Hofbauer cells morphology and density in placentas from normal and pathological gestations

PURPOSE: In placentas from uncomplicated pregnancies, Hofbauer cells either disappear or become scanty after the fourth to fifth month of gestation. Immunohistochemistry though, reveals that a high percentage of stromal cells belong to Hofbauer cells. The aim of this study was to investigate the changes in morphology and density of Hofbauer cells in placentas from normal and pathological pregnancies. METHODS: Seventy placentas were examined: 16 specimens from normal term pregnancies, 10 from first trimester's miscarriages, 26 from cases diagnosed with chromosomal abnormality of the fetus, and placental tissue specimens complicated with intrauterine growth restriction (eight) or gestational diabetes mellitus (10). A histological study of hematoxylin-eosin (HE) sections was performed and immunohistochemical study was performed using the markers: CD 68, Lysozyme, A1 Antichymotrypsine, CK-7, vimentin, and Ki-67. RESULTS: In normal term pregnancies, HE study revealed Hofbauer cells in 37.5% of cases while immunohistochemistry revealed in 87.5% of cases. In first trimester's miscarriages and in cases with prenatal diagnosis of fetal chromosomal abnormalities, both basic and immunohistochemical study were positive for Hofbauer cells. In pregnancies complicated with intrauterine growth restriction or gestational diabetes mellitus, a positive immunoreaction was observed in 100 and 70% of cases, respectively. CONCLUSIONS: Hofbauer cells are present in placental villi during pregnancy, but with progressively reducing density. The most specific marker for their detection seems to be A1 Antichymotrypsine. It is remarkable that no mitotic activity of Hofbauer cells was noticed in our study, as the marker of cellular multiplication Ki-67 was negative in all examined specimens.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in São Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO², >37ºC). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

The aim of this study was to evaluate alterations to the physiological profile (cortisol, glycaemia, and blood parameters) of Lithobates catesbeianus caused by the stressors density and hypoxia. The organisms were in the prometamorphosis stage and exposed to different tadpole densities: 1 tadpole/L (T1), 5 tadpoles/L (T2), and 10 tadpoles/L (T3) for 12 days. The blood was collected through the rupture of the caudal blood vessel and collected under normoxia (immediate collection) and hypoxia (after 15 minutes of air exposure) conditions. Cortisol levels rose on the fourth and eighth days of treatment and returned to basal levels by the end of the experiment. The stressor mechanisms tested did not affect glycaemia. White blood cells (total number of lymphocytes, neutrophils, and eosinophils) showed a significant difference at the twelfth day of the experiment when compared with the start of the experiment. We concluded that, under controlled conditions, a density of up to 10 tadpoles/L and air exposure for 15 minutes did not cause harmful physiological alterations during the experimental period. The answer to these stressors maybe was in another hormonal level (corticosterone).

Density of primary and secondary epidermal laminae of equine hoof

Differences in the microscopic morphology of the hoof in forelimbs and hindlimbs of horses have been scarcely reported in the literature, especially concerning the distribution of primary and secondary epidermal laminae in the different regions. This study aimed to determine the density of primary and secondary epidermal laminae in the hoof of horses. For this, it was used fore and hindlimbs of 16 adult mixed breed horses. With a cross section 0.5 cm above the sole, it was quantified the primary epidermal laminae in the regions of the toe, and of lateral and medial quarters. Fragments with about 1cm ³ were taken from the proximal, middle and distal thirds of the hooves, in the different regions, subjected to conventional histological techniques and examined with an optical microscope. Data were statistically analyzed in relation to the fore and hindlimbs and between their various regions. The density of primary epidermal laminae varied around the hoof circumference, with greater values in the hoof toe, which gradually decreased towards the bulb of the hoof, without difference between thoracic and pelvic limbs. The average density of the secondary epidermal laminae per primary epidermal lamina does not change around the circumference of the hoof. Our findings indicated that the density of epidermal laminae is not different between fore and hindlimbs. The variation in the density of primary epidermal laminae around the hoof seems to be part of an adaptive response to different stresses in each region. A better understanding of the structural morphology contributes to a better understanding of the diagnosis, pathophysiology, and treatment of disorders that affect the hoof.

