Joint study of the associations of HLA-B and the transmembrane short tandem repeat polymorphism of MICA protein with alopecia areata shows independent associations of both with the disease.

BACKGROUND: Alopecia areata (AA) is a skin disease that produces hair loss in patches of skin. The underlying mechanism of AA is a loss of immune privilege of hair follicles, which are then attacked by natural killer (NK) cells. A previous genome-wide association study linked single nucleotide polymorphisms of the protein MHC class I chain-related A (MICA) to this disease. MICA is the ligand for the activating receptor NKG2D, expressed mainly by NK cells and CD8+ cytotoxic T cells. As the aforementioned study did not include short tandem repeats (STRs) of MICA, we decided to study these in relation to AA. AIM: To study the association of STRs with AA, alongside that of human leucocyte antigen (HLA) locus B, which is closely linked to MICA. METHODS: DNA amplicon size analysis was carried out, and HLA-B locus genomic typing was performed by PCR-sequence-specific oligonucleotide analysis. RESULTS: We observed an association between AA and both MICA*009 and HLA-B14; associations were also observed between HLA-B alleles and MICA alleles, which have both been previously found to be connected with AA, but never studied together. CONCLUSIONS: We conclude that it is important to study HLA-B and MICA together to avoid the influence of their association in experiments in which they are investigated separately.

Residues of the minimal sequence of G0S2 collectively contribute to ATGL inhibition while C-and N-terminal extensions promote binding to ATGL.

The protein encoded by the G0/G1 switch gene 2 (G0S2) is a potent inhibitor of adipose triglyceride lipase (ATGL) and thus an important regulator of intracellular lipolysis. Since dysfunction of lipolysis is associated with metabolic diseases including diabetes and obesity, inhibition of ATGL is considered a therapeutic strategy. G0S2 interacts with ATGL's patatin-domain to mediate non-competitive inhibition, however atomic details of the inhibition mechanism are incompletely understood. Sequences of G0S2 from higher organisms show a highly conserved N-terminal part, including a hydrophobic region covering amino acids 27 to 42. We show that predicted G0S2 orthologs from platypus, chicken and Japanese rice-fish are able to inhibit human and mouse ATGL, emphasizing the contribution of conserved amino acid to ATGL inhibition. Our site directed mutagenesis and truncation studies give insights in the protein-protein interaction on a per-residue level. We determine that the minimal sequence required for ATGL inhibition ranges from amino acids 20 to 44. Residues Y27, V28, G30, A34 G37, V39 or L42 within this sequence play a substantial role in ATGL inhibition. Furthermore, we show that unspecific interactions of the N-terminal part (amino acids 20-27) of the minimal sequence facilitate the interaction to ATGL. Our studies also demonstrate that full-length G0S2 shows higher tolerance to specific single amino acid exchanges in the hydrophobic region due to the stronger contributions of unspecific interactions. However, exchanges of more than one amino-acid in the hydrophobic region also result in the loss of function as ATGL inhibitor even in the full-length protein.

Tropomyosin IgE-positive results are a good predictor of shrimp allergy.

BACKGROUND: Shrimp is a common cause of food allergy. Our aims were to determine the value of IgE antibodies in the diagnosis of shrimp allergy and to study red shrimp (Solenocera melantho) tropomyosin both as a new allergen and as a cross-reactive IgE-binding protein. METHODS: We have studied 45 subjects. Skin prick test (SPT) was carried out in all subjects, and specific IgE (sIgE) to shrimp, recombinant and natural shrimp tropomyosins rPen a 1 and nPen m 1, recombinant Der p 10, and Dermatophagoides pteronyssinus was assessed by fluoroimmunoassay and/or immunoblotting. Double-blind, placebo-controlled food challenges were carried out to confirm diagnosis of shrimp allergy. Also, in vitro inhibition tests were performed to evaluate cross-reactivity. RESULTS: Shrimp allergy was confirmed in 18 shrimp-allergic patients. Skin prick test and IgE antibodies to shrimp were positive in all shrimp-allergic patients; sIgE to rPen a 1 was detected in 98% of these patients. Of the 18 shrimp-tolerant patients, 61% had positive SPT to shrimp, 55% were IgE-positive to shrimp, and 33% showed IgE antibodies to rPen a 1. Determination of IgE to rPen a 1 yielded a positive predictive value of 0.72 and a negative predictive value of 0.91. CONCLUSION: IgE levels to rPen a 1 provided additional value to the diagnosis of shrimp allergy. Some allergens in mite extract are recognized by patients who are allergic to shrimp, though their clinical relevance remains unknown.

