Effects of 2.1 and 3.5x10(6) sex-sorted sperm dosages on conception rates of Holstein cows and heifers.
The objective was to compare conceptions rates of Holstein cows and heifers after artificial insemination (AI) with 2.1 or 3.5x10(6) sex-sorted sperm or 15x10(6) conventional sperm. Ejaculates collected from 7 Holstein sires were cryopreserved conventionally at 15x10(6) sperm per dose or sorted to 90% purity for X-chromosome-bearing spermatozoa using flow cytometry and cryopreserved at either 2.1 or 3.5x10(6) sperm per dose. All treatments were processed in an egg-yolk (20%), Tris, glycerol (7%) extender and packaged in color-coded 0.25-mL French straws. Straws (n=700 straws/dosage per sire) were packaged and distributed in aliquots of 12 (4 straws/sperm dosage) to 69 Holstein herds with an across-herd goal of achieving approximately 50% use in heifers and cows. Straw color was recorded in the on-farm recordkeeping system at the time of AI and retrieved by electronic download. Data for cows and heifers were analyzed separately. Among heifers, 6,268 services were retrieved from 45 herds (298+/-4.2 services/sperm dose per sire; range: 244 to 344). Conception rate of heifers was influenced by the sire by treatment interaction. Conception rate of the 2.1 and 3.5x10(6) sex-sorted sperm dosages were comparable in 6 of 7 sires. Conception rate of both sex-sorted dosages were less than those of conventional semen for 6 of 7 sires. Across sires, heifer conception rates for 2.1 and 3.5x10(6) sex-sorted sperm dosages and 15x10(6) conventional dosages were 44, 46, and 61%, respectively. Among cows, 5,466 services were retrieved from 52 herds (260+/-3.3 services/sperm dose per sire; range: 236 to 289). Conception rates of cows were influenced by herd, sire, and sperm dosage. Conception rates of the 2.1 and 3.5x10(6) sex-sorted sperm dosage were comparable for all 7 sires. Conception rates of 2.1x10(6) sex-sorted sperm dosage were less than those of conventional semen for 4 of 7 sires and conception rates of the 3.5x10(6) sex-sorted sperm dosage were less than those of conventional semen for 2 of 7 sires. Across sires, conception rates for 2.1 and 3.5x10(6) sex-sorted sperm dosages and 15x10(6) conventional dosages in cows were 23, 25, and 32%, respectively. In conclusion, these data could not confirm that a meaningful improvement in conception rates should be expected in cows or heifers from increasing sex-sorted sperm dosage from 2.1 to 3.5x10(6) sperm per dose.


Substantial gains in word learning ability between 20 and 24 months: A longitudinal ERP study.
This longitudinal ERP study investigated changes in children's ability to map novel words to novel objects during the dynamic period of vocabulary growth between 20 and 24 months. During this four-month period the children on average tripled their productive vocabulary, an increase which was coupled with changes in the N400 effect to pseudoword-referent associations. Moreover, productive vocabulary size was related to the dynamics of semantic processing during novel word learning. In children with large productive vocabularies, the N400 amplitude was linearly reduced during the five experimental learning trials, consistent with the repetition effect typically seen in adults, while in children with smaller vocabularies the N400 attenuation did not appear until the end of the learning phase. Vocabulary size was related only to modulation of the N400 to pseudowords, not to real words. These findings demonstrate a remarkable development of fast mapping ability between 20 and 24 months.


[Interhemispheric epidermoid cyst. A clinical case].
The authors report a rare case of epidermoid cyst with interhemispheric growth closely connected to the anterior portion of the corpus callosum. Although extremely sensitive in determining the site, dimensions and relations of the lesion, NMR does not allow the nature of this pathology to be diagnosed owing to the lack of signal specificity. CT, using the measurement of Hounsfield units, allows a differential diagnosis of epidermoid cysts and arachnoid cysts and lipomas to be made in almost all cases. Treatment is surgical and the complete excision of the capsule avoids recidivation.


Thermal and Electronic Fluctuations of Flexible Adsorbed Molecules: Azobenzene on Ag(111).
We investigate the thermal and electronic collective fluctuations that contribute to the finite-temperature adsorption properties of flexible adsorbates on surfaces on the example of the molecular switch azobenzene C_{12}H_{10}N_{2} on the Ag(111) surface. Using first-principles molecular dynamics simulations, we obtain the free energy of adsorption that accurately accounts for entropic contributions, whereas the inclusion of many-body dispersion interactions accounts for the electronic correlations that govern the adsorbate binding. We find the adsorbate properties to be strongly entropy driven, as can be judged by a kinetic molecular desorption prefactor of 10^{24} s^{-1} that largely exceeds previously reported estimates. We relate this effect to sizable fluctuations across structural and electronic observables. A comparison of our calculations to temperature-programed desorption measurements demonstrates that finite-temperature effects play a dominant role for flexible molecules in contact with polarizable surfaces, and that recently developed first-principles methods offer an optimal tool to reveal novel collective behavior in such complex systems.


Anti-inflammatory, gastroprotective, free-radical-scavenging, and antimicrobial activities of hawthorn berries ethanol extract.
Hawthorn [Crataegus monogyna Jacq. and Crataegus oxyacantha L.; sin. Crataegus laevigata (Poiret) DC., Rosaceae] leaves, flowers, and berries are used in traditional medicine in the treatment of chronic heart failure, high blood pressure, arrhythmia, and various digestive ailments, as well as geriatric and antiarteriosclerosis remedies. According to European Pharmacopoeia 6.0, hawthorn berries consist of the dried false fruits of these two species or their mixture. The present study was carried out to test free-radical-scavenging, anti-inflammatory, gastroprotective, and antimicrobial activities of hawthorn berries ethanol extract. Phenolic compounds represented 3.54%, expressed as gallic acid equivalents. Determination of total flavonoid aglycones content yielded 0.18%. The percentage of hyperoside, as the main flavonol component, was 0.14%. With respect to procyanidins content, the obtained value was 0.44%. DPPH radical-scavenging capacity of the extract was concentration-dependent, with EC50 value of 52.04 microg/mL (calculation based on the total phenolic compounds content in the extract). Oral administration of investigated extract caused dose-dependent anti-inflammatory effect in a model of carrageenan-induced rat paw edema. The obtained anti-inflammatory effect was 20.8, 23.0, and 36.3% for the extract doses of 50, 100, and 200 mg/kg, respectively. In comparison to indomethacin, given in a dose producing 50% reduction of rat paw edema, the extract given in the highest tested dose (200 mg/kg) showed 72.4% of its activity. Gastroprotective activity of the extract was investigated using an ethanol-induced acute stress ulcer in rats with ranitidine as a reference drug. Hawthorn extract produced dose-dependent gastroprotective activity (3.8 +/- 2.1, 1.9 +/- 1.7, and 0.7 +/- 0.5 for doses of 50, 100, and 200 mg/kg, respectively), with the efficacy comparable to that of the reference drug. Antimicrobial testing of the extract revealed its moderate bactericidal activity, especially against gram-positive bacteria Micrococcus flavus, Bacillus subtilis, and Lysteria monocytogenes, with no effect on Candida albicans. All active components identified in the extract might be responsible for activities observed.


Identification of the Zn(II) site in the copper-responsive yeast transcription factor, AMT1: a conserved Zn module.
The N-terminal metal-binding domains of the copper-activated yeast transcription factors, ACE1 and AMT1, bind to specific DNA sequences in a Cu-dependent fashion. Recombinant AMT1 and ACE1 metal-binding domains are isolated as Cu4Zn1-protein complexes. Site-directed mutagenesis of AMT1 was used in this study to map the ligands of the Cu(I) and Zn(II) ions. The results are consistent with the N-terminal halves of AMT1 and ACE1 consisting of two independent submodules, one binding a single Zn(II) ion and the second binding the tetracopper cluster. The basis of this conclusion is, first, that mutations of two cysteinyl codons and a histidyl codon in the first 42 residues of AMT1 do not alter DNA binding. In contrast, serine substitutions at four cysteine positions at codons 43, 61, 90, and 98 abolish DNA binding. We demonstrated previously that population of the Zn(II) site in AMT1 does not alter the ability of the protein to bind DNA but bound Cu(I) ions are essential for DNA binding [Thorvaldsen, J. L., et al. (1994) Biochemistry 33, 9566-9577]. Second, mutations in the N-terminal 42 residue segment reduce the Zn(II) content of purified mutant AMT1 molecules. Third, a synthetic peptide consisting of the N-terminal 42 residues in AMT1 forms a stable Zn(II) complex and substitution with Co(II) reveals an electronic spectrum identical to that of the Co-substituted intact Cu4AMT1 protein. 113Cd(II) NMR studies reveal that the divalent metal site consists of ligands provided by three cysteinyl thiolates and a single histydyl imidazole. The sequence homology between AMT1, ACE1, and MAC1 in the N-terminal 42 residues suggests that ACE1 and MAC1 will, likewise, contain N-terminal Zn modules. A 42-residue ACE1 synthetic peptide gives identical metal binding properties to the corresponding AMT1 synthetic peptide. Thus, AMT1 and likely ACE1 consist of two contiguous modules, residues 1-42 forming an independent Zn(II) module and residues 43-110 enfolding a tetracopper cluster.


The p97 ATPase dislocates MHC class I heavy chain in US2-expressing cells via a Ufd1-Npl4-independent mechanism.
The human cytomegalovirus (HCMV) protein US2 hijacks the endoplasmic reticulum (ER)-associated degradation machinery to dispose of MHC class I heavy chain (HC) at the ER. This process requires retrotranslocation of newly synthesized HC molecules from the ER membrane into the cytosol, but the mechanism underlying the dislocation reaction has been elusive. Here we establish an in vitro permeabilized cell assay that recapitulates the retrotranslocation of MHC HC in US2-expressing cells. Using this assay, we demonstrate that the dislocation process requires ATP and ubiquitin, as expected. The retrotranslocation also involves the p97 ATPase. However, the mechanism by which p97 dislocates MHC class I HC in US2 cells is distinct from that in US11 cells: the dislocation reaction in US2 cells is independent of the p97 cofactor Ufd1-Npl4. Our results suggest that different retrotranslocation mechanisms can employ distinct p97 ATPase complexes to dislocate substrates.


[The relationship between the anti-HCV titer and the state of chronic hepatitis C].
By dilution of the serum, we quantitatively analyzed anti-HCV to examine the relation between the anti-HCV titer and the state of chronic hepatitis C, and determined the effectiveness of interferon therapy. The anti-HCV titer was related with the state of chronic hepatitis C, but not as closely as GPT. Cases with blood transfusion had a lower titer of anti-HCV, weaker histological finding and better response to interferon than those without blood transfusion. The anti-HCV titer was not useful for determining the effectiveness of interferon therapy.


