Inter-annual variations in the floristic and population structure of an herbaceous community of "caatinga" vegetation in Pernambuco, Brazil

The influence of climatic variations on the herbaceous component of the "caatinga" vegetation was examined in the state of Pernambuco, Brazil. A total of 105 1 × 1 m-plots were established, of which 35 were in a level micro-habitat, 35 in a riparian micro-habitat, and 35 in a stony microhabitat. During two consecutive years all herbaceous plants in these plots were counted, measured (height and diameter), and collected for identification. The Shannon-Wiener diversity index and the equitability were calculated for each year, as well as the density, frequency, dominance, total basal area and importance index for each species. The total annual pluviometric was 819.5 and 448.8 mm in 2002 and 2003, respectively. The herbaceous flora in the study area was composed of 71 species, of which 58 were sampled in the plots. The families with the greatest species richness were Malvaceae (8 species), Euphorbiaceae (7), Poaceae (6), Convolvulaceae (4), Fabaceae (4), and Portulacaceae (4). The diversity indices were 2.66 and 3.01 nats ind-1 in 2002 and 2003, respectively. The density, frequency, dominance and importance value of herbaceous populations, as well as, the height and diameter of plants were low in the dryer year. The riparian group was the most isolated of the microhabitats examined, both in terms of its floristic and its population structure. Annual seasonal climatic variations greatly modified these populations structure during the course of this study, emphasizing the fact that long-term studies are needed in order to better understand the dynamics of the herbaceous component of the "caatinga" vegetation.

Anomalous somatic embryos in Acca sellowiana (O. Berg) Burret (Myrtaceae)

Somatic embryogenesis represents a valuable tool for the studies on the basic aspects of plant embryo development. Today this process is used as a potencial technique for large-scale plant micropropagation although, so far, it has been applied to only a small number of species. However, when somatic embryos are malformed they are considered economically useless. In Acca sellowiana (O. Berg) Burret, an important fruit-producing crop, large amounts of anomalous somatic embryos (76.3%) were found just after 40 days of culture of explants in a 2,4-D containing medium. Among the anomalous forms found in the cotiledonary stage, 12.2% consisted of fused embryos, 40.4% displayed fused cotyledons, 13.0% presented supernumerary cotyledons, and 10.7% showed absence or poorly developed cotyledons, including those without the shoot apical meristem. Histological analyses indicated that the altered embryos were formed either directly from cotyledons, hypocotyl and radicle of the zygotic embryos used as explants, or indirectly from calli formed from these tissue parts. It is suggested that the formation of anomalous somatic embryos, as well as a low frequency of conversion into emblings reflect physiological and/or genetic disturbances triggered by the presence of 2,4-D in the medium. In vitro experimental alternative approaches are discussed in order to lessen the occurrence of malformed somatic embryos.

Spatial variations at different observational scales and the seasonal distributions of stream macroalgae in a Brazilian subtropical region

We examined the relationships between environmental variations in lotic ecosystems with the seasonal dynamics of macroalgae communities at different spatial scales: drainage basin of two rivers (Rio das Pedras and Rio Marrecas), shading (open and shaded stream segments), mesohabitat (riffles and pools), and microhabitats. Data collections were made on a monthly basis between January and December/2007. A total of 16 taxa were encountered (13 species and 3 vegetative groups). All of the biotic parameters (richness, abundance, diversity, equitability, and dominance) were found to be highly variable at all of the spatial scales evaluated. On the other hand, abiotic variables demonstrated differences only at mesohabitat (in terms of current velocity) and shaded habitat (in terms of irradiance) scales. The seasonality of the macroalgae community structure was strongly influenced by microhabitat variables (current velocity, substrate H', and irradiance), demonstrating their importance over time and at different scales. Regional variables (temperature, oxygen saturation, specific conductance, pH, and turbidity) were found to have little influence on the temporal dynamics of the macroalgae communities evaluated.

