Effect of melatonin on the functional recovery from experimental traumatic compression of the spinal cord

Spinal cord injury is an extremely severe condition with no available effective therapies. We examined the effect of melatonin on traumatic compression of the spinal cord. Sixty male adult Wistar rats were divided into three groups: sham-operated animals and animals with 35 and 50% spinal cord compression with a polycarbonate rod spacer. Each group was divided into two subgroups, each receiving an injection of vehicle or melatonin (2.5 mg/kg, intraperitoneal) 5 min prior to and 1, 2, 3, and 4 h after injury. Functional recovery was monitored weekly by the open-field test, the Basso, Beattie and Bresnahan locomotor scale and the inclined plane test. Histological changes of the spinal cord were examined 35 days after injury. Motor scores were progressively lower as spacer size increased according to the motor scale and inclined plane test evaluation at all times of assessment. The results of the two tests were correlated. The open-field test presented similar results with a less pronounced difference between the 35 and 50% compression groups. The injured groups presented functional recovery that was more evident in the first and second weeks. Animals receiving melatonin treatment presented more pronounced functional recovery than vehicle-treated animals as measured by the motor scale or inclined plane. NADPH-d histochemistry revealed integrity of the spinal cord thoracic segment in sham-operated animals and confirmed the severity of the lesion after spinal cord narrowing. The results obtained after experimental compression of the spinal cord support the hypothesis that melatonin may be considered for use in clinical practice because of its protective effect on the secondary wave of neuronal death following the primary wave after spinal cord injury.

Delivery of the Sox9 gene promotes chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in an in vitro model

SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.

Salvaged single-unit cord blood transplantation for 26 patients with hematologic malignancies not in remission

Treatments for patients with hematologic malignancies not in remission are limited, but a few clinical studies have investigated the effects of salvaged unrelated cord blood transplantation (CBT). We retrospectively studied 19 patients with acute leukemia, 5 with myelodysplastic syndrome (MDS with refractory anemia with excess blasts [RAEB]), and 2 with non-Hodgkin's lymphoma who received 1 CBT unit ≤2 loci human leukocyte antigen (HLA)-mismatched after undergoing myeloablative conditioning regimens between July 2005 and July 2014. All of them were in non-remission before transplantation. The infused total nucleated cell (TNC) dose was 4.07 (range 2.76-6.02)×107/kg and that of CD34+ stem cells was 2.08 (range 0.99-8.65)×105/kg. All patients were engrafted with neutrophils that exceeded 0.5×109/L on median day +17 (range 14-37 days) and had platelet counts of >20×109/L on median day +35 (range 17-70 days). Sixteen patients (61.5%) experienced pre-engraftment syndrome (PES), and six (23.1%) patients progressed to acute graft-versus-host disease (GVHD). The cumulative incidence rates of II-IV acute GVHD and chronic GVHD were 50% and 26.9%, respectively. After a median follow-up of 27 months (range 5-74), 14 patients survived and 3 relapsed. The estimated 2-year overall survival (OS), disease-free survival (DFS), and non-relapse mortality (NRM) rates were 50.5%, 40.3%, and 35.2%, respectively. Salvaged CBT might be a promising modality for treating hematologic malignancies, even in patients with a high leukemia burden.

Assessment of the diagnostic value of diffusion tensor imaging in patients with spinal cord compression: a meta-analysis

We investigated the diagnostic value of the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) of magnetic resonance diffusion tensor imaging (DTI) in patients with spinal cord compression (SCC) using a meta-analysis framework. Multiple scientific literature databases were exhaustively searched to identify articles relevant to this study. Mean values and standardized mean differences (SMDs) were calculated for the ADC and FA in normal and diseased tissues. The STATA version 12.0 software was used for statistical analysis. Of the 41 articles initially retrieved through database searches, 11 case-control studies were eligible for the meta-analysis and contained a combined total of 645 human subjects (394 patients with SCC and 251 healthy controls). All 11 studies reported data on FA, and 9 contained data related to the ADC. The combined SMDs of the ADC and FA showed that the ADC was significantly higher and the FA was lower in patients with SCC than in healthy controls. Subgroup analysis based on the b value showed higher ADCs in patients with SCC than in healthy controls at b values of both ≤500 and >500 s/mm2. In summary, the main findings of this meta-analysis revealed an increased ADC and decreased FA in patients with SCC, indicating that DTI is an important diagnostic imaging tool to assess patients suspected to have SCC.