Avaliação da deposição da calda de pulverização em função da densidade populacional de Brachiaria plantaginea, do volume e do ângulo de aplicação: effects of density, volume and spraying angle

O presente trabalho teve por objetivo avaliar a quantidade de calda de pulverização depositada nas folhas de Brachiaria plantaginea, em aplicações de pós-emergência precoce, em que se variou o volume de calda de pulverização, a densidade de plantas m-2 e o ângulo de posicionamento da ponta de pulverização na barra de aplicação. Para isso, foram conduzidos três experimentos em condições de laboratório. Nestes estudos, o volume de calda de pulverização foi obtido por meio da variação da velocidade de deslocamento de um veículo composto por plataforma e quatro rolamentos tracionados por um motor elétrico. Foi utilizada a ponta de pulverização XR Teejet 8001 EVS, na pressão de 241,4 kPa. O delineamento experimental utilizado foi o inteiramente casualizado, com cinco repetições. No primeiro experimento, os volumes utilizados foram de 1147,57; 860,68; 573,78; 459,02; 344,27; 229,51; 114,75; e 57,37 L ha-1 de calda de pulverização, com densidade de 600 plantas m-2. No segundo experimento foram estudadas as densidades de 300, 600, 900 e 1.200 plantas m-2; neste caso, utilizou-se o volume de 229,51 L ha-1 de calda de pulverização. No terceiro experimento avaliou-se o posicionamento do ângulo da ponta de pulverização na barra de aplicação e utilizou-se a densidade de 600 plantas m-2. Estudaram-se os ângulos de -30º, -15º, 90º, +15º e +30º e os volumes de calda de pulverização de 198,76; 221,69; 229,51; 221,69; e 198,76 L ha-1, respectivamente. Foram adotados sinais negativos para o sentido de deslocamento do veículo e sinais positivos para o sentido contrário ao deslocamento. As avaliações do depósito de calda de pulverização, na planta e no solo, foram feitas utilizando-se condutividade elétrica. A porcentagem de depósito de calda de pulverização nas plantas de B. plantaginea foi incrementada com a redução do volume de calda pulverizada por hectare. O depósito de calda por planta foi maior nas densidades maiores de plantas. O ângulo de posicionamento da ponta de pulverização na barra de aplicação incrementou o depósito de calda nas plantas de B. plantaginea, quando comparado com o ângulo de 90º.

Planting density of gliricidia when intercropped with corn for weed control

Reduced use of herbicides that cause environmental pollution problems is of great interest in modern agriculture. Soil mulching with gliricidia (Gliricidia sepium) branches does not have an allelopathic effect on corn, but decreases weed populations. The objective of this study was to evaluate the effects of gliricidia planting density, when grown as an intercrop, on weed control and corn yield parameters. A randomized block design with split-plots and ten replicates was adopted. Corn cultivars AG 1051 and BM 3061 were grown without hoeing, with two hoes (at 24 and 44 days after planting), and intercropped with gliricidia (planted simultaneously with corn, between crop rows, using two seedlings/pit, spaced at 30, 40, or 50 cm). Twenty-one weed species were found in the experimental area. Increased gliricidia planting density reduced weed biomass, but no difference was found between weed biomass in the intercrop and weed biomass in non-hoed corn. Gliricidia intercropped with corn, planted at a row spacing of 30 cm, did not significantly differ from hoed corn in most characteristics considered to evaluate green corn yield, although mean values were smaller. As to the number and weight of marketable green ears, reductions of 5% and 13%, respectively, were observed. Intercropping caused a 17% reduction in grain yield, reducing the losses (36%) observed in non-hoed corn by more than 50%. The highest green ear yield and grain yield values were obtained with two hoeings, while the lowest values were observed for non-hoed corn. The cultivars did not differ regarding green ear yield and grain yield.

Influence of corn sowing density and gliricidia intercropping on weed control

There is interest in the identification of the best seeding density for new corn hybrids and on reduced use of herbicides for weed control. The objective of this study was to evaluate the effects of seeding density (30, 50, 70, and 90 thousand plants ha-1) and weed control on green ear yield and grain yield in corn cultivar AG 1051. A completely randomized block design was adopted with split-plots (seeding densities assigned to plots) and ten replicates. Weed control was achieved by means of two hoeings and by planting corn intercropped with gliricidia (between corn rows, in pits spaced 0.3 m apart). A "no weeding"treatment was included as well. Increased seeding density increased the total number and weight of marketable green ears and decreased the biomass of both weeds and gliricidia. In non-weeded, intercropped and hoed plots, the maximum grain yield values achieved as seeding density increased were 7,881, 7,021, and 9,213 kg ha-1, respectively, obtained with populations of 67 thousand, 74 thousand, and 67 thousand plants per hectare, respectively. Intercropping did not control weeds (26 species) and provided weed growth, green ear yield, and grain yield (at the lowest densities) similar to those obtained without hoeing, except for total number of green ears, in which no influence of weed control was observed. At densities of 70 thousand and 90 thousand plants per hectare, grain yield with two hoeings was not different from yield values obtained without weeding or in the treatment intercropped with gliricidia, respectively, indicating that increased corn seeding density as well as gliricidiamay help to control weeds.

