Evaluation of myopic cornea lenticules. A histochemical and clinical correlation.

In this work, we have analyzed the main clinical and corneal histological parameters that may be associated to the spherical equivalent (SE), age and gender of individuals with myopic refractive errors. For this purpose, 108 cornea stroma lenticules were obtained from patients subjected to ReLEx-SMILE myopia correction. Histological analyses were carried out and histochemistry and immunohistochemistry were used to quantify key histological components of the cornea stroma, including mature collagen fibers, reticular and elastic fibers, glycoproteins, proteoglycans, type-V collagen and several crystallins. Clinical and histological data were analyzed to determine their association with SE, age and gender. Results showed a significant correlation between the age range of the patients and the expression of crystallins CRY-α-A, CRY-λ1 and type-V collagen and between CRY-λ1 and corneal thickness, spherical diopters (D) and SE, although correlation between CRY-λ1 and SE was non-significant when age was controlled. Comparison of cases with low myopia and high/moderate myopia found statistical differences for D and lenticule thickness and diameter. The binary logistic regression analysis allowed us to construct a model using two clinical parameters (D and lenticule thickness). Parameters showing significant correlation with the age were the corneal radius, keratometry reading (K), OZ, CRY-α-A and type-V collagen, whereas SE, lenticule thickness, OZ, CRY-λ1 and type-V collagen showed statistically significant differences between the youngest and the oldest patients. A binary logistic regression analysis model was generated including 3 variables (D, cornea radius and OZ). No gender differences were found. The specific clinical and histological modifications found to be associated to the SE and age could be useful for a better understanding of the mechanisms involved in the genesis or progression of myopia and could establish the basement for future therapeutic options.

TNFR1 mRNA expression level and TNFR1 gene polymorphisms are predictive markers for susceptibility to develop invasive pulmonary aspergillosis.

Tumour necrosis factor (TNF) is primarily secreted by monocytes/macrophages and activated T lymphocytes in response to fungal infections. TNF acts through TNF receptor 1 (TNFR1) triggering a pro-inflammatory response, and therefore plays a pivotal role in immune regulation and host immune responses. We hypothesized that single nucleotide polymorphisms (SNPs) in TNFR1 gene may influence the innate immune response against Aspergillus. Three SNPs were genotyped in 275 individuals (144 immunocompromised haematological patients with high-risk of developing IPA and 131 healthy controls): TNFR1(-383(A/C)) (rs2234649) and TNFR1(-609(G/T)) (rs4149570) in the 5 prime UTR region, and TNFR1(+36(A/G)) SNP (rs767455) in the first exon of the gene. Of the 144 haematological patients, 77 patients developed Invasive Pulmonary Aspergillosis (IPA) infection and the remaining 67 patients were not infected. TNFR1(+36(A/G)) and TNFR1(-609(G/T)) were associated with IPA susceptibility (p=0.033 and p=0.018, respectively). A role of TNFR1 genetic variants in the susceptibility of patients to develop IPA was also supported by the significantly lower TNFR1 mRNA expression level in IPA than in IPA-resistant patients and the strong correlation between the TNFR1(-609) genetic variant and the expression levels of TNFR1. There was also a tendency for a higher frequency of galactomannan (GM) positivity in patients with TNFR1(-609G/G) genotype than in patients with TNFR1(-609G/T) (p=0.0909) or TNFR1(-609T/T) (p=0.0913) genotype. Predictive sequence analysis of the effects of TNFR1(-609) promoter polymorphism revealed that this SNP might play a critical role in modifying the affinity of ICSBP/IRF-8, a transcription factor that is involved in the TNFR1-mediated activation of NFkappaB signalling pathway. Taken together, these data suggest that TNFR1 polymorphisms influence the risk of IPA disease and might be useful for risk stratification strategies. These findings need to be confirmed in validation studies with larger samples of haematological patients.

Zero Liquid Discharge approach in plating industry: treatment of degreasing effluents by electrocoagulation and anodic oxidation.

