Diagnostic and prognostic significance of flow cytometry immunophenotyping in patients with leptomeningeal carcinomatosis.

Some patients with epithelial-cell cancers develop leptomeningeal carcinomatosis (LC), a severe complication difficult to diagnose and with an adverse prognosis. This study explores the contribution of flow cytometry immunophenotyping (FCI) to the diagnosis and prognosis of LC. Cerebrospinal fluid (CSF) samples from patients diagnosed with LC were studied using FCI. Expression of the epithelial-cell adhesion molecule (EpCAM) was the criterion used to identify the epithelial cells. To test the diagnostic precision, 144 patients (94 diagnosed with LC) were included. The prognostic value of FCI was evaluated in 72 patients diagnosed with LC and eligible for therapy. Compared with cytology, FCI showed greater sensitivity and negative predictive value (79.79 vs. 50%; 68.85 vs. 51.55%, respectively), but lower specificity and positive predictive value (84 vs. 100%; 90.36 vs. 100%, respectively). The multivariate analysis revealed that the percentage of CSF EpCAM+ cells predicted an increased risk of death (HR: 1.012, 95% CI 1.000-1.023; p=0.041). A cut-off value of 8% EpCAM+ cells in the CSF distinguished two groups of patients with statistically significant differences in overall survival (OS) (p=0.018). This cut-off value kept its statistical significance regardless of the absolute CSF cell-count. The FCI study of the CSF improved the sensitivity for diagnosing LC, but refinement of the technique is needed to improve specificity. Furthermore, quantification of CSF EpCAM+ cells was revealed to be an independent prognostic factor for OS in patients with LC eligible for therapy. An 8% cut-off value contributed to predicting clinical evolution before initiation of therapy.

ω-3 Long-Chain Polyunsaturated Fatty Acids and Fatty Acid Desaturase Activity Ratios as Eventual Endophenotypes for ADHD.

OBJECTIVE: Epidemiological studies suggest that long-chain polyunsaturated fatty acids (LC-PUFAs) may be suitable as endophenotypes for ADHD. To be appropriated vulnerability traits, endophenotypes should be altered in unaffected relatives of index cases. Serum profiles of LC-PUFAs in unaffected relatives of ADHD patients remain understudied. The main objective of this study was to compare serum LC-PUFAs in ADHD patients, unaffected relatives of index cases, and general-population unaffected participants. METHOD: LC-PUFA profiles of 72 participants (27 ADHD patients, 27 unaffected relatives, and 18 general-population participants) were obtained by gas chromatography-mass spectrometry (GC-MS). Groups were compared by parametrical statistics. RESULTS: Unaffected females from the general population presented lower Docosapentaenoic acid (DPA; p = .0012) and a-linolenic acid (ALA; p = .0091) levels compared with ADHD females and unaffected relatives. In addition, docosahexaenoic acid (DHA)/ALA and DHA/DPA ratios, addressing desaturase activity, were significantly lower in ADHD patients and unaffected relatives of ADHD patients in the female-subgroup (p = .022 and .04, respectively). CONCLUSION: DHA/ALA, DHA/DPA, serum DPA, and serum ALA may be suitable as endophenotypes for ADHD women.

Proteomic and transcriptomic analysis of rice tranglutaminase and chloroplast-related proteins.

The recently cloned rice transglutaminase gene (tgo) is the second plant transglutaminase identified to date (Campos et al. Plant Sci. 205-206 (2013) 97-110). Similarly to its counterpart in maize (tgz), this rice TGase was localized in the chloroplast, although in this case not exclusively. To further characterise plastidial tgo functionality, proteomic and transcriptomic studies were carried out to identify possible TGO-related proteins. Some LHCII antenna proteins were identified as TGO related using an in vitro proteomic approach, as well as ATPase and some PSII core proteins by mass spectrometry. To study the relationship between TGO and other plastidial proteins, a transcriptomic in vivo Dynamic Array (Fluidigm™) was used to analyse the mRNA expression of 30 plastidial genes with respect to that of tgo, in rice plants subjected to different periods of continuous illumination. The results indicated a gene-dependent tendency in the expression pattern that was related to tgo expression and to the illumination cycle. For certain genes, including tgo, significant differences between treatments, principally at the initiation and/or at the end of the illumination period, connected with the day/night cycling of gene expression, were observed. The tgo expression was especially related to plastidial proteins involved in photoprotection and the thylakoid electrochemical gradient.

