Heterozygous CAPN3 missense variants causing autosomal-dominant calpainopathy in seven unrelated families.

AIMS: Recessive variants in CAPN3 gene are the cause of the commonest form of autosomal recessive limb girdle muscle dystrophy. However, two distinct in-frame deletions in CAPN3 (NM_000070.3:c.643_663del21 and c.598_621del15) and more recently, Gly445Arg and Arg572Pro substitutions have been linked to autosomal dominant (AD) forms of calpainopathy. We report 21 affected individuals from seven unrelated families presenting with an autosomal dominant form of muscular dystrophy associated with five different heterozygous missense variants in CAPN. METHODS: We have used massively parallel gene sequencing (MPS) to determine the genetic basis of a dominant form of limb girdle muscular dystrophy in affected individuals from seven unrelated families. RESULTS: The c.700G> A, [p.(Gly234Arg)], c.1327T> C [p.(Ser443Pro], c.1333G> A [p.(Gly445Arg)], c.1661A> C [p.(Tyr554Ser)] and c.1706T> C [p.(Phe569Ser)] CAPN3 variants were identified. Affected individuals presented in young adulthood with progressive proximal and axial weakness, waddling walking and scapular winging or with isolated hyperCKaemia. Muscle imaging showed fatty replacement of paraspinal muscles, variable degrees of involvement of the gluteal muscles, and the posterior compartment of the thigh and minor changes at the mid-leg level. Muscle biopsies revealed mild myopathic changes. Western blot analysis revealed a clear reduction in calpain 3 in skeletal muscle relative to controls. Protein modelling of these variants on the predicted structure of calpain 3 revealed that all variants are located in proximity to the calmodulin-binding site and are predicted to interfere with proteolytic activation. CONCLUSIONS: We expand the genotypic spectrum of CAPN3-associated muscular dystrophy due to autosomal dominant missense variants.

Long-term administration of the TNF blocking drug Remicade (cV1q) to mdx mice reduces skeletal and cardiac muscle fibrosis, but negatively impacts cardiac function.

Duchenne muscular dystrophy (DMD) is a degenerative skeletal muscle disease caused by mutations in the gene encoding dystrophin (DYS). Tumor necrosis factor (TNF) has been implicated in the pathogenesis since short-term treatment of mdx mice with TNF blocking drugs proved beneficial; however, it is not clear whether long-term treatment will also improve long-term outcomes of fibrosis and cardiac health. In this investigation, short and long-term dosing studies were carried out using the TNF blocking drug Remicade and a variety of outcome measures were assessed. Here we show no demonstrable benefit to muscle strength or morphology with 10mg/kg or 20mg/kg Remicade; however, 3mg/kg produced positive strength benefits. Remicade treatment correlated with reductions in myostatin mRNA in the heart, and concomitant reductions in cardiac and skeletal fibrosis. Surprisingly, although Remicade treated mdx hearts were less fibrotic, reductions in LV mass and ejection fraction were also observed, and these changes coincided with reductions in AKT phosphorylation on threonine 308. Thus, TNF blockade benefits mdx skeletal muscle strength and fibrosis, but negatively impacts AKT activation, leading to deleterious changes to dystrophic heart function. These studies uncover a previously unknown relationship between TNF blockade and alteration of muscle growth signaling pathways.

Impaired calcium calmodulin kinase signaling and muscle adaptation response in the absence of calpain 3.