Near-Infrared II Gold Nanocluster Assemblies with Improved Luminescence and Biofate for In Vivo Ratiometric Imaging of H2S.
Ultrasmall gold nanoclusters (AuNCs) are emerging as promising luminescent nanoprobes for bioimaging due to their fantastic photoluminescence (PL) and renal-clearable ability. However, it remains a great challenge to design them for in vivo sensitive molecular imaging in desired tissues. Herein, we have developed a strategy to tailor the PL and biofate of near-infrared II (NIR-II)-emitting AuNCs via ligand anchoring for improved bioimaging. By optimizing the ligand types in AuNCs and using Er3+-doped lanthanide (Ln) nanoparticles as models, core-satellite Ln@AuNCs assemblies were rationally constructed, which enabled 2.5-fold PL enhancement of AuNCs at 1100 nm and prolonged blood circulation compared to AuNCs. Significantly, Ln@AuNCs with dual intense NIR-II PL (from AuNCs and Er3+) can effectively accumulate in the liver for ratiometric NIR-II imaging of H2S, facilitated by H2S-mediated selective PL quenching of AuNCs. We have then demonstrated the real-time imaging evaluation of liver delivery efficacy and dynamics of two H2S prodrugs. This shows a paradigm to visualize liver H2S delivery and its prodrug screening in vivo. Note that Ln@AuNCs are body-clearable via the hepatobiliary excretion pathway, thus reducing potential long-term toxicity. Such findings may propel the engineering of AuNC nanoprobes for advancing in vivo bioimaging analysis.


A Psychometric Study of the Modified Bridges for Newborns Screening Tool.
Welcome Baby, a program that First 5 Los Angeles (First 5 LA) sponsors, provides new mothers with supportive services intended to create enriching environments for their children. To identify mothers in need of these services, First 5 LA employs hospital liaisons who administer a maternal risk assessment tool, the Modified Bridges for Newborns screening tool, during postpartum interviews of mothers. First 5 LA uses risk assessment scores from the Modified Bridges to classify mothers as low, moderate, or high risk; high-risk mothers are eligible for additional supportive services that are not available to low- and moderate-risk mothers. This article describes RAND Corporation work evaluating the psychometric characteristics of the Modified Bridges.


Cathepsin L promotes secretory IgA response by participating in antigen presentation pathways during Mycoplasma Hyopneumoniae infection.
Cathepsin L (CTSL) has been proved to help contain leishmaniasis and mycoplasma infection in mice by supporting cellular immune responses, but the regulatory functions of CTSL on mucosal immune responses haven't been tested and remain undefined. Here, we investigated the effects of CTSL on SIgA responses and invariant chain (Ii) degradations in the co-cultured swine dendritic cells (DCs) and B cells system in vitro. When the cells system were transfected with vector CTSL-GFP or incubated with recombinant CTSL (rCTSL) before they were infected with Mycoplasma hyopneumoniae (M.hp), SIgA significantly increased and Ii chain was degraded into smaller intermediates, while SIgA decreased when CTSL was knockdown or inhibited with E64. To confirm the SIgA responses promoted by CTSL contribute to the resistance to mycoplasma pneumonia, pigs injected with rCTSL before they were challenged with M.hp, showed milder clinical symptoms and histopathological damage of lungs, less mycoplasma burden together with higher secretion of SIgA, percentages of CD4+ T cells and level of MHC II molecules comparing with the group without rCTSL. Collectively, these results suggested that rCTSL could provide effective protection for piglets against mycoplasma pneumonia by enhancing M.hp-specific mucosal immune responses through its role in antigen presentation by processing the invariant chain.


What Are the Prospects for Treating TP53 Mutated Myelodysplastic Syndromes and Acute Myeloid Leukemia?
TP53 is a key tumor suppressor gene involved in fundamental biological processes of genomic stability and is recurrently mutated in a subgroup of myelodysplastic syndromes and acute myeloid leukemia. These patients have unique clinical and molecular features resulting in dismal outcomes despite standard cytotoxic chemotherapy, and long-term survival is seldom achieved with allogeneic stem cell transplant. Upfront use of hypomethylating agents with or without venetoclax has resulted in a favorable initial response over intensive cytotoxic chemotherapy, albeit responses are nondurable, and the median overall survival is typically less than 6 to 8 months. In this review, we examine the evidence of conventional treatments and focus on the emerging novel therapeutic options, including targeted molecular and immunotherapies for this challenging molecular subgroup. Together, there are still significant unmet needs to improve outcomes of patients with TP53 mutated myelodysplastic syndromes and acute myeloid leukemia, and enrollment in clinical trials should be highly favored whenever they are available.


[Effects of silent nucleostemin gene expression on apoptosis of HL-60 cells in vitro].
This study was aimed to explore whether the apoptosis of leukemia cells can be induced by targeting silencing nucleostemin gene in vitro. HL-60 cells were taken as the model, and were directly transfected with nucleostemin short hairpin RNA (NS-shRNA). Sequences unrelated with NS gene were taken as control. The blocking effect of NS-shRNA was detected by RT-PCR, the morphology changes in living cells were observed under inverted microscope, and the changes of cell shape and nucleus were detected by Wright-Giemsa staining. The amount of apoptotic cells were assayed by flow cytometer (FCM) and TUNEL technique, and the positive rate of apoptosis was determined meanwhile. The results showed that two NS-shRNA were synthesized in vitro, and the more effective one was selected to be transfected into HL-60 cells. The blocking rate of NS-mRNA reached to 74.94%. After transfection for 48 hours, Wright-Giemsa staining showed nuclear fragmentations and "apoptosis body" in cells. The apoptosis rate in transfected group detected by flow cytometry and TUNEL method were (25.32 +/- 3.06)% and (27.3 +/- 3.21)% respectively, but were only (3.12 +/- 0.38)% and (3.30 +/- 1.52)% in control group, the difference between the transfected group and the control group was significant (p < 0.01). It is concluded that the apoptosis of HL-60 leukemia cells can be induced by the silencing NS gene expression in vitro, which provides a theoretical basis for using NS gene as a candidate target gene in therapy of malignant tumor.


Immunogenomic Profiling and Classification of Prostate Cancer Based on HIF-1 Signaling Pathway.
The HIF-1 signaling pathway plays an important role in the pathogenesis of cancer. Many studies have explored the progression of prostate cancer (PCa) under hypoxic conditions based on transcriptome data; few have uncovered the immunogenomic profiling and prostate cancer classification based on the HIF-1 signaling pathway. This pathway may help to identify the optimal subset of PCa patients responsive to immunotherapy/chemotherapy. The immunogenomic PCa subsets were classified based on profiling of the HIF-1 signaling pathway, using four publicly available PCa datasets. Three PCa subtypes that named as HIF-1 High (HIF-1_H), HIF-1 Medium (HIF-1_M), and HIF-1 Low (HIF-1_L) were identified. Functional enrichment was analyzed in each subtype. Several cancer-associated and immune-related pathways were hyperactivated in the HIF-1_H subtypes. In contrast, HIF-1_L subtypes were enriched in cell cycle and cell repair. Compared with other subtypes, HIF-1_H subtypes have greater immune cell infiltration, anti-tumor immune activity, and better survival prognosis. The submap and TIDE algorithm were used to predict the clinical response to immune checkpoint blockade, and GDSC was employed to screen potential chemotherapeutic targets for the treatment of PCa. Several chemotherapy drugs were identified in the GDSC dataset, including ABT 888, Temsirolimus, and EHT 1864. Meanwhile, HIF-1_H was defined as an early PCa marker, which is more likely to respond to immunotherapy. The identification of immunogenomic PCa subtypes based on the HIF-1 signaling pathway has potential clinical implications for PCa treatment. Immunopositive PCa subtypes will help to explore the reasons for the poor response of PCa to immunotherapy, and it is expected that immunotherapy will guide the personalized treatment of PCa patients.


Costs, Staffing, and Services of Assisted Living in the United States: A Literature Review.
Assisted-living facilities (ALFs), which provide a community for residents who require assistance throughout their day, are an important part of the long-term-care system in the United States. The costs of ALFs are paid either out of pocket, by Medicaid, or by long-term-care insurance. Monthly costs of ALFs have increased over the past 5 years on an average of 4.1%. The purpose of this research was to examine the future trends in ALFs in the United States to determine the impact of health care on costs. The methodology for this study was a literature review, and a total of 32 sources were referenced. Trends in monthly costs of ALFs have increased from 2004 to 2014. Within the past 5 years, there has been an increase on average of 4.1% in assisted-living costs. Medicaid is one payer for residents of ALFs, whereas another alternative is the use of long-term-care insurance. Unfortunately, Medicare does not pay for ALFs. Staffing concerns in ALFs are limited because of each state having different rules and regulations. Turnover and retention rates of nurses in ALFs are suggested to be high, whereas vacancy rate for nurses is suggested to be lower. The baby-boomer generation can be one contribution to the increase in costs. Over the years, there has been an increase in Alzheimer disease, which has had also an effect on cost in ALFs.