Rice transglutaminase gene: Identification, protein expression, functionality, light dependence and specific cell location.

Transglutaminases (TGases), that catalyze post-translational modification of proteins, are scarcely known in plants. As part of a project to characterize transglutaminase genes in new plant species, the identification and characterization of a TGase in rice is presented. Using differential primers, a cDNA (tgo) of 1767bp from genomic rice DNA amplification was obtained. The primers were designed from the rice DNA sequence relatively homologous to the gene encoding active maize chloroplast TGase. Amino acid sequence of the deduced rice TGase protein (TGO) indicated that it contains the enzyme catalytic triad (Cys-His-Asp), three repeats, myristoylation domains and a leucine zipper motif. The TGO recombinant protein was characterized, showing specific activity regulation, and indicating that tgo encoded for an authentic TGase. Substrate preference and Ca(2+) dependent activity were also detected. In the rice plant TGO protein was immunolocalized in the grana chloroplasts, in protein vesicles near them, and in the bulliform cells. Immunoblot analyses, tgo mRNA expression, and TGase activity indicated that TGO expression in rice was light dependent and regulated by the illumination period. This work increases significantly our plant TGase understanding. Its functional role in rice, which is a good model system for C3 plants, is discussed.

[Complex distal radius fractures in young patients. Comparison between external fixator and volar plate]. / Fracturas complejas de radio distal en pacientes jóvenes. Comparación entre fijador externo y placa volar.

BACKGROUND: Distal radius fractures in young, working-age patients are frequent and are associated with potential disability in case of inappropriate treatment. Our hypothesis is that there is no difference between patients treated with a volar plate and those treated with closed reduction and external fixation. METHODS: We studied 69 patients with complex distal radius fractures. They were prospectively and randomly assigned to treatment with a volar plate or an external fixator: they were followed-up and assessed at one year using two scales, the Visual Analog Scale (VAS) and the McDermid et al. Functionality Scale. RESULTS: Both groups showed good results at one year. Measurements in the pain Visual Analog Scale and disability measured with the McDermid et al. scale were not statistically significant. Higher scores mean greater disability. Scores above 20 were considered as failures. We found 2 cases with complications in the F group; one with material dislodgement and one with malunion, without significant differences. CONCLUSIONS: Both treatments show good results in the medium-term; no differences were found between both groups regarding pain and residual disability. Complications were few and they were addressed conservatively; they were less frequent with open reduction and internal fixation. When ranges of motion and the complication rate were compared, the volar plate showed a statistically significant difference in a decreased complication risk.

Significance of the volume of fetomaternal hemorrhage after performing prenatal invasive tests.

BACKGROUND: Fetal erythrocytes cross the placenta during gestation, but invasive prenatal procedures might develop into fetomaternal hemorrhage (FMH). We examine whether flow cytometry immunophenotyping might be useful for measuring the volume of FMH after such procedures. METHODS: Fetal erythrocytes (%) were determined in 153 pregnant women after amniocentesis (129) and chorionic villous sampling (24) using a monoclonal antibody against fetal hemoglobin. Fetal erythrocytes were identified for their high expression of fetal hemoglobin (HbF(++) ). Blood samples from two control groups, 53 healthy males and 21 pregnant women not submitted to invasive tests, were used to establish normal values of circulating HbF(++) erythrocytes in adults. RESULTS: The highest percentage of HbF(++) erythrocytes in the control groups was 0.015%. The rate of HbF(++) erythrocytes in samples after invasive tests ranged between <0.01% and 0.15%. Seventy-three women (47%) had ≤0.015% HbF(++) erythrocytes, and this rate was higher in 80. Nine women presented >1 ml of FMH (volume of packed cells corresponding to 0.054-0.15% HbF(++) erythrocytes), but only two had sonographic evidence of bleeding. CONCLUSIONS: Most women in our series had a very low volume of FMH after the invasive tests. Acute bleeding should be thoroughly investigated in women with either more than 1 ml of packed cells or more than 0.05% of HbF(++) erythrocytes. Intermediate values between >0.015% and <0.05%, should be carefully considered depending on the week of gestation. Data obtained before 15 weeks might reflect previous cell trafficking between fetus and mother instead of acute hemorrhage.

