Appearance of levator ani muscle subdivision defects on level III vaginal support structures in women with and without pelvic organ prolapse: an MRI study.

INTRODUCTION AND HYPOTHESIS: Injury of the levator ani muscle (LAM) is a significant risk factor for pelvic organ prolapse (POP). The puborectalis (PRM) and pubovisceral (PVM) subdivisions are level III vaginal support structures. The null hypothesis was that there is no significant difference in patterns of LAM subdivisions in healthy nulliparous women. Secondarily, we evaluated the presence of different LAM injury in a POP-symptomatic cohort. METHODS: This retrospective magnetic resonance imaging study included: 64 nulligravidae without any pelvic floor dysfunction (PFD) and 526 women of various parity with symptomatic POP. Primary outcome was PVM and PRM morphology on the axial planes: the attachment site on the pubic bone, and the visible separation/border between the PVM and PRM. The attachment was scored as "normal" or "abnormal". The "abnormal" attachment was divided in two types: "type I"-loss of the muscle substance, but preservation of the overall muscle architecture-and "type II"-muscle detachment from the pubic bone. RESULTS: The puboanal muscle (PAM) subdivision was evaluated as a representative part of the PVM. The PAM and PRM attachments and separation were distinguished in all asymptomatic nulliparae. PAM and PRM attachments did not significantly differ. POP group characteristics were parity 1.9 ± 0.8, instrumental delivery 5.6%, hysterectomy or POP surgery 60%, all Pelvic Organ Prolapse Quantification (POP-Q) stages, LAM defect 77.6% (PRM: 77.1%; PAM: 51.3%). Type I injuries were more frequent (PRM 54.7%; PAM 53.9%) compared with type II (PRM 29.4%; PAM 42.1%). CONCLUSIONS: A LAM defect was present in 77.6% of women with symptomatic POP. In PRM and PAM subdivisions type I injury was more frequent than type II.

Altered dynamics of lipid metabolism in muscle cells from patients with idiopathic inflammatory myopathy is ameliorated by 6 months of training.

KEY POINTS: Regular exercise improves muscle functional capacity and clinical state of patients with idiopathic inflammatory myopathy (IIM). In our study, we used an in vitro model of human primary muscle cell cultures, derived from IIM patients before and after a 6-month intensive supervised training intervention to assess the impact of disease and exercise on lipid metabolism dynamics. We provide evidence that muscle cells from IIM patients display altered dynamics of lipid metabolism and impaired adaptive response to saturated fatty acid load compared to healthy controls. A 6-month intensive supervised exercise training intervention in patients with IIM mitigated disease effects in their cultured muscle cells, improving or normalizing their capacity to handle lipids. These findings highlight the putative role of intrinsic metabolic defects of skeletal muscle in the pathogenesis of IIM and the positive impact of exercise, maintained in vitro by yet unknown epigenetic mechanisms. ABSTRACT: Exercise improves skeletal muscle function, clinical state and quality of life in patients with idiopathic inflammatory myopathy (IIM). Our aim was to identify disease-related metabolic perturbations and the impact of exercise in skeletal muscle cells of IIM patients. Patients underwent a 6-month intensive supervised training intervention. Muscle function, anthropometric and metabolic parameters were examined and muscle cell cultures were established (m. vastus lateralis; Bergström needle biopsy) before and after training from patients and sedentary age/sex/body mass index-matched controls. [14 C]Palmitate was used to determine fat oxidation and lipid synthesis (thin layer chromatography). Cells were exposed to a chronic (3 days) and acute (3 h) metabolic challenge (the saturated fatty acid palmitate, 100 µm). Reduced oxidative (intermediate metabolites, -49%, P = 0.034) and non-oxidative (diglycerides, -38%, P = 0.013) lipid metabolism was identified in palmitate-treated muscle cells from IIM patients compared to controls. Three days of palmitate exposure elicited distinct regulation of oxidative phosphorylation (OxPHOS) complex IV and complex V/ATP synthase (P = 0.012/0.005) and adipose triglyceride lipase in patients compared to controls (P = 0.045) (immunoblotting). Importantly, 6 months of training in IIM patients improved lipid metabolism (CO2 , P = 0.010; intermediate metabolites, P = 0.041) and activation of AMP kinase (P = 0.007), and nearly normalized palmitate-induced changes in OxPHOS proteins in myotubes from IIM patients, in parallel with improvements of patients' clinical state. Myotubes from IIM patients displayed altered dynamics of lipid metabolism and impaired response to metabolic challenge with saturated fatty acid. Our observations suggest that metabolic defects intrinsic to skeletal muscle could represent non-immune pathomechanisms, which can contribute to muscle weakness in IIM. A 6-month training intervention mitigated disease effects in muscle cells in vitro, indicating the existence of epigenetic regulatory mechanisms.

