Performances of the assay MTBDRplus(®) in the surveillance of rifampicin resistance in Mycobacterium tuberculosis

The purpose of the survey was the routine assessment of the MTBDRplus^® kit performance in the determination and characterization of Mycobacterium tuberculosis resistance to rifampicin. The survey was carried out on a collection of 144 strains (126 of which were resistant to rifampicin) isolated on patients from 15 countries. Sensitivity to antituberculosis drugs was determined by a liquid culture system and the reference method was the amplification and sequencing of a target region of the rpoB gene whose mutations are responsible for rifampicin resistance (codons 507 to 533). The assessed kit was based on a reverse hybridization technique using eight overlapping probes covering the target region and four probes representing the most-frequently observed mutations. The assay performance was found excellent, specificity: 100%, sensitivity: 99.2%; 17 mutations affecting 10 codons were reported, two of which were newly identified.     

Changes in the Nature of Behavior and the Activity of the Hypophyseal-Adrenocortical System in the Offspring of Paternal Rats Subjected to Stress in the Stress–Restress Paradigm before Mating

Neuroscience and Behavioral Physiology - We report here studies of the effects of stressing male rats in the stress–restress paradigm, which is a model of post-traumatic stress disorder...     

Can we expect progress in the treatment of fibrosis in the course of chronic pancreatitis?

Chronic pancreatitis (CP) is a necroinflammatory process characterized by loss of both exocrine and endocrine function. To date, the disease has been treated symptomatically. Real advances in CP management can be expected once the pathophysiology of the disease is elucidated and individual stages of its development are properly managed. A key role in the CP pathogenesis is played by activation of pancreatic stellate cells (PSCs) that cooperate with the remaining pancreatic cells. All these cells produce cytokines, growth factors, angiotensin and other substances, which paracrinally or autocrinally induce further, persistent activation of PSCs. The activated PSCs are capable of producing and modifying the extracellular matrix. An optimal therapeutic preparation should exert beneficial effects on all the above-mentioned phenomena observed in CP. The most promising treatment modalities include blocking of the renin-angiotensin system (RAS), activation of peroxisome proliferator-activated receptors gamma (PPAR-γ), influence on the remaining PSC signaling pathways, blocking of substances produced by activated PSCs, and antioxidants. The findings of many recent experimental studies are highly encouraging; however, their efficacy should be confirmed in well-designed clinical trials.     

On the Morphology of the Synaptic Vesicles of the Axospine Synapses in the Rat Visual Cortex

An electron-microscopoical Observationof the shape and form of the synapticvesicles in the axospine synapses of therat visual cortex fixed with glutaralde-hyde and postfixed with osmium tetro-xide is reported.Each axospine synapse in the visualcortex consists of a presynaptic boutonwith a dendritic spine from which it is     

Role of SIRPα in regulation of mucosal immunity in the intestine

Mononuclear phagocytes such as dendritic cells (DCs) and macrophages in the lamina propria (LP) are thought to be important for both induction of inflammatory responses and maintenance of immunologic tolerance in the mammalian intestine. The molecular mechanisms by which these cells regulate intestinal immunity have remained poorly understood, however. Signal regulatory protein α (SIRPα) is a transmembrane protein that is specifically expressed in DCs, macrophages and neutrophils. Here, we show that SIRPα is abundant in CD11c(+) CD11b(+) LP cells of the mouse intestine. Whereas SIRPα did not appear to be important for the steady-state homeostasis of mucosal immunity in the intestine, the flagellin-stimulated production of IL-17 or interferon (IFN)-γ by LP cells of SIRPα mutant (MT) mice that lack the cytoplasmic region of the protein was markedly decreased compared with that observed with wild-type cells. Moreover, the flagellin-induced production of IL-6 by LP cells from SIRPα MT mice was also greatly reduced. SIRPα MT mice were also resistant to the development of colitis induced by IL-10 deficiency. Our data thus suggest that SIRPα expressed on CD11c(+) LP cells is important for the production of IL-17 or IFN-γ in the LP as well as for the development of colitis induced by IL-10 deficiency.     