Temporal variations in the primary productivity of Eleocharis acutangula (Cyperaceae) in a tropical wetland environment

Wetland vegetation typically includes aquatic macrophytes with high primary production capacities. The present study investigated how hydrological variations affect biomass allocation and primary productivity in the emergent macrophyte Eleocharis acutangula (Roxb.) Schult. Eleocharis acutangula ramets were collected from the Campelo Lagoon flood plain (21°39'S, 41°12'W and 21°37S, 41°11'W) between March/2005 and February/2006. This region experienced an unusually short rainy period between November/2005 and February/2006 that generated atypically high primary production levels (128gDWm-2month-1) and total biomass gains (447gDWm-2) in May and June/2005 respectively. Our data indicated that primary production and biomass allocation were strongly influenced by variations in wetland water levels and that macrophytes quickly invested in biomass accumulation when surface water levels rised.

Modeling temporal variations of Gracilaria Greville and Hypnea J.V. Lamouroux (Rhodophyta) assemblages on a midlittoral reef platform at Piedade Beach, Pernambuco State, Brazil

The diversity of algal banks composed of species out the genera Gracilaria Greville and Hypnea J.V. Lamouroux have been impacted by commercial exploitation and coastal eutrophication. The present study sought to construct dynamic models based on algal physiology to simulate seasonal variations in the biomasses of Gracilaria and Hypnea an intertidal reef at Piedade Beach in Jaboatão dos Guararapes, Pernambuco State, Brazil. Five 20 × 20 cm plots in a reef pool on a midlittoral reef platform were randomly sampled during April, June, August, October, and December/2009 and in January and March/2010. Water temperature, pH, irradiance, oxygen and salinity levels as well as the concentrations of ammonia, nitrate and phosphate were measured at the sampling site. Forcing functions were employed in the model to represent abiotic factors, and algal decay was simulated with a dispersal function. Algal growth was modeled using a logistic function and was found to be sensitive to temperature and salinity. Maximum absorption rates of ammonia and phosphate were higher in Hypnea than in Gracilaria, indicating that the former takes up nutrients more efficiently at higher concentrations. Gracilaria biomass peaked at approximately 120 g (dry weight m-2) in March/2010 and was significantly lower in August/2009; Hypnea biomasses, on the other hand, did not show any significant variations among the different months, indicating that resource competition may influence the productivity of these algae.

Variations in gastric emptying of liquid elicited by acute blood volume changes in awake rats

We have observed that acute blood volume expansion increases the gastroduodenal resistance to the flow of liquid in anesthetized dogs, while retraction decreases it (Santos et al. (1991) Acta Physiologica Scandinavica, 143: 261-269). This study evaluates the effect of blood volume expansion and retraction on the gastric emptying of liquid in awake rats using a modification of the technique of Scarpignato (1980) (Archives Internationales de Pharmacodynamie et de Therapie, 246: 286-294). Male Wistar rats (180-200 g) were fasted for 16 h with water ad libitum and 1.5 ml of the test meal (0.5 mg/ml phenol red solution in 5% glucose) was delivered to the stomach immediately after random submission to one of the following protocols: 1) normovolemic control (N = 22), 2) expansion (N = 72) by intravenous infusion (1 ml/min) of Ringer-bicarbonate solution, volumes of 1, 2, 3 or 5% body weight, or 3) retraction (N = 22) by controlled bleeding (1.5 ml/100 g). Gastric emptying of liquid was inhibited by 19-51.2% (P<0.05) after blood volume expansion (volumes of 1, 2, 3 or 5% body weight). Blood volume expansion produced a sustained increase in central venous pressure while mean arterial pressure was transiently increased during expansion (P<0.05). Blood volume retraction increased gastric emptying by 28.5-49.9% (P<0.05) and decreased central venous pressure and mean arterial pressure (P<0.05). Infusion of the shed blood 10 min after bleeding reversed the effect of retraction on gastric emptying. These findings suggest that gastric emptying of liquid is subject to modulation by the blood volume.