Coincidental finding of pelvic splenosis during gynecological surgery.

OBJECTIVE: To describe a case of coincidental finding of splenosis during gynecological laparoscopic surgery. DESIGN: Case report. SETTING: Department of gynecology and obstetrics, Hospital of Frýdek-Místek. CASE REPORT: We describe the case of coincidental perioperative finding of pelvic tumorous mass that was later histologicaly verified as accsessory spleen or splenosis. CONCLUSION: Ectopic spleen is mostly random finding with no symptoms observed. Rarely it can cause pelvic pain or mimick adnexal malignity or endometriosis. It is neccesery to keep this rare diagnosis in mind, mainly at patients with splenic trauma or splenectomy in their medical history to preserve immunological function of this splenic tissue.

Laparoscopic correction of isthmocele combined with ventrosuspensios of uterus.

OBJECTIVE: To describe a case history of a patient after two caesarean sections, planning another pregnancy. Due to the dehiscent lower uterine segment, surgical correction of the defect was performed. Performance followed by an unplanned pregnancy five weeks after the operation. DESIGN: Case report. SETTING: Department of Obstetrics and Gynaecology, Hospital in Frýdek-Místek. CASE REPORT: We present a case of a 31-year-old third-graders, anamnestically after two caesarean sections, which were performed laparoscopical correction of isthmocoele in our department. Our patient was diagnosed with six weeks old intrauterine pregnancy only eleven weeks after surgery. The gravidity was successfully completed in the 38th week of pregnancy by the planned caesarean section with finding of a solid lower uterine segment. Whole duration of the pregnancy was uncomplicated. CONCLUSION: Women, after previous surgery of the uterus, are exposed to complications such as nidation disorders, placental disorders, risk of uterine rupture etc. during future pregnancy and childbirth. We want to show possible advantage of laparoscopic isthmocoele resection in combination with ventrosuspension of uterus.

Anatomy and biomechanic of the musculus levator ani.

OBJECTIVE: The aim of the rewiew is to provide complex new informations about anatomy and biomechanics features of the musculus levator ani. Described are risk factors leading to it´s injury and options of imaging the muscle complex (ultrasound, magnetic imaging resonance and 3D modeling). DESIGN: Review. SETTINGS: Departement of Obstetrics and Gynaecology, Hospital in Frýdek-Místek, GONA Co. Ltd , Institute for Mother and Child Prague. RESULTS: Musculus levator ani (MLA) has a complex structure composed mainly of striated muscles. Minority of smooth muscle fibres are also found. Particular parts of the MLA hold together different angles. Inervation is provided through somatic and visceral nerve fibres. During delivery, more there three times stretching of the muscle was observed. Less strenght is needed do the same stretching of the muscle in repeating stress situations. In the MRI studies, two types of injury of the MLA, were found. Predisponed to the injury is medial part of the MLA known as pubovisceral muscle (PVM). PVM has three insertions. The most fragile is it´s medial insertion to the pubic bone described as enthesis. During experimental delivery studies was found, that the pressure in this part of the muscle reach almost 36MPa. CONCLUSION: MLA is a difficult muscle. Because of the ethical reasons we don´t have, and probably never will have informations, how structuraly and elasticaly differs muscle, that was damaged during the delivery, compared to muscle without any damage. Promising are computer delivery simulations. In future, they would give us an answer, how risky is vaginal delivery in concrete expectant mother.

[MRI analysis of the musculo-fascial component of pelvic floor in woman before planned vaginal reconstruction procedur for symptomatic pelvic organ prolapse]. / Analýza stavu muskulo-fasciální slozky pánevního dna pomocí MRI u pacientek pred plánovaným vaginálním rekonstrukcním výkonem pro symptomatický sestup pánevního dna.