Mutations in the non-lysosomal, cysteine protease calpain 3 (CAPN3) result in the disease limb girdle muscular dystrophy type 2A (LGMD2A). CAPN3 is localized to several subcellular compartments, including triads, where it plays a structural, rather than a proteolytic, role. In the absence of CAPN3, several triad components are reduced, including the major Ca(2+) release channel, ryanodine receptor (RyR). Furthermore, Ca(2+) release upon excitation is impaired in the absence of CAPN3. In the present study, we show that Ca-calmodulin protein kinase II (CaMKII) signaling is compromised in CAPN3 knockout (C3KO) mice. The CaMK pathway has been previously implicated in promoting the slow skeletal muscle phenotype. As expected, the decrease in CaMKII signaling that was observed in the absence of CAPN3 is associated with a reduction in the slow versus fast muscle fiber phenotype. We show that muscles of WT mice subjected to exercise training activate the CaMKII signaling pathway and increase expression of the slow form of myosin; however, muscles of C3KO mice do not exhibit these adaptive changes to exercise. These data strongly suggest that skeletal muscle's adaptive response to functional demand is compromised in the absence of CAPN3. In agreement with our mouse studies, RyR levels were also decreased in biopsies from LGMD2A patients. Moreover, we observed a preferential pathological involvement of slow fibers in LGMD2A biopsies. Thus, impaired CaMKII signaling and, as a result, a weakened muscle adaptation response identify a novel mechanism that may underlie LGMD2A and suggest a pharmacological target that should be explored for therapy.

[Current state of epidemiologic surveillance of natural foci of plague of North Caucasus]. / Sovremennoe sostoianie épidemiologicheskogo nadzora v prirodnykh ochagakh chumy Severnogo Kavkaza.

Information on the epizootic situation in plague in the natural foci of North Caucasus and on the influence of a number of anthropogenic and natural factors on this situation is presented. The data given in this work indicate that under the conditions of the anthropogenic transformation of landscapes the character of the epizootic manifestations of plague is changed and new factors, capable of aggravating epidemiological situation, appear. In addition, some other factors must be considered, such as the insufficient financing of reliable field surveys at present, the impossibility of making reliable epizootological studies due to causes of the social character (armed conflicts), thus making it impossible to evaluate, with a sufficient degree of reliability, the real epizootic state of a number of territories and, therefore, the risk of human infection. In this connection the necessity to carefully plan prophylactic measures and measures aimed at the localization and liquidation of the probable foci of infection arises.

[Principles of predicting the density of rodent flea populations]. / Printsip prognoza plotnosti populiatsii blokh gryzunov.

The paper gives the indices of abundance of fleas of different species of rodents in their fur and in their burrows and lays emphasis on the general mechanisms by which the density of these blood-sucking insects is a species-specific (populational) sign and does not greatly differ from the mean many-year values. By using Citellophylus tesquorum and other specias as an example, the authors present the results of a study of natural mortality in fleas, which suggest that the mean longevity of imagoes and death in the preimaginal phases of development are a hereditary sign. An example of calculation of the death ratio "K" in the imaginal phases is given and evidence is provided for compiling the signs of the density of fleas in taking individual samples of a field material and calculating the abundance indices of specific species.

Partial purification and biochemical characterization of a heteromeric protein phosphatase 2A holoenzyme from maize (Zea mays L.) leaves that dephosphorylates C4 phosophoenolpyruvate carboxylase.

