Effects of the addition of waxy and normal hull-less barley flours on the farinograph and pasting properties of composite flours and on the nutritional value, textural qualities, and in vitro digestibility of resultant breads.
Hull-less barley (HLB), especially waxy HLB, contains many physiologically active ingredients; however, its poor processing performance and end-product quality are unfavorable. In this study, 80% waxy or normal HLB wholegrain flour (WGF) and 20% wheat flour were used for baking bread. The farinograph and pasting properties of composite powders, and the nutritional value, textural properties, and in vitro hydrolysis of resultant breads were evaluated. The addition of a high proportion of HLB WGFs significantly increased the nutritional value of breads, especially the β-glucan contents of waxy HLB breads. The addition of HLB WGFs and a suitable amount of wheat gluten led to a lower degree of softening of HLB bread flours but improved its farinograph characteristics, such as higher water absorption rate, development time, stability time, and farinograph quality number. Although the sensory profiles of HLB breads were considerably lower than those of wheat bread, they still received a good overall acceptability from a panel of sensory evaluators. HLB breads, particularly the waxy types, exhibited higher hardness, gumminess, chewiness, and lower specific volume, glycemic index and equilibrium concentration in starch hydrolysis. After baking, the starch crystallinity of dough changed from A to V type, and the relative crystallinity decreased. Overall, waxy HLB breads had more nutritional value than normal HLB breads. Higher β-glucan and total dietary fiber content in HLB might have a positive effect on the nutritional value of the resultant breads. However, high β-glucan and total dietary fiber was also accompanied by a negative effect on the sensory quality and processing performance of the end product. PRACTICAL APPLICATION: The composite flour with 80 g hull-less barley wholegrain flour, 20 g wheat flour, and 30 g wheat gluten can be substituted in breadmaking. Compared to wheat bread, hull-less barley bread exhibited different but acceptable sensory properties and had more nutritional value, particularly the waxy one. Therefore, a high proportion of hull-less barley could be recommended for bread production.


Vascular leakage caused by loss of Akt1 is associated with impaired mural cell coverage.
Angiogenesis plays a critical role in embryo development, tissue repair, tumor growth and wound healing. In the present study, we investigated the role of the serine/threonine kinase Akt in angiogenesis. Silencing of Akt1 in human umbilical vein endothelial cells significantly inhibited vascular endothelial growth factor (VEGF)-induced capillary-like tube formation. Mice lacking Akt1 exhibited impaired retinal angiogenesis with delayed endothelial cell (EC) proliferation. In addition, VEGF-induced corneal angiogenesis and tumor development were significantly inhibited in mice lacking Akt1. Loss of Akt1 resulted in reduced angiogenic sprouting, as well as the proliferation of ECs and mural cells. Addition of culture supernatant of vascular smooth muscle cells (VSMCs) in which Akt1 was silenced suppressed tube formation, the stability of preformed tubes and the proliferation of ECs. In addition, attachment of VSMCs to ECs was significantly reduced in cells in which Akt1 was silenced. Mural cell coverage of retinal vasculature was reduced in mice lacking Akt1. Finally, mice lacking Akt1 showed severe retinal hemorrhage compared to the wild-type. These results suggest that the regulation of EC function and mural cell coverage by Akt1 is important for blood vessel maturation during angiogenesis.


Cilostazol induces angiogenesis and regulates oxidative stress in a dose-dependent manner: A chorioallantoic membrane study.
In this study, we aimed to investigate the effects of cilostazol on angiogenesis and oxidative stress using the chorioallantoic membrane model. In this experimental study, the Ross 308 chick embryos were used. The negative control group (n=10) received no intervention. The positive control group (n=10) consisted of eggs treated with epidermal growth factor for inducing angiogenesis. Three cilostazol groups were designed with 10-7 (n=10), 10-6 (n=10), and 10-5 (n=10) M concentrations. Each egg was punctured on the sixth day of incubation, and drug pellets were introduced to the positive control and drug groups at the prespecified doses. Vascular development was evaluated on the eighth day of application. The total oxidant status, total antioxidant capacity, and oxidative stress index levels were determined from albumen liquids obtained with a syringe before and after drug application. Lower oxidative stress index levels were obtained from the positive control and cilostazol groups compared to the negative control albumens (p=0.001). The increments in vascular junctions and newly developed vascular nodules were evaluated in drug-free and drug-applied chorioallantoic membranes. The highest activity was obtained in the 10-7 M concentration cilostazol group. An increased angiogenic activity was detected in all drug groups in each concentration compared to the negative control group (p=0.001). Angiogenic activity was similar in all the cilostazol-treated groups (p=0.43). Cilostazol has a positive stimulant effect on angiogenesis and it seems to suppress oxidative stress during embryonic growth. Cilostazol exerts these effects significantly and similarly at different doses.


Evaluation of the use of intrauterine infusions of oxytetracycline, subcutaneous injections of fenprostalene, or a combination of both, for the treatment of retained fetal membranes in dairy cows.
Dairy cows (n = 207) that had retained fetal membranes for more than 8 hours after parturition were randomly assigned to 1 of 4 treatment groups (group 1, daily intrauterine infusion of oxytetracycline; group 2, daily intrauterine infusion of oxytetracycline and a single SC injection of fenprostalene; group 3, a single SC injection of fenprostalene; and group 4, untreated controls). Cows were monitored daily. Subsequent disease conditions were recorded, and appropriate treatments were initiated. Interval from parturition to expulsion of fetal membranes was unaffected by treatment regimen. The frequency of cows with high rectal temperatures (> or = 39.5 C) was significantly (P < 0.05) less in cows treated with oxytetracycline infusions; however, the frequency of displaced abomasum, ketosis, and mastitis was unaffected by treatment method. Treatment method appeared to influence the development of postpartum uterine pathologic conditions. Treatment with oxytetracycline and fenprostalene (group 2) increased (P < 0.05) the frequency of pyometra. Treatment with fenprostalene (groups 2 and 3) decreased the number of cows with a palpable uterine lumen detectable during per rectal palpation at 28 to 42 days after parturition. Reproductive efficiency, as determined on the basis of the interval from parturition to conception, was unaffected by treatment protocol.


[Multicenter prospective study of informed consent in general surgery].
To understand the level of acceptance, awareness and usefulness of informed consent, a group of 119 patients (59 men and 60 women) from different types of hospitals were given a questionnaire which required only 'YES or NO' answers, both before and after surgery. The questionnaire concerned the patient's knowledge about pathology, operative risks, approval, anxiety caused, understanding of information received and consent given, and also if he would inform a relative in the same condition. From the analysis of the results it was established that: the more information a patient has about his illness and operation risks, the more he will want to have; the less he knows the less he will want to know, and he will also have more faith in the doctors. Some patients would not inform a relative with a similar pathology. To conclude, informed consent, instead of being a right of the patient is progressively becoming more a right of the doctor. It does not have any real effect on the patient's choice but is useful, as it represents a moment of personalised attention from medical personnel, though the patient may not completely understand the information received. There are few advantages in strictly medical terms but informed consent has increased malpractice litigation.


Co-residence of household heads with parents in Japan: a multivariate explanation.
"This paper uses a multivariate logit model to study the factors affecting the Japanese household heads' propensities to co-reside with elderly parents, based on the micro data of a 1986 national survey. Our major findings are as follows. The most important factors are (1) inheritance of house or residential land from parent; (2) parent's spouseless status and age; (3) household head's sibling status; and (4) household head's nativity status. In a multivariate context, the negative effect of the household head's level of education was significant but not very important, whereas the presence of a working wife with child had a very weak positive effect on the co-residence with parent. Our overall conclusion is that the intergenerational co-residence in Japan is strongly affected by cultural norms and has a strong rational (economic) basis."


Antioxidant activity of plant extracts on the inhibition of citral off-odor formation.
Grape seed, pomegranate seed, green tea, and black tea extracts were used to inhibit the off-odor from citral degradation. A 0.1 M citrate buffer (pH 3), containing 100 ppm citral and 200 ppm gallic acid equivalent plant extract, was incubated at 40 degrees C. The reaction mixtures were analyzed by high-performance liquid chromatography (HPLC) at days 0, 6, 10, 13, and 16 to monitor degradation of citral and formation of alpha, p-dimethylstyrene, p-cymene-8-ol, and p-methylacetophenone. The addition of plant phenolic extracts could not inhibit citral degradation, however, all four plant extracts significantly inhibited p-methylacetophenone formation. The samples, with the addition of plant extracts, exhibited higher concentrations of alpha, p-dimethylstyrene and p-cymene-8-ol than the control. This is presumed to be due to the oxygen-scavenging effect of plant extracts blocking the pathway from p-cymene-8-ol to p-methylacetophenone. Our results suggest that these plant extracts act as general antioxidants inhibiting the generation ofp-methylacetophenone regardless of the types of water-soluble phenolic compounds existing in the plant extracts.


Evaluation of resin sealer penetration of dentin following different final rinses for endodontic irrigation using confocal laser scanning microscopy.
The aim of the study was to assess the depth of sealer penetration into dentinal tubules following different final rinses and indirectly evaluate precipitation of irrigating solutions. Methods: 52 extracted maxillary incisors were prepared to size 40/04 with 6% sodium hypochlorite irrigating solution. Following power analysis, teeth were randomly divided into 4 groups (n = 13) according to final rinse. Group 1 - 17% EDTA, Saline and 2% Chlorhexidine solution; Group 2 - SmearOFF™; Group 3 - Qmix® ; and Group 4 - saline control. Obturation was performed with gutta percha and resin sealer mixed with rhodamine-B-isothiocyanate dye using warm vertical compaction. Teeth were sectioned into apical, middle, and coronal thirds using Buehler diamond saw at 3, 6 and 9 mm from the apex. Sections were examined under confocal laser scanning microscopy, and images were exported to Image J software to measure the maximum depth of sealer penetration circumferentially. Data were analysed using a generalised linear model and post hoc multiple comparisons with significance set at P < 0.05. Group 3 had the highest depth of penetration. There were statistically significant differences between Groups 2 and 3 and control for all sections, Group 1 and control in apical section only. Conclusion: Recently developed final rinses produced higher sealer penetration and more patent dentinal tubules than using EDTA and chlorhexidine sequentially.