Magnetic properties of encapsulated nanoparticles in nitrogen-doped multiwalled cabon nanotubes embedded in SiOx matrices.

We report the production, characterization, thermal transformations (400-1000 degrees C), and magnetic properties of nanoparticles encapsulated in nitrogen-doped multiwall carbon nanotubes (CNx-MWNT), which were embedded in silicon oxide (SiOx) matrices via sol-gel techniques. The vapor chemical deposition (CVD) method with ferrocene-benzelamine mixtures was used to synthesize Fe and Fe3C nanoparticles inside CNx-MWNTs. Composites consisting of CNx-MWNTs (filler) and SiOx (matrix) were fabricated and thermally treated to different temperatures and exposure times (t). All samples were characterized using scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM), thermogravimetic analysis (TGA), and magnetometry (vibrating sample). We found that upon thermal treatment, the ferromagnetic nanoparticles modify their morphology, composition and aspect ratio, thus resulting in drastic changes in the magnetic and structural properties. In particular, as produced encapsulated nanoparticles mainly consisting of Fe and Fe3C phases were thermally modified into magnetite (Fe3O4). We have also observed that the hysteresis loops are very sensitive to the thermal treatment of the sample. Thus we can control the magnetic properties of the samples using thermal treatments.

Identification of feto-maternal haemorrhage around labour using flow cytometry immunophenotyping.

OBJECTIVE: To determine the normal range of feto-maternal haemorrhage (FMH) due to labour, and to investigate if the type of delivery might influence the volume of FMH. STUDY DESIGN: In a prospective cohort trial setting we studied 346 women in their 3rd trimester of gestation. Women were classified according to the type of delivery: vaginal, instrumental and caesarean section. Fetal erythrocytes in maternal blood were measured by flow cytometry immunophenotyping using a fluorochrome-conjugated monoclonal antibody against fetal haemoglobin. For each woman, two blood samples were studied; one pre-labour and one post-labour. The difference between FMH values obtained post- and pre-delivery was established as FMH due to delivery. RESULTS: FMH due to labour ranged between <0.01 and 25.19 ml, being <1.15 ml in 96.13% of cases. This value was established as the upper limit of normal FMH due to delivery. No statistical significance was found between the volume of FMH and type of delivery. Analyzing distributions of groups, most data followed a normal distribution, apart from some patients who had higher volume of FMH. Among these patients caesarean sections showed a higher FMH volume, with statistically significant differences between vaginal deliveries and caesarean sections (p=0.001), and between instrumental deliveries and caesarean sections (p=0.008). CONCLUSIONS: FMH due to labour is small. The route of delivery could not be established as a risk factor for FMH but caesarean section increases the risk of suffering a higher amount of transplacental bleeding.

[Use of the Edinburgh Postnatal Depression Scale to detect postpartum depression]. / Mejorando la pesquisa de depresión posparto a través de un instrumento de tamizaje, la escala de depresión posparto de Edimburgo.

