Rock outcrop habitats in the Venezuelan Guayana lowlands: their main vegetation types and floristic components

The Guayana Shield, located in north-eastern South America, consists of a highly complex and composite mosaic of landscape elements. Amongst these, inselbergs are very conspicuous, because of their peculiar shape and their unusual associated vegetation. Geologically, these rock outcrops are part of the underlying ancient igneous-metamorphic basement and occur mainly in the lowlands of the periphery of the shield. As azonal habitats, inselbergs harbour a highly specialized flora. The characteristic vegetation is composed of lithophytic and savanna-like plant communities, as well as low dry forests. As a whole, the vegetation of an inselberg may be interpreted as a mosaic of marginal habitats. Therefore a large number of taxa find suitable niches in a quite condensed space. Gradients of soil depth and water availability are the main factors determining the floristic composition. A preliminary floristic inventory of the Venezuelan inselberg flora comprises 614 vascular plant species. 24% of them are endemic to the Guayana region, 15% are endemic to inselbergs of the Guayana region. The distribution patterns of these latter, eco-endemic species allows to distinguish a northern and a southern inselberg district. The two districts overlap in the Átures area, in the surroundings of Puerto Ayacucho, where a true centre of endemism is located. The distinction into two districts is emphasised by different phytogeographic relations. The southern inselberg district shows connections to the "tepui" flora, whereas the northern district reveals phytogeographic relations to the Caribbean region as well as to the Brazilian Shield. Possible explanations for the floristic interchange across the equator are discussed.

Mitochondrial DNA synthesis studied autoradiographically in various cell types in vivo

It is generally accepted that mitochondria are able to proliferate even in postmitotic cells due to their natural turnover and also to satisfy increased cell energy requirements. However, no detailed studies are available, particularly with respect to specific cell types. Since [3H]-thymidine is incorporated not only into nuclear (n) DNA but also into the DNA of cytoplasmic mitochondria, an autoradiographic approach was developed at the light microscopy level in order to study basic questions of mitochondrial (mt) proliferation in organs of rodents in situ via the cytoplasmic incorporation of [3H]-thymidine injected into the animals 1 h before sacrifice. Experiments carried out on mice after X-irradiation showed that cytoplasmic labeling was not due to a process such as unscheduled nuclear DNA synthesis (nUDS). Furthermore, half-lives of mitochondria between 8-23 days were deduced specifically in relation to cell types. The phase of mtDNA synthesis was about 75 min. Finally, mt proliferation was measured in brain cells of mice as a function of age. While all neurons showed a decreasing extent of mtDNA synthesis during old age, nUDS decreased only in distinct cell types of the cortex and hippocampus. We conclude that the leading theories explaining the phenomenon of aging are closely related, i.e., aging is due to a decreasing capacity of nDNA repair, which leads to unrepaired nDNA damage, or to an accumulation of mitochondria with damaged mtDNA, which leads to a deficit of cellular energy production

GLUT4 protein expression in obese and lean 12-month-old rats: insights from different types of data analysis

GLUT4 protein expression in white adipose tissue (WAT) and skeletal muscle (SM) was investigated in 2-month-old, 12-month-old spontaneously obese or 12-month-old calorie-restricted lean Wistar rats, by considering different parameters of analysis, such as tissue and body weight, and total protein yield of the tissue. In WAT, a ~70% decrease was observed in plasma membrane and microsomal GLUT4 protein, expressed as µg protein or g tissue, in both 12-month-old obese and 12-month-old lean rats compared to 2-month-old rats. However, when plasma membrane and microsomal GLUT4 tissue contents were expressed as g body weight, they were the same. In SM, GLUT4 protein content, expressed as µg protein, was similar in 2-month-old and 12-month-old obese rats, whereas it was reduced in 12-month-old obese rats, when expressed as g tissue or g body weight, which may play an important role in insulin resistance. Weight loss did not change the SM GLUT4 content. These results show that altered insulin sensitivity is accompanied by modulation of GLUT4 protein expression. However, the true role of WAT and SM GLUT4 contents in whole-body or tissue insulin sensitivity should be determined considering not only GLUT4 protein expression, but also the strong morphostructural changes in these tissues, which require different types of data analysis.