Degreasing waste effluents issued from a surface treatment plant were treated by electrochemical techniques in an attempt to reduce COD so that clean water can be returned to the rinse bath. Electrocoagulation, both with iron and aluminium anodes, and anodic oxidation with boron doped diamond (BDD) anodes were tested. In the electrocoagulation tests, the nature of the anodes did not impact significantly the reduction of COD. Electrocoagulation showed good COD removal rates, superior to 80%, but it was not able to reduce COD down to low levels. Anodic oxidation was able to reduce COD down to discharge limits; the oxidation efficiency was superior to 50%. Economical calculations show that anodic oxidation is best used as a polishing step after electrocoagulation. The bulk of the COD would be reduced by electrocoagulation and, then, anodic oxidation would reduce COD below discharge limits. The maximum treatable flow is somewhat hindered by the small sizes of current BDD installation but it would reach 600 m(3)/year if anodic oxidation is coupled with electrocoagulation, the operational cost being 2.90 Euros /m(3).

Human platelet-specific antigens frequencies in the Argentinean population.

The frequencies of several human platelet antigens (HPAs) vary between different populations and are a major determinant for the prevalence of HPA alloimmunization and its clinical associated entities. The aim of this study was to characterize the allele frequencies of seven HPA systems in two different ethnic groups from the Argentinean city of Rosario, the major population and a minority Amerindian group recently arrived from the north of the country, the Tobas. A total of 192 healthy unrelated individuals from blood donors and hospital staff from the Hospital Italiano Garibaldi and 27 unrelated Toba Amerindians were genotyped for HPA-1, -2, -3, -4, -5, -6 and -15 systems by polymerase chain reaction with sequence specific primers (PCR-SSP). The present data showed that the distribution of the HPA alleles among Argentineans from Rosario is quite similar to that reported among Europeans. The frequencies seen in Tobas, although limited by the small number of aboriginal samples studied, are similar to those reported for other Amerindians populations. Statistically significant differences were found for the genotype distribution of HPA-1, -3, -5 and -15 between both groups, indicating important differences in the potential risk of HPA alloimmunization associated to transfusion and pregnancy. The study of these polymorphisms represents the first step in the elucidation of pathological conditions that are underdiagnosed in our population. It allowed us to establish a panel of characterized blood donors necessary for the serological work out and as a source for compatible platelets transfusion.

HNA-1a, HNA-1b and HNA-1c gene frequencies in Argentineans.

Human neutrophil antigens (HNA) are polymorphic structures located in the neutrophil membrane. The neutrophil-specific antigens HNA-1a (NA1), 1b (NA2) and 1c (SH) are well-recognized allotypic forms of FcgammaRIIIb and the most frequent targets of neutrophil alloantibodies. The aim of this study was to determine the gene frequencies of the neutrophil-specific antigens belonging to the HNA-1 system in blood donors and Toba Amerindians from Rosario, Argentina. Two hundred and eighteen unrelated healthy Argentinean blood donors and Toba Amerindians from Rosario were typed for HNA-1a, HNA-1b and HNA-1c using polymerase chain reaction with sequence-specific primers. For the Argentinean blood donors, the HNA-1a and HNA-1b gene frequencies were 0.44 and 0.56 and for the Amerindians Toba were 0.77 and 0.23, respectively. The HNA-1c antigen is present in 4.7% (gene frequency=0.023) of the blood donors but in none of the Amerindian individuals. The present data showed that the HNA-1 allele frequencies in the major population and the Toba Amerindians from Rosario are similar to those described in European and others distant Amerindians populations, respectively.

Genetic variants of IL6 gene promoter influence on C-reactive protein levels but are not associated with susceptibility to invasive pulmonary aspergillosis in haematological patients.

