Frequency of the stages in the cycle of the seminiferous epithelium in the rat

This study determined the optimum number of tubules to be counted per testis cross section, and the number of animals per treatment group, when changes in stage frequencies in the cycle of the seminiferous epithelium are criteria for assessing effects of treatment on spermatogenesis. A data base of 9,672 observed and staged tubules was collected from testicular cross sections of 15 Sprague-Dawley rats. A significant variation between animals was found for the frequencies of Stages I, II, IV, VI, VIII, and XIII. Computer simulation was used to randomly select different combinations of animal and tubule numbers from the observed data. Stage frequency means from each simulation experiment were compared statistically to observed mean frequencies. A model that used data from all 14 stages was analyzed. The following conclusions were made: a) a minimum of 200 tubule cross sections/testis is recommended for estimating stage frequencies; b) for a fixed number of tubules scored, the number of animals sampled is more important than the number of tubules per animal in reducing variance; c) to detect a difference of 2 standard deviations from the mean with a 2% error rate and examining 200 tubules/testis, at least 12 animals must be used per group when assessing all 14 stages; d) when individual stages are examined using 10 animals per group, only Stage VII has 80% or greater power of test (alpha = 0.05) to detect a frequency difference; e) pooling stages into 3-4 groups is recommended to improve the power of detecting a treatment difference.     

Histogenesis of the taste buds of the vallate papilla in the in the rat in the postnatal stages of development

The developing taste buds of vallate papillae were studied with electron microscope in rats during the first 7 days after birth. Two types of cells--light and dark--are identified in the taste buds of a one day old animal. The apical parts of dark cells are characterized by numerous dark granules. A distinguishing feature of light cells is the presence of synaptic contacts with afferent intragemmal nerves. On the 4th day of development on the top of the apical parts of the cell, a microvillar apparatus is seen to form, which does not yet communicate with the oral cavity. On the 7th day, basal cells appear in the taste buds. Some of these cells are seen mitotically dividing. The differentiated microvillar apparatus now communicates with oral cavity. The structure of the taste buds is getting similar to that in the adults. The structural and functional peculiarities of the developing taste buds are discussed in association with the period of ontogenesis under consideration.     

Effects of turn residues in directing the formation of the beta-sheet and in the stability of the beta-sheet

The designed peptide (denoted 20-mer, sequence VFITS(D)PGKTYTEV(D)PGOKILQ) has been shown to form a three-strand antiparallel beta-sheet. It is generally believed that the (D)Pro-Gly segment has the propensity to adopt a type II' beta-turn, thereby promoting the formation of this beta-sheet. Here, we replaced (D)Pro-Gly with Asp-Gly, which should favor a type I' turn, to examine the influence of different type of turns on the stability of the beta-sheet. Contrary to our expectation, the mutant peptide, denoted P6D, forms a five-residue type I turn plus a beta-bulge between the first two strands due to a one amino-acid frameshift in the hydrogen bonding network and side-chain inversion of the first beta-strand. In contrast, the same kind of substitution at (D)Pro-14 in the double mutant, denoted P6DP14D, does not yield the same effect. These observations suggest that the SDGK sequence disfavors the type I' conformation while the VDGO sequence favors a type I' turn, and that the frameshift in the first strand provides a way for the peptide to accommodate a disfavored turn sequence by protruding a bulge in the formation of the beta-hairpin. Thus, different types of turns can affect the stability of a beta-structure.     