An Autograft for Anterior Cruciate Ligament Reconstruction Results in Better Biomechanical Performance and Tendon-Bone Incorporation Than Does a Hybrid Graft in a Rat Model.
The biomechanical and tendon-bone incorporation properties of allograft-augmented hybrid grafts for anterior cruciate ligament (ACL) reconstruction compared with traditional autografts are unknown. Using an autograft for ACL reconstruction yields better results on biomechanical testing, radiographic analysis, and histological evaluation versus using a hybrid graft. Controlled laboratory study. A total of 66 adult male Sprague Dawley rats underwent unilateral ACL reconstruction with an autograft (AT group; n = 33) or a hybrid graft (HB group; n = 33). The grafts used in both groups were harvested from the peroneus longus tendon and were fixed by suturing to the surrounding periosteum. Samples were harvested for biomechanical testing, micro-computed tomography (CT), and histological evaluation at 4, 8, and 12 weeks postoperatively. Bone tunnels on the femoral and tibial sides were divided into 3 subregions: intra-articular (IA), midtunnel (MT), and extra-articular (EA). A cylinder-like volume of interest in the bone tunnel and a tubular-like volume of interest around the bone tunnel were used to evaluate new bone formation and bone remodeling, respectively, via micro-CT. In the AT group, there were significantly higher failure loads and stiffness at 8 weeks (failure load: 3.04 ± 0.40 vs 2.09 ± 0.54 N, respectively; P = .006) (stiffness: 3.43 ± 0.56 vs 1.75 ± 0.52 N/mm, respectively; P < .001) and 12 weeks (failure load: 9.10 ± 1.13 vs 7.14 ± 0.94 N, respectively; P = .008) (stiffness: 4.45 ± 0.75 vs 3.36 ± 0.29 N/mm, respectively; P = .008) than in the HB group. With regard to new bone formation in the bone tunnel, in the AT group, the bone volume/total volume (BV/TV) was significantly higher than in the HB group on the tibial side at 8 weeks (IA: 22.21 ± 4.98 vs 5.16 ± 3.98, respectively; P < .001) (EA: 19.66 ± 7.19 vs 10.85 ± 2.16, respectively; P = .030) and 12 weeks (IA: 30.50 ± 5.04 vs 17.11 ± 7.31, respectively; P = .010) (MT: 21.15 ± 2.58 vs 15.55 ± 4.48, respectively; P = .041) (EA: 20.75 ± 3.87 vs 10.64 ± 3.94, respectively; P = .003). With regard to bone remodeling around the tunnel, the BV/TV was also significantly higher on the tibial side at 8 weeks (MT: 33.17 ± 8.05 vs 15.21 ± 7.60, respectively; P = .007) (EA: 25.19 ± 6.38 vs 13.94 ± 7.10, respectively; P = .030) and 12 weeks (IA: 69.46 ± 4.45 vs 47.80 ± 6.16, respectively; P < .001) (MT: 33.15 ± 3.88 vs 13.76 ± 4.07, respectively; P < .001) in the AT group than in the HB group. Sharpey-like fibers had formed at 8 weeks in the AT group. A large number of fibroblasts withdrew at 12 weeks. In the AT group, the width of the interface was significantly narrower at 4 weeks (85.86 ± 17.49 vs 182.97 ± 14.35 μm, respectively; P < .001), 8 weeks (58.86 ± 10.99 vs 90.15 ± 11.53 μm, respectively; P = .002), and 12 weeks (42.70 ± 7.96 vs 67.29 ± 6.55 μm, respectively; P = .001) than in the HB group. Using an autograft for ACL reconstruction may result in improved biomechanical properties and tendon-bone incorporation compared with a hybrid graft. Augmenting small autografts with allograft tissue may result in decreased biomechanical performance and worse tendon-bone incorporation, increasing the risk of graft failure.


[Biological malignant potential and carbohydrate antigens in breast cancer].
We investigated the association of expression of carbohydrate antigens with the degree of malignancy in breast cancer. When this expression was compared in the cancerous portion and adjacent non-cancerous portion in each of the patients, increased expression of type2 carbohydrate antigens were found in cancerous portions. The prognosis of patients in whom type2 carbohydrate antigens were increased in the cancerous portion was significantly poorer than in patients in whom they were not increased. The experimental studies, using cell lines derived from human breast cancer, resulted that the adhesion of cancer cells to vascular endothelial cells was mediated by sialyl Lewis(x) via E-selectin. Moreover, it was suggested that there were the induction of E-selectin expression on the endothelial cells by cancer cells and secondary adhesional system following the initial adhesion mediated by E-selectin and carbohydrate ligands. Consequently, it appears that the role that carbohydrates play in metastasis as cell adhesion molecules, is the underlying the relationship between the expression of carbohydrates and the prognosis of patients with breast cancer.


RNA Capping by Transcription Initiation with Non-canonical Initiating Nucleotides (NCINs): Determination of Relative Efficiencies of Transcription Initiation with NCINs and NTPs.
It recently has been established that adenine-containing cofactors, including nicotinamide adenine dinucleotide (NAD+), reduced nicotinamide adenine dinucleotide (NADH), and 3'-desphospho-coenzyme A (dpCoA), can serve as 'non-canonical initiating nucleotides' (NCINs) for transcription initiation by bacterial and eukaryotic cellular RNA polymerases (RNAPs) and that the efficiency of the reaction is determined by promoter sequence (Bird et al., 2016). Here we describe a protocol to quantify the relative efficiencies of transcription initiation using an NCIN vs. transcription initiation using a nucleoside triphosphate (NTP) for a given promoter sequence.


Enteroparasite contamination in peridomiciliar soils of two indigenous territories, State of Paraná, southern Brazil.
To determine 1) the degree of soil contamination by parasites; 2) the prevalence of contaminated samples; 3) the overall number of parasitic forms, by distance between the collection site and the domicile; 4) the mean number of parasite forms per peri-household area; and 5) the variables associated with the total number of parasite forms. Soil samples were collected in the peridomicile of 18 (out of a total sample of 63) and 22 (out of 190) residences of the Faxinal and Ivaí indigenous territories (ITs), respectively, from March 2005 through October 2006, and evaluated by the methods of Baermann, modified Faust et al., and Lutz. The overall prevalence of samples contaminated by enteroparasites of humans or of animals with zoonotic potential was 75.7% (84 out of 111) in Faxinal and 96.2% (127 out of 132) in Ivaí. The most prevalent parasites were Ascaris spp., Isospora spp., and Toxocara spp. Positive statistical associations were observed between the number of parasitic forms per domicile and the following variables: time of year (season), the presence of bathrooms in the domicile, and the presence of excrement on peridomiciliar soil. The high level of soil contamination in Faxinal and Ivaí 1) constitutes a potential source of parasitic infection and 2) suggests that the treatment of human and animal excreta continues to be inadequate.


Role of attention in the associative relatedness effect in verbal working memory: Behavioral and chronometric perspectives.
Long-term semantic memory (LTM) is known for affecting recall during working memory (WM) tasks. However, the way LTM intervenes in WM remains unknown. Moreover, the available findings are incongruent concerning how attention modulates the impact of LTM on WM. To examine this issue, the involvement of LTM representations in a complex span task was manipulated through variations of the associative relatedness of memory items, while the attentional demand of the concurrent task was varied. Children and young adults were also compared, because children are less efficient in using refreshing for maintenance than adults. Despite the impact of the three main factors on recall performance, which was better for related than unrelated words, with the low rather than the high demanding concurrent task and for adults than children, there was no interaction between associative relatedness and attentional demand, neither in children nor in adults. We replicated these results in a second experiment with a more attention-demanding concurrent task. Moreover, analyses of recall latency showed that adults were faster than children at recalling words and both age groups were faster for related (vs. unrelated) words, but there was no effect of the concurrent attentional demand on recall latency and no interaction. Finally, errors were mostly omissions and transpositions, both more prevalent under high concurrent attentional demand. The present findings suggest that the availability of attention does not modulate the effect of LTM on WM. We discuss how WM models can account for this finding and how LTM can act on WM functioning. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Long-term treatment of severe familial hypercholesterolemia in children: effect of sitosterol and bezafibrate.
Seven prepubertal children (age range 5.3 to 10.8 years) with severe heterozygous familial hypercholesterolemia (serum cholesterol concentration 416 +/- 85 mg/dL and low-density lipoprotein [LDL] cholesterol concentration 360 +/- 90 mg/dL) were first treated by dietary intervention, second by sitosterol (3 x 2 g/d), and third by bezafibrate (2 x 200 mg/d). Each treatment period lasted 3 months. Subsequently, a treatment combining half the dose of sitosterol and bezafibrate was administered for the following 24 months. Diet alone reduced total and LDL cholesterol values by 4.5% (not significant) and 6.6% (P less than .05), respectively. Sitosterol lowered total and LDL cholesterol values by 17% (P less than .05) when compared with diet alone. Compared with sitosterol, bezafibrate produced a more pronounced effect on total and LDL cholesterol values (-18% and -28%, P less than .05), and high-density lipoprotein cholesterol concentration increased significantly from 48 mg/dL to 55 mg/dL. Combined treatment with half the dose each of sitosterol and bezafibrate was as effective as the higher dose of bezafibrate, and reduction averaged almost 40% and 50% for total and LDL cholesterol values; this lipid-lowering effect persisted for the next 24 months. Laboratory safety parameters and physical examination revealed no obvious side effects. This study indicates that the combination of sitosterol (3 x 1 g/d) plus bezafibrate (1 x 200 mg/d) is an alternate, acceptable, safe, and effective therapeutic approach for treatment of severe hypercholesterolemia in children with high-risk familial hypercholesterolemia.


Synthesis and characterization of silver nanoparticles using fruit extract of Momordica cymbalaria and assessment of their in vitro antimicrobial, antioxidant and cytotoxicity activities.
Plant mediated synthesis of nanoparticles has been considered as green route and a reliable technique for the synthesis of nanoparticles due to its eco-friendly approach. In this study, we report a simple and eco-friendly approach for the synthesis of silver nanoparticles (AgNPs) using methanolic Momordica cymbalaria fruit extract as reducing agent. The fruit extract of M. cymbalaria exposed to AgNO3 solution showed the change in color from green to light yellow at room temperature within 1h of incubation confirms the synthesis of AgNPs. UV-vis spectra analysis revealed that the synthesized AgNPs had a sharp surface plasmon resonance at around 450 nm, while, the X-ray Diffraction (XRD) patterns confirmed distinctive peaks indices to the crystalline planes of the face centered cubic silver. The Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM) analysis results confirmed the presence of spherical shaped AgNPs by a huge disparity in the particle size distribution with an average size of 15.5 nm. The synthesized AgNPs showed strong antibacterial activity against all the tested multidrug resistant human pathogenic bacterial strains and also exhibited highest free radical scavenging activity (74.2%) compared to fruit extract (60.4%). Moreover, both fruit extract and the synthesized AgNPs showed the cytotoxicity towards Rat L6 skeletal muscle cell line at different concentrations, but the highest inhibition percentage was recorded for AgNPs at concentration of 100 μg/ml.


A Morphing [4Fe-3S-nO]-Cluster within a Carbon Monoxide Dehydrogenase Scaffold.
Ni,Fe-containing carbon monoxide dehydrogenases (CODHs) catalyze the reversible reduction of CO2 to CO. Several anaerobic microorganisms encode multiple CODHs in their genome, of which some, despite being annotated as CODHs, lack a cysteine of the canonical binding motif for the active site Ni,Fe-cluster. Here, we report on the structure and reactivity of such a deviant enzyme, termed CooS-VCh . Its structure reveals the typical CODH scaffold, but contains an iron-sulfur-oxo hybrid-cluster. Although closely related to true CODHs, CooS-VCh catalyzes neither CO oxidation, nor CO2 reduction. The active site of CooS-VCh undergoes a redox-dependent restructuring between a reduced [4Fe-3S]-cluster and an oxidized [4Fe-2S-S*-2O-2(H2 O)]-cluster. Hydroxylamine, a slow-turnover substrate of CooS-VCh , oxidizes the hybrid-cluster in two structurally distinct steps. Overall, minor changes in CODHs are sufficient to accommodate a Fe/S/O-cluster in place of the Ni,Fe-heterocubane-cluster of CODHs.