Several lines of evidence indicate that IL6 plays a major role in the pathogenesis of a number of infectious diseases. The purpose of this study was to determine whether IL6 promoter polymorphisms were genetic markers of susceptibility to invasive pulmonary aspergillosis (IPA). To clarify the relationship between IL6 variants and IPA susceptibility, the IL6-174(G/C) and IL6-634(G/C) promoter single nucleotide polymorphisms (SNPs) were defined and plasma concentrations of IL6 and C-reactive protein (CRP) were measured. The study included 130 patients with haematological malignancies and 145 unrelated healthy individuals. No significant genotypic and allelic differences were found between patients and healthy controls. IPA was diagnosed in 71 of 130 patients according to the consensus criteria. CRP values were significantly associated with both IL6-174(G/C) and IL6-634(G/C) polymorphisms. However, IL6 and CRP values were similar between IPA and non-IPA groups. Neither IL6-174(G/C) nor IL6-634(G/C) polymorphisms were associated with IPA infection (p=0.414 and p=0.184, respectively). No evidence of association was found between allelic frequencies of IL6 promoter polymorphisms and IPA infection (p=0.864 and p=0.104, respectively). Further, no association was detected between IL6 genotypes and clinical profiles in IPA patients. Haplotype analysis also revealed that IL6 gene was not associated with IPA susceptibility in a Spanish population (Global haplotype association p value: 0.31). These findings suggest that IL6 polymorphisms influence on CRP circulating levels but are not associated with IPA susceptibility. Because the sample size is relatively small in our series, larger investigations of IL6-174(G/C)/IL6-634(G/C) genotypes and haplotypes are needed to clarify the potential role of this gene in the pathophysiology of IPA infection.

Correlación interlaboratorio en la valoración de la expresión de glicoproteína P en sangre periférica por citometría de flujo / Correlation interlaboratorio in the valoration of the expression of glycoprotein P in blood periferic flow cytometry

La Glicoproteína P (Gp-P) es una proteína transmembranaria con función transportadora, que tiene por misión expeler xenobióticos lipofilíticos. Esta glicoproteína tiene una distribución específica y unos niveles de expresión variables en las células sanguíneas humanas encargadas de la defensa en el organismo. El objetivo de este trabajo fue comparar y establecer si son reproducibles los resultados obtenidos por dos laboratorios distintos en la valoración de la expresión de glicoproteína P (Gp-P) en muestras celulares con baja expresión de esta glicoproteína, como es el caso de las células de sangre periférica. Se valoraron las subpoblaciones celulares de ciento veinticinco (125) muestras de sangre periférica de individuos sanos por citometría de flujo e inmunofluorescencia directa, utilizando el anticuerpo monoclonal JSB-1 conjugando con isotiocianato de fluoresceína (FITC), en dos citómetros de flujo de la misma marca y modelo, situados en dos laboratorios diferentes: Departamento de Anatomía Patológica, Facultad de Medicina, Universidad de Granada y Centro de Transfusión Sanguínea de Granada, España. Los resultados demostraron que la mejor correlación estadística del porcentaje de Positividad (por ciento POS) fue observada en los linfositos (r=0.93), mientras que para la Incorporación Media Fluorescencia Específica (IMFE) se observó en los granulocitos (r=0.94). Los resultados obtenidos con la metodología empleada en la determinación de Gp-P por citometría de flujo e inmunofluorescencia directa, permiten concluir que es una técnica reproducible y fiable para la evaluación del fenómeno de Resistencia Multiple a Drogas (MDR) en las subpoblaciones celulares normales de sangre periférica

Valoración del estudio Doppler de la arteria cerebral media como método diagnóstico de la anemia fetal / Evaluation of middle cerebral artery Doppler assessment in the diagnosis of fetal anemia