Density and sowing season of two Brachiaria species on the soybean culture

The aim of this study was to evaluate the growth and yield of soybean cultivar M-8766 in consortium with Brachiaria brizantha. BRS Piata and Brachiaria ruziziensis at different densities and sowing dates. The experimental design was randomized blocks with treatments arranged in a factorial 2 x 2 x 3 with four replications. Used as factors grass species (Brachiaria brizantha Piata and Brachiaria ruziziensis BRS) intercropped with soybean M-8766, sowing dates (12 and 24 days after soybean emergence) and three seeding rates (0, 5, 10 kg ha-1 of seed). At 71 days after soybean emergence were evaluated plant height, stem diameter, dry mass of leaves, stems and shoots, and 4 months after sowing determined the weight of 100 grains and soybean yield. The results showed that when seeded at a density of 10 kg ha-1 at 12 and 24 DAE soy, Brachiaria brizantha. BRS Piata caused reduction in yield in the order of 6.71% and 3.03% respectively, while the Brachiaria ruziziensis was one that caused a greater reduction in productivity in the order of 13.42 and 16.23%, respectively, of these values expression when considering the price of soybean sack. B. ruziziensis expressed less competitive with soybean. However, the large biomass production of this grass provides deployment system till the next harvest.

Effect of plant density and seed position on mother plant on physiological characteristic of cocklebur (Xanthium strumarium) seeds

Experiments were conducted in 2010 to determine the influence of plant density and seed position on the mother plant on seed physiological characteristics of cocklebur (Xanthium strumarium). Cocklebur burs were collected in fall of 2010 from Research Farm of University of Agricultural Sciences and Natural Resources of Gorgan, Iran. The experiment was established as factorial arrangement using a completely randomized design with three replications. The factors included different densities of cocklebur (0, 2, 4, 6 and 8 plant m-2) and the top and bottom parts of the canopy. Non dormant seeds were used for determining cardinal temperatures and tolerance to salinity and drought stresses. Base, optimum and ceiling germination temperatures were estimated between 7.09 to 12.33, 32 to 35 and 44 to 45 respectively in different treatments. Salinity stress up to 300 Mm and osmotic potential 8 bar inhibited the germination completely. Comparison of base temperatures and sigmoid equation coefficients showed that seeds produced in the top had higher germination than those that produced at the bottom of the mother plant. It seems plant densities through seed position on the mother plant affect seed quality. Likewise changes of light quality and quantity in shade environment increased seed dormancy in matured seeds. Shade environment affect seed germination on mother plant that increased dormancy of seeds maturing under shade be an adaptive response that reduces the probability of germination of offspring under unfavorable (shade, competitive) conditions.

Population density and weed infestation in organic no-tillage corn cropping system under different soil covers

Currently, one of the biggest challenges faced by organic no-tillage farming is weed control. Thus, the use of cropping practices that help in the control of weeds is extremely important. The objective of this study was to evaluate population density and level of weed infestation in an organic no-tillage corn cropping system under different soil covers. The experiment was conducted in a randomized block design with six repetitions and five treatments, consisting of three soil covers in an organic no-tillage system, and an organic and a conventional system, both without soil cover. The treatments with soil cover used a grass species represented by the black oat, a leguminous species represented by the white lupine, and intercropping between both species. Corn was sown with spacing of 1.0 m between rows and 0.20 m between plants, using the commercial hybrid AG 1051. Infestation in corn was evaluated at stages V5 and V10, and weed density was evaluated at stage V5. The use of black oat straw alone or intercropped with white lupine, in the organic no-tillage corn cropping system, reduced the percentage of weed infestation and absolute weed density. Management-intensive systems and systems without soil cover showed higher relative densities for species Oxalis spp., Galinsoga quadriradiata and Stachys arvensis. The species Cyperus rotundus showed the highest relative density on organic no-tillage corn cropping systems. Black oat straw in the organic no-tillage cropping system limited the productive potential of corn.

