Neuro-cognitive functions in patients of major depression.
Earlier studies have shown conflicting results regarding impairment of executive functions in patients of major depression. In the present study, subjects were divided into following groups-Group 1 (19 Males & 11 females, age 18-45 years) consisting of patients of major depression diagnosed on the basis of DSM IV criteria and Group 2 (20 Males & 10 females, age 18-45 years) consisting of healthy subjects who had been screened for any sub-psychiatric illness by standardized hindi version of Goldberg's General Health Questionnaire (G.H.Q.). All the Group 1 & Group 2 subjects were rated on Hamilton Rating Scale for Depression (HAM-D) and then administered Neuro-cognitive test battery consisting of Letter Cancellation test, Trail Making test 'A' & 'B', Ruff Figural Fluency test, Forward & Reverse Digit Span test for assessing following cognitive domains: Attention span, visuo-motor scanning, short term memory and executive functions like working memory, information manipulation, set-shifting strategy analysis etc. Results showed significant impairment on all the studied neuro-cognitive parameters in Group 1 subjects as compared to Group 2 subjects, which demonstrate significant impairment of attention, visuomotor speed, immediate verbal memory, short-term retentive capacity and executive functions (strategic planning, attentional set shifting and working memory) in patients of major depression. It may probably be due to the disruption of dorso-lateral prefrontal cortex (dlpfc) and orbito-frontal networks in patients of major depression.


[A chapter from the history of the pharmacy of the Brothers of Mercy in Spišské Podhradí in the 19th and 20th centuries].
On the basis of archival materials, period publications and literary sources, this paper presents the data concerning the history of the pharmacy of the Hospital Order of St. John of God - the Brothers of Mercy in Spišské Podhradie in Slovakia. The Brothers of Mercy were active in Spišské Podhradie from the second half of the 17th century and in harmony with the main mission of the Order they ran a hospital and a pharmacy. The paper briefly characterizes the activities of the Brothers of Mercy in Spišské Podhradie, describes their pharmacy, its material-technical equipment and personnel carrying out its operation, focusing on the 19th century and the first decades of the 20th century till the take-over of the pharmacy and convent by the Czechoslovak province.


The impact of protein corona on the behavior and targeting capability of nanoparticle-based delivery system.
Once introduced into physiological environment, nanoparticles (NPs) are immediately coated with proteins, resulting in formation of what is known as protein corona. The formation of protein corona can be affected by many factors. Likewise, the addition of protein corona can alter the physicochemical properties and biodistribution of NPs. NPs with the coating protein corona can be considered as a biological identity that recognized by cells in biological system. Thus, to understand and regulate the effect of protein corona on targeting capability of NPs in vivo, it is necessary to elucidate the interaction between the NPs and the biological fluid. In this review, we first elucidate the factors influencing the formation of protein corona, including NPs physicochemical factors, such as NPs composition, size, shape, surface chemistry, etc., and environmental factors, such as environmental temperature, protein origins, etc. Then, we focus on the effect of protein corona on the passive targeting and active targeting, and discuss the probable reasons that causing the discrepant results. Finally, we review the strategies for tuning the protein corona to promote targeting, including reducing protein adsorption and recruiting specific proteins.


Copy number variations of the IL-22 gene are associated with ankylosing spondylitis: A case-control study in Chinese Han population.
IL-22 provides a new insight into the mechanisms of autoimmunity, and copy number variations (CNVs) are associated with autoimmune diseases. This study aims to explore the association of IL-22 gene CNVs with ankylosing spondylitis (AS) susceptibility. The copy numbers of IL-22 gene (2 fragments: IL-22_1, IL-22_2) were examined by AccuCopy™ methods in a cohort of 649 AS patients and 628 controls. Association of IL-22 CNVs and AS susceptibility was analyzed, and AS risk was estimated by Odds Ratio (ORs) and 95% confidence intervals (CIs), and the Benjamini-Hochberg method was applied to regulate the false discovery rate (FDR). We found one copy of IL-22 gene was significantly associated with AS [OR=0.345, 95%CI (0.144, 0.827), P=0.013, PFDR=0.026] in the IL-22_2 fragment, and this association still exist after adjustment of age and sex [OR=0.344, 95%CI (0.143, 0.825), P=0.017, PFDR=0.034]. In the stratification analysis by gender, the statistical difference was detected in males in the IL-22_2 fragment [OR=0.306, 95%CI (0.121, 0.778), P=0.009, PFDR=0.018; adjusted OR=0.306, 95%CI (0.120, 0.777), P=0.013, PFDR=0.026]. We suggest that IL-22 CNVs are associated with AS and that lower copy number might be a protective factor for AS, especially in male patients.


Exercise-induced urticaria and anaphylaxis.
Five patients with exercise-induced anaphylactoid reactions are reported. Because of a growing interest in physical exercise and the severity of the symptoms it is important to recognize this condition, even though rare. All of our 5 patients had a history of urticaria and anaphylaxis in association with physical stress, but it seems difficult to induce anaphylactoid reactions under laboratory conditions. Two different clinical patterns could be distinguished in these patients. Three had the anaphylactoid form with signs of alternative complement pathway activation, while 2 patients had the variant form presenting first as cholinergic urticaria and progressing to angioedema and vascular collapse. The latter patients had elevated plasma histamine levels during challenge, but no sign of complement activation was observed. Our findings suggest differing pathomechanisms for these two forms.


Total synthesis of halicylindramide A.
A rapid and efficient Fmoc solid-phase synthesis of halicylindramide A is described. The strategy comprises resin attachment of the first amino acid via the side chain of aspartic acid, stepwise solid-phase synthesis of the linear peptide analog up to the cysteine residue, on-resin head-to-tail cyclization and linear peptide synthesis of the N-terminal region, and finally cysteine oxidation and formylation. The stereochemistry of the halicylindramide A was confirmed by comparison of NMR and RP-HPLC data with the natural molecule. A distinctive conformational change was observed from the CD spectra of the halicylindramide A in sodium dodecyl sulfate.


Complex chromosomal rearrangement in a woman with multiple miscarriages.
We report on a 23-yr-old healthy female with repeated miscarriages and a de novo complex chromosomal rearrangement (CCR) involving chromosomes 1, 2, 5 and 11 with 5 breakpoints. A review of cases reported in the last five yr is provided.


Downregulation of egg cell-secreted EC1 is accompanied with delayed gamete fusion and polytubey.
One major player known to be essential for successful gamete interactions during double fertilization in Arabidopsis thaliana is the recently identified family of egg cell-secreted EC1 proteins. Both gamete fusion events are affected in EC1-deficient female gametophytes. Here, we show that the number of ovules with unfused sperm cells is considerably higher than the number of undeveloped seeds in the same ec1-RNAi knockdown lines. We found that some sperm cells are able to fuse with the female gametes even 2 to 3 days after pollination, as reflected by delayed embryo and endosperm development, and by polytubey. We propose that the egg cell secretes EC1 proteins upon sperm arrival to promote rapid sperm activation, thereby accelerating gamete fusion and preventing polytubey.


Translocation of constitutively expressed heat shock protein Hsc70 to synapse-enriched areas of the cerebral cortex after hyperthermic stress.
Hsp70 is a multi-gene family composed of stress-inducible members (Hsp70) and other members that are constitutively expressed (Hsc70). The heat shock proteins Hsp70 and Hsc70 exhibit similar molecular structure and biochemical functions. Constitutively expressed Hsc70 is enriched in the mammalian nervous system compared with non-neural tissues and present at high levels in neuronal cell bodies. After thermal stress, Hsc70 is translocated to synapse-enriched areas of the cerebral cortex where it associates with Hsp40 to form a complex that can refold denaturated proteins. These results suggest that the heat shock response in the nervous system involves not only the synthesis of stress-inducible Hsps but also the translocation of constitutively expressed Hsc70 to synapse-enriched areas where it could participate in neuroprotective mechanisms that preserved synaptic function during times of stress.


[Simultaneous content determination of 14 components in Rhei Radix et Rhizoma by high performance liquid chromatography method].
To establish an HPLC (high performance liquid chromatography) method for the simultaneous content determination of gallic acid, (+)-catechin, (-)-epicatechin-3-O-gallate, isolindleyin, 4-(4'-hydroxyphenyl)-2-butanone, emodin, chrysophanol, physcion, aloe-emodin, rhein, lindleyin, 4-(4'-hydroxyphenyl)-2-butanone-4'-O-β-D-(2″-O-galloyl-6″-O-cinnamoyl)-glucopyranoside, sennoside A and sennoside B in Rhei Radix et Rhizoma. The analysis was performed on Agilent Zorbax SB-C₁₈ (4.6 mm×150 mm, 5 μm) with 0.05% phosphoric acid solution (A) - acetonitrile (B) as mobile phase for gradient elution. The flow rate was 1 mL•min⁻¹, with column temperature of 40 ℃ and the wavelength was set at 268 nm. All calibration curves showed good linearity (r > 0.999 9) within the concentration range. Both the intra- and inter-day precision for 14 analytes was less than 3.1%, with the mean recovery at the range of 91.80%-104.1%. Meanwhile, quantitative determination was carried out for 10 qualified samples from Rheum palmatum and 10 qualified samples from R. tanguticum, respectively. It was found that the content of 4-(4'-hydroxyphenyl)-2-butanone and aloe-emodin were higher in the R. tanguticum and R. palmatum, respectively, and the content of all the compounds was different in each sample. The established HPLC method for simultaneous content determination of 14 compounds from Rhei Radix et Rhizoma could be used for quantitative assessment and quality control of Rhei Radix et Rhizoma.