Objetivos: El estudio Doppler de la arteria cerebral media (ACM) se ha propuesto como método no invasivo para el diagnóstico de la anemia fetal en casos de aloinmunización materna. Se evalúa su relación con los valores de hemoglobina fetal, así como su capacidad predictiva para el diagnóstico de la anemia. Sujetos y métodos: Se determinó el valor de hemoglobina mediante cordocentesis y los valores múltiplos de la mediana (MOM) de la velocidad en el pico sistólico, la velocidad media y el índice de pulsatilidad en la arteria cerebral media, en 72 casos de anemia fetal correspondientes a 19 fetos, antes y después de realizar una transfusión intravascular. Se compararon los valores Doppler antes y después de la transfusión, se estudió su grado de correlación con el valor MOM de hemoglobina fetal y se realizó un cálculo de los puntos de corte óptimos para el diagnóstico de anemia y la ausencia de la misma, así como una valoración de su capacidad predictiva. Resultados: Los tres parámetros velocimétricos estudiados presentaron una disminución significativa tras la reposición hematimétrica. Se comprobó una correlación de la velocidad media del ciclo y en el pico sistólico máximo en ACM con el valor de hemoglobina fetal para cada edad gestacional. Los puntos de corte óptimos para la predicción de anemia moderada o grave fueron 1,49 MOM de la velocidad del pico sistólico máximo y 2,54 MOM de la velocidad media del ciclo. Los valores por debajo de 0,93 MOM del pico sistólico máximo y 1,88 MOM de la velocidad media del ciclo descartaron la presencia de anemia moderada o grave en el 100 per cent de los casos. Conclusiones: Los parámetros Doppler de la arteria cerebral media demuestran una buena correlación con el valor de hemoglobina fetal. El valor de velocidad en pico sistólico máximo y velocidad media del ciclo presentan una buena capacidad predictiva de la presencia de anemia fetal, y pueden ser útiles en el estudio de estos fetos, descartar la presencia de anemia moderada o grave, evitar cordocentesis y programar adecuadamente el intervalo intertransfusión (AU)

Allogeneic transplantation of selected CD34+ cells from peripheral blood: experience of 62 cases using immunoadsorption or immunomagnetic technique. Spanish Group of Allo-PBT.

The objective of this study was to analyze CD34+ cell recovery and T cell depletion (TCD) achieved in CD34+ cell grafts using either immunoadsorption or immunomagnetic methods applied to leukapheresis products from healthy donors. We also wanted to determine the kinetics of engraftment and incidence and severity of graft-versus-host disease (GVHD) after allogeneic transplantation of selected CD34+ cells. HLA-identical sibling donors received G-CSF. After leukapheresis, peripheral blood progenitor cells were selected using immunoadsorption (Ceprate SC) (n = 38) or immunomagnetic (Isolex 300) (n = 24) methods. Sixty-two patients, with a median age of 42 years (range 17-60) diagnosed with hematological malignancies were conditioned with either cyclophosphamide and total body irradiation (n = 43) or busulphan and cyclophosphamide (n = 19). GVHD prophylaxis consisted of cyclosporin A (CsA) and prednisone (n = 48), CsA alone (n = 11) and CsA and methotrexate (n = 3). The median yield and purity of CD34+ cells after the procedure was 65 and 66% with immunoadsorption, and 48 and 86% with immunomagnetic method, respectively. The median number (range) of CD34+ cells infused into the patients was 3.5 x 10(6)/kg (1-9.6). The median number (range) of CD3+ cells administered was 0.4 x 10(6)/kg (0.01-2) using immunoadsorption and 0.14 x 10(6)/kg (0.03-2.5) using immunomagnetic methods. Neutrophil recovery >500 and >1000/microl was achieved at a median (range) of 13 days (8-22) and 14 days (9-31), respectively. Platelets recovered to >20000 and >50000/microl at a median (range) of 13 days (0-128) and 18 days (0-180), respectively. Two patients developed graft failure. Acute GVHD in patients at risk was clinical grade 0 (n = 43), I (n = 8), II (n = 4) and III (n = 1). No patient developed acute GVHD grade IV. Chronic GVHD was limited in two cases and extensive in four cases. The actuarial probability of acute GVHD II-IV was 10% (95% CI, 1-19%), and of extensive chronic GVHD was 12% (95% CI, 11-13%). The cumulative incidence of transplant-related mortality was 12.6%, and this figure was 9% at 6 months. In conclusion, with the immunomagnetic procedure, a lower recovery and higher purity of CD34+ cells, and stronger TCD is obtained as compared to immunoadsorption (P = 0.008, P < 0.0001 and P = 0.0002, respectively). Our results also indicate that allogeneic transplantation of selected CD34+ cells is associated with a very rapid engraftment and with a low incidence of severe GVHD.