On the role of the pineal in thermoregulation in the pigeon

Diurnal rhythm in body temperature of pigeons subjected to different experimental conditions (pinealectomized, sham-operated, melatonin-implanted, cold-exposed) was studied under a 12 h light and 12 h dark regimen. The body temperature of pigeons during photophase was higher than that during scotophase in the normal as well as every treatment group studied. Pinealectomized pigeons showed higher body temperature in the photophase as well as scotophase, than that of the normal and sham-operated birds when examined 2 or 3 weeks following the post-surgical acclimatization to 25 or 3 degrees C. However, subcutaneous implantation of melatonin pellets into pinealectomized pigeons nullified or even reversed the hyperthermic effect of pinealectomy. Exposure of pigeons to--18 degrees C for 280 min during photophase as well as scotophase, produced a marked drop in body temperature in pinealectomized, sham-operated and normal pigeons. The pinealectomized pigeons exhibited a higher body temperature than that of the sham-operated and normal ones when exposed to--18 degrees C during the photophase, but not during the scotophase. It was concluded that while the pineal is not necessary for maintaining the daily thermal rhythm in the avian body, it has a thermoregulatory role, in that it prevents rise in body temperature in warm (25 degrees C) acclimatized and chronic cold (3 degrees C) exposed birds. In acute short-term cold (--18 degrees C) exposure however, the temperature regulatory role of the pineal was not effective during the scotophase.     

The role of the kidney in the biosynthesis of carnitine in the rat.

Intravenous administration of L-[methyl-3H]-labeled trimethyllysine to rats results in a very rapid accumulation of radioactivity by the kidneys, while the incorporation of the label into the liver occurs at approximately 1% of this rate when calculated per g of wet tissue. The kidneys convert a substantial portion of the trimethyllysine taken up to butyrobetaine and to beta-hydroxytrimethyllysine, a precursor of butyrobetaine, but fail to synthesize carnitine. Significant amounts of radioactivity are recovered in both carnitine and butyrobetaine of hepatic tissue after longer time periods, while the level of labeled trimethyllysine in this organ remains very low. Bilateral nephrectomy results in a marked decrease in the incorporation of label into the liver. These results indicate that in rats, the initial conversion of trimethyllysine to butyrobetaine occurs predominantly in kidney and that the liver capacity for this transformation is considerably smaller than its capacity to synthesize carnitine from butyrobetaine.     

2-Hydroxylation of estrogens in the brain participates in the initiation of the preovulatory LH surge in the rat

Estradiol-2-hydroxylase, the enzyme responsible for the conversion of estrogens to catechol estrogens was measured in the brain of female rats at specific stages of the estrus cycle. Radiometric measurements of the enzyme activity in microsomal, mitochondrial, and synaptosomal fractions of the brain revealed a sharp increased in activity at proestrus just prior to the preovulatory LH surge. The enzyme activity declined to lower levels at diestrus and metestrus. No comparable fluctuations were noted in the liver enzyme. These changes in brain enzyme activity in conjunction with demonstrated positive feedback of exogenous catechol estrogens on pituitary LH release, suggest that a rise in endogenous catechol estrogen formation in the brain may be responsible for the physiological induction of the preovulatory LH surge.     

Dynamics of the direction of static connections in the electrical activity of the cerebral cortex in the cat

Data have been obtained on three cats for analysis of statistic connections of electrical activity in the visual, parietal and somatosensory cortical zones during development of evoked potentials to photic signal in these areas, and also in periods previous to photic stimulation and on traces of its action. An increase in shown of the influence of activity in the visual area on that of the somatosensory zone in post-stimulus periods and a sharp increase of two-way connections between these areas against the background of a relative equilibrium of direct and backward connections in trace periods.     

Recapitulation in the development of the components of the counterflow system in the vertebrate metanephros

The investigation has been performed on 107 renal preparations obtained from persons of various age (from 5-month-old fetuses up to 45 years of age), certain representatives of other classes of the Vertebrata are also included: fish, amphibia, reptile and mammalia at various stages of pre- and postnatal periods of ontogenesis by means of preparing graphic and plastic reconstructive models, histological investigation and microdissection. The complexity of the intrarenal branching of derivatives of the mesonephric duct diverticulum, development and structure of the canalicular part in nephrons directly depend on the phylogenetic position of the animal. Complexity of the nephron architectonics occurs along the progressive line of taxonomic groups of higher Vertebrata. The nephron loop becomes longer, thin segment of the nephron canalicular part increases in its length and, at last, in mammalia a cone-shaped fasciculus appears as a structural-functional unit of the osmoregulating apparatus of the constant kidney. In the comparative anatomical and comparative embryological aspects recapitulation is observed concerning certain morphological signs of derivatives of the metanephric duct and nephron.     