Norcantharidin enhances TIMP‑2 anti‑vasculogenic mimicry activity for human gallbladder cancers through downregulating MMP‑2 and MT1‑MMP.
Vasculogenic mimicry (VM) is a tumor microcirculation pattern in highly aggressive gallbladder cancers (GBCs). We recently reported the anti‑VM activity of norcantharidin (NCTD) in highly aggressive GBC‑SD cells and xenografts. In this study, we further investigated that NCTD enhanced tissue inhibitor of matrix metalloproteinase‑2 (TIMP‑2) anti‑VM activity for GBCs and the underlying mechanisms. In vivo and in vitro experiments were performed to determine the effects of NCTD in combination with TIMP‑2 on tumor growth, host survival, VM formation, hemodynamic of GBC‑SD xenografts, and VM‑like networks and malignant phenotypes of GBC‑SD cells. Expression of matrix metalloproteinase (MMP)‑2 and membrane type 1‑MMP (MT1‑MMP) among human GBCs, GBC‑SD cells and xenografts were determined, respectively. The results showed that expression of MMP‑2 and MT1‑MMP in human GBCs, GBC‑SD cells and xenografts was significantly related to VM in GBCs; a shorter survival time of VM‑positive patients with high expression of MMP‑2 or MT1‑MMP compared to that of the patients with low expression. After treatment with NCTD+TIMP‑2, tumor growth, VM formation, VM hemodynamic of the xenografts in vivo were significantly inhibited as compared to control, NCTD or TIMP‑2 group, with a prolonged survival time of the xenograft mice (log‑rank test, P=0.0115); and these observations were confirmed by VM‑like networks by 3‑D matrices and showed that proliferation, apoptosis, invasion, migration of GBC‑SD cells in vitro were markedly affected. Furthermore, expression of MMP‑2 and MT1‑MMP in VM formation of the xenografts in vivo and GBC‑SD cells in vitro was downregulated as compared to control, NCTD or TIMP‑2 group. Thus, we concluded that NCTD enhances TIMP‑2 antitumor and anti‑VM activities in GBCs through downregulating MMP‑2 and MT1‑MMP.


Effects of Beclin 1 overexpression on aggressive phenotypes of colon cancer cells.
Beclin 1 is involved in autophagy, differentiation, apoptosis and cancer progression, and functions as a haploinsufficient tumor suppressor gene. The aim of the present study was to elucidate the function of Beclin 1 in colon cancer. A Beclin 1-expressing plasmid was transfected into HCT-15 and HCT-116 cells, and the phenotypes and associated molecules were determined. Beclin 1 transfectants were subcutaneously injected into nude mice to determine tumor growth, and proliferation and apoptosis levels using Ki-67 immunostaining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), respectively. Beclin 1 overexpression inhibited viability as determined using a Cell Counting Kit-8 assay, inhibited migration and invasion as determined using a wound healing assay or Transwell assay, and lamellipodia formation by filamentous actin staining, induced autophagy as determined using electron microscopy, and light chain 3B (LC-3B) expression, and apoptosis as determined using Annexin V staining in the two cell lines (P<0.05). Beclin 1 induced G2 arrest of HCT-15 transfectants as determined using propidium iodide staining (P<0.05), whereas HCT-116 transfectants were arrested in G1 phase (P<0.05). The two transfectants exhibited increased expression of c-Myc, cyclin D1, β-catenin, insulin-response element 1 and 78 kDa glucose-regulated protein compared with the control and mock cells as determined using the reverse transcription-quantitative polymerase chain reaction (P<0.05). Beclin 1 overexpression upregulated LC-3B and cyclin-dependent kinase 4 expression, but downregulated cyclin E expression of the cancer cell lines as determined using western blot analysis (P<0.05). Beclin 1 expression in vivo significantly suppressed the proliferation of colon cancer cells in xenograft models via inducing apoptosis by TUNEL, and inhibiting proliferation by Ki-67 expression (P<0.05). Beclin 1 overexpression may reverse aggressive phenotypes and suppress colon cancer tumor growth, and be employed as a target molecule for gene therapy of patients with colon cancer.


COVID-Nets: deep CNN architectures for detecting COVID-19 using chest CT scans.
In this paper we propose two novel deep convolutional network architectures, CovidResNet and CovidDenseNet, to diagnose COVID-19 based on CT images. The models enable transfer learning between different architectures, which might significantly boost the diagnostic performance. Whereas novel architectures usually suffer from the lack of pretrained weights, our proposed models can be partly initialized with larger baseline models like ResNet50 and DenseNet121, which is attractive because of the abundance of public repositories. The architectures are utilized in a first experimental study on the SARS-CoV-2 CT-scan dataset, which contains 4173 CT images for 210 subjects structured in a subject-wise manner into three different classes. The models differentiate between COVID-19, non-COVID-19 viral pneumonia, and healthy samples. We also investigate their performance under three binary classification scenarios where we distinguish COVID-19 from healthy, COVID-19 from non-COVID-19 viral pneumonia, and non-COVID-19 from healthy, respectively. Our proposed models achieve up to 93.87% accuracy, 99.13% precision, 92.49% sensitivity, 97.73% specificity, 95.70% F1-score, and 96.80% AUC score for binary classification, and up to 83.89% accuracy, 80.36% precision, 82.04% sensitivity, 92.07% specificity, 81.05% F1-score, and 94.20% AUC score for the three-class classification tasks. We also validated our models on the COVID19-CT dataset to differentiate COVID-19 and other non-COVID-19 viral infections, and our CovidDenseNet model achieved the best performance with 81.77% accuracy, 79.05% precision, 84.69% sensitivity, 79.05% specificity, 81.77% F1-score, and 87.50% AUC score. The experimental results reveal the effectiveness of the proposed networks in automated COVID-19 detection where they outperform standard models on the considered datasets while being more efficient.


[Splitting of tablets: small pieces a risk].
For economic reasons physicians prescribe more and more multiunit tablets. Splitting of multiunit tablets depends on the physical-chemical properties of the agents, the galenic of the dosage form, the size and contour of the tablet and the shape of the score. Tablets with one or more scores are prepared to be divided for a single/multiple dose. How easily and exact a tablet can be divided depends heavily on the physical shape, its size and the outfit of the score. The fragments have to fulfil the requirements according to the European Pharmacopoeia: Uniformity of multiunit tablets. Since exact dosing is guaranteed only if tablets are divided properly, information and guidance of the patients by the physician and pharmacist is of critical importance.


[Surgery of primary hyperparathyroidism in 1988. Strategies during primary operations, in case of failure and in case of cancer].
In primary hyperparathyroidism, an operation is indicated when the calcemia exceeds 115 mg/l and phosphoremia is low in several successive instances, regardless of the symptoms, even if PTH levels and cervical ultrasonography are normal. In case of calcemia under 110 mg/l, the diagnosis must be confirmed by titration of the nephrogenic cyclic AMP and symptomatic patients must be operated upon as well as asymptomatic patients with a life expectancy exceeding 10 years. In case of acute hypercalcemia, the procedure must be performed as a semi-emergency without waiting for a definite diagnosis, since the course may rapidly be fatal in spite of all medical treatments. Ultrasonography mostly presents the advantage of detecting intrathyroid parathyroids glands. The experience of the surgeon is essential for the mandatory locating of 4 glands, and the choice of the surgical strategy. In front of a secondary indication for failure or recurrence, one must take into consideration what was seen and done during the first procedure, the calcemia levels and clinical, radiological or biological consequences. Finally, in case of cancer (2 p. cent), the best prognosis rests in wide excision of the thyroid compartment and nodes areas, since medical treatments and radiation therapy are ineffective.


Experimental and modeling studies of competitive Pb (II) and Cd (II) bioaccumulation by Aspergillus niger.
Co-existence of toxic metals causes complex toxicity to microorganisms during bioremediation in water and soil. This study investigated the immobilization of Pb2+ and Cd2+ by fungus Aspergillus niger, which has been widely applied to environmental remediation. Five treatments were set, i.e., CK (no toxic metals), Pb2+ only, Cd2+ only, Pb2+/Cd2+ = 1:1(molar ratio), and Pb2+/Cd2+ = 2:1. Cadmium induced strong toxicity to the fungus, and maintained the high toxicity during incubation. However, as Pb/Cd ratio increased from 0 to 2, the removal rates of Cd2+ by A. niger were raised from 30 to 50%. The elevated activities of pyruvate dehydrogenase (PDH) and citrate synthetase (CS) enzymes confirmed that Pb addition could stimulate the growth of A. niger. For instance, citric acid concentrations and CS activities were 463.22 mg/L and 78.37 nmol/min/g, respectively, during 3-day incubation as Pb/Cd = 1. However, these two values were as low as ~ 50 with addition of only Cd. It was hence assumed that appropriate co-existence of Pb2+ enhanced microbial activity by promoting TCA cycle of the fungus. Moreover, the SEM analysis and geochemical modeling demonstrated that Pb2+ cations were more easily adsorbed and mineralized on A. niger with respect to Cd2+. Therefore, instead of intensifying metal toxicity, the addition of appropriate Pb actually weakened Cd toxicity to the fungus. This study sheds a bright future on application of A. niger to the remediation of polluted water with co-existence of Pb and Cd. KEY POINTS: • Cd2+ significantly inhibited P consumption, suggesting its high toxicity to A. niger. • Pb2+ stimulated the growth of A. niger by promoting TCA cycle in the cells. • Cd2+ removal by A. niger were improved with co-existence of Pb2+.


The limits on sexual dimorphism in vegetative traits in a gynodioecious plant.
Gynodioecious plants exhibit modest sexual dimorphism in vegetative and phenological traits, which stands in stark contrast to pronounced dimorphism in reproductive traits. I evaluate the roles of limited genetic variation, negative genetic covariation (within and between sex morphs), and lack of gender-differential selection in contributing to minimal sexual dimorphism for these traits in Fragaria virginiana. Major findings are as follows. First, selection was sometimes differential but rarely divergent between male and female fertility modes. Second, response to selection was constrained by low genetic variation and extensive genetic covariance. In fact, covariance between traits within sex morphs appears to represent a constraint on par with that of covariance between sex morphs. Third, these constraints combine with different modes of gamete transmission to produce very different gender-specific contributions to the mean phenotypes of the next generation. Finally, predicted responses to selection for several traits are concordant with the degree and direction of dimorphism in a closely related dioecious species. In sum, this work suggests that minimal sexual dimorphism in vegetative and phenological traits is due to similar directional selection via male and female fertility combined with the constraints of low genetic variation and extensive genetic covariance both within and between sex morphs.