BACKGROUND: Postpartum depression is a serious, common, treatable condition, with broad repercussions in mother and family life, but frequently under diagnosed. A screening tool, the Edinburgh Postnatal Depression Scale (EPDS) could improve the detection rate of depressive symptoms. AIM: To assess the efficacy of EPDS in the detection rates for depressive symptoms in the postpartum period. To evaluate the feasibility of routine use of this screening scale in a Family Medicine Center. MATERIAL AND METHODS: The medical records of women attended during 2002, were reviewed to detect the registry of depressive symptoms or request for psychiatric consultations for depression. During 2003, the EPDS was applied to a sample of 110 women. To confirm the postpartum depression diagnosed using the instrument, a structured psychiatric interview was administered to these women. RESULTS: The medical records of 110 women aged 24+/-7 years were reviewed and the instrument was applied to 110 women aged 26+/-7 years. In the first group, the spontaneous detection rate for depressive symptoms was 4.6% (5/110). In 26.4% of women in whom the instrument was applied, depressive symptoms were found and 15% had a postpartum depression. The application rate of the scale was 88% and the rate of acceptance by mothers was 100%. CONCLUSIONS: The EPDS improves significantly the detection rates of postpartum depressive symptoms, and it is well accepted by health professionals and mothers.

Mejorando la pesquisa de depresión posparto a través de un instrumento de tamizaje, la escala de depresión posparto de Edimburgo / Use fo the Edinburgh postnatal depression sea le to detect postpartum depression

Background: Postpartum depression is a serious, common, treatable condition, with broad repercussions in mother and family life, but frequently under diagnosed. A screening tool, the Edinburgh Postnatal Depression Scale (EPDS) could improve the detection rate of depressive symptoms. Aim: To assess the efficacy of EPDS in the detection rates for depressive symptoms in the postpartum period. To evaluate the feasibility of routine use of this screening scale in a Family Medicine Center. Material and methods: The medical records of women attended during 2002, were reviewed to detect the registry of depressive symptoms or request for psychiatric consultations for depression. During 2003, the EPDS was applied to a sample of 110 women. To confirm the postpartum depression diagnosed using the instrument, a structured psychiatric interview was administered to these women. Results: The medical records of 110 women aged 24±7years were reviewed and the instrument was applied to 110 women aged 26±7years. In the first group, the spontaneous detection rate for depressive symptoms was 4.6 percent (5/110). In 26.4 percent of women in whom the instrument was applied, depressive symptoms were found and 15 percent had a postpartum depression. The application rate of the scale was 88 percent and the rate of acceptance by mothers was 100 percent. Conclusions: The EPDS improves significantly the detection rates of postpartum depressive symptoms, and it is well accepted by health professionals and mothers.

Evaluation of CD7 and terminal deoxynucleotidyl transferase (TdT) expression in CD34+ myeloblasts from patients with myelodysplastic syndrome.

There is an emerging use of flow cytometry to evaluate patients with myelodysplastic syndrome (MDS). We have studied CD7 and TdT expression in the CD34+ myeloid blast cell population in 55 bone marrow samples of patients with MDS. CD7 and/or TdT were detected in 38 out of 55 patients (69%). CD7 expression was not related to other bad prognosis data but conversely, we found an association between TdT+ CD34 myeloblasts and high-risk MDS patients according to the International Prognostic Scoring System. Therefore, CD7 and TdT may help to establish the diagnosis of MDS and, TdT expression also seems to be a useful marker in distinguishing risk groups.

Advantages of flow cytometry immunophenotyping for the diagnosis of central nervous system non-Hodgkin's lymphoma in AIDS patients.