Potentiation of the gastro-protective effect of sulglicotide in the presence of cimetidine in the rat

The effects of sulglicotide, alone or combined with cimetidine, have been investigated on mucosal lesions induced in rats by pylorus ligation. In the same animals, the measurement of acid and pepsin output and of soluble and barrier mucus has been performed. Dose-dependent sulglicotide prevented the development of mucosal lesions and its protective effect was achieved without significant modifications in gastric acid secretion. The secretion of pepsin and of mucus was markedly inhibited at every dosage of the compound. Neither the damage to gastric mucosa nor the secretion of acid, pepsin and mucus were affected by cimetidine. The combination of the highest doses of both compounds resulted in a synergistic gastro-protective effect, not dependent on a synergistic effect on the reduction in acid secretion.     

Assessing protein co-evolution in the context of the tree of life assists in the prediction of the interactome

The identification of the whole set of protein interactions taking place in an organism is one of the main tasks in genomics, proteomics and systems biology. One of the computational techniques used by many investigators for studying and predicting protein interactions is the comparison of evolutionary histories (phylogenetic trees), under the hypothesis that interacting proteins would be subject to a similar evolutionary pressure resulting in a similar topology of the corresponding trees. Here, we present a new approach to predict protein interactions from phylogenetic trees, which incorporates information on the overall evolutionary histories of the species (i.e. the canonical "tree of life") in order to correct by the expected background similarity due to the underlying speciation events. We test the new approach in the largest set of annotated interacting proteins for Escherichia coli. This assessment of co-evolution in the context of the tree of life leads to a highly significant improvement (P(N) by sign test approximately 10E-6) in predicting interaction partners with respect to the previous technique, which does not incorporate information on the overall speciation tree. For half of the proteins we found a real interactor among the 6.4% top scores, compared with the 16.5% by the previous method. We applied the new method to the whole E.coli proteome and propose functions for some hypothetical proteins based on their predicted interactors. The new approach allows us also to detect non-canonical evolutionary events, in particular horizontal gene transfers. We also show that taking into account these non-canonical evolutionary events when assessing the similarity between evolutionary trees improves the performance of the method predicting interactions.     

Variations in the arrangement of tonofilaments in the epidermis of teleost fish

Tonofilaments in epithelial cells of teleost skin can be aligned as bundles or skeins of appreciable bilk, or form a pattern of smaller bundles oriented in various directions, or there may be a condition where individual tonofilaments interlace. If sufficiently close together, interwoven tonofilaments can form a basket-like structure, a “capsule”, proximally in the cell. This arrangement, previously known in epithelial cells of Myxinoids, occurs in localised sites in various teleosts of diverse taxonomic position, for instance in clupeids and gadids. A less intimate interlacing of cortical tonofilaments can accompany a modification of the perinuclear cytoplasm previously described, by light microscopy, as “vesiculated”, as in the middle layers of the epidermis in Periophthalmus. In head epidermis of Sprattus, the outer layers of cells contain proximal capsules, but the middle layers consist of flattened cells with a restricted perinuclear cytoplasm, peripheral tonofilaments, and a second population of filaments of a larger calibre. One implication of these results is that the cytoskeleton can undergo profound modification as cells progress from the basal to the superficial layers of the epidermis.     

The role of the methylation in the detoxication of arsenate in the rabbit

The biotransformation, tissue retention, intracellular binding and biokinetics of arsenic were studied in rabbits exposed to [74As]arsenate (0.4 mg As/kg body wt., i.v.). Inhibition of the methyltransferase activity by injection of periodate-oxidized adenosine (PAD) caused a marked decrease of the formation of [74As]dimethylarsinic acid (DMA), which gave rise to 1.5-4 times increased tissue levels of 74As. This is almost the same as reported for rabbits given arsenite in combination with PAD and was due to a rapid reduction of the arsenate to arsenite which bound to the tissues. Only about 30% of the arsenate given was excreted unchanged in the urine, indicating that a large part was reduced to AsIII. Thus the methylation to DMA seems to be almost as important for the detoxication following exposure to arsenate as that following exposure to arsenite. In the rabbits with normal methylating capacity 50-70% of the produced AsIII was methylated to DMA. The liver was the only organ in which DMA was present 1 h after the administration, indicating that this is the main site of the methylation. The DMA was rapidly cleared from all tissues except the thyroid.     