Exposure to Hazardous Neighborhood Environments in Late Childhood and Anxiety.
This investigation examined the relationship between living in disordered neighborhoods during childhood and anxiety 1 year later. Objective measures of neighborhood environment and individual data from a study of mental health in suburban children were utilized. Linear regression models were used to assess relationships between neighborhood hazard and anxiety. Childhood neighborhood disorder was inversely associated with generalized anxiety (β = 0.037, p<0.01) and social phobia (β = -0.33, p=0.03), but not other forms of anxiety including separation anxiety or panic disorder. We suspect that children with early exposure to disordered neighborhoods are either desensitized to toxic environments or that anxiety is not well specified for this population.


Genotype-phenotype correlations in mapped split hand foot malformation (SHFM) patients.
Split hand foot malformation (SHFM) also known as central ray deficiency, ectrodactyly and cleft hand/foot, is one of the most complex of limb malformations. SHFM can occur as an isolated malformation or in association with other malformations, as in the ectrodactyly-ectodermal dysplasia-clefting (EEC) syndrome and other autosomal dominant conditions with long bone involvement, all showing variable expressivity and reduced penetrance. The deficiency in SHFM patients can also be accompanied by other distal limb anomalies including polydactyly and/or syndactyly. This variability causes the phenotypic classification of SHFM to be far from straightforward and genetic heterogeneity, with at least five loci identified to date, further complicates management of affected patients and their families. Although genotypic-phenotypic correlations have been proposed at the molecular level for SHFM4 patients who have mutations in the P63 gene, phenotypic correlations at the chromosomal level have not been thoroughly documented. Using descriptive epidemiology, Chi square and discriminant function analyses, our laboratory has identified phenotypic patterns associated with the mapped genetic SHFM loci. These findings can assist in classification, provide insight into responsible developmental genes and assist in directing mapping efforts and targeted genetic testing, resulting in more accurate information for family members in the clinical setting. Comparison with relevant animal models is discussed.


Mitochondrial microRNA (MitomiRs) in cancer and complex mitochondrial diseases: current status and future perspectives.
Mitochondria are not only important for cellular bioenergetics but also lie at the heart of critical metabolic pathways. They can rapidly adjust themselves in response to changing conditions and the metabolic needs of the cell. Mitochondrial involvement as well as its dysfunction has been found to be associated with variety of pathological processes and diseases. mitomiRs are class of miRNA(s) that regulate mitochondrial gene expression and function. This review sheds light on the role of mitomiRs in regulating different biological processes-mitochondrial dynamics, oxidative stress, cell metabolism, chemoresistance, apoptosis,and their relevance in metabolic diseases, neurodegenerative disorders, and cancer. Insilico analysis of predicted targets of mitomiRs targeting energy metabolism identified several significantly altered pathways (needs in vivo validations) that may provide a new therapeutic approach for the treatment of human diseases. Last part of the review discusses about the clinical aspects of miRNA(s) and mitomiRs in Medicine.


Electron spin-spin coupling from multireference configuration interaction wave functions.
We present the implementation of two-electron spin-spin coupling as a quasidegenerate perturbative treatment of the Breit-Pauli spin-spin Hamiltonian. The evaluation is based on a multireference CI treatment and constitutes one of the first efforts in the calculation of this effect within a highly sophisticated consideration of both nondynamical and dynamical correlation. The extension of existing schemes for efficient calculation, in particular, of the spin-coupling elements necessitated some involved derivations, the outline of which is presented herein. Application of the program to calculations of diagonal as well as off-diagonal spin-coupling elements is illustrated with the test cases O(2) and NH.


[A case of dry-type tuberculous peritonitis due to small intestinal tuberculosis associated with systemic inflammatory response syndrome].
A 30-year-old man was admitted to another hospital because of abdominal fullness, pain, and fever. Laboratory examinations revealed marked inflammation, and abdominal computed tomography showed diffuse thickening of the small intestinal wall and peritoneum, and soft tissue infiltration of the mesentery, without ascites. A chest X-ray showed no abnormal findings. Despite antibiotic and antituberculous therapy, the patient's general condition deteriorated; he subsequently developed systemic inflammatory response syndrome (SIRS). We therefore started steroid pulse therapy, which resulted in rapid improvement of his condition and symptoms. On the basis of enteroscopy, stool culture, and peritoneal biopsy findings, the patient was diagnosed with tuberculous peritonitis caused by small intestinal tuberculosis and was readministered antituberculous therapy.


Similar Myocardial Perfusion and Vascular Stiffness in Children and Adolescents with High Lipoprotein (a) Levels, in Comparison with Healthy Controls.
This study investigated the possible correlation between elevated lipoprotein (a) (Lp(a)) levels and early vascular aging biomarkers in healthy children and adolescents. Twenty-seven healthy children/adolescents, mean age 9.9 ± 3.7 years, with high Lp(a) levels without other lipid abnormalities and 27 age- and sex-matched controls with normal Lp(a) levels, were included in the study. The investigation of possible early vascular aging was assessed by measuring vascular function indices: carotid intima-media thickness (c-IMT), pulse wave velocity (PWV), augmentation index (AIx), and subendocardial viability ratio (SEVR). Although serum lipid values were within normal levels, mean values of total cholesterol and apolipoprotein B were higher in the group of children with high Lp(a) levels than controls (p = 0.006 and p < 0.001, respectively). Vascular function indices did not show significant differences, neither between the 2 groups nor in the subgroups of children with increased Lp(a) levels. These subgroups were defined by the presence or absence of family history of premature coronary artery disease. Lp(a) levels did not show a significant correlation with the other parameters studied, both regarding the whole sample (patients and controls), as well as in the subgroups of elevated Lp(a) levels. However, in the group of children with high Lp(a) levels, c-IMT and PWV were positively correlated with diastolic blood pressure (r = 0.427, p = 0.026 and r = 0.425, p = 0.030, respectively), while SEVR was negatively correlated with AIx (r = -0.455, p = 0.017). Healthy children and adolescents with high Lp(a) levels do not yet have impaired vascular indices, compared to controls. However, in order to prevent early atherosclerosis, it is crucial to early identify and follow up children with high Lp(a) levels and positive family history of premature coronary disease or other cardiovascular risk factors.


The evolution of bourgeois, parasitic, and cooperative reproductive behaviors in fishes.
Among vertebrate classes, fishes exhibit by far the greatest variability in competitive and cooperative behaviors in male reproduction. Scramble competition between reproductive males is one possibility. Another possibility occurs when resources, mates, or locations can be monopolized, in which case males may invest in primary access to fertilizations by adopting a "bourgeois" strategy, or they may employ alternative mating tactics to evade the reproductive monopoly of other males. Adaptations in morphology, physiology, and behavior to bourgeois and alternative phenotypes are highly divergent. Here I review the functional characteristics that differ between bourgeois and parasitic phenotypes, and discuss the variability of alternative reproductive tactics at the levels of plasticity, determination, and selection. Examples will illustrate the importance of ecology, and will suggest that variation in reproductive tactics is largely adaptive. Behavioral solutions to competition for mates and fertilizations often involve agonistic behavior and conflict, but also cooperation among competitors (e.g., when subordinate males pay a price to bourgeois males for gaining access to fertilizable eggs). Application of molecular genetic tools has helped to uncover intricate sexual and social relationships in various fish species, including species that display some of the most complex reproductive and social patterns known among the vertebrates.


Pharmacogenetic angiogenesis profiling for first-line chemotherapy in patients with advanced gastric cancer.
We retrospectively investigated germline polymorphisms in angiogenesis pathway genes (14 SNPs) and their correlation to clinical outcome (progression free survival and overall survival) in 128 patients with unresectable-advanced gastric carcinoma (AGC) treated with first-line chemotherapy. Our analysis revealed that Endostatin +4349 G>A polymorphism exhibited a worse progression free survival (PFS) and overall survival (OS) compared with the GG genotype. Significant OS difference was also observed in the endothelial nitric oxide synthase (eNOS)-786 T>C polymorphism. Hence, common germline variants in Endostatin and eNOS genes have predictive significance for clinical outcome and survivality in AGC patients treated with first-line chemotherapy.


Evaluation of dexamethasone for the treatment of intracerebral hemorrhage using a collagenase-induced intracerebral hematoma model in rats.
Dexamethasone was evaluated for the treatment of intracerebral hemorrhage using a rat model of cerebral hematoma induced by intracerebral injection of collagenase. The treatment group consisted of hematoma rats receiving dexamethasone 1 mg/kg intraperitoneal (i.p.) at 1 and 24 h following surgery. Controls included hematoma rats receiving saline i.p. and sham-operated animals receiving saline i.p. Each animal was evaluated neurologically prior to, as well as 24 and 48 h following surgery. After the last neurological evaluation, animals were deeply anesthetized and the brain was removed following perfusion for microscopic examination and glial fibrillary acidic protein immunohistochemistry. Behavioral scores were significantly improved in the treated group (P < 0.0001). The hematoma volume was significantly smaller (P < 0.02). Neutrophils and astrocytes were less numerous in the hematoma of dexamethasone-treated animals (P < 0.001), however the number of necrotic neurons in the penumbra was not changed by the treatment. The number of necrotic neurons in the cerebral cortex was less in treated than in nontreated animals (P < 0.01). Controls had many vascular changes including necrotic endothelium and fibrin deposits compared with treated animals. In conclusion, dexamethasone administered shortly after an intracerebral hematoma appears beneficial for the treatment of this condition.


Diphenylmethane diisocyanate (MDI)-induced asthma: evaluation of the immunologic responses and application of an animal model of isocyanate sensitivity.
A worker developed two episodes of severe asthma 90 min after cutting a polyurethane plate made of diphenylmethane diisocyanate (MDI) using a rapidly turning carbide blade. Intradermal skin testing with MDI-human serum albumin (MDI-HSA) and p-tolyl isocyanate-HSA (p-TMI-HSA) were positive at 0.002 mg/ml. Control subjects showed no reaction at 2 mg/ml. Bronchial provocations of the worker with MDI-HSA and p-TMI-HSA, made 1 year after the occupational asthmatic episode, were negative at 10 mg/ml. Bronchial reactivity to methacholine decreased toward normal during a 2-year follow-up. RAST using MDI-HSA or p-TMI-HSA were strongly positive when compared with binding by sera from atopic controls which contained the same amount of total IgE. RAST titres decreased during a 1-year follow-up. In the MDI-HSA RAST, inhibition studies indicated specificity of antibodies for MDI-HSA. In the p-TMI-HSA RAST, p-TMI-HSA was a very effective inhibitor whereas MDI-HSA was not. These results indicated the formation of at least two distinct groups of IgE antibodies: those reactive with MDI, and those reactive with p-TMI determinants. Guinea pigs immunized with MDI formed antibodies with specificities similar to those of the patient. We conclude that the worker had occupational asthma accompanied by the formation of specific IgE antibodies of the specificities. The causal relationship of the antibodies to the occupational asthma remains uncertain.