OBJECTIVE: The aim of the study is to analyse the musculo-fascial component of the pelvic floor in symptomatic group of woman with pelvic organ prolapse before planned vaginal reconstruction using synthetic vaginal mesh. DESIGN: Observational cohort study. SETTING: Department of Obstetrics and Gynaecology, Hospital in Frýdek-Místek; GONA Ltd, Prague; Institute for Care of Mother and Child, Prague; 3rd Faculty of Medicine CHU Prague. METHODOLOGY: The study involved 285 female volunteers (6 nulliparous, all other patients gave birth vaginally at least once) that in the period 2008-2015 before the planned reconstructive vaginal operations have undergone a comprehensive urogynaecology examination supplemented by magnetic resonance imaging (MRI) of the pelvic floor. Assessed was musculofascial component of the pelvic floor containing -musculus levator ani (MLA), endopelvic fascia (EF) and sacrouterine ligaments (SUL). MLA and EF were evaluated at two levels. The first level corresponds to the puborectalis muscle (evaluation of MRI trauma stage and avulsion), the second level correspondes to the iliococcygeus muscule (evaluation only avulsion injury to the muscle). RESULTS: Normal appereance of musculus puborectalis (level 1) was captured only in 25 (8.8) women. In 117 (41.1%) of women were present MRI minor trauma, 143 (50,2%) women were present with MRI major trauma. Avulsion of the muscle was captured in 85 cases (29.8%) at level 1 and in 165 cases (57.9%) in level 2. Preserved architecture of the EF was caught only 99 (34.7%) of the cases in level 1 and in 47 cases (16.5%) in level 2. Sacrouterine ligaments showed normal morphology in 100 cases (35.1%).Conslusion: Defects of musculofascial component of the pelvic floor is found frequently in women with symptomatic pelvic organ prolapse. Often a combination of defects MLA, EF and SUL are found. These comprehensive pelvic floor defects require careful urogynecological examination and planing operating methods with a view to minimizing the likelihood of recurrence of the descent. In indicated cases the use of the synthetic vaginal mesh is as a method of first choice.

Beta-hydroxybutyrate in milk as screening test for subclinical ketosis in dairy cows.

Ketosis is a very frequent metabolic disease in dairy cows, resulting in lower milk production, impaired fertility and increased frequency of other diseases. The course of the disease is often subclinical, so early detection is very important. The aim of the study was to investigate the relation between the concentration of beta-hydroxybutyrate in blood and milk and to determine the cut-off value in milk for detection of subclinical ketosis. The study included 94 cows, which were in the first third of lactation. Beta-hydroxybutyrate (BHB) concentrations were measured in blood and milk serum using a biochemical analyser. The average concentration of BHB in the blood serum samples was 1.14 mmol/L while in the milk it was about ten times lower at 0.117 mmol/L. A statistically significant positive correlation between the concentration of BHB in blood and milk (r=0.705, p<0.001) was found. In cows with BHB in blood below 2.0 mmol/L a stronger correlation between blood and milk BHB was established (r=0.658, p<0.001) than in cows with blood BHB above 2.0 mmol/L (r=-0.292, p=0.206). Therefore, BHB in milk is a very suitable indicator in the diagnosis of subclinical ketosis as there is a good correlation between BHB in the blood and milk of cows with subclinical ketosis. The cut-off concentration of BHB in milk set at ≥0.080 mmol/L (AUC=0.91±0.03; p<0.001) is a significant indicator for subclinical ketosis in dairy cows. The sensitivity of the test was 94% and specificity 74%. Beta-hydroxybutyrate in milk is a good indicator of subclinical ketosis in dairy cows and can be measured accurately with a biochemical analyser.

Spectroscopic and laser properties of bulk iron doped zinc magnesium selenide Fe:ZnMgSe generating at 4.5 - 5.1 µm.