Dynamics of the changes in the content of testosterone and estradiol receptors in the hypothalamus of male rats in the course of sexual development

The content of receptors to testosterone and estradiol in hypothalamus of the male rats was studied during their sexual maturation (7, 14, 21, 28, 35 and 42 days). In all the age groups of animals the concentration of receptors to testosterone in the cytoplasmic and nuclear fractions of hypothalamus was at a relatively constant level, except in 7 day old males in which the minimal concentration of cytoplasmic and the maximal concentration of nuclear receptors were noted. The highest values of estradiol-binding sites in cytosol of hypothalamus were observed on the 7th and 14th days and in the nuclear fraction on the 28th, 35th and 42nd days of life. The binding of both the hormones with their receptors is a specific process characterized by a high affinity. A suggestion is put forward that receptors both to androgens and estrogens take part in the brain sexual differentiation.     

Changes in the size and shape of the synaptic vesicles in the sensorimotor cortex of the rat brain in the initial phases of kindling

The sensorimotor area of rat cerebral cortex was subjected to repeated electrical stimulation at 10-min intervals, with resultant formation and progressive lengthening of self-sustained after-discharges (SSAD). One and 60 min after the third SSAD ended, we carried out an electron microscopy morphometric analysis of the agranular synaptic vesicles in type I synapses (after Gray) in the second cortical layer of the homotopic area of the unstimulated hemisphere. One minute after the seizure ended, 5.8% enlargement of the synaptic vesicles compared with the control was demonstrated in zone II of the synapse (0.1-0.2 micron from the active zone of the synapse). Neither the size nor the shape of the synaptic vesicles in the other parts of the synaptic apparatus altered. Sixty min after the seizure ended, a 5.5% enlargement of the synaptic vesicles in zone I (0.0-0.1 micron) and a 5.4% enlargement of those in zone II was found. The synaptic vesicles in zone I in the experimental animals were more oval than in the controls. Our findings support the vesicular theory and testify that hyperfunction, up to temporary exhaustion of the synaptic apparatuses, produces a change in the transmitter content of the synaptic vesicles. A raised amount of transmitter in the synaptic vesicles near the active zone could be one of the factors responsible for continued hyperexcitability of the tissue one hour after the seizure had ended. The results likewise support the concept of two mechanisms of synaptic vesicle formation, and hence of the existence of two different vesicle populations.     

Analysis of changes in the electrical activity of the neocortex in dogs during the formation of the stereotype of food-procuring conditioned reflexes in them

A system of food-procuring conditioned reflexes (dynamic stereotype after I.P. Pavlov) was elaborated in dogs. In the interstimuli periods, 0.6 to 0.8 sec. prior the action of the conditioned stimulus, they exhibited a "state of expectancy" characterized by an increase of frequency (up to 80 per sec.) of potential oscillations of a small amplitude (20 to 30 mcv) and their pronounced sychroneity, predominantly in the anterior parts of the neocortex. The conditioned signal acting against such background produced specific reactions in the form of a limited number of bursts (3 to 4 in 0.5 sec.) of highfrequency (up to 100 per sec.) synchronized activity (HSA) of considerable amplitude (50 to 60 mcv) which always preceded conditioned reactions, being in certain temporal relations with them. A correlation-spectral analysis has shown that in the HSA period, electrical activity became considerably more regular; in the intensity spectra the extreme frequencies of the analyzed band (5 to 7 c/s and 90 to 100 c/s) became prominent, and in most cases high values of the coherence function were due to them. HSA reactions are regarded as a major link in the trigger mechanism of conditioned food-procuring reactions.     

Role of dominant motivation in organizing the functions of the higher divisions of the auditory analyzer in the cat

In chronic experiments on cats, reorganization of initial interrelations between alpha-activity of higher parts of the auditory analyzer during acoustic stimulation was of a selective character and depended on motivational properties of the elicited excitation. A conditioned acoustic signal of alimentary situation, greatly intensified the functional conjugation of the majority of studied auditory projections and a number of non-specific structures, while in a defence situation there was a considerable reduction of correlative dependence between compared characteristics of the auditory centers state and a change in the hierarchy of their functional interrelations.     

