Influence of hormones on the functional activity of the regenerating cortex of the enucleated adrenal in situ]

The functional activity of the regenerating cortex was studied in 90 female albino rats (150-200 g) after ablation of the right adrenal and enucleation of the left one. In the period of active growth of the regenerating tissue (10 days after operation) the functional activity of parenchymatous cells was low which is evidenced by poor content of both lipids and "ketosteroids". In parallel with reparation of the adrenal adrenocortical tissue mass the content of physiologically active substances was also restored (20 days after operation). After injection of hydrocortizone (daily dosage 2,5 mg) the growth and differentiation of the bundle-reticular zone in the regenerating area was inhibited. In the glomerular zone the reactions to lipids and "ketosteroids" were mainly similar to those in the glomerular zone of intact adrenal. After injection of ACTH (daily dosage 5 or 10 mg) during 10 days the regenerating area was functionally better developed than in the control since moderately pronounced reactions to "ketosteroids" and lipids appeared in it. Fairly high content of these substances in the regenerated cortex after 20 days of injections of ACTH (10 units) as well as presence of secondary necrobiotic changes pointed to functional overstrain of the newly formed organ.     

Presence of Virus Neutralizing Antibodies in Cerebral Spinal Fluid Correlates with Non-Lethal Rabies in Dogs

Background

Rabies is traditionally considered a uniformly fatal disease after onset of clinical manifestations. However, increasing evidence indicates that non-lethal infection as well as recovery from flaccid paralysis and encephalitis occurs in laboratory animals as well as humans.

Methodology/Principal Findings

Non-lethal rabies infection in dogs experimentally infected with wild type dog rabies virus (RABV, wt DRV-Mexico) correlates with the presence of high level of virus neutralizing antibodies (VNA) in the cerebral spinal fluid (CSF) and mild immune cell accumulation in the central nervous system (CNS). By contrast, dogs that succumbed to rabies showed only little or no VNA in the serum or in the CSF and severe inflammation in the CNS. Dogs vaccinated with a rabies vaccine showed no clinical signs of rabies and survived challenge with a lethal dose of wild-type DRV. VNA was detected in the serum, but not in the CSF of immunized dogs. Thus the presence of VNA is critical for inhibiting virus spread within the CNS and eventually clearing the virus from the CNS.

Conclusions/Significance

Non-lethal infection with wt RABV correlates with the presence of VNA in the CNS. Therefore production of VNA within the CNS or invasion of VNA from the periphery into the CNS via compromised blood-brain barrier is important for clearing the virus infection from CNS, thereby preventing an otherwise lethal rabies virus infection.     

Relationship between the activity of the kallikrein, plasmin and thrombin systems of the blood during intense physical exertion]

The authors studied the changes in the level of precursors of kallikrein, plasmine, and their inhibitors with the aid of a complex method. Blood plasma of sportsmen was examined--of healthy persons and of those with the syndrome of myocardial overstraining under conditions of rest and intensive bicycle ergometric exercises. The results obtained and correlation analysis data pointed to the functional association of the "Hageman's factor system". Sportsmen with the overstrain syndrome displayed an aggravation of the functional possibilities of humoral systems of the hemovascular regulation, this being expressed in a reduced level of the proenzymatic activity and a reduction of the inhibitor values. Correlation of the mentioned indices was disturbed both at rest and during the exercises.     

Comparative ultrastructural characteristics of the anterior lobe of the hypophysis, thyroid and gonads in mammals during aging

Ultrastructural changes in certain types of adenocytes of old rats, guinea pigs, rabbits and dogs are characterized by specific peculiarities. However, the direction of these changes mainly coincides. The somatotropic function of rats, guinea pigs and rabbits is suppressed, lactotrophic and corticotrophic cells being activated with aging. The phenomenon of the overstrain of gonadotrophic and thyrotrophic cells is registered. Dystrophic changes in the peripheral glands are observed.     