[Report on the activity of the blood bank accreditation program (1987-1995)]. / Balance de la actividad del programa de acreditación de Bancos de Sangre (1987-1995).

PURPOSE: To show the incidence of the deficiencies detected in the Blood Banks for the accreditation by the Transfusion Accreditation Committee (CAT), previously named PABAS. MATERIALS AND METHODS: Analysis of the reports of the accreditation of 85 Blood Banks made by the PABAS during the period 1987-1995. RESULTS: Eighty-five (20.8%) of the 407 Community Blood Centers, Hospital-Based Blood Banks and Transfusional Services of Spain had been surveyed. There were 244 deficiencies, of which 31 (12.7%) were of the equipment, 114 (46.7%) of the procedures used, and 99 (40.6%) of the documentation. The activities with more incidence of faults were: Control of the temperatures of the storage of units 53 (21.7%), label of the components 38 (15.5%), quality system of the institution surveyed 32 (13.1%), transfusional procedures 30 (12.3%), and on the procedure of the selection of donors 29 (11.9%). By contrary, the areas of work with fewer incidences of faults were those related with the collection of the blood and components 10 (4.1%) and the laboratory 14 (5.7%). CONCLUSIONS: Low percentage of the Community Blood Centers, Hospital-Based Blood Banks and Transfusional Services, which ask to be accredited by the Transfusion Accreditation Committee. The 83.7% of the errors detected are of the procedures and documentation, which could be easily corrected by the training and continuous improving of the quality, and without need of new inversions in equipment.

Allogeneic transplantation of purified CD34+ cells from peripheral blood: Spanish experience of 62 cases. Spanish Group of allo-PBT.

This report summarizes the Spanish experience of 62 cases of allogeneic transplantation of purified CD34+ cells from peripheral blood. HLA-identical sibling donors received G-CSF. After leukapheresis, peripheral blood progenitor cells were purified using one of two methods: Ceprate (n = 38), or Isolex 300 (n = 24). Sixty-two patients median age 42 years (range 17-60) diagnosed with hematological malignancies were conditioned with either cyclophosphamide and total body irradiation (n = 43) or busulphan and cyclophosphamide (n = 19). GVHD prophylaxis consisted of cyclosporin A (CsA) and prednisone (n = 48), CsA alone (n = 11), and CsA and methotrexate (n = 3). The median yield and purity of CD34+ cells after the procedure was 65% and 66% with Ceprate, and 48% and 86% with Isolex, respectively. The median number of CD34+ cells infused into the patients was 3.5 x 10(6)/kg (range 1-9.6). The median number of CD3+ cells administered was 0.4 x 10(6)/kg (range 0.01-2) using Ceprate and 0.14 x 10(6)/kg (range 0.03-2.5) using Isolex. Neutrophil recovery >500 and >1000/microl was achieved at a median of 13 days (range 8-22) and 14 days (range 9-31), respectively. Platelets recovered to >20,000 and >50,000/microl at a median of 13 days (range 0-128) and 18 days (range 0-180), respectively. The actuarial probability of acute GVHD II-IV was 10% (95% CI, 1-19%), and of extensive chronic GVHD 12% (95% CI, 11-13%).

Epidemiology, serological markers, and hepatic disease of anti-HCV ELISA-2-positive blood donors.

