Hormonal control of neuron number in sexually dimorphic spinal nuclei of the rat: II. Development of the spinal nucleus of the bulbocavernosus in androgen-insensitive (Tfm) rats.
The spinal nucleus of the bulbocavernosus (SNB) is a sexually dimorphic motor nucleus whose development is under the control of steroid hormones. The SNB contains many more motoneurons in adult male rats than in females, and this sex difference is produced by a sexually dimorphic motoneuron death which is regulated by androgens. To study further the role of androgens in the development of sex differences in SNB motoneuron number, we examined SNB development in males with the testicular feminization mutation (Tfm) which renders them insensitive to androgens. Counts of SNB motoneurons perinatally revealed that SNB development in normal male and female King-Holtzman rats was similar to that reported previously for Sprague-Dawley rats; SNB motoneuron number increased from initially low levels at embryonic day 18 through the day before birth, when motoneuron numbers in both sexes were substantially higher than adult levels. After this prenatal increase, motoneuron number declined in both sexes, until by postnatal day 10 motoneuron numbers were in their adult ranges and the sex difference was fully expressed. Females lost more motoneurons than did males during this period, and this loss was due to motoneuron death as revealed by counts of degenerating cells. SNB development in King-Holtzman Tfm males was similar to that of normal males through embryonic day 20, suggesting that androgens may not be necessary for the initial increase in motoneuron numbers in the SNB. Thereafter, SNB motoneuron numbers in Tfm males declined in a female-typical fashion; Tfm males and normal females did not differ at any postnatal age.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of selective media for Campylobacter isolation when cycloheximide is replaced with amphotericin B.
Laboratory media for the isolation of Campylobacter usually contain various selective agents, designed to allow these bacteria to grow whilst suppressing that of other organisms. For example, cycloheximide has often been incorporated into Campylobacter media to inhibit the growth of fungi. The production and availability of cycloheximide, however, has recently decreased due to concerns relating to its potential toxicity for mammalian cells and the compound has also become more expensive as a result. An alternative antifungal agent is necessary, and to address this, the effect of using amphotericin B in place of cycloheximide in Campylobacter selective broths and agars was examined. The growth of Campylobacter strains and the suppression of potential competitor organisms in selective broths or on selective agars containing either amphotericin B or cycloheximide was quantified, using pure microbial cultures. The results were then confirmed by testing food and water samples that contained high levels of micro-organisms, including Campylobacter. The results of this study indicate that amphotericin B is a suitable replacement for cycloheximide in Campylobacter selective media.


Supplementary prescribing for the elective surgical patient.
Supplementary prescribing is an exciting new development in nursing. After qualifying, the supplementary nurse prescriber may prescribe a range of medications as part of a clinical management plan. The rationale for the choice of medication for inclusion in a clinical management plan for use in elective surgery is described.


Plant extracts such as pine nut shell, peanut shell and jujube leaf improved the antioxidant ability and gas permeability of chitosan films.
In order to develop the antioxidant and gas permeability packaging film, the effects of three plant extracts (pine nut shell, peanut shell and jujube leaf) on the physical, antioxidative and structural properties of chitosan based biodegradable films were studied. The results showed that three plant extracts improved the antioxidant capacity of films. The DPPH radical scavenging activity of chitosan-jujube leaf films increased by 3.8 times compared with the control films. The chitosan-pine nut shell films had the highest water vapor and oxygen permeability, while chitosan-peanut shell films showed greatest increase in CO2 permeability with a value of 1.71 × 104 cm3/(m2·24h) under standard test. In addition, three plant extracts reduced the homogeneity and caused porous structure of chitosan films. Chitosan-peanut shell films had the highest thermal stability compared with the control and other two films. X-ray diffraction and FTIR indicated three plant extracts changed the hydrogen bonding of film matrix and caused the changes of crystal and chemical structure. The study provided a reference for the preparation of polysaccharide-based active films with strong antioxidant and gas permeability.


[A cerebrovascular stroke in a child with an anomalous origin of the left coronary artery].
A two-year-old male child presenting anomalous origin of the left coronary artery diagnosed by Doppler echocardiography and confirmed by angiography developed an ischemic stroke, which is a rare event in this situation. He [correction of She] had good recovery of the neurological deficit and his anomalous artery was successfully reimplanted.