The problem of adaptation and oscillatory processes in the microvascular bed

Healthy people (n = 16), patients with autonomic dystonia syndrome (n = 38), and patients with traumatic rupture of the median nerve before and after nerve suture (n = 28) were examined by laser Doppler flowmetry (LDF) with a computer wavelet analysis of blood flow oscillations. Functional states (FSs) of the microcirculatory bed wеre assessed using energetic and information indices of microvascular blood flow oscillations. The variation coefficient and the information regime (multistable or resonance) were used as key characteristics. Oscillatory processes are an integral part of adaptation and the FS formation in the microvascular bed. FSs were classified as adaptive, hyperadaptive, hypoadaptive, and failure of adaptation. Because supporting the optimal function of nutritive microvessels is a leading component of the adaptation process, FSs of nutritive and nonnutritive microvessels may differ. A selective contribution of the autonomic sympathetic regulatory channel was related to maintaining considerable hyperadaptation in the microvascular bed with overstrain or marked overstrain of regulatory systems, as in emotional stress. Hypoadaptive FSs formed when skin blood flow increased, an excess decrease in flow resistance was unnecessary, and especially when regulatory factors were in deficiency, e.g., in neurodystrophic syndrome.     

Rhythmic Acoustic Stimulation as a Method for Regulating the Functional State of the CNS in Young Schoolchildren

The author studied the dynamics of EEG (spectral power density and coherence functions) and autonomic (electrodermal resistance, EDR) indices in young schoolchildren with changes in the functional state of the CNS caused by rhythmic sensory stimulation (acoustic signals with frequencies of 6 and 9 Hz). Both of these signals influenced the CNS and may be used to achieve relaxation. Their comparison demonstrated both similar postrelaxation shifts (increased coherence of distant connections) and frequency-specific phenomena (more numerous significant changes in the respective frequency ranges). Thus, the signal frequency determines the pattern of physiological shifts, influencing the relaxation quality. Low-frequency (6 Hz) stimulation caused more pronounced changes in the CNS, reflected in enhanced EDR, and a stronger increase in the short-term memory volume. The author discusses the frequency-specific neurophysiological mechanisms of the effect of stimulation on the functional state of the brain.     

Functional state of the frontal cortex and ventromedial hypothalamus during physical overexertion and experimental neurosis in the rabbit

A change of excitability, pO2 and local cortical blood flow under a long-term maximum physical load and in conditions of experimental neurosis, was investigated in rabbits with electrodes implanted in the frontal cortex and ventromedial hypothalamus. It was found that functional activity of these structures under physical orverstrain rises as the blood flow and pO2 increase, and the excitability also increases. In experimental neurosis, a discoordination in functioning of the cortex and hypothalamus is observed. Relative stabilization of the functional state of tested structures, observed during physical overstrain, is absent in conditions of experimental neurosis.     

Laboratory investigations on "postvaccinall rabies" caused by live sheep-brain vaccine.

The viral ethiology of postvaccinal complications among 30 dogs vaccinated by live antirabies vaccine (Umeno-Doi type, sheep brain vaccine) was fully confirmed. Three lots of virulent vaccine were inoculated subcutaneously into groups of "Wistar" rats according to the different schemes. Between the 1st and 12th day after the end of the vaccination there were no isolations of fixed virus in direct and blind i.c. passages of suspensions made from the thalamus area on succkling mice and rats. Also the viral antigen in the CNS of vaccinated but apparently healthy rats was undetectable. The "postvaccinal rabies" with the next isolation of fixed r.v. in the CNS was developed experimentally in rats only following the subcutaneous injection of the crude sheep-brain's and spinal cord's suspensions--composing the materials to production of antirabies vaccine.     

Ventilatory responses to specific CNS hypoxia in sleeping dogs.