3-(Para-fluorobenzoyl)-propionic acid; a metabolite of haloperidol, reversed oestradiol valerate-induced uterine hyperplasia via modulation of oestrogen receptor signalling pathways in female Wistar rats.
Background 3-(Para-fluorobenzoyl)-propionic acid (3PFBPA) is one of the metabolites of haloperidol used in the treatment of psychotic disorders. 3PFBPA is an inhibitor of mitogen-activated protein kinase (MAPK), implicated in the development of uterine fibroids (UFs) and cellular proliferation. In this study, the effect of 3PFBPA on oestradiol valerate (OV)-induced uterine hyperplasia was investigated. Methods Uterine hyperplasia was induced by intraperitoneal (i.p.) injection of OV (3 mg/kg for 12 weeks). Expression of oestrogen receptor (ER) α, β-catenin and E-cadherin were investigated via immunohistochemistry. The histology and fibroblast cell count/μm2 (using histomorphometry) were carried out. Results There was a significant increase in the levels of oestrogen, progesterone and total cholesterol in the OV-treated group when compared with the control, assessed by enzyme-linked immunosorbent assay (ELISA) kits. Oestrogen and total cholesterol were markedly reduced in the OV + 3PFBPAtreated group when compared with the OV-treated group. The OV-induced overexpression of β-catenin and ER were also ameliorated by 3PFBPA. Also, the loss of E-cadherin function in the OV-treated group was restored by 3PFBPA. The histological findings and histomorphometric results revealed the presence of uterine hyperplasia in the OV-treated rats which was significantly reversed by 3PFBPA. Histological studies revealed a protective role against OV-induced uterine damage that was found after OV + 3PFBPA co-administration. Conclusion This study demonstrated that 3PFBPA ameliorates OV-induced uterine hyperplasia in the female Wistar rat model. The findings warrant further investigation of the antifibrotic effects of 3PFBPA in humans.


Defining pathways of loss and secretion of chemical messengers from astrocytes.
It is becoming evident that glia, and astrocytes in particular, are intimately involved in neuronal signaling. Astrocytic modulation of signaling in neurons appears to be mediated by the release of neuroactive compounds such as the excitatory amino acid glutamate. Release of these transmitters appears to be driven by two different processes: (1) a volume regulatory response triggered by hypo-osmotic conditions that leads to the release of osmotically active solutes from the cytoplasm into the extracellular space, and (2) intracellular calcium-dependent vesicle-mediated excytotic release. The regulatory volume decrease may be mediated by any of several different pathways that increase membrane permeability, thus allowing osmolytes to travel down their concentration gradient into the extracellular space. Such pathways include anion channels, hemichannels, P2X receptor channels, and transporters or multidrug resistance proteins. The excytotic release process may use calcium triggered synaptic like vesicle fusion or alterations in constitutive vesicle trafficking to the membrane. Determining the contribution of any of these release mechanisms requires agents that can be used to specifically block pathways of interest. Currently, many of the pharmacological compounds being used exhibit a great deal of cross-reactivity between several of these pathways. For example, the popular anion channel inhibitor 5-nitro-2-(3-phenyl-propylamino)benzoic acid (NPPB) is an efficient blocker of both hemichannels and vesicle loading. This demonstrates the need to more fully characterize the activities of the agents currently available and to choose pathway blockers carefully when designing experiments.


Is there still hope for single therapies: how do we set up experimental systems to efficiently test combination therapies?
Severe asthma, chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF) are chronic lung diseases with a clear need for development of new and more efficient therapy. In preclinical research, the mouse model has been instrumental in advancing our knowledge of the biology and immunology. However, it has been proven rather difficult and time consuming to develop new treatments that can impact on the clinical course of these diseases. Many challenges need to be overcome for upgrading the quality of currently available experimental disease models in order to enhance the translation rate of basic research to clinical practice. Establishment of transgenic mouse overexpressing disease-causing genes may provide tools to discover new pathological pathways and to evaluate the possibility of molecular targeted therapy in chronic lung diseases. Personalized medicine, if developed, might be the solution for 'disease heterogeneity' and for improving clinical outcome.


MAPK-triggered chromatin reprogramming by histone deacetylase in plant innate immunity.
Microbial-associated molecular patterns activate several MAP kinases, which are major regulators of the innate immune response in Arabidopsis thaliana that induce large-scale changes in gene expression. Here, we determine whether microbial-associated molecular pattern-triggered gene expression involves modifications at the chromatin level. Histone acetylation and deacetylation are major regulators of microbial-associated molecular pattern-triggered gene expression and implicate the histone deacetylase HD2B in the reprogramming of defence gene expression and innate immunity. The MAP kinase MPK3 directly interacts with and phosphorylates HD2B, thereby regulating the intra-nuclear compartmentalization and function of the histone deacetylase. By studying a number of gene loci that undergo microbial-associated molecular pattern-dependent activation or repression, our data reveal a mechanistic model for how protein kinase signaling directly impacts chromatin reprogramming in plant defense.


The structure of the sternal bursa of turkeys.
The sternal bursa developed as a microscopically recognizable synovial sac in turkeys at about 4 1/2 to 6 weeks of age. In birds 12 weeks old it was difficult to delineate the bursa grossly although microscopically it was a definite structure. After 12 weeks, the walls of the bursa were thick enough that the limits of the sac could be determined grossly. The synovial membrane was in the subcutis, and microscopically was a vascular band of loose to dense connective tissue of variable thickness. Laterally the membrane was folded so that in cross section it appeared as a tonguelike projection of variable length into the cavity. The lining cells centrally were flat fibroblasts while laterally they were cuboidal, particularly over the lateral folds. In older turkeys the cuboidal lining cells were sometimes stratified.


The essential role of histone H3 Lys9 di-methylation and MeCP2 binding in MGMT silencing with poor DNA methylation of the promoter CpG island.
Silencing of the O (6)-methylguanine-DNA methyltransferase (MGMT) gene, a key to DNA repair, is involved in carcinogenesis. Recent studies have focused on DNA hypermethylation of the promoter CpG island. However, cases showing silencing with DNA hypomethylation certainly exist, and the mechanism involved is not elucidated. To clarify this mechanism, we examined the dynamics of DNA methylation, histone acetylation, histone methylation, and binding of methyl-CpG binding proteins at the MGMT promoter region using four MGMT negative cell lines with various extents of DNA methylation. Histone H3K9 di-methylation (H3me2K9), not tri-methylation, and MeCP2 binding were commonly seen in all MGMT negative cell lines regardless of DNA methylation status. 5Aza-dC, but not TSA, restored gene expression, accompanied by a decrease in H3me2K9 and MeCP2 binding. In SaOS2 cells with the most hypomethylated CpG island, 5Aza-dC decreased H3me2K9 and MeCP2 binding with no effect on DNA methylation or histone acetylation. H3me2K9 and DNA methylation were restricted to in and around the island, indicating that epigenetic modification at the promoter CpG island is critical. We conclude that H3me2K9 and MeCP2 binding are common and more essential for MGMT silencing than DNA hypermethylation or histone deacetylation. The epigenetic mechanism leading to silent heterochromatin at the promoter CpG island may be the same in different types of cancer irrespective of the extent of DNA methylation.


Strict Selection Alone of Patients Undergoing Liver Transplantation for Hilar Cholangiocarcinoma Is Associated with Improved Survival.
Liver transplantation for hilar cholangiocarcinoma (hCCA) has regained attention since the Mayo Clinic reported their favorable results with the use of a neo-adjuvant chemoradiation protocol. However, debate remains whether the success of the protocol should be attributed to the neo-adjuvant therapy or to the strict selection criteria that are being applied. The aim of this study was to investigate the value of patient selection alone on the outcome of liver transplantation for hCCA. In this retrospective study, patients that were transplanted for hCCA between1990 and 2010 in Europe were identified using the European Liver Transplant Registry (ELTR). Twenty-one centers reported 173 patients (69%) of a total of 249 patients in the ELTR. Twenty-six patients were wrongly coded, resulting in a study group of 147 patients. We identified 28 patients (19%) who met the strict selection criteria of the Mayo Clinic protocol, but had not undergone neo-adjuvant chemoradiation therapy. Five-year survival in this subgroup was 59%, which is comparable to patients with pretreatment pathological confirmed hCCA that were transplanted after completion of the chemoradiation protocol at the Mayo Clinic. In conclusion, although the results should be cautiously interpreted, this study suggests that with strict selection alone, improved survival after transplantation can be achieved, approaching the Mayo Clinic experience.


Clinical efficacy of a combined acceptance and commitment therapy, dialectical behavioural therapy, and functional analytic psychotherapy intervention in patients with borderline personality disorder.
Borderline personality disorder (BPD) consists of a persistent pattern of instability in affective regulation, impulse control, interpersonal relationships, and self-image. Although certain forms of psychotherapy are effective, their effects are small to moderate. One of the strategies that have been proposed to improve interventions involves integrating the therapeutic elements of different psychotherapy modalities from a contextual behavioural perspective (ACT, DBT, and FAP). Patients (n = 65) attending the BPD Clinic of the Instituto Nacional de Psiquiatría Ramón de la Fuente Muñíz in Mexico City who agreed to participate in the study were assigned to an ACT group (n = 22), a DBT group (n = 20), or a combined ACT + DBT + FAP therapy group (n = 23). Patients were assessed at baseline and after therapeutic trial on measures of BPD symptom severity, emotion dysregulation, experiential avoidance, attachment, control over experiences, and awareness of stimuli. ANOVA analyses showed no differences between the three therapeutic groups in baseline measures. Results of the MANOVA model showed significant differences in most dependent measures over time but not between therapeutic groups. Three modalities of brief, contextual behavioural therapy proved to be useful in decreasing BPD symptom severity and emotional dysregulation, as well as negative interpersonal attachment. These changes were related to the reduction of experiential avoidance and the acquisition of mindfulness skills in all treatment groups, which may explain why no differences between the three different intervention modalities were observed. Brief adaptations of acceptance and commitment therapy and dialectical behavioural therapy are effective interventions for BPD patients, in combined or isolated modalities, and with or without the inclusion of functional analytic psychotherapy. The reduction of experiential avoidance and the acquisition of mindfulness skills are related with the diminution of BPD symptoms severity, including emotional dysregulation and negative interpersonal attachment.


Cerebellum and personality traits.
Personality traits are multidimensional traits comprising cognitive, emotional, and behavioral characteristics, and a wide array of cerebral structures mediate individual variability. Differences in personality traits covary with brain morphometry in specific brain regions. A cerebellar role in emotional and affective processing and on personality characteristics has been suggested. In a large sample of healthy subjects of both sexes and differently aged, the macro- and micro-structural variations of the cerebellum were correlated with the scores obtained in the Temperament and Character Inventory (TCI) by Cloninger. Cerebellar volumes were associated positively with Novelty Seeking scores and negatively with Harm Avoidance scores. Given the cerebellar contribution in personality traits and emotional processing, we investigated the cerebellar involvement even in alexithymia, construct of personality characterized by impairment in cognitive, emotional, and affective processing. Interestingly, the subjects with high alexithymic traits had larger volumes in the bilateral Crus 1. The cerebellar substrate for some personality dimensions extends the relationship between personality and brain areas to a structure up to now thought to be involved mainly in motor and cognitive functions, much less in emotional processes and even less in personality individual differences. The enlarged volumes of Crus 1 in novelty seekers and alexithymics support the tendency to action featuring both personality constructs. In fact, Novelty Seeking and alexithymia are rooted in behavior and inescapably have a strong action component, resulting in stronger responses in the structures more focused on action and embodiment, as the cerebellum is.


