Collagen type IV-related nephropathies in Portugal: pathogenic COL4A5 mutations and clinical characterization of 22 families.

Alport syndrome (AS) is caused by pathogenic mutations in the genes encoding α3, α4 or α5 chains of collagen IV (COL4A3/COL4A4/COL4A5), resulting in hematuria, chronic renal failure (CRF), sensorineural hearing loss (SNHL) and ocular abnormalities. Mutations in the X-linked COL4A5 gene have been identified in 85% of the families (XLAS). In this study, 22 of 60 probands (37%) of unrelated Portuguese families, with clinical diagnosis of AS and no evidence of autosomal inheritance, had pathogenic COL4A5 mutations detected by Sanger sequencing and/or multiplex-ligation probe amplification, of which 12 (57%) are novel. Males had more severe and earlier renal and extrarenal complications, but microscopic hematuria was a constant finding irrespective of gender. Nonsense and splice site mutations, as well as small and large deletions, were associated with younger age of onset of SNHL in males, and with higher risk of CRF and SNHL in females. Pathogenic COL4A3 or COL4A4 mutations were subsequently identified in more than half of the families without a pathogenic mutation in COL4A5. The lower than expected prevalence of XLAS in Portuguese families warrants the use of next-generation sequencing for simultaneous COL4A3/COL4A4/COL4A5 analysis, as first-tier approach to the genetic diagnosis of collagen type IV-related nephropathies.

Nutritional and inflammatory status influence darbepoetin dose in pre-dialysis elderly patients.

Anaemia is a common finding in elderly patients particularly in those with chronic kidney disease. Effective correction of anaemia improves survival and quality of life. The association between anaemia and a poor nutritional status as well as the presence of inflammation has already been documented. The aim of our study was to assess the impact of the nutritional and inflammatory status on darbepoetin dose requirements of elderly patients followed in a "Chronic Kidney Disease" outpatient clinic. We included 71 elderly patients (age>or=65 years) in a "Chronic Kidney Disease" outpatient clinic. Creatinine Clearance (CrCl) was estimated according to the Cockroft-Gault equation. Nutritional status was evaluated by biochemical and anthropometric parameters. Tumour Necrosis Factor-alpha (TNF-alpha), Interleukin-6 (IL-6) and high sensitivity C-reactive protein (hs-CRP) were used as biomarkers of inflammation. Our patients (56% males) with a mean age of 76.2+/-6.6 years were followed for 33.1+/-43.6 months. Mean eCrCl was 13.5+/-7.2 ml/mn/1.73 m2. All patients were under supplemental iron therapy and 74.7% needed darbepoietin (0.762+/-0.6 (microg/kg/week) to correct anaemia. Among the several variables regressed on darbepoietin dose, in a multiple regression model, only Hb, IL-6 and TNF-alpha levels and SGA score predicted the need for higher doses of darbepoietin. (r=0.677; r2=0.459). In Conclusion, in our pre-dialysis elderly patients, markers of a poor nutritional status (SGA and albumin) and inflammation (IL-6 and TNF-alpha) independently predicted the use of higher doses of darbepoietin to correct anaemia.

Dopamine-beta-hydroxylase activity in normal and sympathectomized carotid bodies.

Normal carotid bodies excised from cats showed DBH values of 5.04 + or - 0.53 nmol [(3)H]octopamine formed/h/100 mg tissue (x + or - S.E. ). Values from sympathectomized carotid bodies were not significantly different. While the ganglio-glomerular nerves, providing the sympathetic innervation of the carotid bifurcation, had a high DBH level, this was low in the carotid sinus nerve, providing its sensory innervation. Glomeral DBH may be contained in a special type of noradrenergic glomus cells, distinct from the most common dopaminergic glomus cells.

Retrograde endopyelotomy using an original home-made diathermy probe.

PURPOSE: To describe an easily home-made diathermy probe for the performance of retrograde endopyelotomies using a small-caliber rigid ureteroscope. METHODS: The diathermy probe is easily built by putting a rigid guidewire through a 5F ureteral catheter, the distal tip being naked and slightly bent. The other end is linked to the electric generator. Among a total of 24 retrograde endopyelotomies performed to treat ureteropelvic junction obstruction in the last 7 years, five were done using this device. RESULTS: Surgery with the probe took an average of 30 minutes. There were no complications, and, as of today, all cases are successes. CONCLUSION: This device allows the performance of retrograde endopyelotomy using a small-caliber ureteroscope. Long-term results will presumably match the good results obtained using the larger-caliber ureteroscopes with the classic cold knife.

