation methodology from soil samples.

Bacillus thuringiensis is a sporulating bacterium, which produces parasporal inclusions toxic to insects. Widely used methodologies to isolate Bt from soil consist of a thermal shock treatment followed by selective germination of spores. The results presented here suggest that a preliminary 5 h dry-heat treatment largely enhance the selectivity of these procedures.

A novel and efficient and low-cost methodology for purification of Macrotyloma axillare (Leguminosae) seed lectin.

The N-acetyl-galactosamine specific lectin from Macrotyloma axillare seeds (LMA) was purified by precipitation and ion exchange chromatography. The LMA 0.2 mol L(-1) fraction showed hemagglutinating activity on erythrocytes A1. The results for molecular mass determinations were about 28 kDa. The LMA pH-dependent assays showed best hemagglutinating activity at pH 6.0-8.0; being decreased at acidic/alkaline conditions and by EDTA treatment. LMA is a tetramer at pH 8.2 and a dimer at pH 4.0. Human erythrocytes from ABO system confirmed the A1 specificity for LMA. This new methodology is useful and easy, with low costs, for lectin purification in large amounts.

Physical-chemical characterization and stability study of alpha-trypsin at pH 3.0 by differential scanning calorimetry.

alpha-Trypsin is a serine-protease with a polypeptide chain of 223 amino acid residues and six disulfide bridges. It is a globular protein with predominance of antiparallel ss-sheet secondary structure and it has two domains with similar structures. In the present work, a stability study of alpha-trypsin in the acid pH range was performed and some physical-chemical denaturation parameters were measured by using differential scanning calorimetry (DSC). The alpha-trypsin has a shelf-life (t(95%)) of about 10 months at pH 3.0 and 4 degrees C and its hydrolysis into the psi-trypsin isoform is negligible during 6 months. The observed ratio DeltaH(cal)/DeltaH(vH) is close to unity, which suggests the occurrence of a two-state transition. At pH 3.0, alpha-trypsin unfolded with T(m) = 325.9 K and DeltaH = 99.10 kcal mol(-1), and the change in heat capacity between the native and unfolded forms of the protein was estimated to be 1.96+/-0.18 kcal mol(-1)K(-1). The stability of alpha-trypsin calculated at 298 K was DeltaG(U)=6.10 kcal mol(-1) at pH 3.0. These values are in the range expected for a small globular protein. These results show that the thermodynamic parameters of unfolding of beta-trypsin do not change substantially after its conversion to alpha-trypsin.

Living-Donor Kidney Transplantation: Donor-Recipient Function Correlation.

BACKGROUND: With the rising prevalence of living-donor kidney transplantation, evaluation of factors correlated with renal function in the donor-recipient pair constitutes a main goal for kidney transplantation clinicians. Our objective was to analyze the more relevant donor characteristics that contribute to donor and recipient estimated glomerular filtration rates (eGFR) after 1 year. METHODS: We evaluated 48 consecutive donor-recipient pairs from our unit. RESULTS: Mean donor age was 46 ± 11 years, with 71% being women. Mean recipient age was 35 ± 12 years, with 54% being men. Mean duration of donor hospitalization was 7 ± 2 days. Donor eGFR was 104 ± 11 mL/min/1.73 m2 before donation and 70 ± 14 mL/min/1.73 m2 at discharge. After 1 year, donor eGFR was 71 ± 12 mL/min/1.73 m2 and recipient eGFR was 69 ± 10 mL/min/1.73 m2. Donor eGFR <100 mL/min/1.73 m2 before donation and age >50 years correlated with 17.7- and 8.9-fold increased risks, respectively, of recipient eGFR <60 mL/min/1.73 m2 after 1 year. Donor being female, although statistically associated with worse graft function, compared with a male donor (P = .020), did not represent a significantly increased risk of recipient eGFR <60 mL/min/1.73 m2. Higher donor body mass index (BMI) also associated with a lower kidney function for donors (P = .048). In multivariate linear regression to predict pairs' eGFRs after 1 year, only donor eGFR before donation and at discharge retained statistical significance (P ≤ .001 and P = .045, respectively). CONCLUSIONS: Excluding unpredictable complications in the post-transplantation period, donor eGFR before donation, eGFR at discharge, and age were the best parameters to predict recipient and donor eGFRs after 1 year and can be used as a tool for managing expectations regarding the post-transplantation period.

Effect of potent antiglucocorticoids on dexamethasone induced enzymes in cultured hepatoma and rat liver cells.

The antagonism by 3 synthetic steroids of the induction of tyrosine aminotransferase and tryptophan dioxygenase by dexamethasone were compared in HTC and FAZA hepatoma cells and in isolated liver cells. It was observed: (i) the sensitivity of liver cells to glucocorticoid agonists and antagonists changed in relation to time of culture; (ii) the extent of the inhibitory effect on enzyme induction depends on the nature of the antagonist; (iii) hepatoma cells, especially HTC cells, appeared more sensitive to glucocorticoid antagonism than liver cells.