Anthropological assessment of changes in living conditions of the rural population in Poland in the period 1967–2001

Background: Poland is considered an ethnically homogeneous country, with no significant national, linguistic, religious or racial minorities. Thus, social differences in rates of maturation, height and weight may be assumed to contain a negligible genetic component and serve as a reflection of environment, i.e. living conditions.

Aim: This study seeks to determine whether changes in economic conditions in Poland, in particular the acute economic crisis of 1977–1989 and the transformation of the political system in 1989, had an effect on the biological status of girls from various categories of the rural population.

Subjects and methods: Rural girls aged 9.5–18.5 years were studied in 1967 (n?=?7889), 1977 (n?=?7771), 1987 (n?=?13?556) and in 2001 (n?=?9599). The stratification of participants (farmers, farm-workers and non-farmers) was based on the source of their family income, parents' education, number of children per family and household appliances. Age at menarche (AM), body height, and weight were used as biological indicators of living conditions.

Results: During the decade 1967–1977, while a relatively good economic situation prevailed in the country, AM decreased by 0.64 years and distinct secular trends in height and weight were noted. During the decade 1977–1987, years of economic crisis, secular trends were arrested and AM increased by 0.11 years. Landless rural families were more strongly affected by food shortages than were farmers who were the food producers. The study, repeated in 2001, showed positive secular trends in body height and a decrease in AM of 0.24 years for decade for daughters of farmers this decrease in AM was twice as high as in non-farmer families. The latter group experienced acute unemployment after the political and economic system transformation (1989). AM was earliest in daughters of non-farmers, and latest in those from farmer families. In 1967, the difference between the mean ages at AM for these groups amounted to 0.53 years, in 1977 to 0.44 years, in 1987 to 0.33 years and to only 0.15 years in 2001.

Conclusion: The categories of the rural population, farmers, farm-workers and landless rural inhabitants were variously affected by the economic crisis, as well as by the process of economic transformation. This shows that living conditions of each of those categories changed in different ways and to a different degree during the years 1967–2001. Farmers' families achieved the highest social advancement, as the AM of girls from those families decreased by 0.98 years compared to those from farmer-worker and landless rural families, which decreased by 0.85 and 0.60 years, respectively.     

Assessing the role of Asp 194 in the transmembrane domains of the α-chain of the high-affinity receptor complex for immunoglobulin E in signal transduction

The high-affinity receptor complex for IgE plays a pivotal role in allergic responses since cross-linking of the high-affinity IgE receptor (Fc?RI) on target cells initiates a signaling cascade facilitating release of inflammatory mediators causing allergic responses. The transmembrane regions of the ligand binding domains of the high-affinity IgE and low-affinity IgG receptors share an invariant motif (LFAVDTGL) containing a polar aspartate within a predominantly non-polar setting. The functional importance of this aspartate residue (D194) in Fc?RI-mediated receptor signaling was assessed by site-directed mutagenesis. Rat basophilic leukemia cells (RBL-2H3) transfected with the human IgE binding subunit (Fc?RIα) incorporating polar substitutions like asparagine (D194N) or threonine (D194T) resulted in the formation of a functional rat/human chimeric receptor complex. When activated via huIgE and antigen, cells transfected with these variant receptor subunits supported mediator release, intracellular calcium mobilisation and tyrosine phosphorylation of γ-chain and Syk kinase while a non-polar substitution (D194L) gave rise to cell surface expression of the mutated receptor subunit but failed to initiate downstream signaling. No cell surface expression of huFc?RIα gene constructs was observed when D194 was replaced with the non-polar Ile (D194I) residue of similar size, the larger positively charged Arg (D194R) or lysine (D194K) residues, or the negatively charged glutamate (D194E) and smaller polar Ser (D194S) non-polar Ala (D194A) and V (D194V). These observations highlight importance of the size and charge of amino acid residue at position 194 in determining IgE receptor subunit interactions, cell surface localization, and initiation of downstream signaling events.     