Allergenicity and cross-reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach.

BACKGROUND: The genus Senecio is the largest genus of the family Asteraceae (Compositae). The allergenicity of Senecio has not been assessed previously. OBJECTIVE: The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance. METHODS: Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one-dimensional electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross-reactivity with other pollen. RESULTS: Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human-allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two-dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross-reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies. CONCLUSION: S. jacobea constitute a newly discovered allergenic source. It shows cross-reactivity with other members of the Asteraceae plant family as well as with P. judaica.

[Detection of occult malignant melanoma by 18F-FDG PET-CT and gamma probe]. / Detección de melanoma maligno oculto mediante PET-TC con 18F-FDG y sonda gammadetectora.

Malignant melanoma has a high relapse rate, and PET is very useful for its detection, above all when its size is small or it is in locations that are difficult to resect. The radioguided surgery being implanted in many sites, new expectations are opening in these situations--the intraoperative location of the lesions by 18F-FDG and detection probe. This is a case report of a 44 years old woman diagnosed of malignant melanoma in her left leg 12 years ago. At present, she has a tumor in her left thigh, which was no longer palpable after a fine needle puncture was performed. This was reported as a mesenchymal tumor. Due to the possibility of recurrent melanoma, need for excision was suggested. In order to locate it, a PET-CT study with 18F-FDG was performed and the lesion was marked on the skin with the help of metallic points. The lesion was located in surgery with a gamma probe, showing relapse of melanoma. The importance of the case is found in the fact that it is the first step to extend the indications of radioguided surgery with the use of PET, a technique that offers a ride range of possibilities.

Tumor recurrence and tumor-related mortality in endometrial cancer: Analysis in 276 patients.

BACKGROUND: In this manuscript, we assessed tumor recurrence and tumor-related mortality in a clinical series of endometrial cancer patients. MATERIALS AND METHODS: A retrospective evaluation of 276 patients (mean age 64 years) with histologically confirmed endometrial cancer treated at a single hospital in Madrid (Spain) was conducted. The median follow-up was estimated using the inverse Kaplan-Meier method. RESULTS: Salient findings were endometrioid carcinoma (84.8% of cases), grade G1 (48.9%) and stages IB (35.1%) and IC (23.2%). Myometrial infiltration >50% was documented in 31.2% of cases and lymphovascular space invasion in 11.9%. After surgery, 52.5% of patients were classified into the low risk group, 21.4% into the intermediate risk group and 26.1% into the high risk group. Tumor recurrence occurred in 14.5% of patients, with an estimated median follow-up of 45 months (95% confidence interval (CI): 41.2-48.8), locoregional recurrence in 42.5% and distant recurrences in 57.5%. Furthermore, 40% of tumor recurrences developed during the first year after primary treatment and 90% over the first 3 years of follow-up. The tumor-related mortality rate was 15.9%. The estimated median follow-up was 46 months (95% CI: 43.0-49.0). Furthermore, 5.07% of death because of tumor developed during the first year after primary treatment and 13.77% over the first 3 years of follow-up. CONCLUSION: The rates of tumor-related death and tumor recurrence in endometrial cancer patients are low, with the highest percentages occurring within 3 years of primary treatment. Most of the recurrences occur outside the pelvis.

Pulmonary diffusing capacity in chronic dialysis patients.