Lack of association between iron status at birth and growth of preterm infants

OBJECTIVE: To assess the association between iron status at birth and growth of preterm infants. METHODS: Ninety-five premature babies (26 to 36 weeks of gestational age) born from July 2000 to May 2001 in a public hospital in Rio de Janeiro, Southeastern Brazil, were followed up for six months, corrected by gestational age. Iron measurements at birth were available for 82 mothers and 78 children: hemoglobin, hematocrit, mean corpuscular volume and plasma iron. All children received free doses of iron supplement (2 mg/kg/day) during the follow-up period and up to two years of age. Multivariate linear regression analyses with repeated measurements were performed to assess factors associated to linear growth. RESULTS: Growth was more pronounced up to 40 weeks of gestational age, increasing about 1.0 cm/week and then slowing down to 0.75 cm/week. The multivariate analysis showed growth was positively associated with birth weight (0.4 cm/100 g; p<0.001) and negatively associated with gestational age at birth (-0.5 cm/week; p<0.001). There was no association between cord iron and mother iron measurements and growth (p>0.60 for all measures). Only two children had anemia at birth, whereas 43.9% of mothers were anemic (hemoglobin <11 g/dl). Also, there was no correlation between anemia indicators of mothers and children at birth (r<0.15; p>0.20). CONCLUSIONS: Maternal anemia was not associated with anemia in preterm infants and iron status of mothers and children at birth was not associated with short-term growth of preterm infants.

Relationship between the iron status of pregnant women and their newborns

OBJECTIVE: To determine the relationship between iron nutritional status of pregnant women and their newborns using a combination of hematological and biochemical parameters for the diagnosis of iron deficiency. METHODS: A cross-sectional study was conducted in Jundiaí, Southeastern Brazil, in 2000. Venous blood samples collected from 95 pregnant women and from their umbilical cord and used for the determination of complete blood count, serum iron, total iron-binding capacity, serum ferritin, zinc protoporphyrin, and transferrin saturation. Women were classified into three groups: anemic, iron deficient and non-iron deficient. Statistical analysis included the Tukey-HSD test, Pearson's correlation coefficient and multiple linear regression analysis. RESULTS: Among pregnant women, 19% were anemic (97.9% mildly anemic and 2.1% moderately anemic) and 30.5% were iron deficient. No significant difference was seen in mean values of any parameter studied between newborns in the three groups (p>0.05). Multiple linear regression analysis showed weak association between neonatal and maternal parameters. CONCLUSIONS: The iron nutritional status of pregnant women with iron deficiency or mild anemia does not seem to have a significant impact on the iron levels of their children.

Tempo de clampeamento e fatores associados à reserva de ferro de neonatos a termo

OBJETIVO : Analisar o impacto do tempo de clampeamento e parâmetros obstétricos, biológicos e socioeconômicos sobre a reserva de ferro de neonatos nascidos a termo. MÉTODOS : Estudo transversal pelo qual foram avaliados os parâmetros hematológicos de neonatos de Viçosa, MG, de outubro de 2011 a julho de 2012. Foram coletados 7 mL de sangue do cordão umbilical de 144 neonatos a termo e sem baixo peso. Os parâmetros investigados foram: hemograma completo, ferro sérico, ferritina e proteína C-reativa. O tempo de clampeamento do cordão umbilical foi mensurado utilizando cronômetro digital sem interferir nos procedimentos do parto. Os dados de nascimento foram coletados nas Declarações de Nascidos Vivos e as demais informações foram obtidas com a mãe do neonato por aplicação de questionário no primeiro mês pós-parto. Realizou-se análise de regressão linear múltipla visando a estimar a influência de variáveis obstétricas, biológicas e socioeconômicas nos níveis de ferritina ao nascer. RESULTADOS : A mediana de ferritina foi 130,3 µg/L (n = 129, mínimo de 16,4 e máximo 420,5 µg/L), a média de ferro sérico foi 137,9 μg/dL (n = 144, dp = 39,29) e de hemoglobina, 14,7 g/dL (n = 144, dp = 1,47). O tempo mediano de clampeamento do cordão foi 36 segundos, variando entre sete e 100. A análise bivariada detectou associação entre os níveis de ferritina e a cor da criança, tempo de clampeamento de 60 segundos, tipo de parto, a presença de diabetes gestacional e a renda per capita da família. Renda per capita, número de consultas pré-natais e o comprimento ao nascer contribuíram com 22,0% da variação dos níveis de ferritina na análise múltipla. CONCLUSÕES : A reserva de ferro ao nascer sofreu influência de características biológicas, obstétricas e sociais. O combate à anemia deve envolver a implementação de um critério de clampeamento tardio do cordão umbilical para as diretrizes de trabalho de parto, bem como a criação de políticas voltadas para a redução das desigualdades sociais e melhoria da qualidade do atendimento pré-natal.