Our study was concerned with the effect of brain hypoxia on cardiorespiratory control in the sleeping dog. Eleven unanesthetized dogs were studied; seven were prepared for vascular isolation and extracorporeal perfusion of the carotid body to assess the effects of systemic [and, therefore, central nervous system (CNS)] hypoxia (arterial PO(2) = 52, 45, and 38 Torr) in the presence of a normocapnic, normoxic, and normohydric carotid body during non-rapid eye movement sleep. A lack of ventilatory response to systemic boluses of sodium cyanide during carotid body perfusion demonstrated isolation of the perfused carotid body and lack of other significant peripheral chemosensitivity. Four additional dogs were carotid body denervated and exposed to whole body hypoxia for comparison. In the sleeping dog with an intact and perfused carotid body exposed to specific CNS hypoxia, we found the following. 1) CNS hypoxia for 5-25 min resulted in modest but significant hyperventilation and hypocapnia (minute ventilation increased 29 +/- 7% at arterial PO(2) = 38 Torr); carotid body-denervated dogs showed no ventilatory response to hypoxia. 2) The hyperventilation was caused by increased breathing frequency. 3) The hyperventilatory response developed rapidly (<30 s). 4) Most dogs maintained hyperventilation for up to 25 min of hypoxic exposure. 5) There were no significant changes in blood pressure or heart rate. We conclude that specific CNS hypoxia, in the presence of an intact carotid body maintained normoxic and normocapnic, does not depress and usually stimulates breathing during non-rapid eye movement sleep. The rapidity of the response suggests a chemoreflex meditated by hypoxia-sensitive respiratory-related neurons in the CNS.     

Deletion of the SELENOP gene leads to CNS atrophy with cerebellar ataxia in dogs

We investigated a hereditary cerebellar ataxia in Belgian Shepherd dogs. Affected dogs developed uncoordinated movements and intention tremor at two weeks of age. The severity of clinical signs was highly variable. Histopathology demonstrated atrophy of the CNS, particularly in the cerebellum. Combined linkage and homozygosity mapping in a family with four affected puppies delineated a 52 Mb critical interval. The comparison of whole genome sequence data of one affected dog to 735 control genomes revealed a private homozygous structural variant in the critical interval, Chr4:66,946,539_66,963,863del17,325. This deletion includes the entire protein coding sequence of SELENOP and is predicted to result in complete absence of the encoded selenoprotein P required for selenium transport into the CNS. Genotypes at the deletion showed the expected co-segregation with the phenotype in the investigated family. Total selenium levels in the blood of homozygous mutant puppies of the investigated litter were reduced to about 30% of the value of a homozygous wildtype littermate. Genotyping >600 Belgian Shepherd dogs revealed an additional homozygous mutant dog. This dog also suffered from pronounced ataxia, but reached an age of 10 years. Selenop^-/- knock-out mice were reported to develop ataxia, but their histopathological changes were less severe than in the investigated dogs. Our results demonstrate that deletion of the SELENOP gene in dogs cause a defect in selenium transport associated with CNS atrophy and cerebellar ataxia (CACA). The affected dogs represent a valuable spontaneous animal model to gain further insights into the pathophysiological consequences of CNS selenium deficiency.     

Morphometric and electron microscopic study of the small intestine following total orthotopical auto- and allotransplantation]

Total orthotopic autotransplantation of the small intestine was performed in 22 dogs, allotransplantation--in 13 dogs. Changes in the wall of the graft resulted from the effect of both nonspecific (ischaemia, blockage of lymph outflow, denervation) and specific factors connected with the reaction of tissue incompatibility. Changes caused by the nonspecific factors are demonstrated as lymphostasis, edema of the wall in the graft, atrophy of the villi with epithelial flattening, hyperplasy of the intestinal gland, macrophagal and plasmocytic reaction in the mucous lamina propria, drop in villous-cryptic coefficient and reduction of the absorbing surface of the villi. At autotransplantation, structural-functional changes in the intestine undergo three stages: the stage of acute morphological changes and pronounced functional disturbances (up to 1,5 months); the stage of relative structural stabilization and gradual functional restoration (up to 6 months); the stage of atrophic-hyperplastic changes in the wall and decrease in functional activity of the intestinal graft (followed-up for 15 months). At allotransplantation, depending on the degree of cellular reaction and vascular changes, as well as on the localization of the process in the wall of the graft, a "graft versus host" reaction and a "recipient versus graft" reaction is demonstrated. Peculiar rosettes of plasmocytes have been described as a possible index of the reaction "graft versus host".     