Total-Body Perfusion Imaging with [11C]-Butanol.
Tissue perfusion can be affected by physiology or disease. With the advent of total-body PET, quantitative measurement of perfusion across the entire body is possible. [11C]-butanol is a perfusion tracer with a superior extraction fraction compared with [15O]-water and [13N]-ammonia. To develop the methodology for total-body perfusion imaging, a pilot study using [11C]-butanol on the uEXPLORER total-body PET/CT scanner was conducted. Methods: Eight participants (6 healthy volunteers and 2 patients with peripheral vascular disease [PVD]) were injected with a bolus of [11C]-butanol and underwent 30-min dynamic acquisitions. Three healthy volunteers underwent repeat studies at rest (baseline) to assess test-retest reproducibility; 1 volunteer underwent paired rest and cold pressor test (CPT) studies. Changes in perfusion were measured in the paired rest-CPT study. For PVD patients, local changes in perfusion were investigated and correlated with patient medical history. Regional and parametric kinetic analysis methods were developed using a 1-tissue compartment model and leading-edge delay correction. Results: Estimated baseline perfusion values ranged from 0.02 to 1.95 mL·min-1·cm-3 across organs. Test-retest analysis showed that repeat baseline perfusion measurements were highly correlated (slope, 0.99; Pearson r = 0.96, P < 0.001). For the CPT subject, the largest regional increases were in skeletal muscle (psoas, 142%) and the myocardium (64%). One of the PVD patients showed increased collateral vessel growth in the calf because of a peripheral stenosis. Comorbidities including myocardial infarction, hypothyroidism, and renal failure were correlated with variations in organ-specific perfusion. Conclusion: This pilot study demonstrates the ability to obtain reproducible measurements of total-body perfusion using [11C]-butanol. The methods are sensitive to local perturbations in flow because of physiologic stressors and disease.


Clinical, etiologic, and histopathologic features of Stevens-Johnson syndrome during an 8-year period at Mayo Clinic.
To examine clinical, etiologic, and histologic features of Stevens-Johnson syndrome and to identify possible correlates of clinical disease severity related to etiologic and histopathologic findings. This is a retrospective review of patients seen at Mayo Clinic between January 1, 2000, and December 31, 2007. Of 27 patients (mean age, 28.1 years), 22 (81%) had involvement of 2 or more mucous membranes, and 19 (70%) had ocular involvement. Medications, most commonly antibiotics and anticonvulsants, were causative in 20 patients. Mycoplasma pneumoniae infection caused 6 of the 27 cases. Corticosteroids were the most common systemic therapy. No patients with mycoplasma-induced Stevens-Johnson syndrome had internal organ involvement or required treatment in the intensive care unit, in contrast to 4 patients each in the drug-induced group. Three patients had chronic ocular sequelae, and 1 died of complications. Biopsy specimens from 13 patients (48%) showed epidermal necrosis (8 patients), basal vacuolar change (10 patients), and subepidermal bullae (10 patients). Biopsy specimens from 11 patients displayed moderate or dense dermal infiltrate. Histologic features in drug-induced cases included individual necrotic keratinocytes, dense dermal infiltrate, red blood cell extravasation, pigment incontinence, parakeratosis, and substantial eosinophils or neutrophils. Our clinical and etiologic findings corroborate those in previous reports. M pneumoniae-induced Stevens-Johnson syndrome manifested less severely than its drug-induced counterpart. The limited number of biopsies precludes unequivocal demonstration of histopathologic differences between drug-induced and M pneumoniae-induced Stevens-Johnson syndrome.


Regulation of ompF porin expression by salicylate in Escherichia coli.
The expression of ompF, the gene encoding a major outer membrane protein of Escherichia coli, is regulated by various environmental factors. The mechanism by which salicylate (SAL) drastically reduces ompF expression was studied here by means of lacZ fusions to ompF, ompC, and micF, by sodium dodecyl sulfate-gel electrophoresis of outer membrane proteins, and by measurements of outer membrane permeability. Growth of E. coli in LB broth containing SAL strongly reduced ompF-specific translation of an ompF-lacZ fusion. The extent of this reduction varied with the SAL concentration from 64% at 0.5 mM to 95% at 2 mM and greater than 99% at 10 mM. ompF-lacZ transcription was not affected by SAL, whereas ompC-lacZ transcription was elevated by 70%. Since the micF transcript is antisense to a portion of the ompF transcript and is capable of decreasing the translation of ompF, the effect of SAL on micF transcription was measured in a micF-lacZ fusion strain. SAL-grown cells contained three- to fourfold more micF transcript during the logarithmic phase of growth than did the control cultures. However, micF was not absolutely required for the response to SAL. In micF-deleted strains, the effects of SAL on ompF translation, on OmpF in the outer membrane, and on outer membrane permeability were diminished but still evident. The effect of SAL on ompF expression was independent of the osmolarity of the medium and was epistatic to certain ompB regulatory mutations: the high levels of ompF expression found in envZ3 and ompR472 strains were greatly reduced by growth in SAL. Unexpectedly, the OmpC- phenotypes of these mutants were suppressed by SAL. Thus, growth in SAL severely decreases the translation of ompF while enhancing the transcription of micF and ompC. In this respect, SAL-grown cells resemble certain marA and tolC mutants that have high levels of micF and ompC transcripts and low levels of OmpF.


Electrical-thermal analytical modeling of monopolar RF thermal ablation of biological tissues: determining the circumstances under which tissue temperature reaches a steady state.
It has been suggested that during RF thermal ablation of biological tissue the thermal lesion could reach an equilibrium size after 1-2 minutes. Our objective was to determine under which circumstances of electrode geometry (needle-like vs. ball-tip), electrode type (dry vs. cooled) and blood perfusion the temperature will reach a steady state at any point in the tissue. We solved the bioheat equation analytically both in cylindrical and spherical coordinates and the resultant limit temperatures were compared. Our results demonstrate mathematically that tissue temperature reaches a steady value in all cases except for cylindrical coordinates without the blood perfusion term, both for dry and cooled electrodes, where temperature increases infinitely. This result is only true when the boundary condition far from the active electrode is considered to be at infinitum. In contrast, when a finite and sufficiently large domain is considered, temperature reaches always a steady state.


Lead poisoning from an Ayurvedic herbal medicine in a patient with chronic kidney disease.
A 60-year-old man with a history of diabetes and hypertension was referred to a nephrology clinic for investigation of his elevated serum creatinine level. Physical examination; laboratory investigations, including measurement of whole-blood lead level, body lead burden and urine albumin:creatinine ratio; history of lead exposure and use of herbal medical products; and renal ultrasonography. Stage 3 chronic kidney disease that was probably worsened by consumption of lead in the form of an Ayurvedic herbal remedy. Cessation of the herbal product, followed by lead-chelation therapy with calcium disodium ethylenediaminetetraacetic acid. The patient's whole-body lead burden and blood lead level decreased to acceptable levels and his serum creatinine value was within the normal range at final follow-up.


Gut microbiota-derived metabolites are novel targets for improving insulin resistance.
The gut microbiota plays a key role in metabolic diseases. Gut-microbiota-derived metabolites are found in different dietary sources, including: Carbohydrate (acetate, propionate, butyrate, also known as short-chain fatty acids, as well as succinate); protein (hydrogen sulfide, indole, and phenylacetic acid); and lipids (resveratrol-, ferulic acid-, linoleic acid-, catechin- and berry-derived metabolites). Insulin resistance, which is a global pandemic metabolic disease that progresses to type 2 diabetes mellitus, can be directly targeted by these metabolites. Gut-microbiota-derived metabolites have broad effects locally and in distinct organs, in particular skeletal muscle, adipose tissue, and liver. These metabolites can modulate glucose metabolism, including the increase in glucose uptake and lipid oxidation in skeletal muscle, and decrease in lipogenesis and gluconeogenesis associated with lipid oxidation in the liver through activation of phosphatidylinositol 3-kinase - serine/threonine-protein kinase B and AMP-activated protein kinase. In adipose tissue, gut-microbiota-derived metabolites stimulate adipogenesis and thermogenesis, inhibit lipolysis, and attenuate inflammation. Importantly, an increase in energy expenditure and fat oxidation occurs in the whole body. Therefore, the therapeutic potential of current pharmacological and non-pharmacological approaches used to treat diabetes mellitus can be tested to target specific metabolites derived from intestinal bacteria, which may ultimately ameliorate the hyperglycemic burden.


The effect of most important medicinal plants on two importnt psychiatric disorders (anxiety and depression)-a review.
Anxiety and depression are highly comorbid psychiatric conditions that the prevalence will be increased to the second greatest risk of morbidity, causing a significant socioeconomic burden. Due to side effects and destructive effects of some chemical drugs, many patients prefer herbal medicines to treat diseases. Although there are key review papers in the area of medicinal plants and psychiatry disorders, they have either covered the area in a relatively cursory manner or focused on a specific plant medicine. In the present study, we tried to present the effect of most important medicinal plants on two important highly comorbid psychiatric conditions-anxiety and depression.


Historical analysis of genetic variation reveals low effective population size in a northern pike (Esox lucius) population.
Effective population size (Ne) of a natural fish population was estimated from temporal changes in allele frequencies at seven microsatellite loci. Use of a historical collection of fish scales made it possible to increase the precision of estimates by increasing the time interval between samples and to use an equation developed for discrete generations without correcting for demographic parameters. Estimates of Ne for the time intervals 1961-1977 and 1977-1993 were 35 and 72, respectively. For the entire interval, 1961-1993, the estimate of Ne was 48 when based on a weighted mean derived from the above two estimates or 125 when calculated from 1961 and 1993 samples only. Corresponding ratios of effective size to adult census size ranged from 0.03 to 0.14. An Ne of 48 over a 32-year period would imply that this population lost as much as 8% of its heterozygosity in that time. Results suggest the potential for using genetic methods based on microsatellite loci data to compare historical trends in Ne with population dynamic parameters. Such comparisons will help to evaluate the relationship between genetic diversity and long-term persistence of natural populations.


Engineering Crystal Packing in RNA-Protein Complexes II: A Historical Perspective from the Structural Studies of the Spliceosome.
Cryo-electron microscopy has greatly advanced our understanding of how the spliceosome cycles through different conformational states to conduct the chemical reactions that remove introns from pre-mRNA transcripts. The Cryo-EM structures were built upon decades of crystallographic studies of various spliceosomal RNA-protein complexes. In this review we give an overview of the crystal structures solved in the Nagai group, utilizing many of the strategies to design crystal packing as described in the accompanying paper.


Apically Extruded Debris following Programmed Over Instrumentation of Curved Canals with Three Nickel Titanium Rotary Instruments.
This in vitro study aimed to compare the amount of mixed and dried debris extruded with three endodontic NiTi (nickel-titanium) systems at three levels beyond the major apical foramen. The null hypothesis is that there would be no differences between the different rotary systems in terms of apically extruded debris. Forty-eight roots of human extracted molars with 20 to 40-degrees curvature were divided randomly into three groups (n = 16) instrumented with ProTaper Next, BT RaCe, and WaveOne Gold, respectively. Instrumentation was performed at the major foramen and over instrumented 0.5 mm, 1.0 mm and 1.5 mm beyond. Irrigation was performed with sodium hypochlorite. Apically extruded debris was collected. Comparison of mixed extruded debris (solution/debris)and dried debris according to group and level of instrumentation showed that WaveOne Gold at the foramen and ProTaper Next beyond the foramen (p < 0.0001) resulted in significantly less extruded debris than other files and within the four levels using repeated measures analysis of variance (p < 0.0001b). All systems exhibit extruded debris during instrumentation but in different amounts.