BACKGROUND: Neurological disorders are common in HIV-infected patients. Central nervous system (CNS) lymphoma should always be considered because it is an important cause of morbidity and mortality. OBJECTIVES: To investigate the clinical utility of flow cytometry immunophenotyping (FCI) in diagnosing or discarding leptomeningeal involvement in HIV-infected patients and to compare its sensitivity with that of conventional cytological methods. METHODS: Fifty-six cerebrospinal fluid (CSF) samples from 29 HIV-infected patients were independently evaluated by flow cytometry and cytology. The description of an aberrant immunophenotype was the criterion used to define the malignant nature of any CSF cell population. RESULTS: FCI and cytology gave concordant results for 48 of the 56 CSF samples studied: 37 were negative for malignancy and 11 had evidence of CNS lymphoma. Discordant results were obtained for eight CSF samples, and the accuracy of the FCI findings could be demonstrated for four CSF samples described as positive for malignancy according to the FCI criteria. CONCLUSIONS: A high level of agreement was found between the results obtained using the two methods, but FCI gave at least 25% higher sensitivity than conventional cytomorphological methods for the detection of malignant cells. This advantage suggests that, in case of negative flow cytometry results, disorders other than non-Hodgkin's lymphoma should be strongly considered.

Induction of a virus-specific antibody response to foot and mouth disease virus using the structural protein VP1 expressed in transgenic potato plants.

We have recently communicated the oral and parental immunogenicity of the structural protein VP1 of foot and mouth disease virus (FMDV) expressed in different transgenic plants. Those results clearly indicated the necessity of increasing the expression of the foreign genes in the transgenic plant to avoid additional steps toward the purification and/or concentration of the antigen of interest. Here, we report the production of transgenic potatoes plants containing the VP1 gene cloned under the regulatory activity of either a single (pRok2) or a double (pRok3) copy of the S35 cauliflower mosaic virus (CaMV 35S) promoter, as a strategy for increasing the level of VP1 gene expression. The presence of the VP1 gene in the plants was confirmed by polymerase chain reaction (PCR) and its specific transcription activity was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that, although the immunized animals presented a FMDV VP1 specific antibody response and protection against the experimental challenge, no significant differences were demonstrated in the immunizing activity of plant extracts obtained from the pRok2 or pRok3 transformed plants. These results confirm those previously obtained using other plant species allowing the possibility of using plants as antigen expression vectors, and demonstrated that at least in the potato system, the use of double CaMV 35S promoter does not cause a significant increase in the level of the VP1 expressed.

Flow cytometry and the study of central nervous disease in patients with acute leukaemia.

Central nervous system (CNS) leukaemia is still a matter of debate and new technologies are required to improve the classic morphological definition. One hundred and sixty-eight cerebrospinal fluid (CSF) samples from 31 patients with acute leukaemia were analysed by flow cytometry and conventional cytology. Concordant positive and negative findings were found in 158 samples but 10 produced discrepant results. Cytology seemed to offer more precise information in one CSF sample and flow cytometric accuracy could be demonstrated in five samples. We conclude that flow cytometry is of great help in confirming CNS leukaemia and eliminating other conditions. Therefore, leukaemic patients can benefit from double cytological and flow cytometric CSF studies.

Hepatitis B virus-specific T-cell proliferation and cytokine secretion in chronic hepatitis B e antibody-positive patients treated with ribavirin and interferon alpha.

Immune elimination of hepatitis B virus (HBV) during antiviral therapy depends on the activation of T-cell responses, which are generally impaired in chronic hepatitis B. HBV-specific T helper (Th)-cell reactivity has been assessed post-treatment in liver and peripheral blood of 18 anti-HBe-positive patients with chronic hepatitis B administered combined ribavirin/interferon alfa (IFN-alpha) therapy. The results showed that patients with undetectable HBV DNA by quantitative polymerase chain reaction under combination therapy were able to mount an HBV-specific CD4(+) Th-cell proliferative response and such T-cell reactivity is detectable 1 year after HBV DNA clearance. Hepatitis B virus core (HBcAg) and e (HBeAg) antigen-specific Th-cell proliferation was found more frequently in the liver and peripheral blood in those patients who sustained the alanine aminotransferase (ALT) normalization together with HBV DNA loss. However, HBV-specific IFN-gamma production in vitro in peripheral blood mononuclear cells augmented in 4 of 5 sustained responders and all 13 nonresponders, interleukin 10 (IL-10) production decreased in all 5 sustained responders but increased in 7 of 13 nonresponders. Furthermore, intrahepatic HBcAg plus HBeAg-specific Th-cell proliferation only occurred in sustained responders (2 of 3, 67%, vs. 0 of 9; P =.045) whose cells showed in vitro significantly increased productions in HBcAg/HBeAg-specific IFN-gamma and IL-12 compared with nonresponders in whom IFN-gamma and IL-12 productions decreased together with increased IL-10 secretion. In conclusion this study indicates that combined therapy with ribavirin and IFN-alpha for chronic hepatitis B not only significantly reduces viremia levels but also induces lasting CD4(+) T-cell proliferation and Th1 cytokine release at the site of infection, which may lead to sustained eradication of the HBV.