Morphofunctional changes in the chief cells of the intrinsic glands of the stomach in the presence of an excess of glucocorticoid and sex hormones in the body

In 260 male white rats electron microscopically, cytochemically with biochemical estimation of proteolitic activity of gastric juice, it has been demonstrated that principle cells of the gastric glands, after injection of small doses of hydrocorticone, respond with a complex of adequate ultrastructural, cytochemical and functional changes directed to realization of the drugs stimulating effect. With increasing time of the experiment, or the doses, progressively developing distrophic processes, involving predominantly the protein-synthesizing apparatus and, to a less degree, the mitochondrial mechanism, are observed in the principle cells. This results in inhibition of the pepsinogene synthesis phase and in decreasing proteolytic activity of gastric juice. Reaction of the principle cells is independent on testosterone doses; the protein-synthesizing and mitochondrial (to a less degree) apparatuses develop; the pepsinogene extrusion phase is suppressed; it results in its deposition in cytoplasm and in decreasing proteolytic activity of gastric juice. When testosterone and hydrocortisone are injected simultaneously in small doses, the stimulating effect of hydrocortisone to the principle cells is abolished and at large doses it is sharply manifested and is not accompanied with destructive changes in the principle cells.     

Changes in the epithelial permeability of the large intestine of mice in the process of carcinogenesis

1,2-dimethylhydrazine (DMH), a colon carcinogen, being injected weekly to BALB/c mice, inhibits an active sodium transport, increases the transepithelial passive ion permeation and decreases ion selectivity in the descending colon. A single DMH injection leads to the same alterations, manifested for a month, followed by normalization of all the parameters to the control value. Distinctive, wavy changes in electrophysiological parameters were noted after a single injection of "non-colon" carcinogen 7,12-dimethyl-benz(alpha)antracen. It is supposed that the prolonged drop in active sodium transport, transepithelial resistance and ion selectivity are specific reactions of the colonic epithelium to carcinogenic treatment with DMH.     

Cranial cartilages: Players in the evolution of the cranium during evolution of the chordates in general and of the vertebrates in particular

The present contribution is chiefly a review, augmented by some new results on amphioxus and lamprey anatomy, that draws on paleontological and developmental data to suggest a scenario for cranial cartilage evolution in the phylum chordata. Consideration is given to the cartilage-related tissues of invertebrate chordates (amphioxus and some fossil groups like vetulicolians) as well as in the two major divisions of the subphylum Vertebrata (namely, agnathans, and gnathostomes). In the invertebrate chordates, which can be considered plausible proxy ancestors of the vertebrates, only a viscerocranium is present, whereas a neurocranium is absent. For this situation, we examine how cartilage-related tissues of this head region prefigure the cellular cartilage types in the vertebrates. We then focus on the vertebrate neurocranium, where cyclostomes evidently lack neural-crest derived trabecular cartilage (although this point needs to be established more firmly). In the more complex gnathostome, several neural-crest derived cartilage types are present: namely, the trabecular cartilages of the prechordal region and the parachordal cartilage the chordal region. In sum, we present an evolutionary framework for cranial cartilage evolution in chordates and suggest aspects of the subject that should profit from additional study.     

Phosphoinositide metabolism in sections of the cortex of the large hemisphere of the brain in anoxia]

Depolarization (an increased concentration of KCL in the medium) has been investigated for its effect on the content and turnover rate of phospholipid phosphorus from the rat brain cortex slice under normal oxygen supply and under anoxia. It is shown that anoxia results in a small increase of phosphatidyl-inositol-4.5-bisphosphate (PIP2) and phosphatidylinositol-4-phosphate (PIP) content and in the depression of the turnover rate of all the phosphoinositides. Depolarization leads to a decrease in PIP2 concentration with a simultaneous increase in their turnover rate, these results being more expressed under anoxia. The development of depolarization by the 5th min. of anoxia in vivo leads, most probably, to the enhanced PIP2 breakdown, that is to a progressive decrease in their content.