Modificações na estrutura vertical de pastos de capim-xaraés sob lotação intermitente por bovinos / Changes in the vertical structure of xaraés palisadegrass pastures under intermittent stocking by cattle

O objetivo deste trabalho foi avaliar os efeitos dos índices de área foliar residual (IAFr), dos anos de avaliação e dos ciclos de pastejo sobre as modificações na estrutura vertical do dossle, bem como a proporção de componentes morfológicos de pastos capim-xaraés submetidos a intensidades de pastejo, durante dois verões. O experimento foi conduzido na Faculdade de Ciências Agrárias e Veterinária da Unesp, Câmpus de Jaboticabal, SP, e as intensidades de pastejos foram definidas por IAFr: 0,8; 1,3; 1,8 e 2,3. Quando o dossel interceptou 95% da luz incidente, os animais foram colocados no piquete para o pastejo e permaneceram até o IAFr alvo ser alcançado. Os pastejos foram realizados por vacas da raça Holandesa (Bos taurus taurus L.) não lactantes, com peso corporal médio de 450kg±50kg, utilizando-se a técnica de mob-stocking. Foram avaliadas a proporção de componentes morfológicos dos pastos e a estrutura vertical do dossel pelo aparelho de ponto inclinado. Índices de área foliar residual de 1,3 a 1,8 são os que melhores respostas apresentam em relação à estrutura vertical do dossel e à proporção dos componentes morfológicos de pastos de capim-xaraés submetidos a intensidades de pastejo.

The aim of this study was to evaluate the effects of residual leaf area index (rLAI), years of evaluation and grazing cycles in the vertical structure of the canopy and the proportion of morphological components of xaraés palisadegrass pastures submitted to grazing intensities in two summers. The experiment was carried out at the Faculdade de Ciências Agrárias e Veterinária of Unesp, Campus of Jaboticabal, SP and the intensities of grazing were defined by four rLAI: 0.8, 1.3, 1.8 and 2.3. When the canopy intercepted 95% of incident light, the animals were placed on the pasture for grazing and kept until the rLAI target had been reached. Pastures were grazed by non-lactating Holstein cows (Bos taurus taurus L.) using the mob-stocking technique. the proportion of the morphological components of the pasture and the vertical structure of the canopy by the inclined point device was evaluated. Residual leaf area index from 1.3 to 1.8 are the best in relation to the vertical structure of the canopy and proportion of the morphological components of the pasture submitted to grazing intensities.

Influência de subprodutos de oleaginosas sobre parâmetros ruminais e a degradação da matéria seca e da proteína bruta / Influence byproducts of oleaginous on ruminal parameters and degradation of dry matter and crude protein

Avaliou-se o efeito da adição de subprodutos de oleaginosas na dieta de ovinos em substituição ao farelo de soja. Foram distribuídos quatro ovinos Santa Inês, com peso corporal de, aproximadamente, 40kg, canulados no rúmen, em quadrado latino (4x4), com quatro dietas e quatro períodos, analisados por regressão para a avaliação da degradação ruminal, dos valores de pH e das concentrações de nitrogênio amoniacal in vivo. Os subprodutos foram as tortas de amendoim, girassol e soja, em dietas isonitrogenadas, com 70% de concentrado e 30% de volumoso (feno de tifton). Não foi observado efeito da interação tempo x dieta para os valores de pH e concentração ruminal de nitrogênio amoniacal (P>0,05). O pH apresentou valor médio de 6,2. As concentrações ruminais de nitrogênio amoniacal não foram afetadas, com valor médio de 29,9mg/dL. Não foram observadas diferenças (P>0,05) nas taxas de degradação da matéria seca e na degradabilidade potencial.

The aim of this work was to evaluate the introduction of oleaginous byproducts in sheep feeding as a replacement for soybean meal. Four sheep weighing 40kg, with cannulated rumen were selected in a Latin Square (4x4) design, with for diets and four periods analyzed by regression for ruminal degradability evaluate, pH values and ammonia nitrogen concentration in vivo. Byproducts were the following cakes: peanut, sunflower and soybean, in isonitrogenous diets with 70% concentrate and 30% roughage (tifton hay). There was no effect of diet x time interaction for pH and ruminal concentrations of ammonia nitrogen (P>0.05). The average pH value was 6.2. Ruminal concentrations of ammonia nitrogen were not affected, and had an average value of 29.9mg/dL. There were no differences (P>0.05) in DM degradation and degradability rates. The use of high degradability protein sources, such as soybean cake and peanut cake, proved to be an interesting alternative to replace soybean meal.

Gene algebra from a genetic code algebraic structure.