The Fe:Zn(1-x)Mg(x)Se (x = 0.19, 0.27, and 0.38) solid solutions spectroscopic properties were investigated and laser oscillations were achieved for the first time. The increase of the magnesium concentration in the Fe:ZnMgSe crystal was shown to result in an almost similar long wavelength shift of both absorption and fluorescence spectra of about 60 nm per each 10% of magnesium. With the Fe:ZnMgSe crystal temperature decrease, the fluorescence spectrum maximum shifts towards shorter wavelength resulting mainly from strong narrowing of the longest wavelength fluorescence line. Laser radiation wavelength dependence on the magnesium concentration as well as on temperature was observed. The Fe:ZnMgSe x = 0.38 laser oscillation wavelength increased from 4780 nm at 80 K to 4920 nm at 240 K using the optical resonator without any intracavity spectrally-selective element. In comparison with the Fe:ZnSe laser operating in similar conditions, these wavelengths at both temperatures were shifted by about 500 nm towards mid-IR region.

Comparison of DNA-based techniques for differentiation of production strains of ale and lager brewing yeast.

AIMS: Brewing yeasts are classified into two species-Saccharomyces pastorianus and Saccharomyces cerevisiae. Most of the brewing yeast strains are natural interspecies hybrids typically polyploids and their identification is thus often difficult giving heterogenous results according to the method used. We performed genetic characterization of a set of the brewing yeast strains coming from several yeast culture collections by combination of various DNA-based techniques. The aim of this study was to select a method for species-specific identification of yeast and discrimination of yeast strains according to their technological classification. METHODS AND RESULTS: A group of 40 yeast strains were characterized using PCR-RFLP analysis of ITS-5·8S, NTS, HIS4 and COX2 genes, multiplex PCR, RAPD-PCR of genomic DNA, mtDNA-RFLP and electrophoretic karyotyping. Reliable differentiation of yeast to the species level was achieved by PCR-RFLP of HIS4 gene. Numerical analysis of the obtained RAPD-fingerprints and karyotype revealed species-specific clustering corresponding with the technological classification of the strains. Taxonomic position and partial hybrid nature of strains were verified by multiplex PCR. Differentiation among species using the PCR-RFLP of ITS-5·8S and NTS region was shown to be unreliable. Karyotyping and RFLP of mitochondrial DNA evinced small inaccuracies in strain categorization. CONCLUSIONS: PCR-RFLP of HIS4 gene and RAPD-PCR of genomic DNA are reliable and suitable methods for fast identification of yeast strains. RAPD-PCR with primer 21 is a fast and reliable method applicable for differentiation of brewing yeasts with only 35% similarity of fingerprint profile between the two main technological groups (ale and lager) of brewing strains. SIGNIFICANCE AND IMPACT OF THE STUDY: It was proved that PCR-RFLP method of HIS4 gene enables precise discrimination among three technologically important Saccharomyces species. Differentiation of brewing yeast to the strain level can be achieved using the RAPD-PCR technique.

The Dark Side:Stigma in purpose-built senior environments.

This paper focuses on stigma in collective living environments for older adults, specifically multi-level campuses. We contrast two design profiles, a purpose-built campus which opened in 1997, and an older setting that grew by accretion over decades. Purpose-built housing is used as originally intended, designed, and constructed; housing built by accretion has been modified over time to meet changing needs and uses. The separation by care levels in both sites is reflected in their cultures as residents and staff relate to physical levels of care through a vocabulary of fear and stigma. Residents of the independent living building on the purpose-built campus refer to the assisted living building, accessed only through a second floor link, as "the other side" or "the dark side." In this setting we observe stigma assigned to a place in the built environment. By contrast, in the older setting built by accretion over the last century, levels of care feature a less-structured clustering of independent living and assisted living, and common areas were shared by residents from multiple care levels. We have observed less stigma associated with levels of care in this older building. Grounding our analysis in data drawn from ongoing ethnographic research, we focus on the built environment as it relates to stigma in the social environment. The paper concludes with a discussion of the importance and pervasiveness of stigma in senior environments.

Evaluation of different vitamin E recommendations and bioactivity of α-tocopherol isomers in broiler nutrition by measuring oxidative stress in vivo and the oxidative stability of meat.