A new evidence for the maintenance of the sarcoglycan complex in muscle sarcolemma in spite of the primary absence of δ-SG protein

delta-Sarcoglycan (delta-SG) is one of the first proteins of the sarcoglycan complex (SGC) to be expressed during muscle development, and it has been considered fundamental for the assembling and insertion of the SGC in the sarcolemma. Studies using heterologous cell systems and co-precipitation have demonstrated that SGC assembly was dependent on the simultaneous synthesis of all four sarcoglycan proteins. Mutations in any one of sarcoglycan genes, including the common disease causing mutation c.656delC in the delta-SG gene, block complex formation and its insertion in the plasma membrane. Failure in complex assembly in patients with this mutation would be therefore expected. In this study, we provide evidence for the possibility of preservation of part of the SG complex in the sarcolemma, even in the absence of delta-SG. This is based on the study of one mildly affected patient with limb-girdle muscular dystrophy type 2F (LGMD2F) due to the homozygous c.656delC mutation in the delta-SG gene. Protein analysis in his muscle biopsy presented a significant deficiency of only delta-SG with retention of the other three SG proteins in the sarcolemma. RNA expression analysis showed that zeta-SG, a functionally homologous to delta-SG, is not atypically upregulated in his muscle and would not replace the absent delta-SG, retaining the complex alpha-beta-gamma-zeta. The patient started clinical manifestation at age 25, with frequent falls, but he is currently able to walk unassisted at age 42. His clinical course is significantly milder when compared to several other affected patients carrying the same mutation associated with a total deficiency of the four SG proteins in the muscle studied by our group and confirmed in other patients. Therefore, our results add a new in vivo evidence that alpha-, beta-, and gamma-SG proteins can be maintained in the sarcolemma without delta-SG. Additionally, LGMD2F, with retention of the part of the SGC, might be associated to a milder clinical course, which has important implications for clinical prognosis and genetic counseling of the family.     

Colocalization of Neurotensin and Corticotropin-Releasing Factor in Oval Nucleus of the Bed Nuclei of Stria Terminalis in the Rat

Ju et al first parcellated in detail the bed nuclei of stria terminalis according to its cyto-and chemoarchitecture. They found that the oval nucleus (OV) was rich in neurotensin (NT)-and corticotropin-releasing factor (CRF)like immunoreactivities. Using indirect immunohistochemical technique it has been shown in the present study that both neuropeptides show quite similar distribution pattern in OV of the Sprague-     

End of life in ICU--care of the dying or 'pulling the plug'?

This study, a modified subsection of the European ETHICUS study on End-of-Life (EOL) Decision Making in the Intensive Care Unit (ICU), examines the pattern of limiting futile life-sustaining therapies in an Irish ICU including the practice of withdrawing mechanical ventilation in anticipation of death. 1146 patients were admitted to the Mater Hospital, Dublin ICU from 1/9/1999 to 30/6/2000 and all 126 patients who died in ICU were included. EOL categories were prospectively defined (by Ethicus methodology) as cardiopulmonary resuscitation (CPR); brain death; withholding (WH); withdrawing (WD) life sustaining therapy and active shortening of the dying process (SDP). Complete data were obtained for 122 of the 126 patients who died during this period. 45 patients (36%) had therapy withheld, 40 (33%) had therapy withdrawn, 26 (21%) had unsuccessful CPR and 11 (10%) were Brain Dead. SDP was not performed. In total, 85 patients had a limitation of life sustaining therapy. CPR was the main therapy withheld (96% of WH/WD patients). Inotropic infusions were limited (WH or WD) in 40/85 (47%) of patients. Fluids, feeding and oxygen were rarely withdrawn (2.4%, 6%, 4.8% respectively). Twenty-two patients had two or more EOL decisions. Tracheal extubation or withdrawal of ventilation was less frequent (16.4%) but more common if a second EOL decision was made. No patient had sedation withdrawn or decreased. Eight patients of 85 (9%) had sedation increased. The study demonstrates that EOL decision making is common (69% of deaths and 7.4% of ICU admissions) in Ireland and demonstrates that the pattern of treatment limitation relates primarily to cardiovascular and other treatments and less to respiratory life sustaining treatment. Artificial nutrition and hydration were rarely withdrawn.     

What is the role of rituximab in the treatment of rheumatoid arthritis?