Influence of rumen escape starch on pancreatic exocrine secretion of goats.
The objective of the experiment was to evaluate the effect of rumen escape starch (RES), accomplished by altering dietary starch concentrations on pancreatic exocrine secretion of goats. Four goats (36.8 +/- 3.2 kg) with common bile duct re-entrant and duodenal catheters were used in a 4 x 4 Latin square. Goats were fed diets containing 20%, 30%, 40% and 50% starch. Periods consisted of 10 day adaptation followed by 3 day of sample collection. Juice was collected in 1-h fractions continuously for 72 h. Total juice secreted was recorded, and 3% sub samples were retained to form a composite sample. The remaining fluid was returned to the duodenum. Juice composite samples were analyzed for activities of alpha-amylase, trypsin, chymotrypsin and lipase. Secretion of pancreatic alpha-amylase was lower (p < 0.05) when comparing lambs fed 20% starch diet with 30%, 40% and 50% starch diets. Lipase secretion was greater (p < 0.05) in lambs fed 40% starch diet compared with the other diets. Total secretion of juice, trypsin and chymotrypin was not affected (p > 0.05) by dietary starch concentration. Rumen escape starch increased with increasing dietary starch concentration (p < 0.05). Regression analysis showed that increasing RES results in a quadratic increase (p < 0.05) in pancreatic alpha-amylase and lipase secretion, and the secretion of alpha-amylase and lipase is maximum when RES is 113 and 83 g/day respectively. These results suggest that optimal RES for pancreatic secretion of alpha-amylase and lipase is 80-110 g/day in adult goats.


Low-Boron Tolerance Strategies Involving Pectin-Mediated Cell Wall Mechanical Properties in Brassica napus.
Boron (B) is an essential micronutrient for the growth and development of plants. Oilseed rape (Brassica napus L.) is a staple oleaginous crop, which is greatly susceptible to B deficiency. Significant differences in tolerance of low-B stresses are observed in rapeseed genotypes, but the underlying mechanism remains unclear, particularly at the single-cell level. Here we provide novel insights into pectin-mediated cell wall (CW) mechanical properties implicated in the differential tolerance of low B in rapeseed genotypes. Under B deficiency, suspension cells of the low-B-sensitive genotype 'W10' showed more severely deformed morphology, lower viabilities and a more easily ruptured CW than those of the low-B-tolerant genotype 'QY10'. Cell rupture was attributed to the weakened CW mechanical strength detected by atomic force microscopy; the CW mechanical strength of 'QY10' was reduced by 13.6 and 17.4%, whereas that of 'W10' was reduced by 29.0 and 30.4% under 0.25 and 0.10 μM B conditions, respectively. The mechanical strength differences between 'QY10' and 'W10' were diminished after the removal of pectin. Further, 'W10' exhibited significantly higher pectin concentrations with much more rhamnogalacturonan II (RG-II) monomer, and also presented obviously higher mRNA abundances of pectin biosynthesis-related genes than 'QY10' under B deficiency. CW regeneration was more difficult for protoplasts of 'W10' than for those of 'QY10'. Taking the results together, we conclude that the variations in pectin-endowed CW mechanical properties play key roles in modulating the differential genotypic tolerance of rapeseed to low-B stresses at both the single-cell and the plant level, and this can potentially be used as a selection trait for low-B-tolerant rapeseed breeding.


Nerve excitability changes in critical illness polyneuropathy.
Patients in intensive care units frequently suffer muscle weakness and atrophy due to critical illness polyneuropathy (CIP), an axonal neuropathy associated with systemic inflammatory response syndrome and multiple organ failure. CIP is a frequent and serious complication of intensive care that delays weaning from mechanical ventilation and increases mortality. The pathogenesis of CIP is not well understood and no specific therapy is available. The aim of this project was to use nerve excitability testing to investigate the changes in axonal membrane properties occurring in CIP. Ten patients (aged 37-76 years; 7 males, 3 females) were studied with electrophysiologically proven CIP. The median nerve was stimulated at the wrist and compound action potentials were recorded from abductor pollicis brevis muscle. Strength-duration time constant, threshold electrotonus, current-threshold relationship and recovery cycle (refractoriness, superexcitability and late subexcitability) were recorded using a recently described protocol. In eight patients a follow-up investigation was performed. All patients underwent clinical examination and laboratory investigations. Compared with age-matched normal controls (20 subjects; aged 38-79 years; 7 males, 13 females), CIP patients exhibited reduced superexcitability at 7 ms, from -22.3 +/- 1.6% to -7.6 +/- 3.1% (mean +/- SE, P approximately 0.0001) and increased accommodation to depolarizing (P < 0.01) and hyperpolarizing currents (P < 0.01), indicating membrane depolarization. Superexcitability was reduced both in patients with renal failure and without renal failure. In the former, superexcitability correlated with serum potassium (R = 0.88), and late subexcitability was also reduced (as also occurs owing to hyperkalaemia in patients with chronic renal failure). In patients without renal failure, late subexcitability was normal, and the signs of membrane depolarization correlated with raised serum bicarbonate and base excess, indicating compensated respiratory acidosis. It is inferred that motor axons in these CIP patients are depolarized, in part because of raised extracellular potassium, and in part because of hypoperfusion. The chronic membrane depolarization may contribute to the development of neuropathy.


Apoptosis and mitosis in gonocytes of the rat testis during foetal and neonatal development.
This study was undertaken to determine the extent of apoptosis and mitosis in the various testicular cell types throughout rat development from foetal day 14.5 to postnatal days 9-10. Apoptotic activity was studied by detecting DNA fragmentation (TUNEL method) in situ. A TUNEL-positive reaction was detected in gonocytes, while none of the other testicular cells were labelled. The morphology of the TUNEL-positive gonocytes was characteristic of apoptotic cells and was different from that observed in experimentally induced necrosis. The percentage of stained gonocytes peaked on day 15.5-16.5 post-conception (dpc), decreased thereafter and no TUNEL-positive gonocytes were found from foetal day 18.5 onwards. On postnatal day 2, apoptosis resumed and increased to reach a maximum on day 7. Mitosis in the gonocytes, as evaluated by the immunodetection of 5-bromo-2'-deoxyuridine (BrdU) incorporation, was present during the same developmental periods but the ratio of BrdU-positive/TUNEL-positive gonocytes was much greater in the foetal period than in the neonatal period. In an organotypic culture system, the changes in the apoptotic and mitotic activities of the gonocytes in testicular explants from foetuses on days 18.5 and 20.5 or from neonates on day 3, cultured for two days were similar to those observed in vivo. Addition of LH or FSH did not influence either apoptosis or mitosis in the germ cells. These results suggest that both apoptosis and mitosis of gonocytes are independent of gonadotrophins and are mainly controlled by intratesticular factors.


Microcatheter contrast injections during intra-arterial thrombolysis increase intracranial hemorrhage risk.
Combined analysis of the Interventional Management of Stroke I and II trials demonstrated a significant association between microcatheter contrast injections and both intracranial hemorrhage (ICH) and contrast extravasation following combined intravenous (IV) and intra-arterial (IA) thrombolysis. The reliability of these observations was tested in our local registry of IA cases. Treatment angiograms and post-procedure CTs of patients treated with combined IV/IA or IA only recombinant tissue plasminogen activator for ICA-T, M1 or M2 occlusions (n=77) were reviewed. The number of microcatheter injections (MCIs) within/distal to the target occlusion was assigned for every case. The association of MCIs to total ICH, total parenchymal hematoma (PH1+PH2) and PH2 after adjusting for significant covariates was tested. MCIs were used in 21 (27%) cases (range MCI 0-6). Any ICH occurred in 38 (49%) cases, including eight (10%) PH1s and eight (10%) PH2s. The use of MCIs was associated with increased PH (p=0.04), PH2 (p=0.07) and total ICH (p=0.03). MCIs were associated with increased contrast extravasation (CEx) (p=0.02). ICH was observed in all CEx cases (n=5, 100% vs 46% non-CEx; p=0.03), and four (80%) CEx cases developed PH2s (p<0.01). MCIs remained associated with total ICH after adjustment for significant covariates of Thromolysis in Cerebral Infarction score, glucose level and presence of atrial fibrillation (OR 3.60; 95% CI 1.12 to 11.49, p=0.03). MCI use was the only significantly associated covariate for total PHs. MCI use was associated with ICH and with clinically significant PHs in this cohort, providing further evidence that MCIs be reduced during IA thrombolysis.


[Definition and clinical characteristics of mild hypertension].
Mild hypertension is defined as blood pressure level of 140-159 mmHg systolic and/or 90-99 mmHg diastolic. The patients with blood pressure level of mild hypertension occupy about 60% of total hypertensive patients in Japan, and most of them are free of subjective symptoms except elevated blood pressure. However, some of the patients with mild hypertension develop cardiovascular events, since thay have occasionally cardiovascular damages on this level of blood pressure and several risk factors of cardiovascular diseases such as diabetes mellitus and hyperlipidemia.


Microbial communities inhabiting the fairy ring of Floccularia luteovirens and isolation of potential mycorrhiza helper bacteria.
Floccularia luteovirens, an important edible mushroom widely distributed in the Qinghai-Tibet plateau, is ecologically important as an ectomycorrhizal fungus and can form the fairy ring. To explore the influence of F. luteovirens fairy ring on soil microbial communities, we compared the soil microbial communities in three different fairy ring zones (inside the fairy ring (IN); beneath the fairy ring (ON); and outside the fairy ring (OUT)). A total of 1.77 million bacterial reads and 1.59 million fungal reads were obtained. Moreover, sequence clustering yielded 519,613 (57,735 per sample) bacterial OTUs, and 513,204 (57,023 per sample) fungal OTUs representing. Microbial diversity was lower in samples from the ON zone compared with the other two zones. Mycorrhiza helper bacteria (MHB) such as Bradyrhizobium and Paenibacillus were more common in the ON zone, and we isolated four potential MHB from rhizosphere soil. Canonical correspondence analysis showed that the soil nutritional condition and physical changes caused by F. luteovirens shaped the microbial communities in the ON zone. This is the first report on the study of soil microbial diversity influenced by fairy ring F. luteovirens, and further studies need to be conducted to study the ecological function influenced by this species.