Effects of the serotonin 2C receptor agonist WAY163909 on the abuse-related effects and mesolimbic dopamine neurochemistry induced by abused stimulants in rhesus monkeys.
Accumulating evidence shows that the serotonergic system plays a major role in psychostimulant abuse through its interactions with the dopaminergic system. Studies indicate that serotonin 5-HT2C receptors are one of the main classes of receptors involved in mediating the influence of serotonin in drug abuse. The aim of the present study was to evaluate the effects of the selective serotonin 5-HT2C receptor agonist WAY163909 on the behavioral neuropharmacology of cocaine and methamphetamine in adult rhesus macaques. Cocaine or methamphetamine self-administration and reinstatement were evaluated under second-order and fixed-ratio schedules of reinforcement, respectively. Cocaine- and methamphetamine-induced increases in dopamine were assessed through in vivo microdialysis targeting the nucleus accumbens. Pretreatment with WAY163909 dose-dependently attenuated cocaine and methamphetamine self-administration and drug-induced reinstatement of extinguished behavior previously maintained by cocaine or methamphetamine delivery. In an additional experiment, WAY163909 induced a dose-dependent attenuation of cocaine- or methamphetamine-induced dopamine overflow in the nucleus accumbens. Our data indicate that selective 5-HT2C receptor activation decreases drug intake and drug-seeking behavior in nonhuman primate models of psychostimulant abuse through neurochemical mechanisms involved in the modulation of mesolimbic dopamine.


Lewis and Fischer 344 rats as a model for genetic differences in spatial learning and memory: Cocaine effects.
Lewis (LEW) and Fischer 344 (F344) rats are considered a model of genetic vulnerability to drug addiction. We previously showed important differences in spatial learning and memory between them, but in contrast with previous experiments demonstrating cocaine-induced enhanced learning in Morris water maze (MWM) highly demanding tasks, the eight-arm radial maze (RAM) performance was not modified either in LEW or F344 rats after chronic cocaine treatment. In the present work, chronically cocaine-treated LEW and F344 adult rats have been evaluated in learning and memory performance using the Y-maze, two RAM protocols that differ in difficulty, and a reversal protocol that tests cognitive flexibility. After one of the RAM protocols, we quantified dendritic spine density in hippocampal CA1 neurons and compared it to animals treated with cocaine but not submitted to RAM. LEW cocaine treated rats showed a better performance in the Y maze than their saline counterparts, an effect that was not evident in the F344 strain. F344 rats significantly took more time to learn the RAM task and made a greater number of errors than LEW animals in both protocols tested, whereas cocaine treatment induced deleterious effects in learning and memory in the highly difficult protocol. Moreover, hippocampal spine density was cocaine-modulated in LEW animals whereas no effects were found in F344 rats. We propose that differences in addictive-like behavior between LEW and F344 rats could be related to differences in hippocampal learning and memory processes that could be on the basis of individual vulnerability to cocaine addiction.


Concurrent Aerobic and Resistance Training Has Anti-Inflammatory Effects and Increases Both Plasma and Leukocyte Levels of IGF-1 in Late Middle-Aged Type 2 Diabetic Patients.
Type 2 diabetes (T2D) is an age-related chronic disease associated with metabolic dysregulation, chronic inflammation, and activation of peripheral blood mononuclear cells (PBMC). The aim of this study was to assess the effects of a concurrent exercise training program on inflammatory status and metabolic parameters of T2D patients. Sixteen male patients (age range 55-70) were randomly assigned to an intervention group (n = 8), which underwent a concurrent aerobic and resistance training program (3 times a week; 16 weeks), or to a control group, which followed physicians' usual diabetes care advices. Training intervention significantly improved patients' body composition, blood pressure, total cholesterol, and overall fitness level. After training, plasma levels of adipokines leptin (-33.9%) and RBP4 (-21.3%), and proinflammatory markers IL-6 (-25.3%), TNF-α (-19.8%) and MCP-1 (-15.3%) decreased, whereas anabolic hormone IGF-1 level increased (+16.4%). All improvements were significantly greater than those of control patients. Plasma proteomic profile of exercised patients showed a reduction of immunoglobulin K light chain and fibrinogen as well. Training also induced a modulation of IL-6, IGF-1, and IGFBP-3 mRNAs in the PBMCs. These findings confirm that concurrent aerobic and resistance training improves T2D-related metabolic abnormalities and has the potential to reduce the deleterious health effects of diabetes-related inflammation.


Intraoperative radiotherapy for extremity soft-tissue sarcomas: can long-term local control be achieved?
Intraoperative electron-beam radiation therapy (IOERT) during limb-sparing surgery has the advantage of delivering a single high boost dose to sarcoma residues and surgical bed area near to radiosensitive structures with limited toxicity. Retrospective studies have suggested that IOERT may improve local control compared to standard radiotherapy and we aimed to demonstrate this theory. Therefore, we performed an observational prospective study to determine (1) if it is possible to achieve high local control by adding IOERT to external-beam radiation therapy (EBRT) in extremity soft-tissue sarcomas (STS), (2) if it is possible to improve long-term survival rates, and (3) if toxicity could be reduced with IOERT MATERIALS AND METHODS: From 1995-2003, 39 patients with extremity STS were treated with IOERT and postoperative radiotherapy. The median follow-up time was 13.2 years (0.7-19). Complications, locoregional control and survival rates were collected. Actuarial local control was attained in 32 of 39 patients (82%). Control was achieved in 88% of patients with primary disease and in 50% of those with recurrent tumors (p = 0.01). Local control was shown in 93% of patients with negative margins and in 50% of those with positive margins (p = 0.002). Limb-sparing was achieved in 32 patients (82%). The overall survival rate was 64%. 13% of patients had grade ≥3 acute toxicity, and 12% developed grade ≥3 chronic toxicity. IOERT used as a boost to EBRT provides high local control and limb-sparing rates in patients with STS of the extremities, with less toxicity than EBRT alone.


Visualization of back pain data--a 3-D solution.
Traditional approaches to gathering and visualizing pain data rely on two-dimensional (2-D) human body models, where different types of sensation are recorded with various monochrome symbols. We propose an alternative that uses a three-dimensional (3-D) representation of the human body, which can be marked in color to visualize and record pain data.


Clinical Predictors of Mortality in Adults with Intellectual Disabilities with and without Down Syndrome.
Background. Mood, baseline functioning, and cognitive abilities as well as psychotropic medications may contribute to mortality in adults with and without Down Syndrome (DS). Methods. Population-based (nonclinical), community-dwelling adults with intellectual disabilities (IDs) were recruited between 1995 and 2000, assessed individually for 1-4 times, and then followed by yearly phone calls. Results. 360 participants (116 with DS and 244 without DS) were followed for an average of 12.9 years (range 0-16.1 years as of July 2011). 108 people died during the course of the followup, 65 males (31.9% of all male participants) and 43 females (27.6% of all female participants). Cox proportional hazards modeling showed that baseline practical skills, seizures, anticonvulsant use, depressive symptoms, and cognitive decline over the first six years all significantly contributed to mortality, as did a diagnosis of DS, male gender, and higher age at study entry. Analysis stratified by DS showed interesting differences in mortality predictors. Conclusion. Although adults with DS have had considerable improvements in life expectancy over time, they are still disadvantaged compared to adults with ID without DS. Recognition of potentially modifiable factors such as depression may decrease this risk.


A novel genetic locus linked to pro-inflammatory cytokines after virulent H5N1 virus infection in mice.
Genetic variation in the human population is a key determinant of influenza disease severity. A single nucleotide polymorphism in the antiviral gene IFITM3 was linked to outcomes during the 2009 H1N1 pandemic. To identify variant host genes associated with increased virus replication and severe disease, we performed a quantitative trait locus analysis on pro-inflammatory cytokine production 48 hours after intranasal infection with highly pathogenic H5N1 influenza virus. Pro-inflammatory cytokines CCL2, TNFα and IFN-α, were measured by ELISA in lung homogenates of DBA/2J (D2), C57BL/6J (B6) and 44 different BXD recombinant inbred mouse strains. Virus titer was also assessed in a subset of these animals. CCL2 (8-fold), TNFα (24-fold) and IFN-α (8-fold) concentrations varied significantly among the different BXD RI strains. Importantly, cytokine concentration correlated very well (r =0.86-0.96, P <0.0001) with virus titer suggesting that early cytokine production is due to increased virus infection and replication. Linkage analysis of cytokine concentration revealed a significant locus on chromosome 6 associated with differences in TNFα, IFN-α and CCL2 cytokine concentration (LRS =26). This locus accounted for nearly 20% of the observed phenotypic variation in the BXD population studied. Sequence and RNA expression analysis identified several candidate host genes containing missense mutations or deletions; Samd9l, Ica1, and Slc25a13. To study the role of Slc25a13, we obtained Slc25a13 knockout line, but upon challenge with H5N1 influenza virus observed no effect on CCL2 production, or morbidity and mortality. A novel genetic locus on chromosome 6 modulates early pro-inflammatory cytokine production and virus replication after highly pathogenic influenza virus infection. Candidate genes, Samd9l and Ica1, may be important for the control of influenza virus infection and pathogenesis.


[Anesthesia and analgesia for lower limb amputation].
This study describes the methods of anesthesia and analgesia used in 296 limb amputations for vascular disease over a fifteen year period (1986-2000). The main type of anesthesia was general in 55%, epidural in 29% and spinal in 14%: there were no significant differences for ASA grade, age, or amputation level, nor any statistical differences in mortality for each method of anesthesia. The main methods of analgesia in the first 48 hours changed between 1986 and 1998, with decreasing intramuscular and oral opioids (from 42% to 1%, and from 14% to 1% respectively) while epidurals became the commonest method (2% in 1986 and 83% in 2000). Thirty-seven percent of patients were prescribed pethidine for phantom pain. There have been substantial changes in postoperative analgesia following amputation, and epidurals are now common practice, despite the controversy about their role in preventing phantom pain.