Adenylyl cyclase types I and VI but not II and V are selectively inhibited by nitric oxide

Adenylyl cyclase (AC) isoforms catalyze the synthesis of 3',5'-cyclic AMP from ATP. These isoforms are critically involved in the regulation of gene transcription, metabolism, and ion channel activity among others. Nitric oxide (NO) is a gaseous product whose synthesis from L-arginine is catalyzed by the enzyme NO synthase. It has been well established that NO activates the enzyme guanylyl cyclase, but little has been reported on the effects of NO on other important second messengers, such as AC. In the present study, the effects of sodium nitroprusside (SNP), a nitric oxide-releasing compound, on COS-7 cells transfected with plasmids containing AC types I, II, V and VI were evaluated. Total inhibition (~98.5%) of cAMP production was observed in COS-7 cells transfected with the AC I isoform and previously treated with SNP (10 mM) for 30 min, when stimulated with ionomycin. A high inhibition (~76%) of cAMP production was also observed in COS-7 cells transfected with the AC VI isoform and previously treated with SNP (10 mM) for 30 min, when stimulated with forskolin. No effect on cAMP production was observed in cells transfected with AC isoforms II and V.

Changes in types of muscle fibers induced by transcutaneous electrical stimulation of the diaphragm of rats

The objective of the present study was to assess the effect of transcutaneous electrical diaphragmatic stimulation (TEDS) on different types of diaphragm muscle fibers. Male Wistar rats (8-12 weeks old) were divided into 2 experimental groups (N = 8 in each group): 1) control, 2) animals submitted to TEDS [frequency = 50 Hz; T ON/T OFF (contraction/relaxation time) = 2/2 s; pulse duration = 0.4 ms, intensity = 5 mA with a 1 mA increase every 3 min for 20 min] for 7 days. After completing this treatment period, the I, IIA, IIB, and IID diaphragm muscle fibers were identified using the mATPase technique. Statistical analysis consisted of the normality, homoscedasticity and t-tests (P < 0.05). There was a 19.6% (P < 0.05) reduction in the number of type I fibers and a 49.7% increase (P < 0.05) in type IID fibers in the TEDS group compared with the control group. An important result of the present study was that electrical stimulation with surface electrodes was efficient in altering the distribution of fibers in diaphragm muscle. This therapeutic resource could be used in the treatment of respiratory muscle alterations.

Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Correlation between total nitrite/nitrate concentrations and monoamine oxidase (types A and B) and semicarbazide-sensitive amine oxidase enzymatic activities in human mesenteric arteries from non-diabetic and type 2 diabetic patients

The aim of this study was to determine the correlation between total nitrite/nitrate concentrations (NOx) and the kinetic parameters of monoamine oxidase enzymes (MAO-A and MAO-B) and semicarbazide-sensitive amine oxidase (SSAO) in human mesenteric arteries. Arteries were from non-diabetic and type 2 diabetic patients with sigmoid or rectum carcinoma for whom surgery was the first option and who were not exposed to neo-adjuvant therapy. Segments of human inferior mesenteric arteries from non-diabetic (61.1 ± 8.9 years old, 7 males and 5 females, N = 12) and type 2 diabetic patients (65.8 ± 6.2 years old, 8 males and 4 females, N = 12) were used to determine NOx concentrations and the kinetic parameters of MAO-A, MAO-B and SSAO by the Griess reaction and by radiochemical assay, respectively. The NOx concentrations in arteries from diabetic patients did not differ significantly from those of the non-diabetic group (10.28 ± 4.61 vs 10.71 ± 4.32 nmol/mg protein, respectively). In the non-diabetic group, there was a positive correlation between NOx concentrations and MAO-B parameters: Km (r = 0.612, P = 0.034) and Vmax (r = 0.593, P = 0.042), and a negative correlation with the SSAO parameters: Km (r = -0.625, P = 0.029) and Vmax (r = -0.754, P = 0.005). However, in the diabetic group no correlation was found between NOx concentrations and the three kinetic parameters of the enzymes. These results suggest an important function of sympathetic nerves and vascular NOx concentrations in arteries of non-diabetic patients. Thus, these results confirm the importance of a balance between oxidants and antioxidants in the maintenance of vascular homeostasis to prevent oxidative stress.