The aim of this study was to compare recommendations for vitamin E supplementation regarding high polyunsaturated fatty acid intake and to compare the bioactivity of RRR- and all-rac-α-tocopherol with respect to oxidative stress in vivo and the oxidative stability of broiler meat. Fifty male broilers were divided into 5 groups. All groups received diets with a high inclusion of fat (7.5%), one with palm fat and the others with linseed oil, which were either unsupplemented or supplemented with vitamin E to contain in total 85 or 200 IU of vitamin E as all-rac-α-tocopherol and 85 IU as RRR-α-tocopherol. Oxidative stress in vivo was studied by measuring the DNA damage; measuring malondialdehyde (MDA) in plasma, liver, and breast muscle; and analyzing the antioxidant capacity of the lipid-soluble compounds, total antioxidant status of plasma, and antioxidant enzyme assays. The tocopherols in plasma, liver, and breast muscle were also analyzed. In vitro oxidative stability was studied by measuring MDA in fresh, stored, and heat-treated breast meat. Linseed oil, as opposed to palm fat, induced DNA fragmentation and MDA formation. Both forms and concentrations of vitamin E reduced DNA damage and breast muscle MDA. The groups receiving 200 IU of all-rac-α-tocopherol and 85 IU of RRR-α-tocopherol had much higher values for antioxidant capacity of lipid-soluble compounds than did the controls. No differences were observed in the values of antioxidant enzymes. The α-tocopherol levels in tissues and plasma were significantly influenced by the level of α-tocopherol supplementation. Malondialdehyde formation in meat from the vitamin E-supplemented groups was decreased in comparison with that from the control linseed oil group. We conclude that both vitamin E concentrations were insufficient to prevent all harmful effects of lipid oxidation in vivo and that both were equally effective. On the contrary, to ensure good stability of meat lipids, higher vitamin E supplementation is needed, especially after heat treatment. The results of in vivo oxidative stress and meat lipid oxidation confirmed the currently accepted bioactivity of the RRR-α- to all-rac-α-tocopherol ratio of 1.39 in in vivo and in vitro systems.

Prenatal developmental toxicity studies of inhaled methyl iodide vapor in rabbits reveal a susceptible window of exposure inducing late gestational fetotoxicity.

Methyl iodide (MeI), an intermediate used in the manufacture of some insecticides and pharmaceuticals, is under review for U.S. registration as a non-ozone-depleting alternative to methyl bromide in the pre-plant soil fumigation market. Guideline (OPPTS 870.3700) developmental toxicity studies in New Zealand White (NZW) rabbits showed dose-dependent increases in the litter proportions of late fetal deaths and postimplantation loss and/or decreased fetal body weight following inhalation exposure of pregnant rabbits to MeI during gestation days (GD) 6-28. A subsequent phased-exposure study was performed to pinpoint the critical window of gestational exposure that produced the rabbit fetotoxicity. Artificially inseminated NZW female rabbits were exposed to 20 ppm MeI vapors by whole-body inhalation (6 h/day) throughout major organogenesis and fetal development (GD 6-28), during early gestation (GD 6-14) or mid-gestation (GD 15-22) only, or during 2-day intervals late in gestation (GD 23-24, 25-26, or 27-28). No maternal or developmental toxicity was elicited from maternal exposure during GD 6-14, 15-22, or 27-28. However, MeI-related fetotoxicity, including increased litter proportions of late fetal deaths with or without corresponding decreases in fetal body weight, were observed for females exposed during GD 6-28 (p < .01), 23-24 and 25-26. Although the increase in late-stage fetal death for each of the 2-day exposures on GD 23-24 and GD 25-26 was not statistically significant, as noted for the combined total of fetal deaths during the GD 6-28 exposure, it can be deduced that the gestational window of GD 23-26 was the most susceptible window of exposure for eliciting developmental toxicity in rabbits exposed to MeI vapors.

Methyl iodide-induced fetal hypothyroidism implicated in late-stage fetal death in rabbits.