[A Case of Renocolic Fistula Caused by Infected Renal Cyst].
A 76-year-old male was previously found to have a renal cyst at the center of the right kidney, on a computed tomography (CT)scan for examination of another disease. The patient was admitted to the hospital because of fever. The CT scan showed an enlarged mass at the center of the right kidney and an increase in the density of peripheral fat tissue, suggesting an infection of the right renal cyst. In spite of conservative treatment with antibiotics, CT scan on the sixth day of admission revealed an increase in the size of the mass, and penetration in the ascending colon was suspected. An ultrasound-guided abscess puncture was performed, and a pigtail catheter (PC)was placed. Injection of contrast agent through the PC showed communication with the colon. The fistula site was identified using colon fiberscopy, and it was clipped. PC was removed after the closure of the fistula was confirmed by imaging. This is a rare case of renocolic fistula caused by an infected renal cyst, which was diagnosed by colon fiberscopy, and was treated by clipping the fistula.


IR spectra of lens crystallins.
The IR technique has been applied to investigate secondary structure of the crystallins from the normal bovine eye. Crystallins have been isolated by column chromatography. IR spectra were recorded for the solid phase of proteins. From these spectra, especially amide I, amide II and amide V bands, the presence of alpha-helix, beta-sheet, beta-chain and unordered structures is stated. It was elucidated that alpha-crystallins are present mainly in a beta-sheet conformation but they also contain a considerable quantity of alpha-helix and a slight quantity of unordered and beta-chain forms. In beta H-crystallins, alpha-helix and, in a lesser percentage, beta-structures predominante. beta-sheet, alpha-helix and a low content of beta-chain forms are present in beta L-crystallins. In gamma-crystallins all forms secondary structure have been found, with predominance of beta-sheet and alpha-helix forms. A satisfactory agreement has been noticed between the forms of secondary structures in crystallins investigated by the IR technique and the results obtained by means of other methods. In conclusion IR spectroscopy has been suggested to be applied to observe crystallin structure during formation and development of a cataract.


Differential IgE reactivity to Der p 1 and Der p 2 allergens of Dermatophagoides pteronyssinus in mite-sensitized patients.
Several studies have shown that the presence of IgE antibodies to house dust mites (HDM), particularly Dermatophagoides pteronyssinus (Dpt), is an important risk factor for asthma. Allergen immunotherapy is indicated for patients with IgE antibodies to clinically relevant allergens. The aims of this study were to analyze the levels of specific serum IgE to Der p 1 and Der p 2 allergens in mite-sensitized atopic patients and to compare them with both in vivo (skin prick test) and in vitro (IgE-ELISA) sensitizations to Dpt crude extract. Forty-seven atopic patients with allergic rhinitis with or without intermittent or persistent mild asthma and positive skin prick test (SPT) to Dpt total extract were studied. Thirty age-matched healthy subjects with negative SPT to HDM were included as controls. Levels of total IgE and Dpt-, Der p 1- and Der p 2-specific IgE were measured by ELISAs in SPT-positive atopic patients and SPT-negative control subjects. Among 47 symptomatic atopic patients, 27 (57.4%) were double positive IgE to Der p 1 and Der p 2 allergens, 3 (6.4%) were single positive IgE to Der p 1, 4 (8.5%) were single positive IgE to Der p 2, and 13 (27.6%) were double negative IgE to both allergens. There was a significant correlation between Der p 1- and Der p 2-specific IgE levels, but not between Der p 1- or Der p 2-IgE levels and SPT results. The double negative IgE patients had the smallest skin test reactions although they showed high mean levels of total serum IgE. Therefore, the knowledge of specific IgE levels to Der p 1 and Der p 2 major allergens might support physicians for indication or follow-up in mite-sensitized patients under allergen-specific immunotherapy. These approaches might be important for obtaining improved safety and efficacy of the current clinical practice of allergen immunotherapy.


Disruption of integrin-fibronectin complexes by allosteric but not ligand-mimetic inhibitors.
Failure of Arg-Gly-Asp (RGD)-based inhibitors to reverse integrin-ligand binding has been reported, but the prevalence of this phenomenon among integrin heterodimers is currently unknown. In the present study we have investigated the interaction of four different RGD-binding integrins (α5β1, αVβ1, αVβ3 and αVβ6) with fibronectin (FN) using surface plasmon resonance. The ability of inhibitors to reverse ligand binding was assessed by their capacity to increase the dissociation rate of pre-formed integrin-FN complexes. For all four receptors we showed that RGD-based inhibitors (such as cilengitide) were completely unable to increase the dissociation rate. Formation of the non-reversible state occurred very rapidly and did not rely on the time-dependent formation of a high-affinity state of the integrin, or the integrin leg regions. In contrast with RGD-based inhibitors, Ca2+ (but not Mg2+) was able to greatly increase the dissociation rate of integrin-FN complexes, with a half-maximal response at ~0.4 mM Ca2+ for αVβ3-FN. The effect of Ca2+ was overcome by co-addition of Mn2+, but not Mg2+. A stimulatory anti-β1 monoclonal antibody (mAb) abrogated the effect of Ca2+ on α5β1-FN complexes; conversely, a function-blocking mAb mimicked the effect of Ca2+. These results imply that Ca2+ acts allosterically, probably through binding to the adjacent metal-ion-dependent adhesion site (ADMIDAS), and that the α1 helix in the β subunit I domain is the key element affected by allosteric modulators. The data suggest an explanation for the limited clinical efficacy of RGD-based integrin antagonists, and we propose that allosteric antagonists could prove to be of greater therapeutic benefit.


What Kind of Signaling Maintains Pluripotency and Viability in Human-Induced Pluripotent Stem Cells Cultured on Laminin-511 with Serum-Free Medium?
Xeno-free medium contains no animal-derived components, but is composed of minimal growth factors and is serum free; the medium may be supplemented with insulin, transferrin, and selenium (ITS medium). Serum-free and xeno-free culture of human-induced pluripotent stem cells (hiPSCs) uses a variety of components based on ITS medium and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 (DMEM/F12) that contain high levels of iron salt and glucose. Culture of hiPSCs also requires scaffolding materials, such as extracellular matrix, collagen, fibronectin, laminin, proteoglycan, and vitronectin. The scaffolding component laminin-511, which is composed of α5, β1, and γ1 chains, binds to α3β1, α6β1, and α6β4 integrins on the cell membrane to induce activation of the PI3K/AKT- and Ras/MAPK-dependent signaling pathways. In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway. The expression levels of α6β1 integrin and E-cadherin on cell membranes are controlled through the activation of insulin receptor/insulin, FGF receptor/FGF2, or activin-like kinase 5 (ALK5)-dependent TGF-β signaling. A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold. In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies. In addition, we suggest the optimum serum-free medium components for use with laminin-511, a new scaffold for hiPSC culture.


The sagittal profile of the cervical and lumbosacral spine in Scheuermann thoracic kyphosis.
The sagittal profiles of the cervical and lumbar spine have not been studied in Scheuermann kyphosis. The purpose of this study was to investigate these profiles. Standing lateral radiographs of the spine in 34 children with Scheuermann kyphosis were reviewed. Cervical lordosis, lumbar lordosis, thoracic kyphosis, sagittal vertebral axis, and sacral inclination were measured. The relations between these variables were explored using the Pearson correlation. The average patient age was 15.5 +/- 1.8 years, thoracic kyphosis was 65 degrees +/- 12 degrees, lumbar lordosis 71 degrees +/- 13 degrees, and cervical lordosis 4 degrees +/- 15 degrees (Cobb angle), and 9 degrees +/- 14 degrees (posterior vertebral body angle [PVBA]). No correlations were noted between cervical lordosis and thoracic kyphosis. Correlations were noted between cervical lordosis and lumbar lordosis (r2 = 0.17, Cobb angle; r2 = 0.16, PVBA) and between cervical lordosis and the residual sagittal difference (thoracic kyphosis minus lumbar lordosis; r2 = 0.32, p = 0.001 [Cobb angle], and r2 = 0.19, p = 0.01 [PVBA]). In Scheuermann kyphosis, the flexible cervical and lumbar spine is linked by the intermediate rigid thoracic segment. As the residual sagittal difference becomes more kyphotic, lordosis of the cervical spine increases as the patient strives to maintain a forward visual gaze.


Parkinson's disease alters the composition of subgingival microbiome.
The current study aimed to test the hypothesis that Parkinson's disease exacerbates periodontitis by altering its microbiome. Clinical periodontal parameters were recorded. Subgingival samples from healthy controls, periodontitis patients (PD), and Parkinson's patients with periodontitis (PA+PD) were analyzed using the checkerboard DNA-DNA hybridization technique for targeting 40 bacterial species typically associated with periodontal disease and health. Next-generation sequencing (NGS) of the 16S ribosomal RNA gene (V1-V3 regions) was performed to analyze the microbiome comprehensively. Parkinson's patients had mild-to-moderate motor dysfunctions. Bleeding on probing was significantly increased in the PA+PD group compared to PD (p < 0.05). With checkerboard analysis, PA was associated with increased Treponema socranskii (p = 0.0062), Peptostreptococcaceae_[G-6] [Eubacterium]_nodatum (p = 0.0439), Parvimona micra (p < 0.0001), Prevotella melaninogenica (p = 0.0002), Lachnoanaerobaculum saburreum (p < 0.0001), and Streptococcus anginosus (p = 0.0020). Streptococcus intermedia (p = 0.0042), P.nodatum (p = 0.0022), P. micra (p = 0.0002), Treponema denticola (p = 0.0045), L.saburreum (p = 0.0267), P.melaninogenica (p = 0.0017), Campylobacter rectus (p = 0.0020), and T.socranskii (p = 0.0002) were higher; Aggregatibacter actinomycetemcomitans (p = 0.0072) was lower in deep pockets in the PA+PD compared to PD. Schaalia odontolytica (p = 0.0351) and A.actinomycetemcomitans (p = 0.002) were lower; C.rectus (p = 0.0002), P. micra (p = 0065), Streptococcus constellatus (p = 0.0151), T.denticola (p = 0.0141), P.melaninogenica (p = 0.0057), and T.socranskii (p = 0.0316) were higher in shallow pockets in the PA+PD. Diversity decreased in PD (p = 0.001) and PA+PD (p = 0.026) compared to control, with minimal differences in alpha and beta diversities among PD and PA+PD based on NGS results. These data demonstrated that Parkinson's disease modifies PD-associated subgingival microbiome.