Quality of minimally processed guava with different types of cut, sanification and packing

The purpose of this project was to evaluate the sanitization effect on the quality of minimally processed guava. Initially, research was carried out with consumers in a supermarket to verify preferences of packaging for guava. Following this, the guava cv. Paluma underwent two sanitization sequences using dehydrated sodium dichloroisocyanurate compound, in 50 ppm concentration, sanitization prior to (S1) and after (S2) being cut; removal of excess water; conditioning in PET packaging and PSPVC and storage at 3 ºC ± 1 ºC. Physicochemical analysis - [pH, total soluble solids (SST), total labeled acidity (ATT), ascorbic acid (AA), total sugars (AT) and reducers (AR)], textural sensorial and microbiological analyses were used to monitor the quality of the products. The consumers preferred the guava cut in halves with pulp and packed in PET, although this packaging promoted condensation of water vapor on the inner surface of the lid, compromising the appearance of the product. The two sanitization sequences and the two kinds of packaging did not significantly affect the pH, SST, ATT, SST/ATT, texture and AA values. The AT and AR tenors increased significantly in the MP guavas stored in the PSPVC package. Both sanitizations were efficient in the bacterial control of the indicators of the hygienicsanitary conditions, although the S1 sanitization proved to be more efficient in the control of autochthonous aerobic microbiota (aerobic mesophylic microorganisms). It can be concluded that guava cv. Paluma packed in PSPVC can be conserved for 6 days when stored at 3 ºC.

Effect of different sweetener blends and fat types on ice cream properties

The development of processed foods requires the understanding of the phenomena that dictate the ingredient interactions normally used in food formulations, as well as the effects of the numerous operations involved in the processing of the final product. In ice creams, sugars are responsible for taste, but they also affect the freezing behavior and viscosity of processed mixes. Components such as fats influence mechanical properties, melting resistance, and palatability of final products. The objective was to study the technological effects of different sugars and fats on the structure of ice cream formulations. Fructose syrup was used as a substitute for glucose syrup in blends with sucrose, and palm fat was employed as an alternative to hydrogenated vegetable fat. The analysis of variance showed significant differences in chemical compositions. Hygroscopicity of fructose syrup increased the solids content in the formulations. Melting rate and overrun were higher in products added with this sugar. Palm fat caused changes in melting ranges of formulations, and higher melting rate was observed in the combination of palm fat and fructose syrup.

Oxidative stability of cereal bars made with fruit peels and baru nuts packaged in different types of packaging

Food industries have been concerned about managing the waste generated by their production processes in order to minimize environmental impacts and also about the development of formulations with different and innovative ingredients such as fruits from the Brazilian savanna. Seeking to meet the expectations of consumers who desire healthy and practical products, this study aimed to evaluate the oxidative stability and the variations in chemical composition and antioxidant potential of cereal bars made with fruit peels and baru nuts packaged in different types of packaging. The bars formulated were packed in four different types of packaging: laminated without vacuum (LWV), transparent without vacuum (TWV), transparent under vacuum (TV), and laminated under vacuum (LV); they were subsequently analyzed for proximate composition, fatty acid profiles, antioxidant activity, and oxidative capacity. The results showed that the cereal bars made with fruit peel and baru are sources of protein, dietary fiber, and fat, especially unsaturated fatty acids such as oleic and linoleic acids. The cereal bars exhibited oxidative stability up to 120 days of storage, and the type of packaging was not significant for the variables evaluated; therefore, they can be stored in low cost packaging such as transparent packaging without vacuum for a period of 120 days.

Effect of an edible crosslinked coating and two types of packaging on antioxidant capacity of castilla blackberries

In order to increase the shelf life and maintain the quality and stability of the biological compounds with antioxidant activity present in Castilla blackberry fruits, a sodium alginate-based edible crosslinked coating was applied, and the fruits were packed in two different plastic containers and stored under refrigeration (3 ± 1 °C). Total antioxidant capacity and its relationship to physicochemical variables such as pH, Brix, and acidity were evaluated in six treatments: uncoated blackberry stored in a macroperforated container (T1) and thermosealed container (T2), without crosslinked coating in a macroperforated container (T3) and thermosealed container (T4), with crosslinked coating (calcium ions) packed in macroperforated container (T5) and thermosealed container (T6). The results indicated that factors such as gas permeability in the coatings, the packaging used, and physicochemical parameters significantly affected the fruit total antioxidant capacity, with the highest level in T1 (0.22 µgEAA/ml) at the end of the essay, which is related to the lowest levels of pH and direct exposure to air. On the other hand, the lowest value was obtained in T6 (0.16 µgEAA/ml) due to the crosslinked coating, packaging in the thermosealed container, and higher pH value. Variations in acidity, Brix, and pH indicate the presence of degenerative processes in the crosslinked coating treatments, which limited the physicochemical changes.