Breast microcalcifications: the lesions in anatomical pathology.
Microcalcifications are actually indirect signs of pathological processes, and only a few of these processes may be correctly correlated to the morphologic pattern of calcifications. This is true of the microcalcifications typically classified as benign by the 4th edition of the BI-RADS Atlas, except for round and punctuate microcalcifications. This is also the case of polymorphous fine and linear fine microcalcifications most often, but not exclusively, associated with DCIS with necrosis. For other types of microcalcifications, other parameters are analyzed in a more global approach: the associated clinical or mammographical signs; the context, especially genetic; the spatial distribution; the number; the evolution over time. The radiologist should compare the images with the anatomy of the terminal ductal-lobular unit, from where most cancers arise, and estimates the risk by taking into account the clinical context and the antecedents.


The selectively bred high alcohol sensitivity (HAS) and low alcohol sensitivity (LAS) rats differ in sensitivity to nicotine.
Studies in rodents selectively bred to differ in alcohol sensitivity have suggested that nicotine and ethanol sensitivities may cosegregate during selective breeding. This suggests that ethanol and nicotine sensitivities may in part be genetically correlated. Male and female high alcohol sensitivity (HAS), control alcohol sensitivity, and low alcohol sensitivity (LAS) rats were tested for nicotine-induced alterations in locomotor activity, body temperature, and seizure activity. Plasma and brain levels of nicotine and its primary metabolite, cotinine, were measured in these animals, as was the binding of [3H]cytisine, [3H]epibatidine, and [125I]alpha-bungarotoxin in eight brain regions. Both replicate HAS lines were more sensitive to nicotine-induced locomotor activity depression than the replicate LAS lines. No consistent HAS/LAS differences were seen on other measures of nicotine sensitivity; however, females were more susceptible to nicotine-induced seizures than males. No HAS/LAS differences in nicotine or cotinine levels were seen, nor were differences seen in the binding of nicotinic ligands. Females had higher levels of plasma cotinine and brain nicotine than males but had lower brain cotinine levels than males. Sensitivity to a specific action of nicotine cosegregates during selective breeding for differential sensitivity to a specific action of ethanol. The differential sensitivity of the HAS/LAS rats is due to differences in central nervous system sensitivity and not to pharmacokinetic differences. The differential central nervous system sensitivity cannot be explained by differences in the numbers of nicotinic receptors labeled in ligand-binding experiments. The apparent genetic correlation between ethanol and nicotine sensitivities suggests that common genes modulate, in part, the actions of both ethanol and nicotine and may explain the frequent coabuse of these agents.


Growth and survival signalling in B16F10 melanoma cells in 3D culture.
The two-way communication between the ECM (extracellular matrix) and the cytoplasm via the integrins has many functions in cancer cells, including the suppression of apoptosis. As cells in a 3D (three-dimensional) architecture resemble the in vivo situation more closely than do cells in more conventional 2D cultures, we have employed a substratum that prevents cell adhesion and induces cell aggregation to determine why highly metastatic B16F10 melanoma cells resist anoikis. We compared the behaviour of B16F10 cells in 2D [on tPS (tissue culture polystyrene)] and 3D culture {on polyHEMA [poly(2-hydroxyethylmethacrylate)]} configurations. For this, we analysed cell morphology, proliferation, apoptosis and the activation status of several proteins involved in cell proliferation and survival [RhoA, FAK (focal adhesion kinase), Akt, ERK1/2 (extracellular-signal-regulated kinase 1/2)]. B16F10 cells in 3D architecture were able to proliferate as cell aggregates for 3 days, after which the number of cells decreased. The normal Swiss 3T3 cells used as an anoikis-sensitive control did not proliferate on the anti-adhesive substratum. Rho A was activated in B16F10 aggregates throughout their time in culture, whereas it was not in Swiss 3T3 aggregates. An absence of apoptotic activity was correlated with the proliferation of B16F10 cells in aggregates: caspase 3 was significantly activated only after 3 days in culture on polyHEMA. FAK and Akt were transiently activated, and their inactivation was correlated with the induction of apoptosis. ERK1/2 were activated throughout the 3D culture. No survival protein was activated in Swiss 3T3 aggregates. Data obtained from cells in 3D culture suggest that B16F10 cells are resistant to anoikis through the activation of the FAK and Akt signalling pathways.


Atelectasis is inversely proportional to transpulmonary pressure during weaning from ventilator support in a large animal model.
In mechanically ventilated, lung injured, patients without spontaneous breathing effort, atelectasis with shunt and desaturation may appear suddenly when ventilator pressures are decreased. It is not known how such a formation of atelectasis is related to transpulmonary pressure (PL ) during weaning from mechanical ventilation when the spontaneous breathing effort is increased. If the relation between PL and atelectasis were known, monitoring of PL might help to avoid formation of atelectasis and cyclic collapse during weaning. The main purpose of this study was to determine the relation between PL and atelectasis in an experimental model representing weaning from mechanical ventilation. Dynamic transverse computed tomography scans were acquired in ten anaesthetized, surfactant-depleted pigs with preserved spontaneous breathing, as ventilator support was lowered by sequentially reducing inspiratory pressure and positive end expiratory pressure in steps. The volumes of gas and atelectasis in the lungs were correlated with PL obtained using oesophageal pressure recordings. Work of breathing (WOB) was assessed from Campbell diagrams. Gradual decrease in PL in both end-expiration and end-inspiration caused a proportional increase in atelectasis and decrease in the gas content (linear mixed model with an autoregressive correlation matrix; P < 0.001) as the WOB increased. However, cyclic alveolar collapse during tidal ventilation did not increase significantly. We found a proportional correlation between atelectasis and PL during the 'weaning process' in experimental mild lung injury. If confirmed in the clinical setting, a gradual tapering of ventilator support can be recommended for weaning without risk of sudden formation of atelectasis.


Unilateral pulmonary oedema of the contralateral lung following transaxillary sympathectomy.
A previously healthy young male developed unilateral pulmonary oedema immediately after re-expansion of an intentionally collapsed lung after transaxillary sympathectomy. The pulmonary oedema was localised to the contralateral, uncollapsed lung.


[Aortic dissection without intimal laceration: a case report and review of the problem].
A 43-year-old hypertensive woman with symptoms of aortic dissection was referred to our hospital 3 days after the initial episode. Transesophageal echocardiography and magnetic resonance showed a large intramural hematoma of the ascending and descending aorta without intimal flap and tears. Aortography confirmed the absence of intimal disruption. Medical therapy with beta and alpha blocking agents was established. After 5 months the patient was asymptomatic. On transesophageal echocardiogram and magnetic resonance, complete reabsorption of the hematoma of descending aorta was evident while a clearly identifiable false lumen and intimal flap (without tear) were present at the aortic arch level. The diagnostic criteria and therapeutic options are discussed and the literature on this topic is reviewed.


Ongoing evaluation of ease-of-use and usefulness of wireless tablet computers within an ambulatory care unit.
This ongoing research is to assess user acceptance of wireless convertible tablet portable computers in their support of patient care within the clinic environment and to determine their impact on workload reduction for the information staff. A previous publication described our initial experience with a limited wireless environment. There, we tested the premise that wireless convertible tablet computers were equivalent to desktop computers in their support of user tasks. Feedback from users demonstrated that convertible tablet computers were not able to replace desktop computers. Poor network access was a weakness as well as the "cognitive overhead" encountered due to technical problems. This paper describes our further experience with a centre-wide wireless implementation while using a new wireless device. The new tablets, which have some unique functions that existing desktop computers do not provide, have been well received by the clinicians.


The first records of quill mites of the family Syringophilidae (Acariformes: Prostigmata: Cheyletoidea) from trogoniform birds (Aves: Trogoniformes).
Two new species of quill mites of the family Syringophilidae parasitising trogoniform birds (Trogoniformes: Trogonidae) are described: Syringophiloidus quetzali sp. nov. from Pharomachrus mocinno Llave and Ph. antisianus (Orbigny); and Syringophilopsis trogoni sp. nov. from Trogon citreolus Gould and T. melanocephalus Gould. These findings are the first records of syringophilids associated with trogoniform birds.


Psychosocial Improvements in Children and Their Parents Following Strabismus Surgery.
To evaluate the psychosocial effects of strabismus surgery, the authors investigated the psychiatric symptoms and quality of life of children with strabismus and their parents. Children with strabismus between 6 and 18 years old (n = 39) and their parents (n = 39) were included. Psychiatric problems and the health-related quality of life were assessed with the Child Behavior Checklist and Pediatric Quality of Life Inventory, respectively. The State-Trait Anxiety Inventory for Children and Beck Anxiety Inventory were administered to children and parents, respectively. All assessments were conducted before and 3 months after strabismus surgery. Compared with preoperative assessments, children with strabismus had significantly fewer psychiatric problems and a better quality of life after the surgery (P < .001). Following surgery, decreased symptom levels of anxiety scores were also obtained from children and their parents (P < .001). The amount of ocular realignment was directly correlated with improvement in social and attention problems among children (r = 0.354 and r = 0.324, respectively; P < .05). Successful strabismus surgery has positive effects on psychosocial functioning and quality of life in children and their families. Surgical interventions seem beneficial for the management of strabismus in improving both physical and mental health. [J Pediatr Ophthalmol Strabismus. 2022;59(4):261-268.].


Recommended Values of the Fundamental Physical Constants: A Status Report.
We summarize the principal advances made in the fundamental physical constants field since the completion of the 1986 CODATA least-squares adjustment of the constants and discuss their implications for both the 1986 set of recommended values and the next least-squares adjustment. In general, the new results lead to values of the constants with uncertainties 5 to 7 times smaller than the uncertainties assigned the 1986 values. However, the changes in the values themselves are less than twice the 1986 assigned one-standard-deviation uncertainties and thus are not highly significant. Although much new data has become available since 1986, three new results dominate the analysis: a value of the Planck constant obtained from a realization of the watt; a value of the fine-structure constant obtained from the magnetic moment anomaly of the electron; and a value of the molar gas constant obtained from the speed of sound in argon. Because of their dominant role in determining the values and uncertainties of many of the constants, it is highly desirable that additional results of comparable uncertainty that corroborate these three data items be obtained before the next adjustment is carried out. Until then, the 1986 CODATA set of recommended values will remain the set of choice.