By considering two important factors involved in the codon-anticodon interactions, the hydrogen bond number and the chemical type of bases, a codon array of the genetic code table as an increasing code scale of interaction energies of amino acids in proteins was obtained. Next, in order to consecutively obtain all codons from the codon AAC, a sum operation has been introduced in the set of codons. The group obtained over the set of codons is isomorphic to the group (Z(64), +) of the integer module 64. On the Z(64)-algebra of the set of 64(N) codon sequences of length N, gene mutations are described by means of endomorphisms f:(Z(64))(N)-->(Z(64))(N). Endomorphisms and automorphisms helped us describe the gene mutation pathways. For instance, 77.7% mutations in 749 HIV protease gene sequences correspond to unique diagonal endomorphisms of the wild type strain HXB2. In particular, most of the reported mutations that confer drug resistance to the HIV protease gene correspond to diagonal automorphisms of the wild type. What is more, in the human beta-globin gene a similar situation appears where most of the single codon mutations correspond to automorphisms. Hence, in the analyses of molecular evolution process on the DNA sequence set of length N, the Z(64)-algebra will help us explain the quantitative relationships between genes.

Allometric algorithms.

The aim of the present study is to emphasize the applicability and versatility of the allometric equation in the biological sciences. This equation (Y = a x Mb) was introduced by Huxley (1932) for intra- and interspecific comparisons of morphological, physiological and ecological variables (Y), when they are expressed as functions of body mass (M). The regression analysis of the experimental data, plotted in a double logarithmic scale, yields a straight line, which is equivalent to the logarithmic form of the above mentioned allometric equation [log Y = log(a) + (b) x log(M)]. Only the exponent (b) can be calculated a priori for a given function, based firstly on the corresponding dimensional analysis in accordance with the MLT-system of physics, and secondly on one of the theories of biological similarity, while parameter (a) is of empirical nature. A relevant feature of the allometric equations is that they can be treated algebraically to obtain allometric ratios, mass independent numbers (MIN), and even dimensionless numbers (M0L0T0), which are valid for all organisms pertaining to the same taxonomic classification.

Duality in physiological time: Euclidean and fractal.

The aim of the present study was to differentiate two modalities of intrinsic time scales: i- the geometric or Euclidean modality, which is based on the constant speed of mass transport or of wave transmission in cylindrical structures (arteries, veins, nerves), whose allometric exponent (TE = aMb) is b = 0.33, where M is body mass (kg) and a the mass coefficient; ii- the fractal time scale (TF), which is characteristic of organs with self-similar branching structures and with volume-specific flows, whose allometric exponent is b = 0.25. The proposed dichotomy could be confirmed by means of the statistical analysis of empirical allometric exponents (b). Our findings demonstrate the need to separate the chronology of bulk transport at long distances (inter-organic) which follows an Euclidean geometry (cylinders), from the fractal time scale, which operates at short distances (intra-organic) and is represented by a self-similar branching system which determines both the morphometric and physiometric characteristics within each organ.

Allometry of ECG waves in mammals.

The present allometric study deals with the duration of three electrocardiographic intervals (PQ, QRS, QT) and their relationships with the corresponding cardiac cycle length (R-R interval) in mammals across a wide body mass range. The numerical values of the different ECG intervals were obtained from Grauwiler's (1965) monograph on the subject. Because the corresponding body masses were not given by this author, Heusner's (1991) data on basal metabolic rate as function of body mass were used to establish the most likely body mass figure for each case, based on the taxonomic identity between the corresponding specimens. On the other hand, in a recent study we established the "duality" of physiological times (Günther & Morgado, 1996) and, therefore, we adopted this novel approach to investigate the ECG intervals and their relationships with the R-R interval (heart rate reciprocal). Considering that the anatomy and physiology of auricles and ventricles are different (spheroids versus quasi-cylinders), and that excitation (sino-atrial node and His-Purkinje's system) and contraction processes can be described either by Euclidean or fractal geometries, only a quantitative analysis of the different ECG waves could resolve the dilemma. From the present preliminary study we can conclude that fractal geometry is prevalent with regard to ECG intervals.

Transport similarity: dimensional analysis of diffusion at cellular level.

Scale dependent variables in animals of different sizes can be studied by means of dimensional analysis and biological similarity criteria. The aim of this report is to demonstrate a transport similarity at the cellular level, based on two postulates derived from Fick's law of diffusion; the constancy of the "concentration gradient" between two compartments separated by a membrane; and the invariance of the "diffusion coefficient" of a given substance in homologous cells. A general equation for a transport similarity was deduced from these two postulates and it was then possible to calculate the corresponding "reduced exponents" as functions of body weight, which, in turn, can be compared with the empirical allometric exponents of Huxley's power equation. The biological meaning of the theoretically predicted reduced exponents of the transport similarity are discussed and the predicted values are compared with the experimental findings.

