HLA-B14 subtyping by semi-nested PCR-SSP and haplotype distribution in a Spanish population

HLA-B14 serological subtyping is very limited probably due to the internal position of the unique amino acid residue that differentiates B64 and B65 molecules. In order to carry out an accurate B14 subtyping we have designed a semi-nested PCR-SSP procedure that can differentiate B*1401 and B*1402 in any HLA-A, -B or -C antigen combination. A panel of 133 B14-positive and 31 B14-negative healthy and unrelated Spanish individuals were studied. Additionally, 45 B14-bearing haplotypes (-A,-B,-C,-DRB1,-DRB3/DRB4/DRB5,-DQA1,-DQB1) were available through family studies. The relative frequencies of HLA-B14 subtypes were 74% for B*1402 and 26% for B*1401, in agreement with those found in other Central European populations, but differing from those in Wales, where the relative presence of B64 goes to 41%. A total of 11/17 and 18/28 different haplotypes for B*1401 and B*1402, respectively, were identified. Both alleles showed the strongest association to Cw8 (43/45), indicating a primary ancestral B14-Cw8 association. However, B14 subtypes evidenced very distinguishable haplotype distributions. B*1401 is strongly associated with the common HLA class II haplotype DRB1*0701-DQA1*0201-DQB1*02 (13/17), while B*1402 is mainly associated to DRB1*0102 (16/28). Three major haplotypes were identified: A32-Cw8-B*1401-DR7-DQ2 (5/17), A33-Cw8-B*1402-DRB1*0102-DQ5 (5/28) and A2-Cw8-B*1402-DRB1*0102-DQ5 (5/28).     

HIV-1 DNA burden dynamics in CD4 T cells and monocytes in patients undergoing a transient therapy interruption

Replication-competent HIV, as well as HIV-1 DNA, has been detected in CD4 T cells and in monocytes during antiretroviral therapy (ART), indicating that these cells could represent an important viral reservoir. We measured HIV-1 DNA in monocytes and CD4 T cells in patients undergoing transient therapy interruption (TTI), to establish the dynamic of HIV-1 DNA burden and to find possible correlations with immune restoration and re-establishment of virological control after ART resumption. In most patients CD4 depletion and viral load rebound followed TTI. Rapid resumption of virological and immunological control was achieved after ART reintroduction. After TTI, in most cases a transient increase of both monocyte and CD4 HIV-1 DNA burden was observed. After ART reintroduction, both CD4 T cell and monocyte HIV-1 DNA copy number decreased, reaching baseline levels at the end of observation. At this time monocyte HIV-1 DNA burden was always undetectable, while CD4 T cell HIV-1 DNA burden was lower than at baseline. As CD4 T cell HIV-1 DNA values are independently associated with CD4 depletion, the increase of HIV-1 DNA burden in these cells after TTI is presumably due to acute infection, causing cell death. This is also supported by the pattern of 2-LTR appearance in these cells after TTI. HIV-1 DNA burden in monocytes and CD4 T cells show high correlation, suggesting reciprocal re-feeding of two cell populations. Repopulation by HIV these cells after TTI is temporary, and no significant changes of HIV-1 DNA burden were observed after ART resumption respect to pre-TTI period.     

MR Imaging Findings in Molecular Subtypes of Breast Cancer According to BIRADS System

To evaluate magnetic resonance imaging (MRI) findings, according to Breast Imaging‐Reporting and Data System (BI‐RADS), and to relate them with molecular subtypes of breast cancer. The MRI findings were reviewed retrospectively in 201 women diagnosed of invasive breast cancer confirmed by surgery and were compared with the molecular subtypes. Following the BI‐RADS, MRI findings included disease type, size, enhancement, morphology and contrast kinetics. In mass‐like lesion types were studied shape, margin and enhancement, and in nonmass‐like lesion types, distribution modifiers and internal enhancement. Chi‐squared analysis showed significant association (p < 0.01) between molecular subtypes and lesion type on MRI and histologic grade. Shape, margin and mass enhancement (p < 0.05) also showed significant association among molecular subtypes. Triple negative were more frequently unifocal and mass‐like lesion, high histologic grade, round shape, smooth margin, and rim enhancement. Luminal‐A were more frequently low grade, mass‐like lesion, irregular shape and spiculated or irregular margin. Luminal‐B were more frequently moderate‐low grade, mass‐like lesion, nonirregular shape and spiculated margin. HER‐2‐enriched were more frequently moderate grade, nonmass‐like lesion and multicentric lesions were more present than in other subtypes. There are significantly different MRI features, according to BI‐RADS, between the molecular subtypes breast cancer.     