Discovery of new selective human aldose reductase inhibitors through virtual screening multiple binding pocket conformations.
Aldose reductase reduces glucose to sorbitol. It plays a key role in many of the complications arising from diabetes. Thus, aldose reductase inhibitors (ARI) have been identified as promising therapeutic agents for treating such complications of diabetes, as neuropathy, nephropathy, retinopathy, and cataracts. In this paper, a virtual screening protocol applied to a library of compounds in house has been utilized to discover novel ARIs. IC50's were determined for 15 hits that inhibited ALR2 to greater than 50% at 50 μM, and ten of these have an IC50 of 10 μM or less, corresponding to a rather substantial hit rate of 14% at this level. The specificity of these compounds relative to their cross-reactivity with human ALR1 was also assessed by inhibition assays. This resulted in identification of novel inhibitors with IC50's comparable to the commercially available drug, epalrestat, and greater than an order of magnitude better selectivity.


In Vitro Evaluation of the Aberdeen Knot for Continuous Suture Patterns with Large Gauge Suture.
To evaluate the strength, size, and holding capacity of the Aberdeen knot compared to surgeon's and square knots using large gauge suture. In vitro mechanical study. Knotted suture. Aberdeen, surgeon's, and square knots were tested using 2 and 3 USP polyglactin 910 and 2 USP polydioxanone under linear tension on a universal testing machine. Mode of failure and knot holding capacity (KHC) were recorded and relative knot security (RKS) was calculated as a percentage of KHC. Knot volume and weight were quantified by a digital micrometer and balance, respectively. Strength between number of throws, suture, suture size, and knot type were compared by ANOVA and post hoc testing. P≤.05 was considered significant. Aberdeen knots had higher KHC and RKS than surgeon's or square knots for all suture types and number of throws (P<.001). For all suture materials, none of the Aberdeen knots unraveled, but a portion of square and surgeon's knots with <7 throws did unravel (P=.101). Aberdeen knots had a smaller volume and weight than both surgeon's and square knots with equal numbers of throws (P<.001). The knot with the combined highest RKS and smallest size and weight was an Aberdeen knot with 4 throws using 3 USP polyglactin 910. The Aberdeen knots were stronger, more secure, and smaller than surgeon's and square knots for ending a continuous suture pattern. Clinically, the Aberdeen knot may be a useful alternative for completion of continuous patterns using large gauge suture, without sacrificing knot integrity.


Endometriosis with a pleural effusion and ascites. Report of a case treated with nafarelin acetate.
Although endometriosis is generally confined to the pelvis, it may occur at remote sites with unusual manifestations. Rare examples include pulmonary endometriosis and endometriosis associated with ascites. These complications represent diagnostic and therapeutic dilemmas, especially when both occur concurrently in the same patient. A 21-year-old nulligravida had a history of endometriosis and complained of severe dyspnea and weight loss. Pleural effusion and pelvic ascites associated with recurrent endometriosis were found. The patient was treated with thoracentesis and nafarelin acetate nasal spray. A remarkable improvement in her condition occurred, and the side effects of the spray were minimal during treatment. Gonadotropin-releasing hormone agonist therapy is recommended as the initial treatment for endometriotic ascites and/or pleural effusion.


Primary hyperparathyroidism diagnosed after surgical ablation of a costal mass mistaken for giant-cell bone tumor: a case report.
Primary hyperparathyroidism is a common endocrine disorder characterized by elevated parathyroid hormone levels, which cause continuous osteoclastic bone resorption. Giant cell tumor of bone is an expansile osteolytic tumor that contains numerous osteoclast-like giant cells. There are many similarities in the radiological and histological features of giant cell tumor of bone and brown tumor. This is a rare benign focal osteolytic process most commonly caused by hyperparathyroidism. We report the unusual case of a 40-year-old Caucasian woman in which primary hyperparathyroidism was diagnosed after surgical ablation of a costal mass. The mass was suspected of being neoplastic and histopathology was compatible with a giant cell tumor of bone. On the basis of the biochemical results (including serum calcium, phosphorous and intact parathyroid hormone levels) primary hyperparathyroidism was suspected and a brown tumor secondary to refractory hyperparathyroidism was diagnosed. Since giant cell tumor is a bone neoplasm that has major implications for the patient, the standard laboratory tests in patients with bone lesions are important for a correct diagnosis.


Follicular stimulating hormone, luteinizing hormone and estrogen levels before and after female sterilization.
A prospective pilot investigation of 14 women subjected to sterilization via laparoscopy, using bipolar electrocoagulation of the Fallopian tubes, did not reveal any significant change in the levels of follicular stimulating hormone (FSH), luteinizing hormone (LH) or estradiol either before or 3 months after the operation. However, the investigation gave evidence of a slight decrease in estradiol levels in the first 3 months after sterilization.


Transdermal fentanyl for cancer pain. Repeated dose pharmacokinetics.
The transdermal therapeutic system (fentanyl), or TTS(fentanyl), continuously delivers fentanyl for up to 72 h. The transdermal therapeutic system (fentanyl)-100 delivers approximately 100 micrograms/h. The repeated dose pharmacokinetics of this drug using the recommended dosing interval have not been evaluated previously and were determined in the present study. Blood samples were obtained from ten opioid-tolerant cancer patients who received five applications of TTS(fentanyl) at 72-h intervals. A sample of venous blood was taken before each dose; multiple samples were taken during and after the fifth application. A gas chromatographic/mass spectrometry method was used to assay fentanyl (limit of detection 0.2 ng/ml). For the fifth dose, the mean (SD) maximum concentration was 2.6 (1.3) ng/ml and the mean (SD) area under the serum fentanyl concentration-time curve (0-72 h) was 116.9 (59.9). Following removal of the system, the mean (SD) apparent half-life was 21.9 (8.9) h. There were no differences among the serum fentanyl concentrations measured before the second through fifth doses. Fentanyl absorption was 47% complete at 24 h, 88% complete at 48 h, and 94% complete at 72 h. The mean (SD) dose delivered during the 72-h period was 4.3 (1.1) mg. A first-dose trough concentration predicted from fifth-dose kinetics and the actual first-dose trough concentration were very similar. Adverse effects ascribed to the transdermal system were minimal. These results suggest that steady-state serum concentrations are approached by the second dose of TTS(fentanyl) and that the kinetics are stable with repeated dosing. The apparent half-life following system removal is relatively long, indicating ongoing absorption from a subcutaneous depot.


Protein acetylation and deacetylation: An important regulatory modification in gene transcription (Review).
Cells primarily rely on proteins to perform the majority of their physiological functions, and the function of proteins is regulated by post-translational modifications (PTMs). The acetylation of proteins is a dynamic and highly specific PTM, which has an important influence on the functions of proteins, such as gene transcription and signal transduction. The acetylation of proteins is primarily dependent on lysine acetyltransferases and lysine deacetylases. In recent years, due to the widespread use of mass spectrometry and the emergence of new technologies, such as protein chips, studies on protein acetylation have been further developed. Compared with histone acetylation, acetylation of non-histone proteins has gradually become the focus of research due to its important regulatory mechanisms and wide range of applications. The discovery of specific protein acetylation sites using bioinformatic tools can greatly aid the understanding of the underlying mechanisms of protein acetylation involved in related physiological and pathological processes.


Pulsed laser deposition with simultaneous in situ real-time monitoring of optical spectroscopic ellipsometry and reflection high-energy electron diffraction.
We present a pulsed laser deposition system that can monitor growth by simultaneously using in situ optical spectroscopic ellipsometry (SE) and reflection high-energy electron diffraction (RHEED). The RHEED precisely monitors the number of thin-film layers and surface structure during the deposition, and the SE measures the optical spectra of the samples simultaneously. The thin-film thickness information obtained from RHEED facilitates the SE modeling process, which allows extracting the in situ optical spectra, i.e., the dielectric functions of thin-films during growth. The in situ dielectric functions contain indispensable information about the electronic structure of thin-films. We demonstrate the performance of this system by growing LaMnO(3+δ) (LMO) thin-films on SrTiO3 (001) substrates. By using in situ SE and RHEED simultaneously, we show that real-time thickness and dielectric functions of the LMO thin-films can be effectively extracted. The simultaneous monitoring of both optical SE and RHEED offers important clues to understand the growth mechanism of atomic-scale thin-films.


Neurophysiology of juvenile myoclonic epilepsy: EEG-based network and graph analysis of the interictal and immediate preictal states.
The neuronal mechanisms of enduring seizure propensity and seizure precipitation in juvenile myoclonic epilepsy (JME) are not known. We investigated these issues, within the framework of the "network concept" of epilepsy. Design1: 19, unmedicated JME patients were compared with nineteen, age-, and sex-matched normal control persons (NC). A total of 120s, artifact-free, paroxysm-free, eyes-closed, resting state EEG background activity was analyzed for each person. Design2: interictal and immediate preictal periods of the JME patients were compared in order to explore interictal-preictal network differences. For both comparison designs, statistically significant differences of EEG functional connectivity (EEGfC), nodal and global graph parameters were evaluated. Design1: maximum abnormalities were: increased delta, theta, alpha1 EEGfC and decreased alpha2 and beta EEGfC in the JME group as compared to the NC group, mainly among cortical areas that are involved in sensory-motor integration. Nodal degree and efficiency of three, medial, basal frontal nodes were greater in JME than in NC, in the alpha1 band. Design2: preictal delta EEGfC showed further increase in the above-mentioned areas, as compared to the interictal state. Increased EEGfC indicates a hypercoupled state among the specified cortical areas. This interictal abnormality further increases in the preictal state. Nodal graph statistics indicates abnormal neuronal dynamics in the cortical area that is the ictal onset zone in JME. Interictal and preictal neuronal dysfunction has been described in terms of network dynamics and topography in JME patients. Forthcoming investigations of seizure precipitation and therapeutic drug effects are encouraged on this basis.


Application of optical imaging to study of extrapulmonary spread by tuberculosis.
The incidence of extrapulmonary tuberculosis is increasing, possibly due to the high frequency of co-infection with HIV. Extrapulmonary infections complicate diagnosis, have higher mortality rates and are more difficult to treat. Insight into the mechanisms involved in extrapulmonary spread of tuberculosis is critical to improving management. We set out to better understand extrapulmonary spread kinetics in mice and guinea pigs as well as the effects of infectious dose. We found that extrapulmonary spread occurs at a discrete time point when infected by low-dose aerosol, but at high-dose aerosol it occurs within the first 24h. The ability to follow tuberculosis in real-time during infection would allow us to better address the mechanisms involved. We found that mycobacteria can be optically imaged after pulmonary infection in the mouse lung, suggesting that this technology could be applied to study of extrapulmonary spread of tuberculosis.