Rituximab is a monoclonal antibody against CD20 that was developed for the treatment of relapsed or refractory B-cell non-Hodgkin's lymphoma (NHL). Recent controlled trials have shown that B cell-targeted therapy with rituximab is effective in RA (which suggest that B lymphocytes may be critical in its pathogenesis of RA) and early exposure data suggest that the tolerability and safety profile of rituximab may be even better in RA than in NHL patients. Rituximab is generally well tolerated, with a low incidence of serious adverse events, including serious infections. Available evidence suggests that its clinical benefits depend on effective B cell depletion, and the fact that its novel mode of action leads to the depletion of B cells makes it distinct from other biological therapies for RA that target T cells and their related cytokines. Although complete peripheral B cell depletion is regularly seen in RA and other autoimmune diseases, especially systemic lupus erythematosus (SLE), incomplete depletion has been reported in a subset of patients, even after full dosing with rituximab.     

Correlation of Aβ deposition in the nasal cavity with the formation of senile plaques in the brain of a transgenic mouse model of Alzheimer's disease

The deposition of β-amyloid peptides (Aβ) is commonly reported in the nasal cavity of Alzheimer's disease (AD) patients, although the pathological significance of this finding is unknown. This study compared Aβ concentrations in the nasal area with those in the brain, blood, and cerebrospinal fluid, respectively. Immunohistochemical analysis identified Aβ deposits in the nasal epithelium of Tg2576 mice. Enzyme-linked immunosorbent assay measurements revealed a correlation between the content of Aβ42 in the nasal area and that in the brain, but not with that in the blood. These results suggest that the highly accessible nasal cavity could be a useful site for diagnostic analysis of AD based on Aβ content.     

The Role of β1,2-Adrenoceptors in the Amygdala in the Behavior of Rats with Different Levels of Freezing in Conditioned Reflex Fear

Neuroscience and Behavioral Physiology - Individual sensitivity to blockade of β1,2-adrenoceptors was studied by dividing rats into groups depending on the manifestation of conditioned reflex...     

Intensity of Proliferative Processes and Degree of Oxidative Stress in the Mucosa of the Ileum in Crohn’s Disease

The intensity of proliferative processes (estimated from Ki-67 expression) and degree oxidative stress (chemiluminescence assay) in biopsy specimens from the terminal portion of the ileal mucosa were studied in patients with Crohn’s disease. Crohn’s disease is characterized by hyper-regenerative processes in the ileal mucosa. The labeling index (Ki-67 expression) in biopsy specimens from the intact ileal mucosa in patients with the irritable bowel syndrome (reference group) was 10.64 ± 0.62%. The corresponding values in patients receiving monotherapy with mesalazine (group 1) and combination therapy with mesalazine and dalargin (group 2) were 24.05 ± 1.17 and 22.90 ± 0.92%, respectively. Analysis of free radical oxidation showed that this state is accompanied oxidative stress. Spontaneous and H₂O₂-induced luminol-dependent chemiluminescence in biopsy specimens from the ileal mucosa was 1.8-2.3-fold higher compared to the reference group. After therapy, the labeling index in groups 1 and 2 decreased to 18.60 ± 1.18 and 14.38 ± 0.81%, respectively. Histologically, normalization of the disease symptoms was more pronounced after combination therapy. The decrease in free radical oxidation in this group of patients was more pronounced than after mesalazine monotherapy. Our results suggest that oxidative stress plays a role in the hyper-regenerative reaction.     

The role of the acetylcholine — Cholinesterase system in the origin of bioelectrical phenomena in skeletal muscle

Summary Poisoning of the isolated sartorius muscle in frog with eserine and strychnine in the 110⁵ and 110⁴ concentrations causes the change of the value and the form of the excitation biocurrent. Strychnine and eserine in the 110⁵, 110⁴ and 110³ concentrations have no effect on the electromotive force of the sartorius muscle injury. These data lead us to presume certain differences in the mechanism of the origin of the excitation and injury biocurrents in the skeletal muscle, at least with relation to the participation of the acetylcholine-cho-linesterase system in this process.Presented by Academician the late L. A. Orbeli