Role of ejection fraction in patients at risk for advanced heart failure: insights from the HELP-HF registry.
Patients with heart failure (HF) with reduced ejection fraction (EF) (HFrEF), mildly reduced EF (HFmrEF), and preserved EF (HFpEF) may all progress to advanced HF, but the impact of EF in the advanced setting is not well established. Our aim was to assess the prognostic impact of EF in patients with at least one 'I NEED HELP' marker for advanced HF. Patients with HF and at least one high-risk 'I NEED HELP' criterion from four centres were included in this analysis. Outcomes were assessed in patients with HFrEF (EF ≤ 40%), HFmrEF (EF 41-49%), and HFpEF (EF ≥ 50%) and with EF analysed as a continuous variable. The prognostic impact of medical therapy for HF in patients with EF < 50% and EF > 50% was also evaluated. All-cause death was the primary endpoint, and cardiovascular death was a secondary endpoint. Among 1149 patients enrolled [mean age 75.1 ± 11.5 years, 67.3% males, 67.6% hospitalized, median follow-up 260 days (inter-quartile range 105-390 days)], HFrEF, HFmrEF, and HFpEF were observed in 699 (60.8%), 122 (10.6%), and 328 (28.6%) patients, and 1 year mortality was 28.3%, 26.2%, and 20.1, respectively (log-rank P = 0.036). As compared with HFrEF patients, HFpEF patients had a lower risk of all-cause death [adjusted hazard ratio (HRadj ) 0.67, 95% confidence interval (CI) 0.48-0.94, P = 0.022], whereas no difference was noted for HFmrEF patients. After multivariable adjustment, a lower risk of all-cause death (HRadj for 5% increase 0.94, 95% CI 0.89-0.99, P = 0.017) and cardiovascular death (HRadj for 5% increase 0.94, 95% CI 0.88-1.00, P = 0.049) was observed at higher EF values. Beta-blockers and renin-angiotensin system inhibitors or sacubitril/valsartan were associated with lower mortality in both EF < 50% and EF ≥ 50% groups. Among patients with HF and at least one 'I NEED HELP' marker for advanced HF, left ventricular EF is still of prognostic value.


Synthesis of a Fluorescently Labeled 68Ga-DOTA-TOC Analog for Somatostatin Receptor Targeting.
Fluorescently labeled imaging agents can identify surgical margins in real-time to help achieve complete resections and minimize the likelihood of local recurrence. However, photon attenuation limits fluorescence-based imaging to superficial lesions or lesions that are a few millimeters beneath the tissue surface. Contrast agents that are dual-labeled with a radionuclide and fluorescent dye can overcome this limitation and combine quantitative, whole-body nuclear imaging with intraoperative fluorescence imaging. Using a multimodality chelation (MMC) scaffold, IRDye 800CW was conjugated to the clinically used somatostatin analog, 68Ga-DOTA-TOC, to produce the dual-labeled analog, 68Ga-MMC(IRDye 800CW)-TOC, with high yield and specific activity. In vitro pharmacological assays demonstrated retention of receptor-targeting properties for the dual-labeled compound with robust internalization that was somatostatin receptor (SSTR) 2-mediated. Biodistribution studies in mice identified the kidneys as the primary excretion route for 68Ga-MMC(IRDye 800CW)-TOC, along with clearance via the reticuloendothelial system. Higher uptake was observed in most tissues compared to 68Ga-DOTA-TOC but decreased as a function of time. The combination of excellent specificity for SSTR2-expressing cells and suitable biodistribution indicate potential application of 68Ga-MMC(IRDye 800CW)-TOC for intraoperative detection of SSTR2-expressing tumors.


MicroRNA control of high-density lipoprotein metabolism and function.
Recent discoveries of microRNAs (miRNAs) that control high-density lipoprotein abundance and function have expanded our knowledge of the mechanisms regulating this important lipoprotein subclass. miRNAs have been shown to regulate gene networks that control high-density lipoprotein biogenesis and uptake, as well as discrete steps in the reverse cholesterol transport pathway. Furthermore, high-density lipoprotein itself has been shown to transport miRNAs selectively in health and disease, offering new possibilities of how this lipoprotein may alter gene expression in distal target cells and tissues. Collectively, these discoveries offer new insights into the mechanisms governing high-density lipoprotein metabolism and function and open new avenues for the development of therapeutics for the treatment of cardiovascular disease.


Variable Bone Density of Scaphoid: Importance of Subchondral Screw Placement.
Background Ideal internal fixation of the scaphoid relies on adequate bone stock for screw purchase; so, knowledge of regional bone density of the scaphoid is crucial. Questions/Purpose The purpose of this study was to evaluate regional variations in scaphoid bone density. Materials and Methods Three-dimensional CT models of fractured scaphoids were created and sectioned into proximal/distal segments and then into quadrants (volar/dorsal/radial/ulnar). Concentric shells in the proximal and distal pole were constructed in 2-mm increments moving from exterior to interior. Bone density was measured in Hounsfield units (HU). Results Bone density of the distal scaphoid (453.2 ± 70.8 HU) was less than the proximal scaphoid (619.8 ± 124.2 HU). There was no difference in bone density between the four quadrants in either pole. In both the poles, the first subchondral shell was the densest. In both the proximal and distal poles, bone density decreased significantly in all three deeper shells. Conclusion The proximal scaphoid had a greater density than the distal scaphoid. Within the poles, there was no difference in bone density between the quadrants. The subchondral 2-mm shell had the greatest density. Bone density dropped off significantly between the first and second shell in both the proximal and distal scaphoids. Clinical Relevance In scaphoid fracture ORIF, optimal screw placement engages the subchondral 2-mm shell, especially in the distal pole, which has an overall lower bone density, and the second shell has only two-third the density of the first shell.


Aspects of family caregiving as addressed in planned discussions between nurses, patients with chronic diseases and family caregivers: a qualitative content analysis.
Caregiving by family members of elderly with chronic conditions is currently intensifying in the context of an aging population and health care reform in the Netherlands. It is essential that nurses have attention for supporting roles of family caregivers of older patients and address family caregiving aspects on behalf of the continuity of care. This study aims to explore what aspects of family caregiving were addressed during planned discussions between nurses, patients and family caregivers in the hospital. Qualitative descriptive research was conducted using non-participant observation and audio-recordings of planned discussions between nurses, older patients and their family caregivers as they took place in the hospital. Through purposive sampling eligible patients (≥ 65 years) with one or more chronic conditions were included. These patients were admitted to the hospital for diagnostics or due to consequences of their chronic illness. Retrospective chart review was done to obtain patient characteristics. Data were collected in November/December 2013 and April/May 2014 in four hospitals. Qualitative content analysis was performed using the inductive approach in order to gain insight into addressed aspects of family caregiving. A total of 62 patients (mean age (SD) 76 years (7.2), 52% male) were included in the study, resulting in 146 planned discussions (62 admission and discharge discussions and 22 family meetings). Three themes were identified regarding addressed aspects of family caregiving. Two themes referred to aspects addressing the patients' social network, and included 'social network structure' and 'social network support'. One theme referred to aspects addressing coordination of care issues involving family caregiving, referred to as 'coordination of care'. During discussions nurses mostly addressed practical information on the patients' social network structure. When specific family caregiving support was addressed, information was limited and nurses did not seem to explore the nature of the family support. Patients discharge and after care needs were addressed occasionally as aspects of coordination of care. Current nursing policies could be evaluated on nursing and family oriented theories. Implications for education could include mirroring study findings with nurses in a group discussion to enhance their awareness on family caregiving aspects.


Clinical genetics of Kallmann syndrome.
The Kallmann syndrome (KS) combines hypogonadotropic hypogonadism (HH) with anosmia. This is a clinically and genetically heterogeneous disease. KAL1, encoding the extracellular glycoprotein anosmin-1, is responsible for the X chromosome-linked recessive form of the disease (KAL1). Mutations in FGFR1 or FGF8, encoding fibroblast growth factor receptor-1 and fibroblast growth factor-8, respectively, underlie an autosomal dominant form with incomplete penetrance (KAL2). Mutations in PROKR2 and PROK2, encoding prokineticin receptor-2 and prokineticin-2, have been found in heterozygous, homozygous, and compound heterozygous states. These two genes are likely to be involved both in autosomal recessive monogenic (KAL3) and digenic/oligogenic KS transmission modes. Mutations in any of the above-mentioned KS genes have been found in less than 30% of the KS patients, which indicates that other genes involved in the disease remain to be discovered. Notably, KS may also be part of pleiotropic developmental diseases including CHARGE syndrome; this disease results in most cases from neomutations in CHD7 that encodes a chromodomain helicase DNA-binding protein.


Prior caloric restriction increases survival of prepubertal obese- and PCOS-prone rats exposed to a challenge of time-limited feeding and physical activity.
We hypothesized that a polycystic ovary syndrome (PCOS) background associated with obese-prone genotype, coupled with preconditioning by caloric restriction, would confer a survival benefit in genetically prepubertal obese/PCOS (O/PCOS)-prone rats faced with an unpredictable challenge of food shortage. Female, juvenile JCR:LA-cp rats, O/PCOS- and lean-prone, were exposed to 1.5 h of daily meals and 22.5 h of voluntary wheel-running, a procedure that leads to activity anorexia (AA). One week before the AA challenge (AAC), O/PCOS-prone rats were freely fed (O/PCOS-FF) or pair fed (O/PCOS-FR) to lean-prone, free-feeding animals (Lean-FF). O/PCOS-FR and lean-prone, food-restricted (Lean-FR) groups were matched on relative average caloric intake. Animals were removed from protocol at 75% of initial body weight (starvation criterion) or after 14 days (survival criterion). The AAC induced weight loss in all rats, but there were significant effects of both genotype and feeding history on weight loss (lean-prone rats exhibited a higher rate of weight loss than O/PCOS-prone; P < 0.001), and rats with prior caloric restriction retained more weight than those free fed previously (90.68 ± 0.59% vs. 85.47 ± 0.46%; P < 0.001). The daily rate of running was higher in lean-prone rats compared with O/PCOS-prone. This difference in running rate correlated with differences in mean days of survival. All O/PCOS-FR rats survived at day 14. O/PCOS-FF rats survived longer (10.00 ± 0.97 days) than Lean-FR (6.17 ± 1.58 days) and Lean-FF (4.33 ± 0.42 days) rats (P < 0.05). Thus preconditioning by caloric restriction induces a substantial survival advantage, beyond genotype alone, in prepubertal O/PCOS-prone rats.


