Distal arthrogryposis type 5 – arthrogryposis with ophthalmoplegia, polyneuropathy. Case Report

A clinical case of a 27-year-old patient with distal arthrogryposis of the 5th type – arthrogryposis with ophthalmoplegia, which was combined in a patient with polyneuropathy is presented. To assess tissue respiration (mitochondrial respiratory chain) and other types of metabolism in mitochondria, cytochemical analysis of lymphocytes in peripheral blood was carried out according to A. Pearse's method modified by R.P. Narcissov. The activity of four mitochondrial enzymes involved in carbohydrate metabolism (lactate dehydrogenase), amino acid metabolism (glutamate dehydrogenase), fatty acid metabolism (α-glycerophosphate dehydrogenase) and the second complex of the mitochondrial respiratory chain (succinate dehydrogenase) was assessed. A slight decrease in the activity of the enzyme succinate dehydrogenase, which is part of the second complex of the mitochondrial respiratory chain, was determined. The activity of the enzyme α-glycerophosphate dehydrogenase was more significantly reduced, an increase in the activity of lactate dehydrogenase was noted. In the presented observation, along with the typical manifestations of the disease (contractures of the hands and feet, ophthalmoplegia, ptosis of the eyelids, visual impairment), polyneuropathy with impaired sensitivity of the polyneuritic type was revealed. Thus, a patient with polyneuropathy had a hereditary type 5 arthrogryposis disease. Along with the typical manifestations of the disease, polyneuropathy with hyporeflexia and impaired sensitivity of the polyneuritic type was revealed. Secondary mitochondrial disorders were identified, which was the basis for the appointment of energotropic therapy.

Fiber Metabolism, Procollagen and Collagen Type III Immunoreactivity in Broiler Pectoralis Major Affected by Muscle Abnormalities

The present study aimed to evaluate the muscle fiber metabolism and assess the presence and distribution of both procollagen and collagen type III in pectoralis major muscles affected by white striping (WS), wooden breast (WB), and spaghetti meat (SM), as well as in those with macroscopically normal appearance (NORM). For this purpose, 20 pectoralis major muscles (five per group) were selected from the same flock of fast-growing broilers (Ross 308, males, 45-days-old, 3.0 kg live weight) and were used for histochemical (nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) and alpha-glycerophosphate dehydrogenase (α-GPD)) and immunohistochemical (procollagen and collagen type III) analyses. When compared to NORM, we found an increased proportion (p < 0.001) of fibers positively stained to NADH-TR in myopathic muscles along with a relevant decrease (p < 0.001) in the percentage of those exhibiting a positive reaction to α-GPD. In addition, an increased proportion of fibers exhibiting a positive reaction to both stainings was observed in SM, in comparison with NORM (14.3 vs. 7.2%; p < 0.001). After reacting to NADH-TR, SM exhibited the lowest (p < 0.001) cross-sectional area (CSA) of the fibers (−12% with respect to NORM). On the other hand, after reacting to α-GPD, the CSA of WS was found to be significantly larger (+10%) in comparison with NORM (7480 vs. 6776 µm2; p < 0.05). A profound modification of the connective tissue architecture involving a different presence and distribution of procollagen and collagen type III was observed. Intriguingly, an altered metabolism and differences in the presence and distribution of procollagen and collagen type III were even observed in pectoralis major muscle classified as NORM.

Deficient Activity of FAD-Linked Glycerophosphate Dehydrogenase in the Mitochondria of Insulin-Producing Pancreatic Islets B-Cells: A Possible Cause of the Metabolic Syndrome

A deficient activity of mitochondrial FAD-linked glycerophosphate dehydrogenase in insulin-producing pancreatic islet B-cells was recently identified in selected animal models of non-insulin-dependent diabetes mellitus. The incidence of this enzymatic defect remains however to be unambiguously documented in type-2 diabetic patients. The present review mainly aims at speculating that the altered recognition of nutrients, especially D-glucose, by pancreatic B-cells, as resulting from the abovementioned enzymatic perturbation could also eventually lead to the development of the metabolic syndrome (MetS).

Experimental substantiation of the possibility of using the cytochemical parameters of peripheral blood to assess the functional state of the organism of the pilot

The activity of succinate (mitochondrial), lactate, α-glycerophosphate, glucose-6-phosphate peripheral blood lymphocytes in rats under the conditions of modeling acute and chronic gravitational stress was studied by quantitative spectrophotometry of cytochemical reaction products to reveal the localization of oxidative enzymes. The dynamics of morphofunctional transformations is determined, the severity of which depends on the duration of the effect (the rotation time of the animals on the centrifuge). For acute gravitational stress, changes in the activity of mitochondrial succinate dehydrogenase are characteristic, and in the various stages of chronic gravitational stress, glycolysis (increase in lactate dehydrogenase and α-glycerophosphate dehydrogenase activity) and plastic metabolism (increase in glucose-6-fosfata dehydrogenase activity) are characteristic. These changes are explained by the inclusion of neurohumoral regulatory mechanisms that ensure synchronization of oxidative metabolic processes in the cells of organs of various body systems under unfavorable conditions of their functioning. Significant correlation links between the activity of lymphocyte enzymes with the same indices of neurons of the celiac sympathetic node, adenocytes of the cortical and medulla of the adrenal glands have been established. This suggests that the activity of lymphocyte enzymes can be used as indirect indicators of the processes occurring in the organs of the sipato-adrenomedular and pituitary-adrenocortical systems under conditions of acute and chronic gravitational stress.