Qualitative Research Process Applied to Organ Donation

The family interview for organ donation continues to be one of the main concerns of transplant coordinators. We approach the family because we need to know the opinion of the person who has just died about the donation of his or her organs. The objective of this article is to describe how the application of the qualitative methodology has been followed to perceive what the relatives live in the moment in which they are informed of the death of their relative and they are offered the option of organ donation. Phenomenology is a philosophy that deals with the granting of meanings. It can simply describe or try to interpret them. Our informants were selected from among donor families until the 11 families were obtained. We use the Giorgi method because it allows us to identify the essence of a phenomenon and transform the lived experience into a textual expression of its essence. The unit of analysis was, each of the statements of the discourse that describes and interprets a reality, what we call “essence” or “meaning units.” It must be the minimum expression of an episode and the experience in relation to it. We have detected 24 themes of the meaning units are most important for families in our area. Families agree on talking about donation, mourning, death, consolation from the donation, and the importance of the support received. Phenomenology is adequate to deepen into the feelings and interests of families in the process of donation.     

Pediatric renal transplantation: a single center experience over 14 years

Between 1989 and 2003, 100 transplants were performed in 96 patients at the pediatric nephrology unit of the Calvo Mackenna Children's Hospital. Mean age 10.9 +/- 3.9 yr (1-17.6), 30% from LD. Donors were younger than 5 yr in five patients and all recipients received an 'en bloc' graft. Original disease was hypo/dysplasia 27%, reflux nephropathy 22 and 17% chronic glomerulonephritis. The immunosuppressive protocol during the first period (n = 56, 1989-2000): Cyclosporine, steroids and azathioprine, and during the second period (n = 44, 2001-2003): FK, steroids, MMF and anti-CD25 antibody (mAbs). AR was reported in 22 patients, 11% in LD, 31% in DD (p < 0.01). The AR rate decreased from 40 to 8% after anti-CD25 monoclonal induction. Patient actuarial survival rate at 1, 3 and 5 yr was 100% for LD and 96% for DD. The overall actuarial graft survival at 1,3, and 5 yr was 96.7, 96.7 and 71% for LD and 89, 76 and 73% for DD donors. Graft survival rate improved from the first period (1989-2000) to the second period (2001-2003; p = 0.05). No difference in graft survival rate with HLA-A,B,DR matching was found. Graft survival rate was better when cold ischemia time was <24 h (p < 0.01). CMV infections increased from 19 to 40% when MMF and anti-CD25 Ab were introduced (p < 0.01). The height/age Z score at 1, 3 and 5 yr post-transplant was -2.2, -2.1, -2.2, respectively, for children older than 7 yr and -1.8, -1.9, -2.1 for those transplanted younger than 7 yr of age who were switched to alternate day steroids (p < 0.01). The cause of graft lost was: chronic rejection eight, non-adherence four, AR four and vascular thrombosis two. The cause of death in two patients was fungus septicemia and accelerated rejection. Pediatric renal transplantation can be performed in our group with acceptable morbidity, low mortality and graft survival rates similar to other reports in North America and Western Europe. Graft survival rate improved with newer immunosuppression and greater experience at the center. Management of non-adherence and chronic rejection remain the major challenges.     

Periodontal disease and nitric oxide levels in low risk women with preterm labor

Objectives

The aims of the study were to evaluate the possible association between periodontal disease (PD) and preterm labor (PTL), to assess whether the presence of periodontal disease in women with PTL increases the risk of preterm birth (PTB), and to evaluate the role of nitric oxide (NO) in this possible association.

Study design

This study included 820 nulligravid women with low risk socioeconomic status: 400 cases with PTL and 420 controls with normal pregnancy, between 25 and 33 weeks + 6 days of weeks’ gestation. At enrollment, periodontal examination and detection of plasma, gingival and cervical NO levels were performed. We compared the periodontal status of the two groups; we detected the presence of PD and compared NO levels. We then followed the outcome of women with PTL and compared obstetric status and PTB rate between patients with and without PD.

Results

Logistic regression analysis revealed a strong association between PD and PTL (adjusted odds ratio: 2.83, 95% confidence interval (CI) 1.86-4.23; P < 0.0001). Non parametric Mann-Whitney U-test demonstrated significant differences in gingival and cervical NO levels between women with PTL and controls (respectively, median 85.1 μmol/L, interquartile range (i.r.) 51.2-177 vs median 50.5 μmol/L, i.r. 34.5-109.65, P < 0.0001; and median 102 μmol/L, i.r. 53.05-182.7 vs median 38.9 μmol/L, i.r. 32.87-46.1, P < 0.0001). The ability of mean-gingival NO levels to predict PTL was examined by ROC curve analysis: the area under the curve was 0.817 (95% CI 0.774-0.854; P < 0.0001). The cut-off level for the greatest sensitivity and specificity for mean-gingival NO levels was 116.04 μmol/L (sensitivity 0.57, specificity 0.94). Positive and negative predictive values were, respectively, 90.65% and 69.73%. Among women with PTL, the risk of PTB increased in women with PD (adjusted relative risk: 3.85, 95% CI 2.11-6.06; P < 0.0001).

Conclusions

In this tested population, PTL is associated with PD. The presence of PD in women with PTL increases the risk of premature delivery. NO may be a new marker to explain this association.     