On the tissular parasitism of Trypanosoma cruzi y strain in swiss mice

A review of the tissular parasitism of Trypanosoma cruzi Y strain in Swiss mice was carried out. This strain parasitized preferentially smooth, skeletal and cardiac muscle fibers, with low transitory spleen and liver parasitism, as previously found by some Authors, although differing from other reports. These results can be related to the host genetical constitution and/or the degree of the strain virulence at the time of this study. Furthermore, we discuss that the high macrophagotropism reported for this strain in some instances could be an artificially induced condition resulting from its serial maintenance in mice, either for a longer time and/or by using young animals. The heavy parasitism and inflammation observed in the bladder, pancreas and spermatic duct of some inoculated mice, as well as the testis parasitization, were also noteworthy findings.

Congenital cytomegalovirus infection: occurrence in two socioeconomically distinct populations of a developing country

In São Paulo, Brazil, between November 1980 and July 1982, 1614 newborns of middle socioeconomic background and 1156 newborns of low socioeconomic background were examined for the occurrence of congenital cytomegalovirus (CMV) infection by isolation of virus from urine samples or detection of specific anti-CMV IgM in umbilical cord serum tested by immunofluorescence. In the low socioeconomic population prevalence of CMV complement-fixing antibodies in mothers was 84.4%(151/179) and the incidence of congenital infection assessed by virus isolation 0.98% (5/508), as compared with 0.46% (3/648) in the group of newborns tested by detection of specific anti-CMV IgM in umbilical cord-serum. In middle socioeconomic level population prevalence of CMV complement-fixing antibodies in mothers was 66.5% (284/427) and the incidence of CMV congenital infection was 0.39% (2/518) in the group of newborns screened by virus isolation and 0.18% (2/1096) in the group tested by detection of specific anti-CMV IgM. In the present study none of the 12 congenitally infected newborns presented clinical apparent disease at birth.

Comparison of indirect immunofluorescence test for measles antibodies with haemagglutination inhibition and plaque neutralization tests

Indirect Immunofluorescence (IFA), Plaque Reduction Neutralization (PRN) and Haemagglutination Inhibition (HI) tests for measles antibodies were carried out in 197 sera obtained from umbilical cord and vaccinated children. The IFA was also applied to blood samples collected with filter paper. IFA results demonstrated that the test is relatively simple to perform, with good reproducibility for different antigen lots. Good correlation was obtained between IFA, PRN and HI antibody titers. Better correlation was demonstrated with IFA and PRN than with HI and PRN tests. Sensitivity of IFA in detecting antibody was less effective than PRN, however more effective than HI using rhesus monkey red blood cells. PRN antibody titers over 100 were detected by IFA but not by HI (9.7% with negative results). IFA may be of considerable practical use and able to substitute HI in Seroepidemiological surveys and to evaluate vaccine efficacy. It also can be simplified by employing filter paper collected samples.