Somatic gene therapy for hypertension

Gene therapy for hypertension is needed for the next generation of antihypertensive drugs. Current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). Gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). We are currently using two strategies: a) antisense oligodeoxynucleotides (AS-ODN) and b) antisense DNA delivered in viral vectors to inhibit genes associated with vasoconstrictive properties. It is not necessary to know all the genes involved in hypertension, since many years of experience with drugs show which genes need to be controlled. AS-ODN are short, single-stranded DNA that can be injected in naked form or in liposomes. AS-ODN, targeted to angiotensin type 1 receptors (AT1-R), angiotensinogen (AGT), angiotensin converting enzyme, and ß1-adrenergic receptors effectively reduce hypertension in rat models (SHR, 2K-1C) and cold-induced hypertension. A single dose is effective up to one month when delivered with liposomes. No side effects or toxic effects have been detected, and repeated injections can be given. For the vector, adeno-associated virus (AAV) is used with a construct to include a CMV promoter, antisense DNA to AGT or AT1-R and a reporter gene. Results in SHR demonstrate reduction and slowing of development of hypertension, with a single dose administration. Left ventricular hypertrophy is also reduced by AAV-AGT-AS treatment. Double transgenic mice (human renin plus human AGT) with high angiotensin II causing high blood pressure, treated with AAV-AT1-R-AS, show a normalization of blood pressure for over six months with a single injection of vector. We conclude that ODNs will probably be developed first because they can be treated like drugs for the treatment of hypertension with long-term effects. Viral vector delivery needs more engineering to be certain of its safety, but one day may be used for a very prolonged control of blood pressure.

Detection of somatic mutations of the PIG-A gene in Brazilian patients with paroxysmal nocturnal hemoglobinuria

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal syndrome characterized by intravascular hemolysis mediated by complement, thrombotic events and alterations in hematopoiesis. Basically, the molecular events which underlie the complexity of the syndrome consist of the absence of the glycosylphosphatidylinositol (GPI) anchor as a consequence of somatic mutations in the PIG-A gene, located on the X chromosome. The GPI group is responsible for the attachment of many proteins to the cytoplasmic membrane. Two of them, CD55 and CD59, have a major role in the inhibition of the action of complement on the cellular membrane of blood cells. The absence of GPI biosynthesis can lead to PNH. Since mutations in the PIG-A gene are always present in patients with PNH, the aim of this study was to characterize the mutations in the PIG-A gene in Brazilian patients. The analysis of the PIG-A gene was performed using DNA samples derived from bone marrow and peripheral blood. Conformation-sensitive gel electrophoresis was used for screening the mutation and sequencing methods were used to identify the mutations. Molecular analysis permitted the identification of three point mutations in three patients: one G->A transition in the 5' portion of the second intron, one T->A substitution in the second base of codon 430 (Leu430->stop), and one deletion deltaA in the third base of codon 63. This study represents the first description of mutations in the PIG-A gene in a Brazilian population.

Variations in gastric compliance induced by acute blood volume changes in anesthetized rats

The impact of acute volume imbalances on gastric volume (GV) was studied in anesthetized rats (250-300 g). After cervical and femoral vessel cannulation, a balloon catheter was positioned in the proximal stomach. The opposite end of the catheter was connected to a barostat with an electronic sensor coupled to a plethysmometer. A standard ionic solution was used to fill the balloon (about 3.0 ml) and the communicating vessel system, and to raise the reservoir liquid level 4 cm above the animals' xiphoid appendix. Due to constant barostat pressure, GV values were considered to represent the gastric compliance index. All animals were monitored for 90 min. After a basal interval, they were randomly assigned to normovolemic, hypervolemic, hypovolemic or restored protocols. Data were compared by ANOVA followed by Bonferroni's test. Mean arterial pressure (MAP), central venous pressure (CVP) and GV values did not change in normovolemic animals (N = 5). Hypervolemic animals (N = 12) were transfused at 0.5 ml/min with a suspension of red blood cells in Ringer-lactate solution with albumin (12.5 ml/kg), which reduced GV values by 11.3% (P<0.05). Hypovolemic rats (N = 12) were bled up to 10 ml/kg, a procedure that increased GV values by 15.8% (P<0.05). In the restored group (N = 12), shed blood replacement brought GV values back to basal levels in bled animals (P>0.05). MAP and CVP values increased (P<0.05) after hypervolemia but decreased (P<0.05) with hypovolemia. In conclusion, blood volume level modulates gastric compliance, turning the stomach into an adjustable reservoir, which could be part of the homeostatic process to balance blood volume.