Methyl iodide (MeI) induces fetotoxicity in New Zealand White (NZW) rabbits when maternal exposure occurs during a susceptible window late in gestation (gestation days [GD] 23-26). To identify the possible mode of action, comprehensive maternal and fetal bioanalysis and thyroid structure/function assessments were conducted in MeI-exposed (25 ppm by whole-body inhalation) and unexposed time-mated NZW rabbits (10/group) during GD 21-27. Key developmental events were observed within this window in unexposed fetuses, including the appearance of colloid in the thyroid follicular lumen and the detection of serum T(3) beginning on GD 22. Fetal T(4) and T(3) levels were diminished following maternal MeI exposure compared to baseline values. Fetal TSH was significantly increased following 4 days of maternal MeI exposure. MeI-induced changes in the fetal thyroid included reduced colloid formation, epithelial follicular hypertrophy, and epithelial cytoplasmic vacuolation. Time-course investigations using 20 ppm MeI revealed highly concentrated levels of iodide in fetal versus maternal serum. Direct maternal administration of sodium iodide by intravenous infusion during GD 23-26 induced similar effects on fetal thyroid structure and function as MeI, identifying iodide as the putative agent. Elevated S-methylcysteine adduct concentrations were noted in fetal hemoglobin, indicating that some unreacted MeI may be delivered directly to the fetus. However, the weight of evidence from these studies suggests that late-stage fetal death following maternal exposure to MeI during GD 23-26 is the result of preferential accumulation of iodide in the fetal compartment causing disruption of the fetal hypothalamic-pituitary-thyroid axis at a critical time in the development of the rabbit fetal thyroid.

Effects of running on adiponectin, insulin and cytokines in cerebrospinal fluid in healthy young individuals.

Exercise can prevent the sedentary lifestyle-related risk of metabolic and cognitive decline, but mechanisms and mediators of exercise effects on human brain are relatively unexplored. We measured acute exercise-induced changes in adiponectin, insulin and other bioactive molecules in cerebrospinal fluid (CSF) and serum from young lean individuals. Samples of serum and CSF were obtained before and 1-h after the 90-min run (75-80% HRmax; maximal heart rate), additional serum was taken at finish-line. Body composition, physical fitness, metabolic rate, cognitive functions, food preference, glucose, insulin and albumin were measured. The spectrum of 174 cytokines was assessed by protein arrays, adiponectin was also determined by ELISA and immunoblotting. CSF adiponectin decreased post-exercise by 21.3% (arrays) and 25.8% (ELISA) (p < 0.009). Immunoblotting revealed reduction in a low-molecular-weight-adiponectin (p < 0.005). CSF adiponectin positively correlated with CSF/serum albumin ratio (p < 0.022), an indicator of blood-brain-barrier permeability. CSF and serum adiponectin were positively associated with memory and running-induced changes in insulinemia and CSF insulin. Additionally, running modulated CSF levels of 16 other cytokines. Acute running reduced CSF adiponectin and modulated insulin and albumin in CSF and serum. Associations of adiponectin with memory and metabolism indicate the potential role of this bioactive molecule in mediating exercise-induced adaptive response in human brain.

Ribotyping and biotyping of Staphylococcus epidermidis isolated from hospital environment.

The Staphylococcus strains acquired from scrapings from hospital environments were identified to the species level based on their biochemical properties. From the monitored sample the Staphylococcus epidermidis strains were selected for more accurate typing and tested on their virulence factor and ribotyped. The biotyping of S. epidermidis did not show any considerable intraspecific variation of these isolates and there were no atypical reactions, with the exception of three strains (out of 33). In contrast, the results of ribotyping showed greater heterogeneity of strains and unequivocally demonstrated the relation between the ribotype and the place of sample drawing. In addition to this fact, the found ribotypes repeat in the same environment in the long-term which suggests the occurrence and persistence of the same strains of conditionally pathogenic bacteria in hospital environment. We showed that ribotyping is a suitable method for precise and reliable detection of some coagulase-negative staphylococci.

Recent Development of Augmented Reality in Surgery: A Review.

INTRODUCTION: The development augmented reality devices allow physicians to incorporate data visualization into diagnostic and treatment procedures to improve work efficiency, safety, and cost and to enhance surgical training. However, the awareness of possibilities of augmented reality is generally low. This review evaluates whether augmented reality can presently improve the results of surgical procedures. METHODS: We performed a review of available literature dating from 2010 to November 2016 by searching PubMed and Scopus using the terms "augmented reality" and "surgery." Results. The initial search yielded 808 studies. After removing duplicates and including only journal articles, a total of 417 studies were identified. By reading of abstracts, 91 relevant studies were chosen to be included. 11 references were gathered by cross-referencing. A total of 102 studies were included in this review. CONCLUSIONS: The present literature suggest an increasing interest of surgeons regarding employing augmented reality into surgery leading to improved safety and efficacy of surgical procedures. Many studies showed that the performance of newly devised augmented reality systems is comparable to traditional techniques. However, several problems need to be addressed before augmented reality is implemented into the routine practice.