Patients with end-stage renal disease treated by hemodialysis with bioincompatible membranes are exposed during the dialysis period to acute effects on lung microcirculation, which may result in pulmonary fibrosis and diffusion defects in long-standing dialysis. To investigate the occurrence of these possible chronic pulmonary alterations, we determined lung function in patients with chronic renal failure not undergoing hemodialysis and in patients who had been receiving regular hemodialysis both for short and long periods of time. Forty-three patients divided into three groups were studied: 17 patients before dialysis with a mean (SD) creatinine clearance of 14.1 (6.8) ml/min 11.73 m2, 10 patients receiving regular hemodialysis for a period of less than 12 months (mean 6.4 +/- 3.5 months), and 16 patients receiving regular hemodialysis for more than 5 years (mean 8.3 +/- 3.6 years). First-use bioincompatible cellulosic dialysis membranes were used in all the cases. The following parameters were recorded: forced vital capacity (FVC), forced expiratory volume in 1s (FEV1), total lung capacity (TLC), residual volume (RV), carbon monoxide transfer factor (TLCO), accessible lung volume (VA), carbon monoxide transfer factor/accessible lung volume (KCO- that is, TLCO/VA), and arterial blood gases. Patients receiving regular hemodialysis for more than 5 years showed significantly lower values of TLCO and KCO than patients before dialysis and patients receiving regular hemodialysis for less than 12 months. Seventy-five percent of patients on long-term hemodialysis had markedly reduced TLCO or KCO values (below 80% of the reference value) as compared with 17% of patients before dialysis and 10% of patients dialyzed for less than 12 months (P < 0.001). Differences among groups for the remaining parameters were not observed. In conclusion, patients undergoing long-term regular hemodialysis with a bioincompatible membrane showed a selective reduction in pulmonary diffusing capacity possibly due to chronic pulmonary fibrosis.

Serum copper in institutionalized elderly subjects: relations with dietary intake of energy, specific nutrients and haematological parameters.

The concentrations of serum copper were determined as an indicator of corporal status of Cu and its relation to aging in 93 institutionalized elderly subjects (24 men and 69 women) in Granada (Spain). We found that aging does not affect serum Cu levels in the two age groups included in the study: Group I < 80 years and Group II > or = 80 years. Application of linear regression analysis to the serum Cu levels and the intake of energy, fibre, iron and magnesium showed a statistically significant negative correlation (P < 0.05) among them in all the subjects. However, no correlation was observed when the serum Cu levels were related to the intake of zinc or vitamin C. Therefore, at the daily intake levels of these two nutrients, they do not interfere in the absorption of copper and hence do not affect the corporal status of this element in the elderly. Blood erythrocyte and haemoglobin concentrations do not show any significant correlation with the serum Cu concentrations (P > 0.05).

Serum concentration and dietary intake of Mg and Ca in institutionalized elderly people.

We determined the serum levels and the dietary intake of Mg and Ca in 93 institutionalized elderly people (24 men and 69 women) in Granada (Spain). The serum samples had mean concentrations of 21.39 +/- 2.08 mg/l magnesium and 93.39 +/- 9.24 mg/l calcium. No significant differences were found in the serum levels of the two minerals as regards the sex of the subjects. Application of linear regression analysis to the serum Mg and Ca levels showed a statistically significant positive correlation (P < 0.05). A low vitamin D intake of these subjects (51.8% of the recommended dietary allowance) was observed. The intakes of Ca and Mg correlated positively together, as well as with the intakes of energy, protein, fibre (in the case of Mg only), iron and phosphorus (P < 0.05). The Ca and energy intake were adequate to their current recommendations, as too was the Ca:P ratio, whereas the diet had a poor supply of fibre, vitamin D and Mg.

Serum concentration and dietary intake of Zn in healthy institutionalized elderly subjects.

We determined the serum concentrations and dietary intake of zinc, as indicators of Zn status, in 44 healthy institutionalized elderly subjects in Granada (Spain) (mean age 81.4 +/- 7.9 years). Determination of Zn in serum was carried out by electrothermal atomic absorption spectrophotometry. Serum samples had a mean Zn concentration of 10.49 +/- 3.5 mumol/1. No significant differences were found in the serum levels of Zn as regards the sex of the subjects. However, concerning Zn intake, determined by a 7-day weighted food record, a significant statistical difference (P < 0.001) was found between men and women, with mean values of 10.01 +/- 1.76 mg/day and 7.33 +/- 1.33 mg/day, respectively. Application of regression analysis to the serum concentration of Zn and other nutritional parameters shows a statistically significant correlation (P < 0.05) between serum levels of Zn and the body mass index. The lack of statistical correlation between the serum concentration of zinc and its intake indicates that this index cannot be used as an indicator of zinc status in the elderly.