Neonatal administration of citalopram delays somatic maturation in rats

We investigated the somatic maturation of neonate rats treated during the suckling period with citalopram, a selective serotonin reuptake inhibitor. Groups with 6 male neonates were randomly assigned to different treatments 24 h after birth. Each litter was suckled by one of the dams until the 21st postnatal day. Body weight, head axis and tail length were measured daily from the 1st to the 21st postnatal day. Time of ear unfolding, auditory conduit opening, incisor eruption, and eye opening was determined. Pups received 5 mg (Cit5), 10 mg (Cit10) or 20 mg/kg (Cit20) citalopram sc, or saline (0.9% NaCl, w/v, sc). Compared to saline, body weight was lower (24.04%, P < 0.01) for Cit10 from the 10th to the 21st day and for Cit20 from the 6th to the 21st day (38.19%, P < 0.01). Tail length was reduced in the Cit20 group (15.48%, P < 0.001) from the 8th to the 21st day. A reduction in mediolateral head axis (10.53%, P < 0.05) was observed from the 11th to the 21st day in Cit10 and from the 6th to the 21st day in Cit20 (13.16%, P < 0.001). A reduction in anteroposterior head axis was also observed in the Cit20 group (5.28%, P < 0.05) from the 13th to the 21stday. Conversely, this axis showed accelerated growth from the 12th to the 21stday in the Cit5 group (13.05%, P < 0.05). Auditory conduit opening was delayed in the Cit5 and Cit20 groups and incisor eruption was delayed in all citalopram groups. These findings show that citalopram injected during suckling to rats induces body alterations and suggest that the activity of the serotoninergic system participates in growth mechanisms.

Somatic cytogenetic and azoospermia factor gene microdeletion studies in infertile men

The objective of the present study was to determine the frequency of somatic chromosomal anomalies and Y chromosomal microdeletions (azoospermia factor genes, AZF) in infertile males who seek assisted reproduction. These studies are very important because the assisted reproduction techniques (mainly intracytoplasmic sperm injection) bypass the natural selection process and some classical chromosomal abnormalities, microdeletions of AZF genes or some deleterious genic mutations could pass through generations. These genetic abnormalities can cause in the offspring of these patients male infertility, ambiguous external genitalia, mental retardation, and other birth defects. We studied 165 infertile men whose infertility was attributable to testicular problems (60 were azoospermic, 100 were oligospermic and 5 were asthenospermic). We studied 100 metaphases per patient with GTG banding obtained from temporary lymphocyte culture for chromosomal abnormality detection and performed a genomic DNA analysis using 28 Y chromosome-specific sequence-tagged sites for Y AZF microdeletion detection. Karyotyping revealed somatic anomalies in 16 subjects (16/165 = 9.6%). Of these 16, 12 were in the azoospermic group (12/60 = 20%) and 4 were in the oligospermic group (4/100 = 4%). The most common chromosomal anomaly was Klinefelter syndrome (10/165 = 6%). Microdeletions of AZF genes were detected in 12 subjects (12/160 = 7.5%). The frequencies detected are similar to those described previously. These results show the importance of genetic evaluation of infertile males prior to assisted reproduction. Such evaluation can lead to genetic counseling and, consequently, to primary and secondary prevention of mental retardation and birth defects.