Intrinsic times in biology.

The meaning of time in the physical sciences (absolute vs. relational) is compared with the "intrinsic" times in the biological sciences. Since all organisms can be considered as "mixed regimes", the corresponding intrinsic times will be different in a mechanical similarity (organism and environment), in a biological similarity (morphometry and physiometry inside the organism), and in a transport similarity (diffusion processes at the cellular level). These three similarity criteria are based on specific postulates, and it is possible -through dimensional analysis- to obtain quantitative predictions of the numerical value for the reduced exponents of the body weight ratio (w), which can then be compared with the empirically found allometric exponent (b) of Huxley's power equation, which is expressed as a function of body weight (W). The correlation between the theoretically predicted values and the empirical findings of numerous chronological functions pertaining to living beings is satisfactory.

On the hidden physical dimensions of the allometric equation.

The aim of the present study was to submit Huxley's allometric equation (Y = aMb) to a dimensional analysis; in this equation Y is any biological variable, a is the mass-coefficient, M represents body mass, and b the mass-exponent. The dimensions of each of its components is thoroughly analyzed by means of the MLT-system of physics, as is the dimensionality of the whole equation. The relationship between the dimensional analysis and the postulates of some theories of biological similarity is discussed. In conclusion, parameter a of the allometric equation is always dimensionless, while the physical dimensions of the dependent variable Y can be defined by means of the power function Mb.

Physical bases for a triad of biological similarity theories.

The dimensional analysis of physics, based on the MLT-system (M = mass, L = length, T = time), can be applied to the living world, from mycoplasmas (10(-13) g) to the blue whales (10(8) g). Body mass (M), or body weight (W), are utilized as convenient reference systems, since they represent the integrated masses of all elementary particles--at the atomic level--which conform an organism. A triad of biological similarities (mechanical, biological, transport) have been previously described. Each similarity was based on two postulates, of which the first was common to all three, i.e., the constancy of body density; whereas the second postulates were specific for each of the three theories. In this study a physical foundation for these second postulates, based on three universal constants of nature, is presented, these are: 1) the acceleration of gravity (g = LT-2); 2) the velocity of light (c = LT-1); and 3) the mass-specific quantum (h/m = L2T-1). The realm of each of these biological similarities is the following: 1) the gravitational or mechanical similarity (where g = constant), deals mainly with the relationship between a whole organism and its environment, particularly with locomotion. The acceleration of gravity (g) is also one of the determining factors of the "potential" energy (E = m.g.H), where m is the mass, and H is the height above the reference level; 2) the electrodynamic similarity (formerly biological similarity), (c = constant), is able to quantitatively define the internal organization of an organism from both a morphological and a physiological point of view.(ABSTRACT TRUNCATED AT 250 WORDS)

Oxidative metabolism and body weight: inactive, active, and mitochondrial volumes.

In homeotherms, the standardized (basal) metabolic rate should not be expressed per kilogram of body weight (specific metabolic rate), nor per unit of body surface (square meters of body-ambient interface), since both mitochondrial thermogenesis and heat-loss mechanisms (radiation, conduction, convection, evaporation) are not uniform processes. On the contrary, each organism is an heterogeneous bioreactor, which is composed at least of two compartments: 1) a metabolically active volume (aV), where oxidative phosphorylation takes place; and 2) a metabolically inactive volume (iV), where oxygen consumption is negligible. The ratio (aV/iV) is not invariant, since iV increases disproportionately with the scaling up of body size, and as shown by us, when the three main components of iV, i.e., skeleton, fat deposits, and blood volume, are added together, a similar disproportionality is found. The aV was determined by subtracting the iV from the total volume (V) of an organism, or by estimating the volume occupied by all mitochondria, or mitochondrial volume (mtV). For this purpose two procedures are discussed: 1) the stereological or morphometric method; and 2) the oxygen consumption per unit time or physiometric method. The latter procedure is based on the equivalence between an VO2 = 3 ml O2.min-1 and a mtV of 1 ml, whose oxidative phosphorylation yields an approximate power output of 1 watt. The correspondence between oxygen consumption, heat production, and electron flux at the respiratory chain of the mitochondrial cristae, is discussed. From a physical point of view, the metabolic rate is a "power" function (P = M L2T-3), where M = mass, L = length, and T = time. The dimensional analysis and the statistical treatment of the corresponding numerical values of more than 200 allometric equations yields the 3/4 power, law established by Kleiber (1961), for the relationship between basal metabolism and body weight. Instead of expressing the metabolic rate per unit body weight (kg-1) or per unit body surface (m-2) structural and functional criteria should be taken into account as, for instance, the distinction between iV and aV, and particularly by emphasizing the paramount importance of the mtV where oxidative phosphorylation takes place. An allometric equation relating mtV and body weight (W) could be tentatively established for interspecies comparisons.