Homeostatic role of the active transport in elimination of [3H]benzylpenicillin out of the cerebrospinal fluid system

Cerebral acidic metabolites and penicillin are organic anions which can be carried by active transport into capillaries of the central nervous system (CNS). However, it is generally believed that these metabolites are mainly delivered from CNS to cerebrospinal fluid (CSF) and eliminated by CSF circulation over cortex and its absorption into dural venous sinuses. To test this hypothesis we studied fate of penicillin ([3H]benzylpenicillin) in the CSF under control conditions and when its active transport was blocked by probenecid. After application of penicillin into cisterna magna of control dogs, it is distributed only in traces to lumbar, ventricular and cortical CSF. However, when active transport of penicillin across capillary wall is blocked by probenecid, its disappearance from cisterna is slowed down and its distribution is greatly enhanced so that at 300 min penicillin concentrations in cisternal, lumbar and cortical CSF approach or equal each other. Disappearance of penicillin from cisternal CSF shows a single exponential course (half-time 30 min) in control, while in probenecid pretreated dogs this is a slow multiexponential process. The results indicate that the active transport across capillary wall in CNS, but not generally postulated unidirectional CSF circulation over cortex and its absorption into dural venous sinuses, is instrumental in elimination of cerebral acidic metabolites and in such a way homeostasis in brain and cerebrospinal fluid is maintained.     

"Smouldering" and symptomatic forms of central nervous system involvement in the course of non-Hodgkin's lymphoma in children

34 children with Non-Hodgkin's lymphoma (NHL) were treated with LSA2L2 protocol from 1978 to 1981. In 12 cases (31.4%) the central nervous system (CNS) was involved, including 3 cases at the onset of the disease. CNS involvement was always diagnosed by the presence of blast cells in cerebro-spinal fluid, also in cases with normal pleocytosis and no neurological symptoms. Such cases were called "smouldering" CNS involvement. Four children had a smouldering form of CNS involvement. 3 of them are in continuous complete remission with cessation of therapy, while all those 6 patients with symptomatic CNS involvement died. "Smouldering" CNS involvement seems to have a better prognosis than the symptomatic one.     

Expression of microRNAs in cerebrospinal fluid of dogs with central nervous system disease

In this pilot study we investigated the expression of 14 microRNAs in the cerebrospinal fluid (CSF) of dogs with neoplastic, inflammatory and degenerative disorders affecting the central nervous system (CNS). CSF microRNA (miRNA) expression profiles were compared to those from dogs with neurological signs but no evidence of structural or inflammatory CNS disease. Seven miRNAs were easily detected in all samples: miR-10b-5p, miR-19b, miR-21-5p, miR-30b-5p, miR-103a-3p, miR-124, and miR-128-3p. Expression of miR-10b-5p was significantly higher in the neoplastic group compared to other groups. There was no relation between miRNA expression and either CSF nucleated cell count or CSF protein content. Higher expression of miR-10b-5p in the neoplastic group is consistent with previous reports in human medicine where aberrant expression of miR-10b is associated with various neoplastic diseases of the CNS.     

Morphological changes in the filtration-reabsorption barrier of the remaining kidney after its denervation and disordered lymph outflow

An electron microscopical investigation on combined effect of denervation and disturbance of lymphatic outflow to structural elements of the FRB in the preserved kidney after contralateral nephrectomy has been performed on 20 mature white rats in 3, 30, 180, 360 days. In 3 days in the cellular components of the FRB cytoplasmic structures are at the state of functional strain and overstrain. Hemostasis is observed in glomeruli and in peritubular capillaries, as well as edema of the interstitial connective tissue. In 30 days certain signs of hyperplasia and hypertrophy of intracellular structures appear in the components of the FRB. However, later (in 180, 360 days) certain tendency to predominance of processes of functional strain in the intracellular structures with their certain destruction and reaction of the connective stroma is observed.     