Severe forms of status epilepticus.
Severe forms of status epilepticus needing ventilation with or without the need for a general anaesthetic were studied among the children admitted to the paediatric department at Assir Central Hospital, Saudi Arabia. From November 1988 to April 1990, 255 children were admitted because of fits, of whom 12 (4.7%) developed severe status epilepticus and required admission to the intensive care unit for ventilation. This group was studied in detail with regard to past history, course in the hospital and outcome. Five children recovered fully, four were left with neurological disability, and three died. The need to ventilate a patient or give a general anaesthetic for status epilepticus rarely arises in ordinary clinical practice. This study demonstrates the importance of these measures to reduce the risks of permanent neurological deficit and, possibly, death in severe status epilepticus.


VdThit, a Thiamine Transport Protein, Is Required for Pathogenicity of the Vascular Pathogen Verticillium dahliae.
Verticillium dahliae causes a serious wilt disease of important crops and is difficult to control. Few plasma-membrane transport proteins for nutrient acquisition have been identified for this fungus, and their involvement in the disease process is unknown. Here, a plasma-membrane protein, the V. dahliae thiamine transporter protein VdThit, was characterized functionally by deletion of the VdThit gene in V. dahliae. Disruption strains were viable, but growth and conidial germination and production were reduced and virulence was impaired. Interestingly, by supplementing exogenous thiamine, growth, conidiation, and virulence of the VdΔThit mutants were partially restored. Stress-tolerance assays showed that the VdΔThit mutant strains were markedly more susceptible to oxidative stress and UV damage. High-pressure liquid chromatography-mass spectrometry (HPLC-MS) and gas chromatography-mass spectrometry (GC-MS) analyses showed low levels of pyruvate metabolism intermediates acetoin and acetyl coenzyme A (acetyl-CoA) in the VdΔThit mutant strains, suggesting that pyruvate metabolism was suppressed. Expression analysis of VdThit confirmed the importance of VdThit in vegetative growth, reproduction, and invasive hyphal growth. Furthermore, a green fluorescent protein (GFP)-labeled VdΔThit mutant (VdΔThit-7-GFP) was suppressed in initial infection and root colonization, as viewed with light microscopy. Together, these results showed that VdThit plays an indispensable role in the pathogenicity of V. dahliae.


Using databases.
Through a relatively simple computer technique known as online searching, practitioners can use computers to "reach out" beyond their practice walls to expand their knowledge base. By bringing information sources closer to veterinarians, feelings of isolation, frustration, and ineptitude can be replaced with feelings of certainty, confidence, and knowledge. This article explores the equipment and the software needed, and by giving examples of a few of the veterinary databases, illustrates some techniques employed in signing up and signing on to the power of online searching.


Prevalence and barriers to HIV testing among mothers at a tertiary care hospital in Phnom Penh, Cambodia. Barriers to HIV testing in Phnom Penh, Cambodia.
One-third of all new HIV infections in Cambodia are estimated to be due to mother-to-child transmission. Although the Ministry of Health adopted a policy of provider-initiated HIV testing and counseling (PITC), nearly a quarter of pregnant mothers were not tested in 2007. Greater acceptance of HIV testing is a challenge despite Cambodia's adoption of the PITC policy. A hospital-based quantitative and cross-sectional survey was conducted to assess the prevalence of and barriers to HIV testing among mothers after delivery at the National Maternal and Child Health Center in Phnom Penh. The Center is one of the largest maternal and child care hospitals in the country to offer PITC services. All 600 eligible mothers who were admitted to the hospital after delivery from October to December 2007 were approached and recruited. Data were collected via a semi-structured questionnaire. The prevalence of HIV testing among women who delivered at the hospital was 76%. In multivariate logistic regression, factors such as the perceived need to obtain a partner's permission to be tested (OR=0.27, 95% CI=0.14-0.51, p<0.01), the lack of knowledge about HIV prevention and treatment (OR=0.38, CI=0.22-0.66, p<0.01), and the lack of access to ANC services (OR=0.35, 95% CI=0.21-0.58, p<0.01) were found to be the main barriers to HIV testing. To achieve greater acceptance of HIV testing, counseling on HIV prevention and treatment must be provided not only to mothers but also to their partners. In addition, utilization of non-laboratory staff such as midwives to provide HIV testing services in rural health facilities could lead to the greater acceptance of HIV testing.


Hormone- and light-regulated nucleocytoplasmic transport in plants: current status.
The gene regulation mechanisms underlying hormone- and light-induced signal transduction in plants rely not only on post-translational modification and protein degradation, but also on selective inclusion and exclusion of proteins from the nucleus. For example, plant cells treated with light or hormones actively transport many signalling regulatory proteins, transcription factors, and even photoreceptors and hormone receptors into the nucleus, while actively excluding other proteins. The nuclear envelope (NE) is the physical and functional barrier that mediates this selective partitioning, and nuclear transport regulators transduce hormone- or light-initiated signalling pathways across the membrane to mediate nuclear activities. Recent reports revealed that mutating the proteins regulating nuclear transport through the pores, such as nucleoporins, alters the plant's response to a stimulus. In this review, recent works are introduced that have revealed the importance of regulated nucleocytoplasmic partitioning. These important findings deepen our understanding about how co-ordinated plant hormone and light signal transduction pathways facilitate communication between the cytoplasm and the nucleus. The roles of nucleoporin components within the nuclear pore complex (NPC) are also emphasized, as well as nuclear transport cargo, such as Ran/TC4 and its binding proteins (RanBPs), in this process. Recent findings concerning these proteins may provide a possible direction by which to characterize the regulatory potential of hormone- or light-triggered nuclear transport.


Heavy atom nitroxyl radicals. IV. Experimental and theoretical studies of the F(2)P=S free radical in the gas phase.
The difluorothiophosphoryl (F(2)PS) free radical has been produced in a supersonic discharge jet from a precursor mixture of F(3)PS and high pressure argon and detected by laser-induced fluorescence and single vibronic level emission spectroscopy. With the aid of high level ab initio predictions of the properties of the ground and first two excited doublet states, the observed band system has been positively identified as B(2)A(')-X(2)A('). The electronic transition involves promotion of an electron from the pi to the pi( *) orbital with concomitant increases in the out-of-plane angle and PS bond length on excitation. The observed vibrational structure, Franck-Condon profile, rotational band contours, T(0) energy, and ground and excited state vibrational frequencies are all in accord with expectations based on our theoretical predictions.


Healthcare utilisation and costs associated with adherence to antipsychotics among people living with HIV/AIDS and schizophrenia: a population-based cohort study in British Columbia, Canada.
Non-adherence to antipsychotics is the greatest obstacle to treating schizophrenia. We assessed the economic and clinical impacts of adherence to antipsychotics among people living with HIV/AIDS (PLWH) and schizophrenia in British Columbia, Canada. A population-based cohort study in British Columbia, Canada. Eligible PLWH were enrolled in the Seek and Treat for Optimal Prevention HIV/AIDS population-based cohort during 2001-2016, diagnosed with schizophrenia, on antipsychotics for ≥1 day, and followed for ≥1 year from schizophrenia diagnosis date or 1 January 2001, whichever occurred last. A two-part model assessed the marginal effect of adherence on healthcare costs (in 2016 Canadian dollar), while logistic regression examined the effect on virological failure, and generalised linear mixed models examined the effect on hospital readmissions within 30 days and length of hospital stay. Among 726 PLWH with schizophrenia, ≥80% adherence to antipsychotics increased from 25% (50/198) in 2001 to 41% (225/554) in 2016. In most years, we observed no difference in adherence to antipsychotics among those who used only injectables, only non-injectables, and a combination of both, or among those who have ever consumed typical/first-generation antipsychotics and who consumed only atypical/second-generation antipsychotics. Overall healthcare costs were higher in the non-adherent group ($C2185), driven by the average annual hospitalisation costs ($C5517), particularly among women ($C8806) and people who ever injected drugs (PWID) ($C5985). Non-adherent individuals also experienced higher hospital readmissions (adjusted odds ratio (aOR) 1.48, 95% CI 1.23 to 1.77), and longer hospital stays (adjusted mean ratio 1.23, 95% CI 1.13 to 1.35) in comparison to adherent individuals. We found no difference in virological failure by adherence groups, except when we stratified by gender where the aOR for women was 2.48 (95% CI 1.06 to 5.82). Our results showed that implementing strategies and interventions to increase antipsychotic adherence, particularly among women and PWID, will be critical in addressing this public health challenge.


Complexity of tumor vasculature in clear cell renal cell carcinoma.
Clear cell renal cell carcinoma (CCRCC) is a highly vascularized cancer resistant to conventional chemotherapy and radiotherapy. Antiangiogenic therapy has achieved some effectiveness against this unique malignancy. The complexity of the tumor vasculature in CCRCC has led to differences in correlating tumor microvessel density with patient prognosis. The authors' recent findings demonstrated that there were at least 2 major categories of tumor vessels in CCRCC-namely, undifferentiated and differentiated-correlating with patient prognosis in contrasting ways, with higher undifferentiated vessel density indicating poorer prognosis, and higher differentiated vessel density correlating with better prognosis. Furthermore, the presence of pericytes supporting the differentiated vessels varied in CCRCC. The distributions of pericyte coverage and differentiated vessels in CCRCC were uneven. The tumor margin had a higher pericyte coverage rate for differentiated vessels than did the inner tumor area. The uneven distributions of pericyte coverage and differentiated vessels in CCRCC prompted the authors to revisit the mechanism of tumor central necrosis, which was also known to be a prognostic indicator for CCRCC. The discrepancy of prognostic correlation between protein and messenger RNA levels of vascular endothelial growth factor in CCRCC was discussed. The complexity of the tumor vasculature in CCRCC also led the authors to begin to re-evaluate the therapeutic effects of antiangiogenic agents for each type of tumor vessel, which will in turn significantly broaden understanding of tumor angiogenesis and improve therapeutic effect.