Nutritional status of vitamin A and E in institutionalized elderly people in Granada (Spain).

A study has been carried out on the nutritional status of vitamin A and E in 93 institutionalized elderly people in Granada (Spain) by studying their vitamin intake and the serum values of retinol and alpha-tocopherol. The influence of lipid intake and lipids in plasma has been also discussed. The vitamin A intake is higher than the recommended amount and represent 209% DR in men and 217% DR in women. The vitamin E intake is deficient in 33% of the men and 27% of the women. The serum values of retinol indicate that 11% of the women have a deficiency (< 0.7 microM), and that 39% of the men and 32% of the women have low values (0.7-1.22 microM). As for the serum values of alpha-tocopherol, 6% of the women have deficient values, and 82% of the men and 37% of the women have low values (11.6-23.2 microM).

[Value of Pet-18FDG in lung cancer]. / Valor de la tomografía por emisión de positrones (PET)-18FDG en el cáncer de pulmón.

INTRODUCTION: 18FDG-PET was studied in the diagnosis of malignancy of the solitary pulmonary nodule and in the early staging of non-small-cell lung cancer. PET results were compared with thoracic-abdominal computed tomography (CT) and brain magnetic resonance (MR). PATIENTS AND METHODS: Fifty-five patients with a radiologically detected solitary pulmonary nodule (54 CT, 1 plain radiography), were studied following an intravenous injection of 370 MBq 18FDG. Attenuation corrected emission data were acquired and analyzed qualitatively and semi-quantitatively. 30 non-small-cell lung cancer underwent MR. Biopsies were obtained in 48 non-small-cell lung cancer and 7 were controlled by follow-up (18 months). The staging of 43 non-small-cell lung cancer was confirmed by surgery (n = 13), mediastinoscopy (n = 9) and follow-up (n = 21). RESULTS: PET correctly diagnosed 52 solitary pulmonary nodules with 3 false positives (100% sensitivity and 75% specificity). In the mediastinal staging (N), CT and PET demonstrated a sensitivity and specificity of 46% vs. 100%, and 59.3% vs. 93.3%, respectively. In 6 patients, some visceral metastases detected by PET were not detected by CT (including 3 adrenals), whereas 2 brain metastases in MR were not diagnosed by PET. PET was considered decisive in the treatment and follow-up of 17 patients (32.7%). CONCLUSIONS: Whole body PET imaging is a cost-effective diagnostic technique that simplifies the malignant characterization of solitary pulmonary nodule and improves the early staging of non-small-cell lung cancer. In combination with CT, PET makes an outstanding contribution to the correct assessment of therapeutical decisions in these patients.

[Hepatitis C virus infection in patients treated with hemodialysis]. / Infección por el virus de la hepatitis C en enfermos tratados con hemodiálisis.

The prevalence of hepatitis C infection was evaluated (Ortho HCV Antibody ELISA Test) in 64 patients with chronic renal failure treated in a single hemodialysis unit. None of these patients was a carrier of hepatitis B virus nor of antibodies against human immunodeficiency virus. Antibodies against hepatitis C virus were detected in 11 patients (17%). The prevalence was higher in the 13 previously diagnosed of non A, non B hepatitis (77%) than in the 51 without previous hepatitis (2%) (p less than 0.001). A relationship between the infection rate and the number of previous blood transfusions was also observed: 5% in the patients without previous transfusions, 13% in the 30 patients who had received between 1 and 10 blood units and 40% in the 15 who had received more than 10 blood units (p less than 0.05). These data suggest that the hepatitis C virus may be responsible for most episodes previously diagnosed as non A, non B hepatitis, and that blood transfusions are the major risk factor.

Acute cardiovascular effects of intravenous cyclosporine.