Biological similarity theories: a comparison with the empirical allometric equations.

Twelve biological variables were submitted to dimensional analysis in accordance with the MLT-system of physics (M, mass; L, length; T, time). Each of these variables has a characteristic numerical value for the exponents alpha for mass, beta for length, and gamma for time. By means of Newton's reduction coefficient (chi), the three dimensions (MLT) can be expressed as power functions of body mass (Mb); the exponent (b) is the result of the combination of the three dimensional exponents (alpha, beta, gamma). By linear regression analysis of 203 allometric exponents (betaE) obtained from the literature, the following equation was found for the regression exponent (bR) (equation: see text). The estimated numerical coefficients (ki) for the three exponents (alpha, beta, gamma) of the basic dimensions (MLT) do not agree with those of the prevailing theories of biological similarity.

[Polychaetes as bioindicators of organic pollution on the beaches of São Paulo]. / Poliquetas bioindicadores de poluição orgânica em praias paulistas.

This study consisted of an analysis of the occurrence of bioindicator polychaete species of organic pollution along beaches of the north coast of São Paulo State, southeastern Brazil, taking into account the importance of benthic species as indicators of organically enriched environments. A comparison was established between the frequence and abundance of these species in areas qualified as impacted and improper for bathing. The study area comprehends 17 beaches of the Caraguatatuba Bay and the Channel of São Sebastião. The data about the macrobenthos were collected from March, 1988 to March, 1991 and the data on bathing conditions came from reports published by CETESB. The biological samples were obtained with an iron core of 0.025 m2 surface area. The results show markedly the presence of species considered as indicators of organically enriched places, such as Capitella capitata, Heteromastus filiformis, Laeonereis acuta, and Scolelepis squamata. The high density of Capitella and Heteromastus in beaches considered improper for bathing consist in a strong indication of the degree of damage to these areas.

The suturoscope. A preliminary report on the first clinical applications.

One of the authors has developed a special device--the suturoscope--which allows endoscopic sutures with conventional suture material. After several modifications, in order to improve the prototype, the authors began its clinical application in 1992. Five bladder operations were performed--two antireflux operations, two meatotomies with meatoplasty, and an intramural ureteric ligation. The results indicate that this device is suitable for clinical use.

Low activity of the yeast cAMP-dependent protein kinase catalytic subunit Tpk3 is due to the poor expression of the TPK3 gene.

Three genes TPK1, TPK2 and TPK3 encode in Saccharomyces cerevisiae distinct catalytic subunits of cAMP-dependent protein kinase (cAPK). We have measured cAPK activity in vitro and, indirectly, in vivo in yeast strains carrying only one of the three TPK genes. The strain containing TPK3 as the only intact TPK gene showed nearly undetectable phosphorylating activity and no TPK3 mRNA could be detected, although the cells grow normally. Overexpression of TPK3 in a high copy vector or under the control of the inducible GAL1 promoter did not by itself result in a corresponding increase in activity but coexpression of BCY1, the gene coding for the regulatory subunit, was necessary in both cases to achieve high levels of phosphorylating activity. Moreover, BCY1 overexpression not only increased Tpk3 catalytic activity but also increased the amount of TPK3 mRNA detected in Northern blots.

Three-dimensional morphometry of mammalian cells. II. Areas, volumes, and area-volume ratios.

From three-dimensional diameter measurements of eleven kinds of cells pertaining to five different organs, which were excised from eleven adult mammals (nine species) whose body weight range was 40 g to 450 kg, we calculated the corresponding cell soma areas (A), volumes (V), and finally their area-volume ratios (A/V). The dissimilarities among these eleven cell types could be established quantitatively by means of a cluster analysis. The dendrograms for cell areas (A), volumes (V), and their corresponding area-volume ratios (A/V), yielded similar groupings when cell areas and volumes were compared, yet the grouping of the area-volume ratios (A/V) for the eleven types of cells was different. These results were corroborated by means of the principal components analysis, where five distinct cell groupings could be established. The relationship between cellular morphometry, oxidative metabolism, and body mass, was established by means of the fractal geometry of the transport systems (respiration and circulation), which provides the tools for the scale-dependent analysis of the surfaces across which the transport of metabolites is performed.