Genome-Wide Association Analysis Identifies a Mutation in the Thiamine Transporter 2 (SLC19A3) Gene Associated with Alaskan Husky Encephalopathy

Alaskan Husky Encephalopathy (AHE) has been previously proposed as a mitochondrial encephalopathy based on neuropathological similarities with human Leigh Syndrome (LS). We studied 11 Alaskan Husky dogs with AHE, but found no abnormalities in respiratory chain enzyme activities in muscle and liver, or mutations in mitochondrial or nuclear genes that cause LS in people. A genome wide association study was performed using eight of the affected dogs and 20 related but unaffected control AHs using the Illumina canine HD array. SLC19A3 was identified as a positional candidate gene. This gene controls the uptake of thiamine in the CNS via expression of the thiamine transporter protein THTR2. Dogs have two copies of this gene located within the candidate interval (SLC19A3.2 – 43.36–43.38 Mb and SLC19A3.1 – 43.411–43.419 Mb) on chromosome 25. Expression analysis in a normal dog revealed that one of the paralogs, SLC19A3.1, was expressed in the brain and spinal cord while the other was not. Subsequent exon sequencing of SLC19A3.1 revealed a 4bp insertion and SNP in the second exon that is predicted to result in a functional protein truncation of 279 amino acids (c.624 insTTGC, c.625 C>A). All dogs with AHE were homozygous for this mutation, 15/41 healthy AH control dogs were heterozygous carriers while 26/41 normal healthy AH dogs were wild type. Furthermore, this mutation was not detected in another 187 dogs of different breeds. These results suggest that this mutation in SLC19A3.1, encoding a thiamine transporter protein, plays a critical role in the pathogenesis of AHE.     

Hemispheric specialization in domestic dogs (Canis familiaris) for processing different types of acoustic stimuli

Lateralization is considered to be a fundamental feature of vertebrate brains. The aim of the present study was to examine the impact of functional cerebral asymmetry on processing of auditory stimuli in the domestic dog (Canis familiaris) during the orientation reaction. The experiment was conducted on 46 dogs (25 females and 21 males). Four types of auditory stimuli were used in the experiment (three meaningful stimuli: cat meowing, dog barking, the "sit" command ("siad" in Polish), and a neutral word ("wir", meaning "whirl" in Polish). It was predicted that the orientation reaction (turning the head towards the stimuli) would take place only in the case of meaningful sounds. It was also expected that dogs would show consistent lateralization. As predicted, all three meaningful stimuli elicited the orientation reaction. The response of the examined dogs to cat meowing showed significant lateralization with dominant leftwards movement, which hints towards activation of the right cerebral hemisphere and may be related to strong emotions evoked by this stimulus. Contrary to results of previous studies, dogs reacting to dog barking turned their heads leftwards more often, which suggests activation of the right cerebral hemisphere, probably related to the emotional meaning of the stimulus. The "sit" command consistently evoked the orientation reaction but there was no significant lateralization of this movement.     

Human immunity system under conditions of 105-day isolation and confinement in an artificial environment

The study of the adaptive immunity system of six test subjects volunteered for a 105-day isolation and confinement in an artificial environment showed activation of the immune system and overstrain and depletion of its functional reserve. Significant differences in the adaptability of the immune system of the test subjects indicate individual susceptibility to disorders in immunological reactivity.     

Clathrin-mediated endocytic signals are required for the regeneration of, as well as homeostasis in, the planarian CNS

Planarians have a well-organized central nervous system (CNS), including a brain, and can regenerate the CNS from almost any portion of the body using pluripotent stem cells. In this study, to identify genes required for CNS regeneration, genes expressed in the regenerating CNS were systematically cloned and subjected to functional analysis. RNA interference (RNAi) of the planarian clathrin heavy chain (DjCHC) gene prevented CNS regeneration in the intermediate stage of regeneration prior to neural circuit formation. To analyze DjCHC gene function at the cellular level, we developed a functional analysis method using primary cultures of planarian neurons purified by fluorescence-activated cell sorting (FACS) after RNAi treatment. Using this method, we showed that the DjCHC gene was not essential for neural differentiation, but was required for neurite extension and maintenance, and that DjCHC-RNAi-treated neurons entered a TUNEL-positive apoptotic state. DjCHC-RNAi-treated uncut planarians showed brain atrophy, and the DjCHC-RNAi planarian phenotype was mimicked by RNAi-treated planarians of the mu-2 (micro2) gene, which is involved in endocytosis, but not the mu-1 (micro1) gene, which is involved in exocytosis. Thus, clathrin-mediated endocytic signals may be required for not only maintenance of neurons after synaptic formation, but also axonal extension at the early stage of neural differentiation.