The acute effects of intravenous (i.v.) cyclosporine A (CsA) on blood pressure and other haemodynamic parameters were examined in 8 patients with end-stage renal disease on haemodialysis (HD). The study was performed after the mid-week haemodialysis session, when the patients were on their dry body weight. Each patient received an i.v. infusion of 5 mg/kg of CsA in 120 ml of 5% dextrose in water during 2 hours. Heart rate, systolic and diastolic blood pressure (SBP, DBP) were monitored by the Holter system. An echocardiogram (M-mode 2-dimensional and Doppler sonography) was performed using an automatic device (Ultramark 6) before CsA administration, at 30, 60 and 120 minutes during CsA infusion, and at 30 minutes thereafter. SBP, DBP and calculated peripheral vascular resistance (CPVR) increased significantly in respect to basal values at 120 and 150 minutes (SBP: basal 130 +/- 21, 120 min: 136 +/- 20, 150 min: 140 +/- 18, p < 0.05 and p < 0.01, respectively. DBP: basal 80 +/- 9, 120 min: 86 +/- 13, 150 min: 88 +/- 13, p < 0.05 and p < 0.01, respectively. CPVR: basal 1000 +/- 228, 120 min: 1178 +/- 305, 150 min: 1236 +/- 270 dyne/s/cm5, p < 0.01). However, systolic volume (SV) and cardiac output (CO) showed significant decreases from the basal values (SV: basal 103 +/- 29, 120 min: 85 +/- 22, 150 min: 85 +/- 17, p < 0.05. CO: basal 8.2 +/- 2, 120 min: 7.3 +/- 1.1, 150 min: 7 +/- 1.2 l/min, p < 0.05). In conclusion, CsA infusion produces a significant elevation of blood pressure, which seems to be mediated by a direct action on peripheral vascular resistance.

[Correlations between brain SPECT and neuropsychology assessments in mild and moderate stages of Alzheimer's disease]. / Correlaciones entre la SPECT cerebral y la evaluación neuropsicológica en los estadios leve y moderado de la enfermedad de Alzheimer.

UNLABELLED: Thirty-four patients with a probable Alzheimer's Disease (AD) (mild AD = 16; moderate AD = 18) and 12 matched controls were evaluated using semiquantitative SPECT-99mTc-HMPAO and neuropsychological tests (CERAD). RESULTS: Both temporal hypoperfusion (p < 0.01) and memory tests (p < 0.001) made it possible to differentiate the controls from mild AD patients. In these patients, significant correlations (p < 0.05) were also found between: 1) delayed recall test/temporal hypoperfusion, 2) learning memory test/temporoparietal and frontal hypoperfusion and 3) visual constructive praxis/posterior temporal hypoperfusion. In contrast to mild AD, moderate AD patients showed higher temporal (p < 0.01) parietal and frontal (p < 0.05) hypoperfusion along with worsening of praxis (p < 0.001) and memory tests (p < 0.05). CONCLUSIONS: SPECT imaging and neuropsychology evaluation can distinguish controls from AD patients with mild an moderate grades of dementia, showing a strong correlation from the early stages of AD.

[Use of a cyclotron in the production of positron emitting radionuclides]. / Utilización de un ciclotrón para la producción de radionucleidos emisores de positrones.

The experience acquired by our center during the first two years of using cyclotron 18/9 (IBA) dedicated to the production of clinical positron emission radionuclides is described. The cyclotron performance characteristics, production yields, quality control and synthesized radiotracers are analyzed. Cyclotron makes it possible to produce up to 3,300 mCi of 18F-, 270 mCi of 18F2, 3,100 mCi of 11C, 502 mCi of 13N (in 120, 60, 35 and 20 minutes respectively) and 540 mCi/min of 15O. In our center, about 85% of the PET studies are performed with 18F-FDG, whereas the remaining are done with 15O-water, 11C-bicarbonate, 11C-methionine, 13N-ammonia or 18F-. Cyclotron is included in the Radiopharmacy Unit of our PET facility and is subjected to a global quality control program. Follow-up of the bombardment parameters and periodic verifications of the cyclotron performance have made it possible to prevent equipment functioning problems, increase mean time between stoppage and decrease downtime. We conclude that cyclotron has high production capabilities and allows enough flexibility for a clinical and research positron emission tomography center; furthermore, it can also be used for regional distribution of 18F-FDG to satellite PET centers.