Control of phenylalanine and tyrosine metabolism by phosphorylation mechanisms.

A system for the parallel determination of enzyme phosphorylation and expressed activity in rat liver cells, and its application to studies of phenylalanine hydroxylase and tyrosine aminotransferase, is described. Phenylalanine hydroxylase is phosphorylated by agents which stimulate cyclic AMP- and Ca2+-dependent protein kinase activity. The phosphorylation site(s) appear to be the same for both kinases. Phosphorylation is accompanied by increased metabolic flux at low, physiologically relevant, substrate concentrations. Insulin and spermine both inhibit the phosphorylation of the enzyme, possibly by increasing dephosphorylation. Tyrosine aminotransferase is phosphorylated in liver cell incubations but the rate is slow and insensitive to additions to the medium. No parallel changes in flux could be detected. Both enzymes are subject to complex regulatory mechanisms, short- and long-term. Their activities may be coordinated in vivo by control exerted at the level of the plasma membrane where both amino acids share the same transport processes. Determination of the control coefficients for the several components indicates that membrane transport may be a major limitation on flux.

Gliomatosis cerebri: findings with computed tomography and magnetic resonance imaging.

Gliomatosis cerebri (GC), is a rare neoplastic disease (less than 150 cases reported in the literature) with a diffuse, widespread proliferation of neoplastic glial cells in the brain, generally affecting both hemispheres and involving the gray and white matter [1-3]. Less commonly, the cerebellum, the brain stem and the medulla can be affected. Histologic evaluation reveals neoplastic astrocytes with varying levels of differentiation. Perineuronal and perivascular spread of tumor infiltration is observed. Demyelination can be found in the affected areas. A well-preserved underlying neuroanatomic architecture is considered characteristic [2]. Clinical signs vary and are non-specific, including changes in the mental state and headaches, followed by focal motor deficits and convulsive episodes [4]. The prognosis is poor, ranging from weeks to some years after the manifestation of the symptoms. Steroids may be useful in the short term, but chemotherapy is of little value and radiotherapy of questionable benefit. The literature was reviewed and the radiological pattern of three new cases of GC is reported. In two cases the diagnosis was achieved ante-mortem.

Hyperalgesic effect induced by barbiturates, midazolam and ethanol: pharmacological evidence for GABA-A receptor involvement.

The involvement of GABA-A receptors in the control of nociception was studied using the tail-flick test in rats. Non-hypnotic doses of the barbiturates phenobarbital (5-50 mg/kg), pentobarbital (17-33 mg/kg), and thiopental (7.5-30 mg/kg), of the benzodiazepine midazolam (10 mg/kg) or of ethanol (0.4-1.6 g/kg) administered by the systemic route reduced the latency for the tail-flick response, thus inducing a 'hyperalgesic' state in the animals. In contrast, non-convulsant doses of the GABA-A antagonist picrotoxin (0.12-1.0 mg/kg) administered systemically induced an increase in the latency for the tail-flick response, therefore characterizing an 'antinociceptive' state. Previous picrotoxin (0.12 mg/kg) treatment abolished the hyperalgesic state induced by effective doses of the barbiturates, midazolam or ethanol. Since phenobarbital, midazolam and ethanol reproduced the described hyperalgesic effect of GABA-A-specific agonists (muscimol, THIP), which is specifically antagonized by the GABA-A antagonist picrotoxin, our results suggest that GABA-A receptors are tonically involved in the modulation of nociception in the rat central nervous system.

[Neurosyphilis: forms of presentation and clinical management]. / Neurosífilis: formas de presentación y manejo clínico.

INTRODUCTION AND AIMS: Neurosyphilis results from the infection of the central nervous system by Treponema pallidum. It causes diverse clinical pictures which are occasionally similar to other, better known neurological diseases. In this paper our aim is to offer a global clinical vision of this entity by reviewing the different forms it can take and its diagnostic and therapeutic management. DEVELOPMENT: The forms of presentation of neurosyphilis can be grouped in two categories: early (asymptomatic, meningeal and meningovascular neurosyphilis) and late (progressive general paralysis and tabes dorsalis). Other less important forms, such as gummas, ocular forms, syphilitic amyotrophy or hypoacusis, have also been described. Diagnosis is complex and is based on the study of the cerebrospinal fluid. Given the difficulty involved in performing an accurate diagnosis, different criteria have been developed in which T. pallidum serology plays a key role. The most effective treatment is penicillin, although on occasions it may have no effect and we therefore recommend clinical and fluid analysis follow ups. Lastly, we describe the changes in incidence and clinical presentation, and the complications that may arise in diagnosis when HIV carrying patients also suffer from this disease. CONCLUSIONS: Neurosyphilis is a disease that still occurs nowadays and, due to its clinical polymorphism, must be borne in mind as a differential diagnosis in a number of neurological and psychiatric illnesses. This, together with the fact the serological tests are difficult to interpret and its irregular response to the usual treatment, makes it difficult to manage and means that the neurologist must have a thorough knowledge of the disorder.