Modified autotransplantation technique for surgical resection of complex pericardial synovial sarcoma.
Technical details for complex cardiac tumor resection are sparse. We describe the operative technique of modified autotransplantation for resection of a complex pericardial synovial sarcoma in a 63-year-old, Caucasian female. Surgical exposure demonstrated tumor origin at the superior cavoatrial junction and invasion of the aorta, main pulmonary artery, superior pulmonary veins, and left atrial roof. Full macroscopic surgical resection was achieved. The patient received adjuvant radiation for microscopic positive margins and remains alive and with no tumor progression at one year postoperatively. We conclude that modified autotransplantation is a challenging but effective surgical technique when performed with careful patient selection and availability of skilled, cardiothoracic surgeons at a cardiac center of excellence.


Comparison of the variability in the levels of pesticide residue observed in Japanese cabbage and grape units.
To estimate variations in pesticide residue levels in crops, the variability factors (VFs, the 97.5th percentile of the residue levels in the sample divided by the average residue levels in the lot) in residue levels of acetamiprid and cypermethrin applied to cabbage and grapes were investigated, respectively. The VFs in the residue levels of both pesticides in cabbage (2.00 and 2.39, respectively) were clearly higher than those in grapes (1.82 and 1.63, respectively). Although the residue levels of both pesticides in grapes showed a normal distribution, those values in cabbage were slightly skewed at lower residue levels. Individual residue levels in grapes had a good agreement between acetamiprid and cypermethrin. In contrast, the distribution of cypermethrin residue levels in cabbage was slightly skewed at higher residue levels as compared to that of acetamiprid. These results indicate that the difference in the relative distribution of the two pesticides between cabbage and grapes might be due to the influence of various factors such as differences in crop species, plant cultivation methods, and physicochemical properties of the pesticides.


Menopausal experiences and bone density of Mayan women in Yucatan, Mexico.
This cross-sectional study reports on the menopausal transition of Mayan women from Yucatan, Mexico. A total of 228 women completed the study, and 118 women were classified by history as postmenopausal; the others were premenopausal. Demographic information, reproductive history, physical examination, hormone concentrations, radial bone density, food samples, and history of physical activity were obtained. The average age at which menopause occurred by history was 44.3 +/- 4.4 years; this is reflected in the distribution of FSH levels by age. None of the women reported symptoms of hot flashes, and none recalled any history of significant symptoms associated with their menopausal transition. Hormone levels were similar to U.S. reference values with elevated FSH (66.6 +/- 29.1 mIU/ml), low estradiol (9.4 +/- 8.3 pg/ml) and estrone (13.3 +/- 7.8 pg/ml), E1 > E2, and normal levels of testosterone and androstenedione. BMD declined with age, and values were lower than reference values for United States women. Clinical evidence of fracture was not detected by history or physical examination even for those who were 20 years postmenopausal. The endocrine characteristics of menopause among Mayan women in Yucatan are similar to hormonal changes reported for women in the United States, but signs, symptoms, and apparent consequences are different in the two populations.


The ARS309 chromosomal replicator of Saccharomyces cerevisiae depends on an exceptional ARS consensus sequence.
Autonomously replicating sequence (ARS) elements, which function as the cis-acting chromosomal replicators in the yeast Saccharomyces cerevisiae, depend upon an essential copy of the 11-bp ARS consensus sequence (ACS) for activity. Analysis of the chromosome III replicator ARS309 unexpectedly revealed that its essential ACS differs from the canonical ACS at two positions. One of the changes observed in ARS309 inactivates other ARS elements. This atypical ACS binds the origin recognition complex efficiently and is required for chromosomal replication origin activity. Comparison of the essential ACS of ARS309 with the essential regions of other ARS elements revealed an expanded 17-bp conserved sequence that efficiently predicts the essential core of ARS elements.


Clavanin permeabilizes target membranes via two distinctly different pH-dependent mechanisms.
The pH dependence of the antimicrobial and membrane activity of clavanin A, a peptide antibiotic that is rich in histidines and glycines, was analyzed in growth and membrane leakage experiments. Clavanin A more effectively inhibited the growth of the test organism Lactobacillus sake when the pH of the medium was lowered. Whereas the wild-type peptide efficiently released fluorophores from unilamellar vesicles at neutral pH according to a nonspecific permeabilization mechanism, it did not permeabilize model bilayers at low pH. It was therefore suggested that this peptide uses a distinct mode of action under acidic conditions different than that used around neutral pH. However, at low pH, the membrane is still the target for clavanin A, as the peptide collapsed both vital transmembrane proton gradients and ion gradients under these conditions. Clavanin A did not act as a ionophore across phospholipid bilayers, indicating that membrane constituents other than membrane phospholipids are involved in the dissipation of transmembrane ion gradients. Membrane proteins that generate transmembrane ion gradients are suggested to be the targets for clavanin A at low pH. In addition to the histidines, the three glycine residues of clavanin A are shown to play an important role in the specific mode of interaction with these membrane targets. These residues may induce a flexible hydrophobic conformation that allows the peptide to exert different membrane activities. This study demonstrates that clavanin A is a special membrane-active peptide that has access to two markedly distinct pH-dependent modes of actions.


Efficacy of behavioral versus triazolam treatment in persistent sleep-onset insomnia.
This study compared differential effects of behavioral therapy and triazolam in a clinical population with sleep-onset insomnia. Triazolam was hypothesized to decrease sleep latency and frequency and duration of awakening, with some effects during the first night's administration. But at follow-up, sleep measures were predicted to return to baseline levels. Behavioral treatment was hypothesized to effect sleep after 2 or more weeks of training which persisted at follow-up. Thirty patients with average sleep latencies of 81.48 minutes, who reported chronic insomnia for an average of 2.6 years, were randomly assigned to one of two treatment groups: behavioral stimulus control/relaxation training and triazolam. Both treatments decreased sleep latency but differentially. Triazolam was effective immediately but maintained only some gains at follow-up. Behavioral treatment decreased sleep latency beginning the second week, when subjects expected no improvement, with gains maintained at follow-up. Comparisons showed that triazolam group latencies returned toward baseline, while behavioral group gains were maintained at follow-up. Triazolam treatment showed superior immediate treatment effects, while behavioral treatment showed superior treatment effects at follow-up, effects that accrued during the training period and differentially persisted at follow-up. One treatment strategy implied by these results would be to combine these two interventions concurrently. This would seem to use the immediate effects produced by the medication until the behavioral skills were learned, at which point medication would be terminated. This strategy could offer immediate relief and sustained effects at drug termination.


In-situ measurements of low-level mercury vapor exposure from dental amalgam with zeeman atomic absorption spectroscopy.
Alongside food, emissions from amalgam fillings are an essential contribution to man's mercury burden. Previous methods for the determination of intraoral mercury vapor (Hg degrees ) release used principally some form of preconcentration of Hg on gold (film or wool), allowing relatively few measurements with unknown precision and sensitivity at selected times. Recently available computer-controlled Hg detectors operating on Zeeman atomic absorption spectroscopy (ZAAS) facilitate the direct real-time measurement of Hg degrees concentrations. It was the aim to adapt this method for a comparative investigation of emission processes from fillings in situ and from amalgam specimens in vitro. In addition to the ZAAS instrument, the apparatus consisted of a pump, magnetic valves, an electronic flow controller and a handle with a disposable mouth piece for aspiration of oral air. A programmable timer integrated the computer-controlled instrument operation and the data collection into a standard sampling protocol. A fast exponential decay of the emission was found after stimulation of amalgam specimens and of fillings in situ (halftimes 8.6 and 10.7 min). Precision was evaluated by a series of measurements on a single patient which indicated a consistently low coefficient of variation between 18% and 25%. After insertion of a few new fillings, sensitivity was high enough to detect a significant increase in emission against the background emission from the majority of old fillings. Zeeman-AAS in connection with a semi-automated sampling protocol and data storage provides precise in-situ measurements of Hg degrees emission from dental amalgam with real-time resolution. This facilitates the detailed exploration of the Hg degrees release kinetics and the applicability to large-scale studies.


[Structural changes in wild-type Cry3A delta-endotoxin and its mutants in ethyl alcohol solutions at pH 2-2.5].
The methods of circular dichroism, scanning microcalorimetry, electron microscopy, and proteolysis were used to study the ability of wild-type Cry3A-delta-endotoxin and three mutant toxins with cysteine substitutions in helices alpha3 and alpha4 (domain I) to form oligomeric structures in acidic alcohol solutions that reproduce the premembrane environment. At pH 2-2.2 and 20% ethanol, the mutant toxins with single substitutions E132C (alpha3) and E160C (alpha4), as well as the double mutant E132C/S160C with a cysteine bridge connecting helices alpha3 and alpha4, form short linear oligomers specific for Cry3A with a high content of the beta-structure and enhanced sensitivity to proteolysis with pepsin. The data obtained show that the formation of oligomeric structures of this type does not require any divergence of helices alpha3 and alpha4 in domain I of the Cry3A toxin. It has been demonstrated that, at higher pH values in 20% solution of ethanol, the proteins studied are in a metastable state, and their ability to form oligomeric structures depends on temperature.


STAT3-Dependent Gene TRIM5γ Interacts With HBx Through a Zinc Binding Site on the BBox Domain.
Owing to its broad-spectrum antivirus activities, interferon (IFN) is an important alternative agent for use in the treatment of hepatitis B virus (HBV)-infected patients; however, the mechanism involved in the inhibition of HBV infection and replication by IFN remains unclear. We previously reported that the induction of TRIM5γ is important in the IFN treatment of HBV patients as it promotes the degradation of the HBx protein, while the manner in which TRIM5γ is induced by IFN and how TRIM5γ interacts with HBx remain unestablished until date. Our present findings confirmed the TRIM5γ-HBx-DDB1 interactions in the HBV-infected Primary human hepatocytes (PHH), and we further found that STAT3, and not STAT1, was responsible for the induction of TRIM5γ upon IFN stimulation and that the zinc binding site His123 on the BBOX domain was a decisive site in the interaction between TRIM5γ BBOX and HBx. In addition, based on the BBOX domain, we detected a 7-amino acid peptide with the potential of promoting HBx degradation and inhibiting HBV replication. On the other hand, we noted that the TRIM5γ expression was inhibited by HBV in chronically HBV infected patients. Thus, our study identified the crucial role of STAT3 in the induction of TRIM5γ, as well as proposed a 7-amino acid, small peptide as a potential candidate for the development of therapeutic agents targeting HBx.