THE DIAGNOSTICS OF CELL ENERGETIC DEFICIT IN CHILDREN WITH CHRONIC BRONCHOPULMONARY PATHOLOGY

The aim of the research is to improve the diagnosis of disturbances of cell energy in children with chronic nonspecific pulmonary diseases (CNPD) in the remission stage by the methods of detection of cytochemical changes of the energy status and recording the percentage of immune-competent cells of blood with the low membrane potential of mitochondria (MPM). 77 children were examined, including 66 (83%) with lung malformations, and 11 children (17%) with chronic nonspecific lung diseases as an outcome of acute/chronic lung diseases. The presence of energy deficient states was assessed by the level of activity of oxidative-reduction enzymes of succinate dehydrogenase, alpha-glycerophosphate dehydrogenase, lactic dehydrogenase in lymphocytes of the peripheral blood and percent ratio of lymphocytes with low MPM. It was found out that in 53.2% of the examined children even in the remission stage there were identified the signs of mitochondrial insufficiency when the parameters of metabolic activity of enzymes in lymphocytes were significantly lower than the norm. The cluster analysis of the obtained data allowed identifying the parameters typical for energy deficient states of the organism of the child that demand the correction by energotropic medications.

A computational study about the mechanism of action of metformin on hepatic gluconeogenesis, focused on its ability to create stable pseudo-aromatic copper complexes

Metformin is the best therapeutic choice for treating type 2 Diabetes.Despite this, and the fact it has been prescribed worldwide for decades, its mechanism of gluconeogenesis inhibition is still unknown.In the following work a novel mechanism of inhibition is suggested: that metformin performs its action on the target enzyme not as a pure molecule but, after sequestering endogenous cellular copper, as a copper complex.This result was obtained using chemoinformatics methods including homology modeling for the creation of the target enzyme’s tridimensional virtual structure, molecular docking for both the determination of the movement of the prosthetic group inside its cavity and for the identification of the best ligand poses for the metformin copper complexes, and eventually pharmacophore modeling and virtual screening to find alternative virtual leads that could achieve similar effects.The simulations show the complex binding as a non competitive inhibitor to the large exit of the mitochondrial glycerophosphate dehydrogenase enzyme’s FAD cavity, preventing FAD movement inside the cavity and/or quinone interaction and therefore its electron transfer function.The proposed mechanism seems to be successful at explaining a wide range of existing experimental results, both regarding measurements of metformin non-competitive inhibition of GPD2 and the role of copper and pH in its action.The virtual screening outcome of at least two similarly active purchasable molecule hints to an easy way to experimentally test the proposed mechanisms.In fact, the virtual leads are very similar to the copper complex but quite different from metformin alone, and a laboratory confirmation of their activity should plausibly imply that metformin acts in synergy with copper, giving us the ability to design new antidiabetic drugs in a novel and more rational fashion, with significant savings in research costs and efforts.

SOME INDICES OF MORPHOFUNCTIONAL ACTIVITY OF LYMPHOCYTES IN THE PERIPHERAL BLOOD OF WOMEN WITH COMPLICATED PREGNANCY

The aim of the research was to study some indices of morphofunctional state of lymphocytes in the peripheral blood under a complicated course of pregnancy in women of Khabarovsk. 96 women were examined in different trimesters of gestation. Two main groups depending on the type of complications were formed: the first group consisted of pregnant women with vaginitis, the second group had women whose pregnancy was complicated with clinical manifestations in the form of edemas, proteinuria and hypertensive disorders. The group of comparison included the women without complications. In the peripheral blood of the women under study with the method of high tech automated digital system of blood smear test VISION HEMA there was done a morphometric assessment of lymphocytes and with histochemical methods there was found a level of fermentation activity of succinate dehydrogenase (SDH), α-glycerophosphate dehydrogenase (α-GPDH) and lactate dehydrogenase (LDH). By morphometric studies of lymphocytes of the peripheral blood in pregnant women of the first group in the III trimester of gestation the indices of the square of the cell, of cytoplasm, cytoplasm brightness, the square of the nucleus and the brightness of the nucleus were 102.33±9.83 mсm², 36.84±4.88 mсm², 181.39±6.94 units of optical density, 65.69±6.87 mсm², 97.44±6.88 units of optical density, respectively (in the group of the comparison these data were 121.31±4.44 mсm²; 46.14±2.52 mсm²; 167.74±3.44 units of optical density; 75.33±2.66 mkm², 88.00±3.36 units of optical density, respectively). In the second group of women the same data in the same period of pregnancy corresponded to the following values: 128.44±4.89 mсm², 57.15±3.87 mсm², 173.53±4.65 units of optical density, 70.95±3.39 mсm² and 91.13±4.93 units of optical density (in the group of comparison there were the following data: 109.92±3.57 mсm², 39.63±1.53 mсm², 158.96±2.63 units of optical density, 75.37±3.23 mсm², 82.16±2.58 units of optical density, respectively). By the results of histochemical studies the level of activity of enzymes SDH, α-GPDH, LDH in the first group of pregnancy was 15.38±2.10, 10.72±0.58 and 20.53±2.36 pellet/cell, respectively. And just the other way round in pregnant women of the second group there was revealed a statistically reliable suppression of fermentation activity of these enzymes: 9.11±0.75 (p&#60;0.05); 9.61±1.06 (p&#62;0.05); 8.31±1.20 (p&#60;0.05) pellet/cell in regard to the data in the group of comparison (12.84±1.21; 12.18±1.13 and 13.09±0.78 pellet/cell, respectively). Thus the changes of morphometric indices characterizing parameters of the cell, the nucleus and lymphocytes cytoplasm in a complex study with their fermentation activity can be extra diagnostic criteria of the formation of different complications during pregnancy. The obtained data allow to justify in-time application of medications for the correction of metabolic disorders of immune competent cells with the aim to decrease unfavorable perinatal outcomes.