[Massive hepatobiliary fascioliasis]. / Fasciolosis hepatobiliar masiva.

UNLABELLED: Massive hepatobiliary fascioliasis. Report of two cases. INTRODUCTION: Liver fascioliasis is the most frequent parasitic disease of the biliary tract in Mexico. Massive infestation is common in sheep and bovines and rarely occurs in humans. Only two previous cases of massive fascioliasis has been reported and we present here in two additional cases. Case reports. Both cases presented in alcohol-related liver disease patients who had eaten watercress Nasturium officinalis. Upper abdominal pain, obstructive jaundice, and choledochal dilatation were the salient manifestations. Diagnosis was made by surgical exploration and adult fluke identification. One patient died due to progressive liver failure and the other, is alive and received praziquantel treatment. CONCLUSIONS: Since 1935, approximately fifty cases of massive hepatobiliary fascioliasis have been reported in Mexico, most misdiagnosed as cholelithiasis; however, a history of watercress ingestion has been recorded in most of these cases.

[Study of anxiety levels in neonatal unit staff]. / Estudio de los niveles de ansiedad en los profesionales de una unidad neonatal.

INTRODUCTION: The professionals who routinely work in neonatal units become under stress due to the working conditions and the nature of the tasks carried out. As a consequence of this, they may have high levels of anxiety. Anxiety is defined as an emotional response or response patterns that include cognitive, physiological and behavioural aspects. METHODS: A prospective cross-sectional study was conducted on all neonatologist, nurses and care assistants who were given two self-administered questionnaires under baseline conditions, to assess Sate Anxiety and Trait Anxiety. (IDDA-EA; STAI, State-Trait Anxiety Inventory). RESULTS: The response rate was 88.5%, and 36% scored between 30 and 70 percentiles on State Anxiety, with 11.8% above the 70 percentile and 51% below the 30 percentile. There were no significant differences in relation to occupation or age. There were also no significant differences between State and Trait Anxiety. Regarding gender, it has obtained A statistically significant difference of 8 points higher was obtained for women. CONCLUSIONS: For the majority professionals everyday situations in which are involved do not significantly raise the burden of anxiety, as half of them have levels below the average. They perceive themselves as competent and able to cope with their tasks. They have a high degree of interest and attention in the activities performed.

The stigma of being named "AIDS carriers" on Haitian-American women.

AIDS-related stigma has impacted the lives of many individuals, including Haitians. Little, however, is known about the long-term effects AIDS-stigmatization has had on Haitian women living in the United States. A pilot study was conducted to explore Haitian-American women's perceptions of the impact of the AIDS epidemic on their lives. The long-term effects of AIDS-stigmatization fell into five categories: rejection by the dominant society, self-doubt, effect on self-esteem, effect on intimate relationships, and rejection by Haitians within their community. More extensive research is needed to explore the long-term effects of the AIDS epidemic on Haitian women. Additionally, to counter the negative effects of the AIDS epidemic, intervention strategies need to be developed and tested.

Isoelectric focusing of cassava protoplasts.

Cassava (Manihot esculenta Crantz) protoplast was analyzed by using isoelectric focusing techniques. Two populations, representing 68 and 32% of the total sample, with mean isoelectric points of 4.48 and 4.60, were obtained using mesophyll protoplasts. The use of this technique allows demonstration of a discontinuous distribution of protoplast isoelectric point from one species according to their surface potential.

What it takes to become an effector T cell: the process, the cells involved, and the mechanisms.

When activated, CD4(+) T cells differentiate into two major sub-populations differing in their profiles of secreted cytokines. Type One, or TH1, cells secrete IL-2, IFNgamma, and TNFbeta and mediate a cellular immune response. Type Two, or TH2, cells secrete IL-4, IL-5, IL-6, IL-10, and IL-13 and potentiate a humoral response. The nature of any specific immune response depends on the interaction of antigen-presenting cells and T cells. The role of antigen-presenting cells is to respond to the nature of the immune challenge and signal differentiation of CD4(+) T cells. A number of factors are involved in the effector phenotype of T cells-nature and affinity of antigen, co-receptors signals, and cytokine environment. T-cell differentiation is a complex process comprising four defined developmental stages: activation of particular cytokine genes, commitment of the cells, silencing of the opposing cytokine genes, and stabilization of the phenotype. In each of these stages, the cells respond to the products of many signaling cascades from many membrane-bound receptors. The stages in development are mediated by different molecular mechanisms, involving control of gene expression and chromatin remodeling. This review centers on the factors, cellular interactions, and molecular mechanisms involved in the maturation of naïve CD4(+) T lymphocytes into fully effector cells.