Peer deviance, social support, and symptoms of internalizing disorders among youth exposed to Hurricane Georges.
This study examined the influence of peers in meeting DSM-IV symptom criteria for an internalizing disorder in adolescents exposed to Hurricane Georges. Participants included a representative community sample of 905 youth (n = 476 boys) ages 11-17, residing in Puerto Rico. Data were gathered on hurricane exposure, symptoms of internalizing disorders, peer social support, peer violence, and peer substance use through in-person structured interviews with adolescents and caretakers from 1999 to 2000 in Puerto Rico, 12-27 months after Hurricane Georges. Hurricane exposure, peer violence, and peer substance use predicted whether adolescents met DSM-IV symptom criteria for a measured internalizing disorder. An interaction was found between hurricane exposure and peer violence, which indicated that hurricane exposure was significantly related to meeting DSM-IV symptom criteria for an internalizing disorder among adolescents who do not report associating with violent peers. However, for participants who reported high levels of peer violence, hurricane exposure did not convey additional risk for meeting DSM-IV symptom criteria for an internalizing disorder. With the increasing role peers play in adolescents' lives, understanding the influence of peers on the development of internalizing symptoms following hurricane exposure may assist in planning developmentally sensitive response plans.


A causal model of health status and satisfaction with medical care.
Patients with better health status have often been shown to be more satisfied with their medical care, but the causal factors in this relation have not been determined. In this study, a longitudinal assessment of these two constructs was undertaken in which older patients in a health maintenance organization were interviewed at baseline (N = 590) and again 1 year later (N = 526) about their health status and satisfaction with their medical care. Structural equation modeling using LISREL procedures revealed that the predominant direction of causation went from earlier self-perceived overall health and functional ability to later levels of satisfaction. There was no evidence for causal paths going from satisfaction to later health. In addition, a test of spuriousness indicated that for self-perceived overall health, the significant longitudinal path was unlikely to be explained by unmeasured confounding variables.


Creatine kinase alterations after acute malignant hyperthermia episodes and common surgical procedures.
Skeletal muscle may release creatine kinase (CK) during a malignant hyperthermia (MH) episode; however, muscle damaged during surgery may also release CK. This study examined the overlap between peak plasma CK levels in patients suspected of having had a MH episode (data obtained from North American MH Registry) and previously published CK changes occurring after common surgeries. For patients who were subsequently proven to be MH positive by muscle biopsy, there was considerable overlap. This was most significant with surgeries having substantial tissue damage, such as major vascular surgery and abdominal surgery. Overlap was much less with minimally invasive surgery, such as cystoscopy. Approximately 30% of MH positive patients treated with dantrolene had peak CK in the range of most surgical procedures, and approximately 50% of MH positive patients not given succinylcholine had peak CK similar to those of most surgical procedures. Dantrolene did not significantly alter peak CK in MH positive patients; however, succinylcholine was associated with significantly higher peak CK. These data suggest that patients who have had an acute MH episode during a surgical procedure may have peak CK values within the range of CK values expected from the procedure itself.


LC-MS/MS Analysis and Pharmacokinetics of Sodium (±)-5-Bromo-2-(α-hydroxypentyl) Benzoate (BZP), an Innovative Potent Anti-Ischemic Stroke Agent in Rats.
A rapid, sensitive and selective liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of sodium (±)-5-Bromo-2-(α-hydroxypentyl) benzoate (BZP) and its active metabolite 3-butyl-6-bromo-1(3H)-isobenzofuranone (Br-NBP) in rat plasma using potassium 2-(1-hydroxypentyl)-benzoate (PHPB) and l-3-n-butylphthalide (NBP) as internal standards (IS). Chromatographic separation was achieved on a Hypersil GOLD C18 column using a gradient elution of ammonium acetate and methanol at a flow rate of 0.2 mL/min. Good linearity was achieved within the wide concentration range of 5-10,000 ng/mL. The intra-day and inter-day precision was less than 8.71% and the accuracy was within -8.53% and 6.38% in quality control and the lower limit of quantitation samples. BZP and Br-NBP were stable during the analysis and the storage period. The method was successfully applied to pharmacokinetic studies of BZP in Sprague-Dawley rats for the first time. After a single intravenous administration of BZP at the dose of 0.75 mg/kg, the plasma concentration of BZP and Br-NBP declined rapidly and the AUC0-t of BZP was significantly greater in female rats compared to male rats (p < 0.05). The data presented in this study serve as a firm basis for further investigation of BZP in both preclinical and clinical phases.


Variability of Seed Germination and Dormancy Characteristics and Genetic Analysis of Latvian Avena fatua Populations.
Avena fatua is an economically detrimental weed found in cereal growing areas worldwide. Knowledge about the variation of dormancy and germination characteristics, as well as of the genetic diversity and structure can provide additional information about different populations within a region. Identification and development of potential indicators or markers of seed germination behavior, would be useful for modelling emergence and seed bank dynamics. This study aimed to describe the initial germination, dormancy, and morphological characteristics of seeds from different Latvian populations of A. fatua and to investigate the genetic structure of local populations. Seed samples from 26 to 41 locations in different regions of Latvia were collected over three years. Seed morphology, initial germination, and the effect of dormancy-breaking treatments were determined. Seeds from 18 Latvian populations with contrasting seed dormancy characteristics were genotyped. Although morphological differences between seeds from different regions were revealed, genetic analysis of the selected populations indicated that the population structure was not related to geographical location, which probably reflects the impact of anthropogenic dispersal of A. fatua seeds. The effect of dormancy-breaking treatments varied between years, between climatic zones and between populations, indicating environmental influences as well as inherited dormancy.


Antibodies to rotavirus outer capsid glycoprotein VP7 neutralize infectivity by inhibiting virion decapsidation.
The rotavirus capsid is composed of three concentric protein layers. Proteins VP4 and VP7 comprise the outer layer. VP4 forms spikes, is the viral attachment protein, and is cleaved by trypsin into VP8* and VP5*. VP7 is a glycoprotein and the major constituent of the outer protein layer. Both VP4 and VP7 induce neutralizing and protective antibodies. To gain insight into the virus neutralization mechanisms, the effects of neutralizing monoclonal antibodies (MAbs) directed against VP8*, VP5*, and VP7 on the decapsidation process of purified OSU and RRV virions were studied. Changes in virion size were followed in real time by 90 degrees light scattering. The transition from triple-layered particles to double-layered particles induced by controlled low calcium concentrations was completely inhibited by anti-VP7 MAbs but not by anti-VP8* or anti-VP5* MAbs. The inhibitory effect of the MAb directed against VP7 was concentration dependent and was abolished by papain digestion of virus-bound antibody under conditions that generated Fab fragments but not under conditions that generated F(ab')(2) fragments. Electron microscopy showed that RRV virions reacted with an anti-VP7 MAb stayed as triple-layered particles in the presence of excess EDTA. Furthermore, the infectivity of rotavirus neutralized via VP8*, but not that of rotavirus neutralized via VP7, could be recovered by lipofection of neutralized particles into MA-104 cells. These data are consistent with the notion that antibodies directed at VP8* neutralize by inhibiting binding of virus to the cell. They also indicate that antibodies directed at VP7 neutralize by inhibiting virus decapsidation, in a manner that is dependent on the bivalent binding of the antibody.


Modeling of chemical control of human respiratory system.
This paper addresses the fundamental understanding of the chemical control of respiratory system that would be needed to improve the efficiency of artificial ventilators and implement appropriate controllers. Hence, non-linear model for simulation of chemical control of human respiratory system under different physiological conditions is presented. The paper also presents the non-linear dissociation behaviors for oxygen and carbon dioxide solutions in the blood and the Bohr-Haldane effects as well as a new controller model. Tests were conducted on the model under different conditions such as hypoxia, hypercapnia, hypoventilation, and hyperventilation with some combination tests, in order to verify the validity of the assumptions that were made. Results indicate a very close agreement between the responses obtained from the present model and other published experimental and theoretical results. This model can be used to design a novel artificial respirator controller meeting the bodily requirements of the patient under mechanical ventilation.


Risk factors for HIV-1 transmission from HIV-seropositive male blood donors to their regular female partners in northern Thailand.
To describe risks for HIV transmission from male blood donors to their regular female sex partners in Chiang Mai, Thailand. Cross-sectional study. From March 1992 through September 1995, 405 HIV-seropositive male blood donors (index cases) and their regular female partners were enrolled in the study. Women with risk factors for HIV infection other than sexual contact with the index male were excluded. Couples were interviewed and examined; specimens were collected for laboratory analysis. Overall, 46% of the 405 women enrolled were HIV-positive. Ninety-eight per cent of male index cases had a history of sex with a female prostitute; 1.5% reported always using condoms with their regular partner. History of sexually transmitted disease (STD) and swollen inguinal lymph nodes in the female partner were associated with an increased risk of HIV infection in the female. History in the female of genital herpes [odds ratio (OR), 3.46; 95% confidence interval (CI), 1.50-8.78], gonorrhea or chlamydia infection (OR, 2.71; 95% CI, 1.39-5.53), and stable relationship of longer than 24 months (OR, 2.28; 95% CI, 1.02-5.09) were associated with an increased risk of HIV infection in the female. Consistent condom use in the past 2 years (OR, 0.10; 95% CI, 0.01-0.79) was associated with a decreased risk of HIV infection in the female. Married women in northern Thailand who appear otherwise to be at low risk for HIV infection may be exposed to this virus by their husbands. High rates of sex with commercial sex workers among men and low use of condoms within stable relationships may be important factors promoting the transmission of HIV in married couples. Programs to increase the regular use of condoms among married couples could be an important public health intervention to prevent transmission of HIV and other types of STD in northern Thailand.


Egg quality of Japanese quail depends on layer age and storage time.
The aim of the study was to analyse egg quality changes of Japanese quail during the reproductive season and after several days of storage. A significantly higher egg weight was recorded in week 9 than in week 25 and week 31 of age ofbirds. The egg specific gravity value of eggs also decreased with the age of quails. The smallest yolk weight and its percentage proportion in egg mass were observed in week 25. Eggs laid in weeks 9 and 25 were characterised by similar and lower (P < or = 0.05) white weight in comparison to those examined on the last date. The highest white proportion in egg weight was recorded during the 25th week. The value of this trait was similar or smaller on the remaining dates. Similar and lower values of the white index and Haugh units, in comparison with the first date, were determined in eggs laid on weeks 25 and 31, whereas similar and greater eggshell thickness than in the 9th week of age was recorded on the last two dates of examination. Lower (P < or = 0.05) egg weight was observed after 5 and 8 than after 0 (measured on the day of laying) days of storage. Likewise, smaller egg specific gravity was recorded after three consecutive periods of storage in comparison with the eggs examined on the day of laying. Eggs examined after 3, 5 and 8 days of storage, in comparison with those stored for 0 days, were characterised by lower yolk index. Significantly smaller weight and white index than after 0 days were determined in eggs stored, respectively, for 5 and 8 as well as for 3, 5 and 8 days. Similarly, smaller numbers of Haugh units were determined after 3 consecutive storage periods in comparison with the eggs examined on the day of laying. Eggs stored for 0 and 3 days were characterised by a greater eggshell weight than those examined after 5 and 8 days. The weight of Japanese quail eggs decreased significantly from day 5 onwards, whereas the egg specific gravity-- from the 3rd day of storage. Yolk and white quality began to deteriorate already after 3 days of egg storage. It appears, therefore, that egg storage of Japanese quails before hatching lasting more than 3 days in the analysed conditions deteriorated the eggshell quality and internal egg traits.