Enzyme-linked immunosorbent assay (ELISA) for measles antibody: a comparison with haemagglutination inhibition, immunofluorescence and plaque neutralization tests

An enzyme-linked immunosorbent assay (ELISA) for measles antibodies was compared with Plaque Neutralization (PRN), Haemagglutination inhibition (HI) and Fluorescent antibody (IFA) tests in 181 sera from vaccinated children and umbilical cord. Of 179 positive samples by the sensitive PRN, only two, with titers of 8, were negative by ELISA (copositivity of 98.9%). IFA and HI presented, respectively, copo-sitivities of 93.3% and 82.7%. The ELISA presented a high sensitivity as well as a good reproducibility and represents an alternative for the time consuming PRN for detection of low measles antibodies.

Embryology of Triatoma infestans (KLUG), (Hemiptera, Reduviidae), a Chagas' disease vector

This study reports the embryogenesis of T. infestans (Hemiptera, Reduviidae). Morphological parameters of growth sequences from oviposition until hatching (12-14 d 28ºC) were established. Five periods, as percent of time of development (TD), were characterized from oviposition until hatching. The most important morphological features were: 1) formation of blastoderm within 7% of TD; 2) germ band and gastrulation within 30% of TD; 3) nerve cord, limb budding, thoracic and abdominal segmentation and formation of body cavity within 50% of TD; 4) nervous system and blastokinesis end, and development of embryonic cuticle within 65% of TD; 5) differentiation of the mouth parts, fat body, and Malphigian tubules during final stage and completion of embryo at day 12 to day 14 around hatching. These signals were chosen as appropriate morphological parameters which should enable the evaluation of embryologic modifications due to the action/s of different insecticides

Prevalence of rubella antibodies in a non-immunized urban population, São Paulo, Brazil

The prevalence of rubella antibodies was evaluated through a ramdom Seroepidemiological survey in 1400 blood samples of 2-14 year old children and in 329 samples of umbilical cord serum. Rubella IgG antibodies were detected by ELISA, and the sera were collected in 1987, five years before the mass vaccination campaign with measles-mumps-rubella vaccine carried out in the city of São Paulo in 1992. A significant increase in prevalence of rubella infection was observed after 6 years of age, and 77% of the individuals aged from 15 to 19 years had detectable rubella antibodies. However, the seroprevalence rose to 90.5% (171/189) in cord serum samples from children whose mothers were 20 to 29 years old, and reached 95.6% in newborns of mothers who were 30 to 34 years old, indicating that a large number of women are infected during childbearing years. This study confirms that rubella infection represents an important Public Health problem in São Paulo city. The data on the seroprevalence of rubella antibodies before the mass vaccination campaign reflects the baseline immunological status of this population before any intervention and should be used to design an adequate vaccination strategy and to assess the Seroepidemiological impact of this intervention.

A CASE REPORT OF INTRASPINAL PARACOCCIDIOIDOMYCOSIS

The authors report a case of paraplegia caused by a lumbar intraspinal paracoccidioidomycosis (PCM) granuloma. Clinical neurological diagnosis of a compressive spinal cord lesion was confirmed by spinal magnetic resonance imaging (MRI). Patient was submitted to surgery with total excision of the lesion. Histopathological analysis confirmed the diagnosis of PCM. Patient is on sulfamethoxazole/trimethoprim combined with fluconazole and is experiencing positive neurological recovery.

COMPARISON OF A COMMERCIAL ENZYME IMMUNOASSAY WITH PLAQUE REDUCTION NEUTRALIZATION FOR MATERNAL AND INFANT MEASLES ANTIBODY MEASUREMENT

The most practicable assay for measurement of measles IgG (mIgG) in large numbers of sera is an enzyme immunoassay (EIA). To assess how EIA results would agree with those by the gold standard method of plaque reduction neutralization (PRN) we compared the results from the two methods in 43 pairs of maternal and umbilical cord sera, and sera from the corresponding infants when aged 11 - 14 months. In maternal-cord sera, the differences between mean antibody levels by EIA or PRN were not statistically significant, though in individual sera, differences could be large. However, agreement was less good for infants sera, in which levels of mIgG were very low. The conclusions of a study of transplacental transport of mIgG would not be affected by the use of either technique. When studying waning immunity in infants, PRN should be the method of choice, while results from studies using EIA should be interpreted with caution.