Association between Cystatin C and MRI Measures of Left Ventricular Structure and Function: Multi-Ethnic Study of Atherosclerosis.
Introduction. Reduced kidney function, approximated by elevated cystatin C, is associated with diastolic dysfunction, heart failure, and cardiovascular mortality; however, the precise mechanism(s) that account for these relationships remains unclear. Understanding the relationship between cystatin C and subclinical left ventricular (LV) remodeling, across ethnically diverse populations, may help explain the mechanisms underlying the association of kidney dysfunction with heart failure and cardiovascular mortality. Methods. Measures of cystatin C and LV parameters were obtained from the multi-ethnic study of atherosclerosis (MESA) cohort at baseline (N = 4, 970 with complete data on cystatin C and LV parameters). LV parameters; LV end-diastolic (LVEDV) and end-systolic volumes (LVESV), LV mass (LVM), concentricity (LV mass/LV end-diastolic volume), and LV ejection fraction (LVEF) were measured using magnetic resonance imaging. Nested linear models were used to examine the relationship between higher quartiles of cystatin C and LV parameters, with and without adjustment for demographics, height, and weight, and traditional cardiovascular risk factors. Similar analyses were performed stratified by ethnicity and gender. Results. A fully adjusted model demonstrated a linear relationship between higher quartiles of cystatin C and lower LVEDV, (Mean ± SE, 128 ± 0.7, 128 ± 0.7, 126 ± 0.7, 124 ± 0.8 mL; P = 0.0001). Associations were also observed between higher quartiles of cystatin C and lower LVESV (P = 0.04) and concentricity (P = 0.0001). In contrast, no association was detected between cystatin C and LVM or LVEF. In analyses stratified by race and gender, the patterns of association between cystatin C quartiles and LV parameters were qualitatively similar to the overall association. Conclusion. Cystatin C levels were inversely associated with LVEDV and LVESV with a disproportionate decrease in LVEDV compared to LVM in a multi-ethnic population. This morphometric pattern of concentric left ventricular remodeling, may in part explain the process by which kidney dysfunction leads to diastolic dysfunction, heart failure and cardiovascular mortality.


Self-assembling peptide-based nanoparticles enhance cellular delivery of the hydrophobic anticancer drug ellipticine through caveolae-dependent endocytosis.
A special class of self-assembling peptide (EAK16-II) has been found to stabilize the hydrophobic anticancer agent ellipticine (EPT) in aqueous solution. In this study, the mechanism of such peptide-EPT complexes to enhance cellular delivery and anticancer activity was evaluated. Results revealed that EAK16-II can form nanoparticles with EPT, having an average size of ∼100 nm. This nanoformulation had cytotoxicity to human lung carcinoma A549 cells that was comparable to EPT dissolved in dimethyl sulfoxide. It enhanced EPT uptake drastically when compared to the microformulation. Such enhanced uptake was significantly reduced by inhibitors specifically for the caveolae-dependent pathway. We also found both protonated and neutral forms of EPT present in the cells. Interestingly, both were found in the cytoplasm, co-localized with LysoTracker, whereas only protonated EPT was seen in the nucleus. The promising therapeutic efficacy, specific delivery pathway, and intracellular distribution pattern discovered in this work may help further develop EPT as a nanoformulation for clinical applications. A special class of self-assembling peptide (EAK16-II) has been found to stabilize ellipticine in aqueous solution. The authors demonstrate therapeutic efficacy, describe specific delivery pathways, and effective intracellular distribution pattern, which will aid the development of this technology for future clinical applications.


Thromboelastographic Evaluation of Dogs with Acute Liver Disease.
Given the liver's pivotal role in hemostasis and fibrinolysis, the coagulopathy accompanying hepatic disease is complex. To prospectively evaluate kaolin-activated thromboelastography (TEG) in dogs with acute liver disease (ALD) and compare with plasma-based coagulation tests. Twenty-one dogs with a diagnosis of ALD based on recent onset of clinical signs accompanied by increases in serum bilirubin concentration and alanine aminotransferase activity. Clinical presentation, CBC, serum biochemistry, platelet count, prothrombin time (PT), activated partial thromboplastin time (aPTT), and TEG analysis were evaluated in 21 dogs with a subset also having fibrinogen, antithrombin (AT) activity, protein C (PC) activity, d-dimers, and von Willebrand's factor (vWF) activity analyzed. A PT >1.5 times the upper limit of normal defined acute liver failure (ALF). Dogs with ALD had mean increases in R, K, LY30, PT, aPTT, and vWF activity, and decreases in angle, maximal amplitude (MA), G, AT activity, and PC activity. The TEG results defined dogs as hypocoagulable (11/21), normocoagulable (8/21), or hypercoagulable (2/21). Increases in LY30 defined 8/21 dogs as hyperfibrinolytic. Hypocoagulable and hyperfibrinolytic dogs had lower fibrinogen and PC activity than dogs without these abnormalities. Overall, ALF dogs had greater increases in K and LY30, and decreases in MA, G, and PC activity than dogs with less severe hepatic impairment. Results for MA and LY30 were positively correlated with serum bilirubin concentration and white blood cell count, and negatively correlated with serum cholesterol concentration. ALD dogs have a range of coagulation abnormalities that trend toward hypocoagulability and hyperfibrinolysis as functional impairment occurs.


Contraceptive failure in the United States: a critical review of the literature.
The overall goal of this paper is to provide for the first time a comprehensive critical review of the literature on contraceptive failure in developed countries, primarily the United States. The first two sections of our paper lay the groundwork for a critical assessment of the extensive body of studies on this subject, by systematically exploring the concepts and measurement of contraceptive efficacy and the methodological pitfalls that snare many investigators and compromise their results. The next two sections focus on results in the literature. First we provide a method-by-method critique of the available studies and then we summarize our conclusions in a single table that provides efficacy information necessary for women and couples to make an informed choice of a method of contraception. We close with a set of substantive observations and also a set of methodological recommendations intended to improve the quality and comparability of findings from future research.


Coupled electron-nuclear dynamics: charge migration and charge transfer initiated near a conical intersection.
Coupled electron-nuclear dynamics, implemented using the Ehrenfest method, has been used to study charge migration with fixed nuclei, together with charge transfer when nuclei are allowed to move. Simulations were initiated at reference geometries of neutral benzene and 2-phenylethylamine (PEA), and at geometries close to potential energy surface crossings in the cations. Cationic eigenstates, and the so-called sudden approximation, involving removal of an electron from a correlated ground-state wavefunction for the neutral species, were used as initial conditions. Charge migration without coupled nuclear motion could be observed if the Ehrenfest simulation, using the sudden approximation, was started near a conical intersection where the states were both strongly coupled and quasi-degenerate. Further, the main features associated with charge migration were still recognizable when the nuclear motion was allowed to couple. In the benzene radical cation, starting from the reference neutral geometry with the sudden approximation, one could observe sub-femtosecond charge migration with a small amplitude, which results from weak interaction with higher electronic states. However, we were able to engineer large amplitude charge migration, with a period between 10 and 100 fs, corresponding to oscillation of the electronic structure between the quinoid and anti-quinoid cationic electronic configurations, by distorting the geometry along the derivative coupling vector from the D6h Jahn-Teller crossing to lower symmetry where the states are not degenerate. When the nuclear motion becomes coupled, the period changes only slightly. In PEA, in an Ehrenfest trajectory starting from the D2 eigenstate and reference geometry, a partial charge transfer occurs after about 12 fs near the first crossing between D1, D2 (N(+)-Phenyl, N-Phenyl(+)). If the Ehrenfest propagation is started near this point, using the sudden approximation without coupled nuclear motion, one observes an oscillation of the spin density--charge migration--between the N atom and the phenyl ring with a period of 4 fs. When the nuclear motion becomes coupled, this oscillation persists in a damped form, followed by an effective charge transfer after 30 fs.


Association of class II human histocompatibility leukocyte antigens with rheumatic fever.
The association of class I and II HLA antigens with rheumatic fever and its manifestations was examined in 72 patients, including 48 blacks and 24 Caucasians. No significant association was found between class I antigens and rheumatic fever. In contrast, HLA-DR2 and HLA-DR4 phenotypes were encountered in a significantly higher frequency in black and Caucasian patients with rheumatic fever, respectively, compared with the control populations (P less than 0.005). The most significant association (P less than 0.005) of these DR antigens with a major manifestation of rheumatic fever was found for mitral insufficiency. In addition, a significant association was encountered between persistent elevation of antibody to the group A streptococcal carbohydrate and HLA-DR4 in Caucasian patients (P less than 0.04) or HLA-DR2 in the black patients (P less than 0.001). The frequency of HLA-DR2/4 heterozygotes among patients with rheumatic fever did not differ significantly from controls. These findings support the concept of a genetically determined susceptibility to rheumatic fever and, particularly, to rheumatic heart disease. The association of the clinical manifestations of rheumatic fever and the immune hyperresponsiveness to a streptococcal antigen could be ascribed to a disease-associated immune-response gene which is in linkage disequilibrium with the DR2 and DR4 alleles of HLA-DR locus on chromosome six.


Perioperative Concerns for Profound Metabolic Alkalosis During Kidney Transplantation: A Case Report.
Profound metabolic alkalosis is an uncommon consideration for the anesthetic management of kidney transplantation. Serum total carbon dioxide content and complex electrolyte abnormalities might be important diagnostic clues for the presence of metabolic alkalosis in the absence of arterial blood gas analysis. A 34-year-old female visited Gachon University Gil Medical Center, Incheon, South Korea during year 2015. She experienced aggravated renal function due to chronic hypokalemia and severe hypochloremic metabolic alkalosis, induced by laxative abuse, and underwent ABO incompatible kidney transplantation. Serum total carbon dioxide content remained high (about 60 mEq/L) over eight months of monthly follow-up prior to kidney transplantation. The authors described their anesthetic experience of profound metabolic alkalosis with complex electrolyte abnormalities and provided a review of relevant literature.


Four nitrobenzaldehyde isonicotinoylhydrazones at 120 K: four different supramolecular structures in two and three dimensions.
The molecules of 2-nitrobenzaldehyde isonicotinoylhydrazone, C13H10N4O3, (I), are linked into a three-dimensional framework by a combination of one N-H...N and three C-H...O hydrogen bonds. In the isomeric compound 3-nitrobenzaldehyde isonicotinoylhydrazone, (II), the molecules are linked into complex sheets by a combination of three hydrogen bonds, one each of the N-H...N, C-H...N and C-H...O types. The molecules of the third isomer, namely 4-nitrobenzaldehyde isonicotinoylhydrazone, (III), are linked into bilayers by a combination of one N-H...N, one C-H...N and two C-H...O hydrogen bonds. 2,4-Dinitrobenzaldehyde isonicotinoylhydrazone, (IV), crystallizes as a stoichiometric monohydrate, C13H9N5O5.H2O, and the molecular components are linked into a three-dimensional framework by a combination of O-H...O, O-H...N, N-H...O and three independent C-H...O hydrogen bonds.


Ingeneue: a versatile tool for reconstituting genetic networks, with examples from the segment polarity network.
Here we describe a software tool for synthesizing molecular genetic data into models of genetic networks. Our software program Ingeneue, written in Java, lets the user quickly turn a map of a genetic network into a dynamical model consisting of a set of ordinary differential equations. We developed Ingeneue as part of an ongoing effort to explore the design and evolvability of genetic networks. Ingeneue has three principal advantages over other available mathematical software: it automates instantiation of the same network model in each cell in a 2-D sheet of cells; it constructs model equations from pre-made building blocks corresponding to common biochemical processes; and it automates searches through parameter space, sensitivity analyses, and other common tasks. Here we discuss the structure of the software and some of the issues we have dealt with. We conclude with some examples of results we have achieved with Ingeneue for the Drosophila segment polarity network.