Pattern recognition in tufted capuchin monkeys (Cebus apella): the role of the spatial organisation of stimulus parts

We report four experiments aimed at characterising the role played by the encoding of the spatial relationship between stimulus parts in pattern recognition in capuchin monkeys, as assessed by a matching to sample task. The results of the first experiment, which were also reliably replicated at different stages in the course of the study, indicated that the simultaneous rotation and/or translation of the four parts into which the stimuli were divided, but not a global rotation of the entire stimulus, impaired matching performance in capuchin monkeys. Experiments two and three showed that matching performance was not impaired following similar manipulations of a subset of one, two or three parts. In experiment four, the same task was presented to human subjects. The same pattern of results emerged for humans and monkeys in trials where all the four stimulus parts were presented. However, the matching performance of humans was affected more than that of capuchin monkeys when only a subset of stimulus parts was featured in the task. These results support the conclusion that the matching performance of capuchin monkeys is affected by the rearrangement of stimulus parts and, as such it seems to rely on global properties of the stimulus such as the spatial relationships of the component parts. However, the remarkable ability of capuchin monkeys to identify a stimulus on the basis of a subset of parts suggests that the reliance on the global properties of the stimuli may not be pervasive as it is in humans.     

Hepatocyte growth factor protects rat RINm5F cell line against free fatty acid-induced apoptosis by counteracting oxidative stress

Type 2 diabetes is characterized by peripheral insulin resistance, pancreatic beta-cells dysfunction, and decreased beta-cell mass with increased rate of apoptosis. Chronic exposure to high levels of free fatty acids (FFAs) has detrimental effects on beta-cell function and survival. FFAs have adverse effects on mitochondrial function, with a consequent increase in the production of reactive oxygen species. Hepatocyte growth factor (HGF) plays a critical role in promoting beta-cell survival. In the present study, we investigated whether HGF was capable of protecting beta-cells from death induced by prolonged exposure to FFAs. RINm5F cell line was cultured in the presence of FFAs (oleate:palmitate 2:1) for 72 h in order to induce apoptosis. Simultaneous administration of HGF and FFAs significantly suppressed the impaired insulin secretion and FFA-induced apoptosis. Specifically, HGF exerted its protective effect by counteracting: (i) the overproduction of either hydrogen peroxide and superoxide anion, (ii) the reduction of intracellular gamma-glutamylcysteinylglycine level, and (iii) the depolarization of mitochondrial membrane, induced by prolonged FFAs exposure. These effects appear to be mediated by bcl-2 and phosphatidylinositol 3 kinase (PI3K)/Akt pathways. Indeed, HGF increased mRNA and protein expression of bcl-2 downregulated by FFAs-treatment; moreover, pre-treatment with the specific PI3-kinase inhibitor LY294002, significantly abolished the protective effect of HGF. In conclusion, in rat insulin-producing RINm5F cells, HGF exerts its prosurvival effect by counteracting the increased intracellular oxidative stress and, consequently, by inhibiting apoptosis induced by chronic exposure to FFAs.     

Effects of pattern redundancy and hierarchical grouping on global-local visual processing in monkeys (Cebus apella) and humans (Homo sapiens)

Using a Matching-To-Sample (MTS) procedure we assessed the effects of stimulus redundancy, defined on the basis of the information-theory approach to shape goodness proposed by Garner (1974) [20], and grouping on the processing of hierarchical visual patterns in capuchin monkeys and humans. In a first experiment, the MTS performance of both capuchin monkeys and humans benefitted from stimulus redundancy. Moreover, a local advantage in capuchins was observed with visual patterns that required grouping at both global and local level. In a second experiment we eliminated the requirement to group at the local level. This was done to determine if the effects of redundancy would have been evident in condition more similar to those used in previous studies of global-local processing in a comparative context. The benefits of stimulus redundancy emerged again in both species but were confined to local processing in monkeys and to global processing in humans. A local advantage was observed in both species. In a third experiment, the reduction of the size of the stimuli and the increase of the quantity of the local elements produced a shift to global dominance in humans but the local dominance in monkeys was preserved. The implications of these results are discussed in relation to other similarities and differences in higher visual functions in humans and monkeys.     

Progressive subaortic stenosis caused by accessory mitral valve tissue]

Accessory mitral valve tissue is a rare cause of subaortic obstruction. The reported case correspond to a two days old patient diagnosed by 2D-Echocardiography. Serial doppler flow analysis showed progressive left ventricular outflow obstruction. A cardiac catheterization at the age of 9 months confirmed the obstruction without defining the cause. Five months later, transesophageal echocardiography clearly defined the intraventricular connections of the accessory mitral tissue. At the age of 18 months the patient suffered from cardiac failure and underwent surgery for removing the accessory tissue, assisted by intraoperative transesophageal echocardiography pre and post by-pass. Follow-up echocardiography at 25 months showed no left ventricle outflow obstruction and the patient is now asymptomatic.