Conserved domains and lack of evidence for polyglutamine length polymorphism in the chicken homolog of the Machado-Joseph disease gene product ataxin-3.

Ataxin-3 is a protein of unknown function which is mutated in Machado-Joseph disease by expansion of a genetically unstable CAG repeat encoding polyglutamine. By analysis of chicken ataxin-3 we were able to identify four conserved domains of the protein and detected widespread expression in chicken tissues. In the first such analysis in a non-primate species we found that in contrast to primates, the chicken CAG repeat is short and genetically stable.

Automated CBED processing: sample thickness estimation based on analysis of zone-axis CBED pattern.

An automated processing of convergent beam electron diffraction (CBED) patterns is presented. The proposed methods are used in an automated tool for estimating the thickness of transmission electron microscopy (TEM) samples by matching an experimental zone-axis CBED pattern with a series of patterns simulated for known thicknesses. The proposed tool detects CBED disks, localizes a pattern in detected disks and unifies the coordinate system of the experimental pattern with the simulated one. The experimental pattern is then compared disk-by-disk with a series of simulated patterns each corresponding to different known thicknesses. The thickness of the most similar simulated pattern is then taken as the thickness estimate. The tool was tested on [0 1 1] Si, [0 1 0] α-Ti and [0 1 1] α-Ti samples prepared using different techniques. Results of the presented approach were compared with thickness estimates based on analysis of CBED patterns in two beam conditions. The mean difference between these two methods was 4.1% for the FIB-prepared silicon samples, 5.2% for the electro-chemically polished titanium and 7.9% for Ar(+) ion-polished titanium. The proposed techniques can also be employed in other established CBED analyses. Apart from the thickness estimation, it can potentially be used to quantify lattice deformation, structure factors, symmetry, defects or extinction distance.

Monoclonal antibodies against transferrin. Precipitating mixtures and lack of inter-species cross-reactivity.

Five stable hybridoma lines were prepared using the myeloma cell line P3-X63-Ag.653 and spleen cells of mice hyperimmunized by pig transferrin. All hybridomas grew well in mouse peritoneal cavity and produced antibodies of the IgG1 subclass. Antibody preparations obtained from ascitic fluids tested for their capacity of antigen precipitation. No precipitation was obtained with single antibodies and with pairs of antibodies. Three out of 10 possible triads gave clear and sharp precipitation zones and rings in immunodiffusion tests performed in agar gel. All 5 antibodies were shown by quantitative enzyme-immunoassay to be specific for pig transferrin: no cross-reaction was obtained with mouse, human, horse and sheep transferrins.

Developmental toxicity assessment of arsenic acid in mice and rabbits.

To evaluate potential effects of exposure to inorganic arsenic throughout major organogenesis, CD-1 mice and New Zealand White rabbits were gavaged with arsenic acid dosages of 0, 7.5, 24, or 48 mg/kg/d on gestation days (GD) 6 through 15 (mice) or 0, 0.19, 0.75, or 3.0 mg/kg/d on GD 6 through 18 (rabbits) and examined at sacrifice (GD 18, mice; GD 29, rabbits) for evidence of toxicity. Two high-dose mice died, and survivors at the high and intermediate doses had decreased weight gains. High-dose-group fetal weights were decreased; no significant decreases in fetal weight or increases in prenatal mortality were seen at other dosages. Similar incidences of malformations occurred in all groups of mice, including controls. At the high dose in rabbits, seven does died or became moribund, and prenatal mortality was increased; surviving does had signs of toxicity, including decreased body weight. Does given lower doses appeared unaffected. Fetal weights were unaffected by treatment, and there were no effects at other doses. These data revealed an absence of dose-related effects in both species at arsenic exposures that were not maternally toxic. In mice, 7.5 mg/kg/d was the maternal No-Observed-Adverse-Effect-Level (NOAEL); the developmental toxicity NOAEL, while less well defined, was judged to be 7.5 mg/kg/d. In rabbits, 0.75 mg/kg/d was the NOAEL for both maternal and developmental toxicity.