Estimated frequency of B-chromosomes and population density of Astyanax scabripinnis paranae in a small stream

A study was made of the frequency of B-chromosomes and the population density of Astyanax scabripinnis paranae (Pisces, Characidae, Tetragonopterinae) from three stretches of the Cascatinha stream (Botucatu, SP). In the first stretch the population was estimated to be about 212 individuals and among 35 karyotyped specimens, 23 carried one macro B-chromosome; in the second stretch the population was estimated to be about 650 individuals and among 20 specimens karyotyped, two possessed one macro B-chromosome; in the third stretch the population was estimated to be about 107 individuals and among 10 specimens karyotyped, one carried one macro B-chromosome. A significant difference was observed in the frequency of macro B-chromosomes in females (57%) and males (8.7%) (P = 0.0001). These data suggest that the B-chromosome frequency and the populational density are not directly related. The hypothesis of the existence of some adaptive effect conferred by the B-chromosomes to the specimens from the first stretch of the Cascatinha stream is presented and discussed

Vitamin D receptor alleles and bone mineral density in a normal premenopausal Brazilian female population

Studies on the association between vitamin D receptor (VDR) polymorphism and bone mineral density (BMD) in different populations have produced conflicting results probably due to ethnic differences in the populations studied. The Brazilian population is characterized by a very broad genetic background and a high degree of miscegenation. Of an initial group of 164, we studied 127 women from the city of São Paulo, aged 20 to 47 years (median, 31 years), with normal menses, a normal diet and no history of diseases or use of any medication that could alter BMD. VDR genotype was assessed by PCR amplification followed by BsmI digestion of DNA isolated from peripheral leukocytes. BMD was measured using dual energy X-ray absorptiometry (Lunar DPX) at the lumbar site (L2-L4) and femoral neck. Most of the women (77.6%) were considered to be of predominantly European ancestry (20.6% of them reported also native American ancestry), 12.8% were of African-Brazilian ancestry and 9.6% of Asian ancestry, 41.0% (52) were classified as bb, 48.8% (62) as Bb and 10.2% (13) as BB. The BB, Bb and bb groups did not differ in age, height, weight, body mass index or age at menarche. Lumbar spine BMD was significantly higher in the bb group (1.22 ± 0.16 g/cm²) than in the BB group (1.08 ± 0.14; P<0.05), and the Bb group presented an intermediate value (1.17 ± 0.15). Femoral neck BMD was higher in the bb group (0.99 ± 0.11 g/cm²) compared to Bb (0.93 ± 0.12) and BB (0.90 ± 0.09) (P<0.05). These data indicate that there is a significant correlation between the VDR BsmI genotype and BMD in healthy Brazilian premenopausal females.

Risk factors for decreased bone density in premenopausal women

Osteoporosis is a major health problem. Little is known about the risk factors in premenopause. Sixty 40-50-year old patients with regular menses were studied cross-sectionally. None of the patients were on drugs known to interfere with bone mass. Patients answered a dietary inquiry and had their bone mineral density (BMD) measured. The Z scores were used for the comparisons. A blood sample was taken for the determination of FSH, SHBG, estradiol, testosterone, calcium and alkaline phosphatase. Calcium and creatinine were measured in 24-h urine. A Z score less than -1 was observed for the lumbar spine of 14 patients (23.3%), and for the femur of 24 patients (40%). Patients with a Z score less than -1 for the lumbar spine were older than patients with a Z score ³-1 (45.7 vs 43.8 years) and presented higher values of alkaline phosphatase (71.1 ± 18.2 vs 57.1 ± 14.3 IU/l). Multiple regression analysis showed that a lower lumbar spine BMD was associated with higher values of alkaline phosphatase, lower calcium ingestion, a smaller body mass index (BMI), less frequent exercising, and older age. The patients with a Z score less than -1 for the femur were shorter than patients with a Z score ³-1 (158.2 vs 161.3 cm). Multiple regression analysis showed that a lower femoral BMD was associated with lower BMI, higher alkaline phosphatase and caffeine intake, and less frequent exercising. A lower than expected BMD was observed in a significant proportion of premenopausal women and was associated with lower calcium intake, relatively lower physical activity and lower BMI. We conclude that the classical risk factors for osteoporosis may be present before ovarian failure, and their effect may be partly independent of estrogen levels.