Oral immunogenicity of the plant derived spike protein from swine-transmissible gastroenteritis coronavirus.

Transgenic plants represent an inexpensive alternative to classical fermentation systems for production of recombinant subunit vaccines. Transgenic potato plants were created that express the N-terminal domain of the glycoprotein S (N-gS) from Transmissible gastroenteritis coronavirus (TGEV), containing the major antigenic sites of the protein. Extracts from potato tubers expressing N-gS were inoculated intraperitoneally to mice, and the vaccinated mice developed serum IgG specific for TGEV. Furthermore, when potato tubers expressing N-gS were fed directly to mice, they developed serum antibodies specific for gS protein, demonstrating the oral immunogenicity of the plant derived spike protein from TGEV.

Estadística al día / Updated statistics

Es importante para el pediatra y médico general conocer las variaciones experimentadas por los parámetros estadísticos, que reflejan en parte la evolución de la medicina y lo que le va ocurriendo a la población de su país. Este artículo analiza lo ocurrido en los últimos años con las tasas de natalidad y mortalidad en las diferentes edades, y así como las causas de morbilidad a partir de los egresos hospitalarios y lo observado en la evaluación del estado nutritivo. El conocimiento y análisis de las estadísticas son necesarios para la planificación de políticas de salud y para evaluar el impacto de los programas y del desarrollo de una región o país

Immunization with potato plants expressing VP60 protein protects against rabbit hemorrhagic disease virus.

The major structural protein VP60 of rabbit hemorrhagic disease virus (RHDV) has been produced in transgenic potato plants under the control of a cauliflower mosaic virus 35S promoter or a modified 35S promoter that included two copies of a strong transcriptional enhancer. Both types of promoters allowed the production of specific mRNAs and detectable levels of recombinant VP60, which were higher for the constructs carrying the modified 35S promoter. Rabbits immunized with leaf extracts from plants carrying this modified 35S promoter showed high anti-VP60 antibody titers and were fully protected against the hemorrhagic disease.

Study of the DNA content by flow cytometry and proliferation in 281 brain tumors.

In this paper we investigate the distribution of DNA ploidy as well as proliferation rate (S phase of the cell cycle Ki-67 staining) in 281 tumors of the central and peripheral nervous system (87 meningiomas, 75 astrocytomas, 44 nerve sheath tumors, 25 brain metastases, 18 pituitary adenomas, 17 ependymomas, 12 oligodendrogliomas and 3 medulloblastomas) and their correlation with the histopathological grade. Considering all 281 tumors, aneuploidy is the most frequent finding present: 52%. This percentage increases with malignancy: 69% of malignant tumors are aneuploidy. Levels of aneuploidy decrease from brain metastases to pituitary adenoma (92% in brain metastases, 83% in oligodendrogliomas, 59% in nerve sheath tumors, 47% in ependymomas, 40% in astrocytomas, 35% in meningiomas and 33% in pituitary adenomas), but aneuploidy is also found in many benign tumors. With respect to proliferation rate of tumors, S phase above 20% were recorded in the more malignant tumors: brain metastases, oligodendrogliomas, high-grade ependymomas, high grade astrocytomas, and in atypical and malignant meningiomas, but this parameter is not able to distinguish between low and high grade tumors. However, Ki-67 reactivity was equivalent in all histologies with significant differences between low and high grade tumors.