The activity and allosteric properties of plant phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) are controlled posttranslationally by specific reversible phosphorylation of a strictly conserved serine residue near the N-terminus. This up/down-regulation of PEPC is catalyzed by a dedicated and highly regulated serine/threonine (Ser/Thr) kinase (PEPC-kinase) and an opposing type-2A Ser/Thr phosphatase (PP2A). In marked contrast to PEPC-kinase, the PP2A holoenzyme from photosynthetic tissue has been virtually unstudied to date. In the present investigation, we have partially purified and characterized the native form of this PP2A from illuminated leaves of maize (Zea mays L.), a C4 plant, using maize [32P]PEPC as substrate. Various conventional chromatographic matrices, together with thiophosphorylated C4 PEPC-peptide and microcystin-LR affinity-supports, were exploited for the enrichment of this PP2A from soluble leaf extracts. Biochemical and immunological results indicate that the C4-leaf holoenzyme is analogous to other eukaryotic PP2As in being a approximately 170-kDa heteromer comprised of a core PP2Ac-A heterodimer (approximately 38- and approximately 65-kDa subunits, respectively) complexed with a putative, approximately 74-kDa B-type regulatory/targeting subunit. This heterotrimer lacks any strict substrate specificity in that it dephosphorylates C4 PEPC, mammalian phosphorylase a, and casein in vitro. This activity is independent of free Me2+, insensitive to levamisole and the Inhibitor-2 protein that targets PP1, activated by several polycations such as protamine and poly-L-lysine, and highly sensitive to inhibition by microcystin-LR and okadaic acid (IC50 approximately 30 pM), all of which are diagnostic features of yeast and mammalian PP2As. In addition, this C4-leaf PP2A holoenzyme (i) is inhibited in vitro by physiological concentrations of certain C4 PEPC-related metabolites (L-malate, PEP, glucose 6-phosphate, but not the activator glycine) when either 32P-labeled maize PEPC or rabbit muscle phosphorylase a is used as substrate, suggesting a direct effect on this Ser/Thr phosphatase; and (ii) displays, at best, only modest light/dark effects in vivo on its apparent molecular mass, component core subunits and activity against C4 PEPC, in marked contrast to the opposing activity of PEPC-kinase in C4 and Crassulacean acid metabolism leaves. This report represents one of the few studies of a heteromeric PP2A holoenzyme from photosynthetic tissue that dephosphorylates a known target enzyme in plants, such as PEPC, sucrose-phosphate synthase or nitrate reductase.

[Role of some wild birds and mammals in the natural foci of Crimean haemorrhagic fever in Stavropol' region]. / Rol' nekotorykh dikikh ptits i mlekopitaiushchikh v prirodnoi ochagovosti krymskoi gemorragicheskoi likhoradki v Stavropol'skom krae.

The results of the study on the role of some wild mammals and birds as feeding sources of for ticks Hyalomma marginatum, the main vectors of Crimean haemorrhagic fever virus in the Stavropol Territory, in the preimago phases of their development are presented. These phases (larvae and nymphs) were found on rooks, hooded crows, partridges, European brown hares and eared hedgehogs. The examination of domestic fowl resulted in finding larvae and nymphs in small amounts on turkeys. According to the data of the epizootological survey carried out in summer and autumn of 2000 in the Stavropol Territory, rooks and, to a lesser extent, hares were found to be the main feeding sources for ticks in the preimago phases. Rodents seemed to be of minimal importance as feeding sources under the conditions of the Stavropol Territory. Of all animals, rooks must be the main object in the epizootological survey of the territory.

[Main positions with respect to the formation of the planning and norm-setting basis of medical provision]. / Osnovnye polozheniia po formirovaniiu planovo-normativnoi basy meditsinskogo obespecheniia.

Concept on the formation of the design and norm-setting basis of medical service has been developed in order to improve planning and balance between the scope of rendered services and their financial provision.

[Isolation and characteristics of a new trypsin and chymotrypsin inhibitor from potato tubers]. / Vydelenie i kharakteristika novogo ingibitora tripsina i khimotripsina iz klubnei kartofelia.

A novel trypsin and chymotrypsin inhibitor has been isolated from potato (Solanum tuberosum L.) tubers. The isolation procedure included ammonium sulfate precipitation, gel-chromatography on Sephadex G-75 and ion-exchange chromatography on DEAE-cellulose. The inhibitor interacts with trypsin and chymotrypsin at a molar ratio of 1:1. The substrate-dependent dissociation of the enzyme-inhibitor complexes is observed. The inhibitor displays no activity towards subtilisin and pancreatic elastase. The ability of the inhibitor to form a ternary complex containing simultaneously both trypsin and chymotrypsin molecules testifies to the presence of two independent reactive sites for these enzymes.

[Effectiveness of various methods of the surgical treatment in cervix rupture during labor]. / Effektivnost' razlichnykh metodov operativnogo lecheniia razryvov sheiki matki v rodakh.

Long-term results of wound healing in patients with cervical ruptures were presented. The use of two-rowed and one-rowed catgut sutures in 384 and 98 parturients, respectively, was discussed. Employment of two-rowed sutures resulted in 95.3 per cent of primary wound healing. The technique mentioned was a success in all the cases observed.