Active immunization against hepatitis A in dialysis patients.
This study investigated the feasibility, immunogenicity, and reactogenicity of hepatitis A vaccination in end-stage renal failure patients who were on chronic intermittent haemodialysis. Forty-three subjects were vaccinated with an inactivated hepatitis A vaccine according to a 0-, 1-, and 6-month immunization schedule. Two groups were established who received the vaccine either intramuscularly (group A, n=30) or subcutaneously (group B, n=13). All patients in group A and 12/13 in group B developed antibodies against hepatitis A. The geometric mean titres (GMT) were high and similar to those observed in healthy subjects. There was a tendency to higher GMT in the group who received the vaccine subcutaneously. No clinically significant adverse events were observed, and the liver enzyme profile showed no abnormalities. We showed that hepatitis A vaccination of dialysis patients is feasible, well tolerated and immunogenic and that the vaccine can be given subcutaneously in those patients where intramuscular administration is contra-indicated.


Successful repair of recurrent rectovaginal fistula by stratified suture using transanal endoscopic microsurgery: A CARE-compliant case report.
Rectovaginal fistulas (RVFs) are abnormal connections between the rectum and vagina. Although many surgical approaches to correct them have been attempted, management of RVFs still remains a challenge, especially for recurrent RVFs. In the present study, we report a case in a 22-year-old female with a chief complaint of obvious passages of flatus or stool through the vagina for 10 years. She had suffered a vaginal trauma from a violent accident 10 years prior, and gradually noticed the uncontrollable passage of gas or feces from the vagina 2 weeks later.The patient underwent a transvaginal direct repair surgery at local hospital 9 years ago, but the symptoms recurred 1 month after the surgery. After 2-years monitoring, the patient underwent another transvaginal repair surgery (fistulectomy followed by direct suture) at another hospital, but the fistula recurred again. We initially performed a temporary protective transversostomy upon admission. After 8-months of observation, a methylene blue test was conducted and the diagnosis of recurrent RVF was confirmed. Subsequently, we performed a successful repair by stratified suture using transanal endoscopic microsurgery (TEM). The scar tissue on the posterior wall of the vagina and the anterior wall of the rectum were meticulously excised until the margin of the excisional line showed healthy tissue. In addition, the fistulous tract was completely removed. The edges of the fistula on the posterior wall of the vagina were closed by simple continuous suturing, and the rectal anterior wall was sutured in the same manner. During a 1-year follow-up period, the fistulae were not recurrent and no complication such as incontinences or rectal bleeding were found. The latest Wexner score was 3. We present a case of successful treatment with stratified suture using TEM throughout the procedure. We strongly recommend this efficient and minimally invasive procedure for recurrent RVFs.


Clinical course of nontraumatic, nonaneurysmal subarachnoid hemorrhage: a single-institution experience.
Angiogram-negative subarachnoid hemorrhage (SAH) accounts for 15% of nontraumatic SAH and has been reported with low morbidity and mortality rates. We report on a large series of patients with angiogram-negative SAH who experienced an atypical nonbenign clinical course. Between December 2001 and November 2006, 95 patients with spontaneous nonaneurysmal SAH and negative initial angiographic evaluation were treated at the University of Florida. The authors retrospectively reviewed the patients' medical records and radiological images to determine associated morbidity and mortality. Aneurysms were found in 6 of the 95 patients on follow-up imaging after an initial negative angiogram (6.3% false negative rate); these patients were excluded leaving 89 patients as the study group. Hydrocephalus necessitating temporary CSF diversion developed in 22 of these patients (25%); 12 (13%) ultimately required permanent CSF diversion. Clinically significant vasospasm developed in 4 patients (4%), and 2 (2%) had cerebral infarctions. Three patients (3%) died. The authors' experience with a large series of angiogram-negative SAH patients who had an atypical nonbenign clinical course associated with hydrocephalus, vasospasm, stroke, and mortality differs significantly from previously published case series of angiogram-negative SAH.


Supplementation of eicosapentaenoic acid-rich fish oil attenuates muscle stiffness after eccentric contractions of human elbow flexors.
This study aimed to investigate the effect of supplementation of fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the damage of the biceps brachii after eccentric contractions (ECCs) of the elbow flexors, particularly focusing on muscle stiffness. Sixteen men were included in this double-blind, placebo-controlled, parallel design study and the participants were randomly assigned to the EPA and DHA supplement group (EPA, n = 8) and placebo group (PL, n = 8). They consumed either EPA 600 mg and DHA 260 mg per day or placebo supplement for 8 weeks prior to exercise. Moreover, they performed six sets of 10 ECCs at 100% maximal voluntary contraction (MVC) using a dumbbell. Changes in MVC torque, range of motion (ROM), upper arm circumference, muscle soreness, muscle echo intensity, and muscle stiffness were assessed before exercise; immediately after exercise; and 1, 2, and 5 days after exercise. MVC torque and ROM were significantly higher in the EPA group than in the PL group after ECCs (p < 0.05). Muscle soreness, upper arm circumference, and muscle echo intensity were significantly higher in the PL group than in the EPA group after ECCs (p < 0.05). In addition, muscle stiffness at 150° was significantly higher in the PL group than in the EPA group immediately after ECCs (p < 0.05). The present study showed that EPA and DHA supplementation has a positive role in inhibiting muscle stiffness after ECCs. This trial (UMIN000028165) was registered on 10th/July/2017.


Language decline across the life span: findings from the Nun Study.
The present study examines language samples from the Nun Study. Measures of grammatical complexity and idea density were obtained from autobiographies written over a 60-year span. Participants who had met criteria for dementia were contrasted with those who did not. Grammatical complexity initially averaged 4.78 (on a 0-to-7-point scale) for participants who did not meet criteria for dementia and declined .04 units per year; grammatical complexity for participants who met criteria for dementia initially averaged 3.86 and declined .03 units per year. Idea density averaged 5.35 propositions per 10 words initially for participants who did not meet criteria for dementia and declined an average of .03 units per year, whereas idea density averaged 4.34 propositions per 10 words initially for participants who met criteria for dementia and declined .02 units per year. Adult experiences, in general, did not moderate these declines.


Mono- and multisite solid catalysts in cascade reactions for chemical process intensification.
Catscade reactions: Heterogeneous catalysis can enable sustainability by carrying out multistep reactions in one step. It is shown that single- and multisite solid catalysts can be designed to carry out cascade reactions for the synthesis of fine chemicals.


Comparison of bioimpedance techniques to detect changes in fluid status in hemodialysis patients.
Bioimpedance (BI) is maturing as a clinical technique for assessing fluid volume status. The aim of this study was to compare the sensitivity of four BI methods to detect changes in fluid status in hemodialysis patients. Forty-five patients were studied twice in the same week, i.e. once after the long and short interdialytic intervals, respectively. The four BI methods used were: (a) calf normalized resistivity (CNR) at a 5-kHz frequency, (b) whole-body multifrequency BI spectroscopy (MF-BIS) to estimate the normal hydration weight (NHWWBM), (c) whole-body MF-BIS to estimate the ratio of extracellular volume to total body water (wECV/wTBW), and (d) whole-body single-frequency (50 kHz) BI analysis to compute the ratio of ECV (sfECV) to TBW (sfTBW). The relationship (slope of the regressive line) between relative changes (%) in the above mentioned four BI parameters and differences in weight (kg) was most pronounced with CNR (5.2 ± 1.6%/kg), followed by wECV/wTBW (1.7 ± 0.7%/kg) and NHWWBM (0.73 ± 0.2%/kg). Changes in sfECV/sfTBW and differences in weight were not correlated. CNR is more sensitive than whole-body BIS for detecting differences in fluid status.


Probing stability and dynamics of proteins by protease digestion. II: Identification of the initial chymotryptic cleavage sites of homologous cytochromes c.
Three homologous cytochromes c from horse, rabbit and tuna were subjected to chymotryptic digestion and their initial cleavage sites were identified. The sites in oxidized cytochromes c are the COOH-terminal sides of Tyr-48, Phe-46 and Tyr-46 for horse, rabbit and tuna cytochromes c, respectively. The results show that the chymotrypsin attacks a single site in each protein; the sites are located at the almost identical position on the polypeptide chain. Through the time-course studies of digestion, it was found that the three cytochromes c have different chymotrypsin-susceptibility at the initial cleavage site in the order of horse less than rabbit less than tuna. Studies on chymotryptic digestion of tuna cytochrome c in the reduced form revealed that the haem-reduction does not alter the initial cleavage site but increases the resistance to the proteolysis at the site. The uniqueness of the initial cleavage site in each cytochrome c species suggests that the protease susceptibility reflects some overall properties of the protein. At the same time, it was clarified that the initial cleavage site is also affected by a neighboring region by the fact that another potential cleavage site is located near the site in question. In order to elucidate the initial cleavage site, several physical properties of tuna cytochrome c molecule deduced from the X-ray 3D structure, accessible surface area, temperature factor, effective hydrophobicity and electrostatic potential, were compared with the experimental results and it was concluded that these properties given by a residue have no direct relationship with the chymotrypsin susceptibility.


[A case of interstitial pneumonitis induced by FOLFIRI+bevacizumab combination therapy for liver and lung metastasis of colon cancer].
A 64 -year-old female received oral S-1 chemotherapy followed by mFOLFOX6 chemotherapy for postoperative liver and lung metastasis of sigmoid colon cancer. The tumor progression was observed after twelve courses of mFOLFOX6 chemotherapy, and then FOLFIRI+bevacizumab chemotherapy was performed. After two courses of FOLFIRI+bevacizumab chemotherapy, leucopenia was observed. The chemotherapy was then discontinued and G-CSF was administered. Two days later she complained of high fever and dry cough, and was admitted to the hospital. A diffuse ground-glass appearance of bilateral lung was observed on chest X-ray and CT. Drug-induced interstitial pneumonitis was suspected, and Pneumocystis carini pneumonia was considered in the differential diagnosis. Oral administration of prednisolone and sulfamethoxazole/trimethoprim did not improve the symptoms, so steroid pulse therapy was performed. Steroid pulse therapy improved respiratory symptoms, but CT findings did not change remarkably. After nine weeks in the hospital, she was discharged with home oxygen therapy. Interstitial pneumonitis induced by FOLFIRI+bevacizumab chemotherapy is rare, but the number of cases may increase with increased use of this regimen. The possibility of interstitial pneumonitis should always be considered when the patient presents with a respiratory disorder while receiving systemic chemotherapy.