Lipoprotein lipase PvuII polymorphism is associated with variations in serum lipid levels in non-diabetic pregnant women

The aim of the present study was to determine if there is an association between the single nucleotide polymorphisms (SNPs) of the lipoprotein lipase (LPL) and apolipoprotein E (apo E) genes and the serum lipid profile in pregnancy and puerperium. Non-diabetic women of European descent in the third semester of pregnancy (N = 120) were selected. Those with diseases or other condition that could modify their lipid profile were excluded from the study (N = 32). Serum lipids were measured by routine laboratory procedures and genomic DNA was extracted by a salting out method. LPL (PvuII and HindIII) and apo E (HhaI) SNPs were detected by the polymerase chain reaction and restriction fragment length polymorphism. Categorical and continuous variables were compared by the chi-square test and Student t-test or ANOVA, respectively. Women carrying the LPL P1P1 genotype had higher serum LDL cholesterol (N = 21; 155 ± 45 mg/dL) than women carrying the P1P2/P2P2 genotypes (N = 67; 133 ± 45 mg/dL; P = 0.032). During the puerperium period, serum levels of triglycerides and VLDL cholesterol were significantly reduced in women carrying the P1P1 (73%, P = 0.006) and P1P2 (51%, P = 0.002) genotypes but not in women carrying the P2P2 genotype (23%, P > 0.05). On the other hand, serum concentrations of lipids did not differ between the LPL HindIII and apo E genotypes during pregnancy and after delivery. We conclude that LPL PvuII SNP is associated with variations in serum lipids during pregnancy and the puerperal period in non-diabetic women.

Genetic variations of EBV-LMP1 from nasopharyngeal carcinoma biopsies: potential loss of T cell epitopes

To find Epstein-Barr virus (EBV) strains with genetic variations of EBV latent membrane protein 1 (EBV-LMP1) from nasopharyngeal carcinoma (NPC), the full-length DNA of LMP1 genes from 21 NPC biopsies obtained in Hunan province in southern China was amplified and sequenced. Our sequences were compared to those previously reported by the Clustal V method. Results showed that all 21 sequences displayed two amino acid changes most frequently in LMP1 of CD4+ T cell epitopes at codons 144 (F®I, 21/21) and 212 (G®S, 19/21) or (G®N, 2/21). We also show that type A EBV strain is prevalent in the cases of NPC from Hunan province with a 30-bp 18/21 deletion, and we highlight that this deletion resulted in loss of one of the CD4+ T cell-restricted epitopes. The other 3 sequences without this deletion all had a change at codon 344 (G®D). Furthermore, in the major epitope sequence of CD8+ T cells restricted by HLA-A2, all 21 sequences showed changes at codons 126 (L®F) and 129 (M®I). Our study discovered that one of the 21 sequence variations harbored a new change at codon 131 (W®C), and 5/21 specimens showed another novel change at codon 115 (G®A) in the major epitope sequence of CD8+ T cells restricted by HLA-A2. Our study suggests that these sequence variations of NPC-derived LMP1 may lead to a potential escape from host cell immune recognition, protecting latent EBV infection and causing an increase in tumorigenicity.

Liver oxidative stress of the grey mullet Mugil cephalus presents seasonal variations in Ennore estuary

The objective of this study was to determine the liver oxidative stress status of grey mullets living in heavy-metal-rich polluted Ennore estuary compared with unpolluted Kovalam estuary. Fish were collected from both estuaries during the monsoon and summer seasons from October 2004 to September 2006. Fish liver homogenate (N = 20 per group) was prepared for evaluating oxidative stress parameters. Fish living in the polluted estuary had significantly higher lipid oxidation products, conjugated dienes (0.346 ± 0.017 vs 0.141 ± 0.012 DA233/mg protein), lipid hydroperoxides (0.752 ± 0.032 vs 0.443 ± 0.03 nmol/mg protein), and lipid peroxides (3.447 ± 0.14vs 1.456 ± 0.096 nmol MDA/mg protein) than those of the unpolluted estuary during the summer. In contrast, significantly lower levels of superoxide dismutase (20.39 ± 1.14 vs 53.63 ± 1.48 units/mg protein) and catalase (116 ± 6.87vs 153 ± 8.92 units/mg protein) were detected in the liver of fish from the polluted estuary (Ennore) compared to fish from the unpolluted estuary (Kovalam) during the summer. Variations in most of the oxidative stress parameters were observed between the summer and monsoon seasons, indicating the importance of seasonal variation for estuaries and their inhabitants.