Practicalities and costs of adhesions.
In spite of postoperative adhesions being common there appears to be a reluctance to use anti-adhesion products routinely. This article compares the incidence of adhesions with other conditions in order to identify the level of risk. The health economics surrounding adhesion-related disease are described. This combined information may be of help to convince health practitioners of the need to take a more active role in adhesion prevention. The SCAR project has identified the risk of adhesion-related disease. This is compared with published risks of other common clinical situations. An economic model first described by the author in 2002 has been revised with 2006 costs [1]. The SCAR data demonstrates a directly related risk of re-admission in certain groups of 9.4% over 5 years [2]. The frequency of including this fact in the consenting process is low (<15%) [3]. Legal precedent has identified a risk of >2% warrants inclusion in the consent process; failure to do so could be considered negligent [4]. Use of an anti-adhesion product with a cost of 130 euros with an efficacy of 25% in 1 year in the UK could save over 40 million euros over a 10-year period. Adhesion risk is frequent enough to include in consent. Failure to do this and avoidance of treatment, which may reduce adhesions will have major financial consequences on healthcare systems.


Truck traffic related air pollution associated with asthma symptoms in young boys: a cross-sectional study.
The aim of this study was to evaluate the influence of intensity of truck traffic on asthma symptomatology, and its relationship with age and gender. A cross-sectional study was conducted on children and adolescents from Galicia (North-West Spain). Following the methodology of the International Study of Asthma and Allergies in Childhood (ISAAC): children from schools randomly selected, answered a self-administered questionnaire included questions on asthma symptoms and some risk factors. The association between self-reported truck traffic on the street of residence and symptoms of asthma were investigated by logistic regression adjusted for body mass index, maternal education and parental smoking. Almost 40% of children in Galicia are exposed to the frequent and constant passing of heavy goods vehicles. The odds of 6-7 year-old boys having severe or exercise induced asthma is tripled when they live in streets with the constant passing of heavy goods vehicles, compared with those living in streets where these vehicles never pass. In adolescents and 6-7 year-old girls, no relationship was observed between truck traffic and asthma symptoms. The results of this study appear to support a distinct effect of truck traffic on asthma symptoms depending on the age and sex of the exposed population, being more harmful for young males.


Crystallization and preliminary crystallographic data for acetohydroxy acid isomeroreductase from Spinacia oleracea.
Acetohydroxy acid isomeroreductase (EC 188.8.131.52) is one of the enzymes involved in branched-chain amino acid biosynthesis. The enzyme from spinach (Spinacia oleracea) leaves has been crystallized using the hanging drop vapour diffusion method. The free enzyme crystallized from polymethylene glycol solutions, but these crystals were unsuitable for X-ray diffraction analysis. In the presence of NADPH, Mg(2+) and a reaction intermediate analogue (2-dimethylphosphinoyl-2-hydroxy acetic acid (Hoe 704) or N-hydroxy-N-isopropyloxamate (IpOHA)), much better crystals were obtained. Crystals grown from ammonium sulphate belong to space group P2(1) with cell dimensions a + 193.78(7) A, b = 63.69(2) A, c = 112.84(1) A and beta = 121.22(1) degrees. The molecular mass of the protein, the volume of the unit cell, and crystal density measurements indicated that the asymmetric unit contains two dimers. X-ray diffraction patterns showed measurable reflections to beyond 2.5 A.


[Bioethics and care: from human rights to quality of care].
The pluralist nature of bioethics and the fact that it spans various branches of science places it in a privileged position to tackle ethical problems that accompany human choices in situations that influence the conditions of living beings. Clearly, issues such as health and care form part of the bioethical sphere of enquiry and it is obviously of fundamental importance that these subjects receive a suitable degree of attention within a branch of science that has always drawn on the contributions of other human sciences. Bioethics looks at the great questions posed by moral principles and seeks to apply them to present concerns.


Impacts of climate change and habitat loss on the distribution of the endangered crested capuchin monkey (Sapajus robustus).
The crested capuchin monkey (Sapajus robustus) is endemic to the Atlantic Forest and its transition areas within Cerrado in Brazil. The species is currently threatened by habitat loss and has been classified as endangered by the IUCN Red List of Threatened Species since 2015. We used ecological niche models built with MaxEnt to predict the potential impact of climate change on the distribution of this species. The models were projected onto the reference climate, considering six climate scenarios (three Global Climate Models and two Representative Concentration Pathways) from IPCC for 2050 and 2070. We showed that while the amount of suitable area is expected to change little across the species' range in most evaluated climate scenarios, climatic conditions may significantly deteriorate by 2070 in the pessimistic scenario, especially in currently warmer and dryer areas to the west. As seen on other capuchin monkeys, the potential use of tools by crested capuchins may increase the chances of the species adaptation to novel harsher environmental conditions. The major negative impacts across the species range also include habitat loss and fragmentation so that the conservation of the species relies on the protection of the forest remnants in the center of its distribution, which can harbor populations of the species in current and future climate scenarios.


Too Big, Too Small, or Just Right? Cost-Efficiency of Environmental Inspection Services in Connecticut.
To assess optimal activity size/mix of Connecticut local public health jurisdictions, through estimating economies of scale/scope/specialization for environmental inspections/services. Connecticut's 74 local health jurisdictions (LHJs) must provide environmental health services, but their efficiency or reasons for wide cost variation are unknown. The public health system is decentralized, with variation in organizational structure/size. We develop/compile a longitudinal dataset covering all 74 LHJs, annually from 2005 to 2012. We estimate a public health services/inspections cost function, where inputs are translated into outputs. We consider separate estimates of economies of scale/scope/specialization for four mandated inspection types. We obtain data from Connecticut Department of Public Health databases, reports, and other publicly available sources. There has been no known previous utilization of this combined dataset. On average, regional districts, municipal departments, and part-time LHJs are performing fewer than the efficient number of inspections. The full-time municipal departments and regional districts are more efficient but still not at the minimum efficient scale. The regional districts' elasticities of scale are larger, implying they are more efficient than municipal health departments. Local health jurisdictions may enhance efficiency by increasing inspections and/or sharing some services.


Decoding brain calcifications: A single-center descriptive case series and examination of pathophysiological mechanisms.
Brain calcifications, previously known as Fahr's disease, is a rare neurological disorder marked by various clinical symptoms, including movement disorders, cognitive impairment, and psychiatric disturbances. Despite its clinical importance, its pathophysiology is unclear and there are no specific treatments. We present four cases of brain calcifications from our tertiary care center, with three female patients (75%) and an average age of 63 years. Our cohort featured both genetic and endocrine etiologies, including one primary familial brain calcification case with a c.852del frameshift mutation in the SLC20A2 gene, and two endocrinopathy-related cases. One patient had an acute stroke which may have been contributed by brain calcifications. Computerized tomography and magnetic resonance imaging scans revealed basal ganglia and dentate nucleus calcifications. Treatment involved physical and occupational therapy in all patients. Hypoparathyroidism-related brain calcifications were treated with oral supplementation with calcitriol, calcium, and vitamin D. Three patients showed improvement or stability of their symptoms. This case series underscores the diverse clinical presentations and etiologies of brain calcifications. The complex pathophysiology involves disrupted Ca+2-PO43- homeostasis, deficient cellular PO43- transport, and vascular irregularities in genetic etiologies. Future research should focus on identifying novel genetic mutations, understanding molecular pathways, and refining diagnostic techniques. Integrating multidisciplinary approaches may improve diagnosis, management, and prognosis for patients with this intricate neurological disorder.


Ethical issues surrounding the evaluation of oxidative phosphorylation disorders in children.
Genetic testing is becoming increasingly frequent as a part of a pediatric evaluation. Oxidative phosphorylation disorders are likewise becoming part of the differential diagnosis for many children with undiagnosed neurologic problems. The complexity of the oxidative phosphorylation pathway and its dual genetic control by nuclear and mitochondrial DNA raise a number of ethical issues dealing mainly with the informed consent process, the respect for a patient's autonomy, the best interest of the patients and their families, as well as issues of genetic discrimination and genetic privacy laws. Open communication with families in pediatric medicine remains the cornerstone of good ethics, along with the skill needed to avoid having ethical issues arise.


Transcranial Acoustic Metamaterial Parameters Inverse Designed by Neural Networks.
Objective: The objective of this work is to investigate the mapping relationship between transcranial ultrasound image quality and transcranial acoustic metamaterial parameters using inverse design methods. Impact Statement: Our study provides insights into inverse design methods and opens the route to guide the preparation of transcranial acoustic metamaterials. Introduction: The development of acoustic metamaterials has enabled the exploration of cranial ultrasound, and it has been found that the influence of the skull distortion layer on acoustic waves can be effectively eliminated by adjusting the parameters of the acoustic metamaterial. However, the interaction mechanism between transcranial ultrasound images and transcranial acoustic metamaterial parameters is unknown. Methods: In this study, 1,456 transcranial ultrasound image datasets were used to explore the mapping relationship between the quality of transcranial ultrasound images and the parameters of transcranial acoustic metamaterials. Results: The multioutput parameter prediction model of transcranial metamaterials based on deep back-propagation neural network was built, and metamaterial parameters under transcranial image evaluation indices are predicted using the prediction model. Conclusion: This inverse big data design approach paves the way for guiding the preparation of transcranial metamaterials.


Constitutive signaling by the phototaxis receptor sensory rhodopsin II from disruption of its protonated Schiff base-Asp-73 interhelical salt bridge.
Sensory rhodopsin II (SRII) is a repellent phototaxis receptor in the archaeon Halobacterium salinarum, similar to visual pigments in its seven-helix structure and linkage of retinal to the protein by a protonated Schiff base in helix G. Asp-73 in helix C is shown by spectroscopic analysis to be a counterion to the protonated Schiff base in the unphotolyzed SRII and to be the proton acceptor from the Schiff base during photoconversion to the receptor signaling state. Coexpression of the genes encoding mutated SRII with Asn substituted for Asp-73 (D73N) and the SRII transducer HtrII in H. salinarum cells results in a 3-fold higher swimming reversal frequency accompanied by demethylation of HtrII in the dark, showing that D73N SRII produces repellent signals in its unphotostimulated state. Analogous constitutive signaling has been shown to be produced by the similar neutral residue substitution of the Schiff base counterion and proton acceptor Glu-113 in human rod rhodopsin. The interpretation for both seven-helix receptors is that light activation of the wild-type protein is caused primarily by photoisomerization-induced transfer of the Schiff base proton on helix G to its primary carboxylate counterion on helix C. Therefore receptor activation by helix C-G salt-bridge disruption in the photoactive site is a general mechanism in retinylidene proteins spanning the vast evolutionary distance between archaea and humans.