The epidemiology associated with hepatitis C virus (HCV) infection, serologic reactivity, and hepatic disease related to anti-HCV-positive donors of Granada were researched. From 1990 through 1993, medical and epidemiological information and anti-HCV and HCV RNA testing were evaluated in 46,741 blood donors. Serum samples were obtained for anti-HCV ELISA and RIBA and HCV RNA determination. A liver biopsy was conducted in all anti-HCV positives by confirmatory second-generation RIBA to analyze the hepatic lesion and the presence of HCV RNA. The anti-HCV prevalence was 1.12%. A total of 228 anti-HCV second-generation ELISA positive blood donors were analyzed. Intrafamiliar transmission rate was 1.7%. Transfusion and intravenous drug abuse (IVDA) antecedents were associated with a higher risk of seroconversion. A RIBA-positive result was related to high second- and third-generation ELISA ratios (90%), HCV RNA positivity (89%), and elevated alanine aminotransferase (ALT) levels (88%). Approximately 50% of donors with normal ALT levels had high ELISA ratios and second-generation RIBA and HCV RNA positive results. Of the second-generation RIBA indeterminate results, 42% and 82% of the c22 and 33% and 100% of the c100 reactivities were third-generation RIBA and HCV RNA positive, respectively. Liver biopsy was conducted in 85 donors, 74% of whom had a chronic hepatitis and 83% had detectable HCV RNA levels. Chronic hepatitis was diagnosed in 88% vs 43% of donors with elevated and normal alanine aminotransferase levels, respectively. ELISA and confirmatory HCV RNA determinations should be routinely employed in donor screening. A liver biopsy should be conducted in patients with elevated ALT levels and normal ALT levels when viremic.

Use of rhG-CSF in the collection of peripheral blood stem cells for autologous blood stem cell transplantation in acute myeloblastic leukemia.

The use of recombinant granulocyte colony-stimulating factor (rhG-CSF) alone is attractive for the collection of peripheral blood stem cells (PBSC) in several malignancies. In acute myeloblastic leukemia (AML), not enough experience has been gained with rhG-CSF and leukapheresis is more common after recovery from antileukemia chemotherapy. This is the case report of a patient who received rhG-CSF alone to mobilize stem cells where the cells collected in the leukapheresis bag had a blastic immunophenotype.

[Prospective study of the incidence of post-transfusion hepatitis after the exclusion of donors positive for anti-HCV ELISA 2]. / Estudio prospectivo de la incidencia de hepatitis postransfusional después de la exclusión de los donantes anti-VHC ELISA 2 positivos.

BACKGROUND: The incidence of post transfusional hepatitis (PTH) after the exclusion of anti-HCV ELISA 2 positive donors is not well known. The aim of this study was to determine the incidence and type of PTH in 113 post transfused patients. METHODS: A post transfusional follow up was performed for at least one year with periodic controls of transaminase levels. When an increase in GPT level compatible with PTH was demonstrated investigation of all the virus related with the transfusion was carried out in both the donor and the transfused subject: HAV, HCV, HEV, HBV and CMV. RESULTS: Four cases (3.5%) were detected which fulfilled the PTH criteria with the following characteristics: short period of time between transfusion and the increase in GPT level, moderate GPT increase, moderate clinical expression and good evolution. In all the cases the viral study was negative and other non viral possibilities were eliminated. CONCLUSIONS: Transfusions are currently relatively safe and the increase in transaminases may not be related with transfusion.

[Prevalence of antibodies against CMV in Grenada]. / Prevalencia de anticuerpos frente a CMV en Granada.

The prevalence of anti-CMV antibodies was studied on 1,552 serum samples by means of latex passive haemagglutination method. Of all the sera studied, 1,084 were positive (69.8%). Amongst them, 523 samples came from women and 561 from men, which represent 74.7% and 65.8%, respectively. Regarding to age, 62.3% of the positive samples were from people under 30 years and 91.3% from subjects over that age. The screening of anti-CMV antibodies is especially important in blood and organ donors, where the high percentage of positivity makes it difficult to select negative blood donations. Thus, taking into account the cost-effectiveness, and the results of this study, the search for CMV-negative blood must be exerted preferentially on the group of blood donors under 30 years of age, regardless of sex.