A capillary monolithic trypsin reactor for efficient protein digestion in online and offline coupling to ESI and MALDI mass spectrometry.
We describe the preparation of a capillary trypsin immobilized monolithic enzyme reactor (IMER) for a rapid and efficient digestion of proteins down to the femtomole level. Trypsin was immobilized on a poly(glycidyl methacrylate-co-acrylamide-co-ethylene glycol dimethycrylate) monolith using the glutaraldehyde technique. Digestion efficiencies of the IMER were evaluated using model proteins and protein mixtures as well as chemically cross-linked lysozyme regarding the addition of denaturants and increasing digestion temperature. The trypsin IMER described herein is applicable for the digestion of protein mixtures. Even at a 1000-fold molar excess of one protein, low-abundance proteins are readily identified, in combination with MS/MS analysis. An online setup of the IMER with reversed phase nano-HPLC separation and nano-ESI-MS/MS analysis was established. The great potential of the trypsin IMER for proteomics applications comprise short digestion times in the range of seconds to minutes, in addition to improved digestion efficiencies, compared to in-solution digestion.


Cytotoxic Activity and Phytochemical Analysis of Artemisia haussknechtii Boiss.
Cytotoxic activity of crude extract and fractions (petroleum ether, dichloromethane, and n-butanol) of Artemisia haussknechtii aerial parts was investigated by MTT assay. Dichloromethane fraction showed the highest cytotoxic effect on MCF-7 cell line (IC50 = 297.17 ± 7.99 µg/mL). Phytochemical analysis of the most effective fraction was carried out using normal phase column chromatography (CC) to get eight sub-fractions (A-H). Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) were used for further purification. Four known compounds with cytotoxic effects on cancer cell lines were isolated from the most active fraction, including 5-Hydroxy-3',4',6,7-tetramethoxyflavone (eupatilin 7-methyl ether), 5-hydroxy 3,3',4',6,7-pentamethoxy-flavone (artemetin), 6-methoxy-7-hydroxycoumarin (scopoletin), and methyl caffeate. Structure elucidation of isolated compounds was done using spectroscopic techniques, including ESIMS and 1D-NMR (1H and 13C). Cytotoxic activity of A. haussknechtii is probably due to coumarin and flavonoid compounds.


[Clinical applications of hair analysis in the Netherlands].
Hair analysis has been known for its role in forensic science. However, it may also have an important role in the clinical approach of various exposures, for example drugs and heavy metals. In the Netherlands, it is used infrequently probably because clinicians are not familiar with this diagnostic possibility. In this article, we present two cases in which hair analysis proved to be crucial in the diagnosis of two different exposures (tobacco smoking and arsenic). We provide an overview of the various clinical applications of hair analysis and important factors affecting the results of hair analysis.


Main phenolic compounds of the melanin biosynthesis pathway in bruising-tolerant and bruising-sensitive button mushroom (Agaricus bisporus) strains.
Browning is one of the most common postharvest changes in button mushrooms, which often results in economic losses. Phenolic compounds, which are associated with browning, were extracted from the nonbruised and bruised skin tissue of various button mushrooms with a sulfite-containing solution and analyzed with UHPLC-PDA-MS. In total, 34 phenolic compounds were detected. Only small differences in the total phenolic content between bruising-tolerant and -sensitive strains were observed. The contents of γ-L-glutaminyl-4-hydroxybenzene (GHB) and γ-L-glutaminyl-3,4-dihydroxybenzene (GDHB) correlated with bruising sensitivity; for example, R(2) values of 0.85 and 0.98 were found for nonbruised brown strains, respectively. In nonbruised skin tissue of the strains with brown caps, the GHB and GDHB contents in sensitive strains were on average 20 and 15 times higher, respectively, than in tolerant strains. GHB and GDHB likely participate in the formation of brown GHB-melanin, which seemed to be the predominant pathway in bruising-related discoloration of button mushrooms.


Locally delivered doxycycline hyclate: case selection, preparation, and application.
This article presents the perspective of a private practice clinician who participated in the phase III clinical trials of Atridox (doxycycline hyclate) 10%. The selection of periodontal cases most likely to benefit from treatment with Atridox as part of overall comprehensive case management is discussed. Atridox may be used either before or after scaling and root planing or, in more rare circumstances, as a stand-alone therapy. Preparation of the material is discussed, and guidance for optimal application is provided.


Prevalence of low bone mass and endocrine disorders in hip fracture patients in Southern Germany.
Low bone mass and conditions leading to falls are considered to determine the risk of hip fracture. Bone mass is influenced by underlying diseases and risk factors such as malnutrition and life style habits. In order to evaluate the clinical significance of osteopenia and risk factors in the pathogenesis of hip fracture, we have studied 146 consecutive patients (27 males, 119 females, aged 53-95 years) undergoing rehabilitation shortly after hip fracture (4 cases of hip fracture resulting from traffic accidents had been excluded). We measured bone mineral density (BMD) at the hip (DXA, Hologic QDR 2,000+), evaluated vitamin D status (serum 25-OH-Vitamin D) and nutritional calcium intake (clinical dietician), and searched for endocrine disorders (physical examination and serum hormones). Femoral neck BMD was 0.543 +/- 0.084 g/cm2 in females and 0.635 +/- 0.087 g/cm2 in males (p < 0.01). For both sexes, the values were significantly lower compared to age-matched controls. 88% of the females and 69% of the males had T-Scores below -2.5, thus fulfilling the densitometric WHO criteria of osteoporosis. Nutritional calcium intake was similar in both sexes (804 +/- 330 mg/day in women, 738 +/- 295 mg/day in men), but lower compared to coxarthrosis patients (1080 +/- 436 mg/day). Vitamin D deficiency was prevalent in 69% of the women, and in 55%, of the men with hip fracture. In female hip fracture patients, the serum alkaline phosphatase was significantly higher (194 +/- 82 U/I) as compared to patients with surgery because of coxarthrosis (142 +/- 46 U/I), supporting the view that some degree of osteomalacia and high turnover was present. Primary hyperparathyroidism (pHPT) was newly detected in 1% and hyperthyroidism in 4.3% of the cases. Hip fracture occurs at higher BMD values in men compared to women suggesting different fracture thresholds. Vitamin D deficiency and low calcium intake are common in hip fracture patients. However, before initiation of vitamin D treatment pHPT should be excluded. Determination of TSH is recommendable in all hip fracture patients.


Measurement of Reactive Oxygen and Nitrogen Species in Living Cells Using the Probe 2',7'-Dichlorodihydrofluorescein.
Reactive oxygen species and reactive nitrogen species (RONS) are involved in programmed cell death in the context of numerous degenerative and chronic diseases. In particular, the ability of cells to maintain redox homeostasis is necessary for an adaptive cellular response to adverse conditions that can cause damage to proteins and DNA, resulting in apoptosis and genetic mutations. Here, we focus on the 2',7'-dichlorodihydrofluorescein diacetate (DCFH2-DA) assay to detect RONS. Although this fluorescence-based assay is widely utilized due to its high sensitivity to detect changes in cellular redox status that allow measuring alterations in RONS over time, its validity has been a matter of controversy. If correctly carried out, its limitations are understood and results are correctly interpreted, the DCFH2-DA assay is a valuable tool for cell-based studies.


Isolation and sequence of complementary DNA encoding human extracellular superoxide dismutase.
A complementary DNA (cDNA) clone from a human placenta cDNA library encoding extracellular superoxide dismutase (EC-SOD; superoxide:superoxide oxidoreductase, EC 126.96.36.199) has been isolated and the nucleotide sequence determined. The cDNA has a very high G+C content. EC-SOD is synthesized with a putative 18-amino acid signal peptide, preceding the 222 amino acids in the mature enzyme, indicating that the enzyme is a secretory protein. The first 95 amino acids of the mature enzyme show no sequence homology with other sequenced proteins and there is one possible N-glycosylation site (Asn-89). The amino acid sequence from residues 96-193 shows strong homology (approximately 50%) with the final two-thirds of the sequences of all known eukaryotic CuZn SODs, whereas the homology with the P. leiognathi CuZn SOD is clearly lower. The ligands to Cu and Zn, the cysteines forming the intrasubunit disulfide bridge in the CuZn SODs, and the arginine found in all CuZn SODs in the entrance to the active site can all be identified in EC-SOD. A comparison with bovine CuZn SOD, the three-dimensional structure of which is known, reveals that the homologies occur in the active site and the divergences are in the part constituting the subunit contact area in CuZn SOD. Amino acid sequence 194-222 in the carboxyl-terminal end of EC-SOD is strongly hydrophilic and contains nine amino acids with a positive charge. This sequence probably confers the affinity of EC-SOD for heparin and heparan sulfate. An analysis of the amino acid sequence homologies with CuZn SODs from various species indicates that the EC-SODs may have evolved from the CuZn SODs before the evolution of fungi and plants.


[Cardiotoxic effect of the tocolytic agent fenoterol (partusisten) in the neonate (author's transl)].
Forty-two children of mothers treated with beta-adrenergic drugs were investigated cardiologically during a period of 3 1/2 years. Eleven neonates were seen to suffer from severe toxic myocardiopathy associated with heart failure. The mothers had in part been given excessive fenoterol doses for 13 weeks on the average. One child died of regressive myocardial alterations as confirmed by autopsy. Suspicion of toxic myocardiopathy was suggested in another 16 children. Related to the very large number of mothers treated with tocolytic agents, fenoterol-induced myocardiopathy so far appears to be rare. In agreement with other reports, however, our observations suggest the possibility of severe, even perilous myocardial damage. Therefore, the indication and dosage of fenoterol should be subjected to more rigid criteria than so far applied.