Comparative Study on Volatile Compounds of Alpinia japonica and Elettaria cardamomum.
The volatile compounds obtained from the ether extracts, headspace gases and steam distillates of Alpinia japonica and Elettaria cardamomum were analyzed by GC/MS. Both species were rich sources of naturally rare fenchane-type monoterpenoids, fenchene, fenchone, fenchyl alcohol and its acetate, together with 1,8-cineole. The distributions of volatile sesquiterpenoids were very poor in both species. Chiralities of fenchone in A. japonica and E. cardamomum were 99% of (1S,4R)-(+)-form. Camphor in A. japonica is composed of a mixture of (1R,4R)-(+)-form (94.3%) and (1S,4S)-(-)-form (5.7%). On the other hand, E. cardamomum produced only (1R,4R)-(+)-camphor (99%).


Pneumothorax complicating "in-out-in" thoracic pedicle screw placement for kyphotic deformity correction in a child.
The authors describe a rare case of pneumothorax as a complication of thoracic pedicle screw placement in an 11-year-old girl undergoing posterior segmental instrumentation for a kyphotic deformity. Spontaneous pneumothorax after posterior fusion for adolescent idiopathic scoliosis has been reported in the orthopedic literature; however, to the best of the authors' knowledge, pneumothorax directly related to pedicle screw placement for spinal deformity has not been previously described. The authors discuss the anatomical and technical aspects leading to this complication and the lessons learned from it.


Effectiveness of superselective chemoembolization and radiofrequency ablation of hepatocellular carcinoma assessed in 21 explanted livers.
The efficacy of superselective transcatheter chemoembolization (TACE) and chemoembolization taking advantage of pharmacologic (noradrenaline) flow manipulation was assessed on 21 explanted livers with hepatocellular carcinoma (HCC). There was a high concentration of chemoembolizate in the target area. Correlation of gross anatomical and immunohistochemical findings (gold standard) of tumor volume, necrosis and residual viable tumor to predicted results by multiphasic helical computed tomography (CT) was poor. Tumor markers in explanted livers found free of tumor were low or absent; in livers with residual or recurrent tumor high. However, there did not appear to be a correlation between the degree of tumor necrosis and level of tumor markers. The recurrence-free post-orthotopic liver transplant (OLT) survival in our 21 patients appears to be substantially lower than that reported in the literature. The goal of retaining patients on the liver transplant list by repeated TACE or multimodality therapy (MMT) was achieved (drop out rate of only 16%).


A Modular Flow Design for the meta-Selective C-H Arylation of Anilines.
Described herein is an effective and practical modular flow design for the meta-selective C-H arylation of anilines. The design consists of four continuous-flow modules (i.e., diaryliodonium salt synthesis, meta-selective C-H arylation, inline copper extraction, and aniline deprotection) which can be operated either individually or consecutively to provide direct access to meta-arylated anilines. With a total residence time of 1 hour, the desired product could be obtained in high yield and excellent purity without the need for column chromatography, and the residual copper content meets the standards for parenterally administered pharmaceutical substances.


Spatial attention during saccade decisions.
Behavioral measures of decision making are usually limited to observations of decision outcomes. In the present study, we made use of the fact that oculomotor and sensory selection are closely linked to track oculomotor decision making before oculomotor responses are made. We asked participants to make a saccadic eye movement to one of two memorized target locations and observed that visual sensitivity increased at both the chosen and the nonchosen saccade target locations, with a clear bias toward the chosen target. The time course of changes in visual sensitivity was related to saccadic latency, with the competition between the chosen and nonchosen targets resolved faster before short-latency saccades. On error trials, we observed an increased competition between the chosen and nonchosen targets. Moreover, oculomotor selection and visual sensitivity were influenced by top-down and bottom-up factors as well as by selection history and predicted the direction of saccades. Our findings demonstrate that saccade decisions have direct visual consequences and show that decision making can be traced in the human oculomotor system well before choices are made. Our results also indicate a strong association between decision making, saccade target selection, and visual sensitivity.NEW & NOTEWORTHY We show that saccadic decisions can be tracked by measuring spatial attention. Spatial attention is allocated in parallel to the two competing saccade targets, and the time course of spatial attention differs for fast-slow and for correct-erroneous decisions. Saccade decisions take the form of a competition between potential saccade goals, which is associated with spatial attention allocation to those locations.


Time-resolved 3D quantitative flow MRI of the major intracranial vessels: initial experience and comparative evaluation at 1.5T and 3.0T in combination with parallel imaging.
Exact knowledge of blood flow characteristics in the major cerebral vessels is of great relevance for diagnosing cerebrovascular abnormalities. This involves the assessment of hemodynamically critical areas as well as the derivation of biomechanical parameters such as wall shear stress and pressure gradients. A time-resolved, 3D phase-contrast (PC) MRI method using parallel imaging was implemented to measure blood flow in three dimensions at multiple instances over the cardiac cycle. The 4D velocity data obtained from 14 healthy volunteers were used to investigate dynamic blood flow with the use of multiplanar reformatting, 3D streamlines, and 4D particle tracing. In addition, the effects of magnetic field strength, parallel imaging, and temporal resolution on the data were investigated in a comparative evaluation at 1.5T and 3T using three different parallel imaging reduction factors and three different temporal resolutions in eight of the 14 subjects. Studies were consistently performed faster at 3T than at 1.5T because of better parallel imaging performance. A high temporal resolution (65 ms) was required to follow dynamic processes in the intracranial vessels. The 4D flow measurements provided a high degree of vascular conspicuity. Time-resolved streamline analysis provided features that have not been reported previously for the intracranial vasculature.


Benefits Associated With Early Diagnosis and Treatment of Soft Tissue Sarcomas of the Foot and Ankle.
Soft tissue sarcomas arising in the foot and ankle are often misdiagnosed, resulting in excision without adequate preoperative staging or surgical margins. The goal of the present study was to review a case series of soft tissue sarcomas of the foot and ankle with attention directed at unplanned excisions of sarcomas. An unplanned excision means that a patient either underwent an inadequate preoperative workup or the preoperative workup indicated a benign entity, resulting in surgical resection. We retrospectively analyzed the medical records of 10 patients with sarcomas of the foot and ankle treated at our institution. All soft tissue sarcomas were excised with the widest margin possible without any major bone or neurovascular resection to allow for preservation of the foot. All patients were followed up for a minimum of 8 months to assess pain, function, and complications. The patients were followed up for an average of 22.4 (range 8 to 44) months. Of the 10 patients, 6 had undergone preoperative magnetic resonance imaging without contrast and the sarcomas were read as benign or cystic masses. Two patients had undergone preoperative magnetic resonance imaging with contrast, and these scans were also misread. Despite having undergone previous unplanned surgery, none of these sarcomas had recurred after repeat resection with a wider margin at a mean follow-up of 22.4 (range 8 to 44) months. At the last follow-up point, 8 patients were alive without evidence of disease. Minor complications included lymph edema in 1, stress fracture in 1, and wound infection in 1 that resolved. Preoperative MRI with contrast is recommended before resecting any soft tissue masses of the foot and ankle. In the event of an unplanned excision of a soft tissue sarcoma, the patient should be referred to an orthopedic oncologist for definitive surgery to optimize the oncologic and functional results. In the present retrospective analysis, previous intervention did not seem to affect the prognosis, including local recurrence, distant metastasis, disease-free interval, and functional outcomes.


[Current Clinical Assessment Scales and Cognitive Tests in Global Clinical Studies on Alzheimer's Disease].
To quantify the severity of clinical symptoms and cognitive dysfunction, clinical assessment scales and cognitive tests are used in clinical studies involving patients with Alzheimer's disease (AD). As an increasing number of clinical studies involving individuals with preclinical AD and mild cognitive impairment are being conducted, newly developed scales are also utilized in those studies. Scales and tests must be translated into Japanese and validated, with the level of difficulty kept equal to the original, in order to obtain data comparable to those of other countries, in global studies. Moreover, the uniformity of procedures in test administration is crucial to procuring high-quality data in multicenter clinical studies. Such background efforts, which support novel scientific findings, as well as clinical assessment scales and cognitive tests which are used in recent studies are described in this section.


Site-bond percolation in two-dimensional kagome lattices: Analytical approach and numerical simulations.
The site-bond percolation problem in two-dimensional kagome lattices has been studied by means of theoretical modeling and numerical simulations. Motivated by considerations of cluster connectivity, two distinct schemes (denoted as S∩B and S∪B) have been considered. In S∩B (S∪B), two points are connected if a sequence of occupied sites and (or) bonds joins them. Analytical and simulation approaches, supplemented by analysis using finite-size scaling theory, were used to calculate the phase boundaries between the percolating and nonpercolating regions, thus determining the complete phase diagram of the system in the (p_{s},p_{b}) space. In the case of the S∩B model, the obtained results are in excellent agreement with previous theoretical and numerical predictions. In the case of the S∪B model, the limiting curve separating percolating and nonpercolating regions is reported here.


A Case of Oxalate Nephropathy: When a Single Cause Is Not Crystal Clear.
Hyperoxaluria can result in oxalate nephropathy with intratubular calcium oxalate crystallization and acute tubular injury. Primary inherited enzymatic deficiency or secondary causes such as excessive dietary intake, enteric increased absorption, or high doses of vitamin C, which is metabolized to oxalate, may underlie hyperoxaluria and oxalate nephropathy. We report a case of acute kidney injury due to oxalate nephropathy in a patient using chelating therapy with oral ethylenediamine tetra acetic acid (EDTA), intravenous supplementation with vitamin C, and chronic diarrhea and discuss the potential kidney damage these factors can cause in particular settings. To our knowledge, this is the first report suggesting an association between oral EDTA and oxalate nephropathy.


Do aquaporins have a role in nocturnal enuresis?
Aquaporins are proteins that mediate transmembrane water transport in a variety of tissues including the kidney. Vasopressin plays an important role in regulation of water metabolism, and under normal conditions the kidney collecting duct is extremely sensitive to vasopressin. Vasopressin stimulates the synthesis of aquaporin 2 (AQP2) in kidney collecting duct principal cells. Studies in Brattle Boro rats which are vasopressin deficient, revealed low levels of AQP2 in association with extreme polyuria. After vasopressin treatment for 5 days AQP2 levels increased threefold. Using rat models with nephrogenic diabetes insipidus (NDI) we have demonstrated that AQP2 expression is down regulated in association with polyuria, suggesting that reduced levels of AQP2 may be a general factor in acquired forms of NDI from a variety of reasons. The polyuria and urinary concentrating defects associated with an abnormal nightly-increase in AVP in patients with nocturnal enuresis may partly be due to a lack of vasopressin-mediated AQP2 expression since treatment with desmopressin in these patients have normalised their nocturnal urine production.


Tubulointerstitial nephritis and vasculitis.
In the past few years, advances in molecular and cellular immunology have greatly enhanced our understanding of the immunopathogenesis of tubulointerstitial nephritis and vasculitis. This review focuses on recent observations regarding nephritogenic tubulointerstitial antigens, their expression and presentation by tubular epithelial cells, the subsequent immune response, and the factors that serve to modulate this response. The latter part of this review summarizes recent findings regarding the role of autoantibodies in the immunopathogenesis of vasculitis.


Virtual overdose monitoring services and overdose prevention technologies: Opportunities, limitations, and future directions.
Overdose mortality has continued to rise in North America and across the globe in people who use drugs. Current harm reduction strategies such as supervised consumption sites and naloxone kit distribution have been important public health strategies implemented to decrease the harms associated with illicit drug use however have key limitations which prevent their scalability. This is represented in statistics which indicate that the vast majority of overdose mortality occur in individuals who use drugs by themselves. To address this, virtual overdose monitoring services and overdose detection technologies have emerged as an adjunct solution that may help improve access to harm reduction services for those that cannot or choose not to access current in-person services. This article outlines the current limitations of harm reduction services, the opportunities, challenges, and controversies of these technologies and services, and suggests avenues for additional research and policy development.


[Classification of surgical complications: analysis of the group of consecutive patients].
The aim of the work was to evaluate the implementation of Clavien - Dindo classification of surgical complications into a routine clinical praxis and to evaluate the results achieved at the university department of surgery. Prospectively collected data of patients hospitalized at the Clinic of Surgery of the University Hospital in Hradec Králové between January 2010 and September 2012 were retrospectively evaluated. Incidence, spectrum and severity of postoperative complications were evaluated according to individual surgical specializations. 9039 patients were operated and enrolled into the database during the time period from January 2010 to October 2012. A surgical complication was recorded in 1248 (12.9%) patiens, grade I. in 284 (3.4%) cases, grade II. in 384 (4.3%) cases, grade III in 370 (4.1%) cases, grade IV. in 67 (0.7%) patients. Death,i.e. grade V., occurred in 143 (1.43%) patients. Clavien - Dindo classification of surgical complications was successfully implemented into a routine clinical praxis at the department of surgery, University Hospital Hradec Králové. Wider use of this classification system would improve conditions for evaluation and comparison of results between different surgical approaches, surgeons or departments.


Quantitative aspects of preoptic thermosensitivity in the conscious ox.
An attempt has been made to study the quantitative relationship between the temperature of the anterior hypothalamic/preoptic (AH/PO) region and heat balance in the conscious ox (Bos taurus) at a thermoneutral air temperature of 20 degrees C. Heat balance was measured by combined gradient-layer and open-circuit calorimetry and AH/PO temperature was controlled using a water-perfused thermode. Continuous warming or cooling of the AH/PO region caused a transient heat imbalance which shifted core temperature to a new stable level dependent upon the level of stimulation. The heat balance responses to continuous AH/PO warming included peripheral vasodilation and increased respiratory and cutaneous evaporative heat loss; those to AH/PO cooling consisted mainly of decreased heat loss. Heat production was largely unaffected by both AH/PO warming and cooling. The heat balance responses developed only slowly, reaching a peak up to 50 min after the start of stimulation and taking several hours to complete. The change to stability in core temperature during continuous AH/PO stimulation and the additional heat loss prior to that stability were the most sensitive indicators of response of all the variables measured. A significant response was observed in these parameters even at the smallest stimulus level (less than 0.33 degrees C) and correlation of response with stimulus level was precise with a zero threshold for response. This suggests a continuity of operation of the AH/PO mechanism about zero. Several responses showed a curved relationship between slope of response and stimulus level, such that the increment in response per unit increase in stimulus level became greater as stimulus level increased. The results suggest a two-tier form of thermoregulation in the ox with AH/PO sensitivity to warming greater than its sensitivity to cooling. It appears also that AH/PO temperature is at least as dominant in the thermoregulatory mechanisms in the ox as it is in smaller animals.


Nutritional knowledge and attitudes of dental students.
The nutritional knowledge and attitudes of 230 dental students were studied. The students answered 68.6% of the nutritional knowledge questions currectly, but the test scores were low because they were adversely affected by the degree of certainty. First-year students scored significantly higher than fourth-year students. There was no significant difference between scores of men and women. Knowledge scores were highest for questions on nutrition and oral health and lowest for those on nutritional assessment. Dental students generally expressed favorable attitudes toward nutrition and nutritional care of patients. They agreed that dentists were vital members of the health team and had a responsibility to become involved in health screening and nutrition education of patients. Dietitians were seen as valuable resources to be consulted about nutrition education of the dental patient. More first-year students supported the idea that dentists should prescribe nutritoinal supplements for patients, whereas more fourth-year students were undecided about this matter. There were no differences in nutrition attitude scores attributable to gender of the student or year in dental school. In this study, nutritional knowledge scores did not correlate with nutrition attitude scores.


Pharmacokinetics and drug interactions of cholinesterase inhibitors administered in Alzheimer's disease.
Cholinesterase inhibitors are the first agents to be successfully developed specifically for the treatment of cognitive decline associated with Alzheimer's disease. Basic knowledge of their pharmacokinetics is important to their appropriate administration. Their pharmacokinetics help determine the magnitude and duration of their pharmacologic effects, and also the manner in which they affect the degree of cholinesterase inhibition and recovery. The clinical utility of measuring these values in daily practice awaits further research. Drug interactions with cholinesterase inhibitors may occur by pharmacokinetic or pharmacodynamic mechanisms. For the most part, interactions that are mediated by the hepatic cytochrome P-450 system have been inadequately evaluated.


[A prospective study comparing the efficacy of early administration of terlipressin and somatostatin for the control of acute variceal bleeding in patients with cirrhosis].
Terlipressin and somatostatin decrease portal venous pressure and they are used for the treatment of variceal bleeding. However, only a few studies have compared the efficacy of these drugs in combination with other procedures for hemostasis. Therefore, we performed a prospective study to compare the efficacy of terlipressin and somatostatin for controlling acute variceal bleeding when used in combination with other procedures for hemostasis. A total of 98 patients, who presented with variceal bleeding from September 2003 to May 2005, were randomly divided into the somatostatin group or terlipressin group. We compared the 5-day failure rate (defined as failure to control bleeding, rebleeding or death within 5 days of admission) and the 6-week mortality. The prognostic factors for 5-day failure and 6-week mortality were also evaluated. There were no differences in baseline characteristics between the two groups. The overall 5-day failure rate and the cumulative 6-week mortality were 16.3% and 15.8%, respectively. The five-day failure rate and the cumulative 6-week mortality were not significantly different between the somatostatin and terlipressin groups. Hepatocellular carcinoma, the baseline serum creatinine level and endoscopic treatment for hemostasis were the significant predictors of 5-day failure; the baseline serum creatinine level was the predictor of 6-week mortality. Both somatostatin and terlipressin were effective and showed comparable efficacy for the control of the acute variceal bleeding in the setting of a combined therapeutic approach. The baseline serum creatinine level may be a significant predictor for patient failure at 5 days and the 6-week mortality.


Delta protein kinase-C in the rat ovary: estrogen regulation and localization.
The present studies were undertaken to examine the cellular distribution and regulation of delta protein kinase-C (PKC) at different stages of luteal differentiation in the rat. Results from in situ hybridization studies with a delta PKC-specific probe demonstrated that delta PKC was localized specifically in granulosa cells of healthy preantral, small antral, and preovulatory follicles and corpora lutea from the second half of pregnancy. Northern and Western blot analyses showed that levels of delta PKC protein and messenger RNA were elevated 25- and 35-fold, respectively, in the second half of pregnancy over levels in preovulatory follicle-enriched ovaries, peaking between days 18-20 of pregnancy. Levels of alpha PKC, beta PKC, and zeta PKC remained unchanged during luteal differentiation. In rats hypophysectomized and hysterectomized on day 12 of pregnancy and in immature rats containing corpora lutea induced with PMSG followed by hCG injections, exogenous estrogen caused 3- and 7-fold increases in delta PKC protein, respectively. In the immature rat model, delta PKC messenger RNA levels were not altered by exogenous estrogen. Although exogenous testosterone also elevated delta PKC protein levels, exogenous dihydrotestosterone and progesterone were ineffective. These results suggest that estrogen may participate, at a posttranscriptional level, in the dramatic induction of delta PKC seen at the end of pregnancy in rat corpora lutea.


Zebrafish slc30a10 deficiency revealed a novel compensatory mechanism of Atp2c1 in maintaining manganese homeostasis.
Recent studies found that mutations in the human SLC30A10 gene, which encodes a manganese (Mn) efflux transporter, are associated with hypermanganesemia with dystonia, polycythemia, and cirrhosis (HMDPC). However, the relationship between Mn metabolism and HMDPC is poorly understood, and no specific treatments are available for this disorder. Here, we generated two zebrafish slc30a10 mutant lines using the CRISPR/Cas9 system. Compared to wild-type animals, mutant adult animals developed significantly higher systemic Mn levels, and Mn accumulated in the brain and liver of mutant embryos in response to exogenous Mn. Interestingly, slc30a10 mutants developed neurological deficits in adulthood, as well as environmental Mn-induced manganism in the embryonic stage; moreover, mutant animals had impaired dopaminergic and GABAergic signaling. Finally, mutant animals developed steatosis, liver fibrosis, and polycythemia accompanied by increased epo expression. This phenotype was rescued partially by EDTA- CaNa2 chelation therapy and iron supplementation. Interestingly, prior to the onset of slc30a10 expression, expressing ATP2C1 (ATPase secretory pathway Ca2+ transporting 1) protected mutant embryos from Mn exposure, suggesting a compensatory role for Atp2c1 in the absence of Slc30a10. Notably, expressing either wild-type or mutant forms of SLC30A10 was sufficient to inhibit the effect of ATP2C1 in response to Mn challenge in both zebrafish embryos and HeLa cells. These findings suggest that either activating ATP2C1 or restoring the Mn-induced trafficking of ATP2C1 can reduce Mn accumulation, providing a possible target for treating HMDPC.