Nanoprecipitation through solvent-shifting using rapid mixing: Dispelling the Ouzo boundary to reach large solute concentrations.
The addition of a non-solvent to a solute in good solvent solution leads to nanoprecipitation, which is the spontaneous formation of nanodomains. Yet, increasing solute concentration usually leads to the formation of macrodomains that quickly separate into a bulk phase, which is a severe process limitation. The corresponding concentration threshold, often termed as the Ouzo boundary, remains a mystery that could find its origin in the complex interplay between nanoprecipitation and mixing. We performed a systematic investigation of nanoprecipitation thermodynamics and kinetics as well as its interplay with mixing hydrodynamics for the hexadecane-acetone-water system, in the presence of the non-ionic C16EO8 surfactant. The binodal curve and its underlying tie-lines were obtained using Raman spectroscopy, allowing the computation of the spinodal curve. Kinetics were probed using a continuous flow setup that combines two sequential rapid mixers. The impact of mixing efficiency was probed systematically by varying the oil concentration for respectively slow and rapid mixing, while the uncoupling from mixing and nanoprecipitation was quantified by modifying systematically the flow rate in a continuous flow approach. We elucidate the nature of the Ouzo boundary that marks the maximal solute concentration leading to nanoobjects. Rather than a thermodynamic boundary, as evidenced by its uncorrelation to the spinodal curve, it results from the coupling of nanoprecipitation and mixing when both processes occur within the same time range, leading to heterogeneous conditions and the escape of some objects to the macroscale. Increasing the solute concentration speeds up nanoprecipitation and thus requires increasingly faster mixing times to uncouple both processes. Accordingly, if the mixing efficiency is large enough, it is possible to dispel the Ouzo boundary and reach very large solute concentrations. Implementing rapid mixing strategies in continuous flow approaches is thus the solution to overcome the most stringent condition of nanoprecipitation and open the way to scale-up, while also providing efficient means to probe its fast mechanism. Overall, the simultaneous control of hydrodynamics and physical chemistry is thus key to boost up the Ouzo effect.


Structure and function of membrane IL-1.
Both interleukin-1 alpha (IL-1 alpha) and IL-1 beta are initially translated as approximately Mr 30,000 polypeptides, lacking hydrophobic or signal sequence that could facilitate transmembrane translocation and release of mature IL-1 (Mr 17,500). The current study utilizes an antiserum specific for murine IL-1 alpha in order to investigate membrane associated IL-1 alpha polypeptides and possible postsynthetic modifications of the IL-1 alpha precursor, that might account for its intracellular transport. Cell surface iodination of endotoxin stimulated murine macrophages allowed the detection of IL-1 molecules in size similar to the IL-1 alpha precursor (Mr 33,000). Membrane bound IL-1 alpha was sensitive to degradation by serine esterase activity to yield IL-1 peptides of Mr 16,000 to 18,000. Endotoxin stimulated macrophages, but not unstimulated cells, incorporated 32PO4 into the IL-1 alpha precursor. The phosphate label of the IL-1 alpha precursor is resistant to hydroxylamine and alkaline phosphatase treatment. Released IL-1 is not phosphorylated. Approximately 10% of the phosphorylated IL-1 alpha precursor is membrane bound and associated with fractions enriched in lysosomal vesicles. These data are consistent with a model for mIL-1 expression, in which pro IL-1 alpha is post-synthetically modified to achieve intracellular transport and further suggest that mIL-1 may be a prerequisite for the release of IL-1.


Cognitive load during training for out-of-department emergency responses.
Emergency medicine (EM) physicians sometimes respond to critical events outside the emergency department. To prepare for these complex cases-typically called "rapid responses" (RRs)-EM residents receive simulation-based training involving four practice tasks and three exam tasks during a 1-day session. Cognitive load (CL) theory describes how humans function with limited working memories to perform complex tasks. RRs are expected to generate high levels of CL, but the profile of CL across providers and RR cases is not well understood. In this study, we analyzed resident's CL during RR training. We hypothesized variations in CL across individual and case and that exam cases would cause higher CLs than practice cases. Residents anonymously self-reported CL levels after each case using the Paas scale, a single-item, 9-point scale from "very, very low CL" to "very, very high CL." To examine case-based differences in CL, data were rescaled by individual residents. "High CL" was defined as a score of 9/9. Among 18 residents participating, CLs ranged from 4 to 9, with median of 7 and interquartile range of 7-8. While many cases showed bell curve-like distributions of CLs, one case-a bleeding tracheostomy-showed a rightward skew reflecting higher levels of CL. No significant difference was found in CL between practice and exam cases. There were 20 reports (16.5%) of "high" CL with variation across residents (0/7 [0%] to 5/6 [83.3%] cases) and across cases (1/18 [5.6%) to 8/18 [44.4%]). The CL that EM residents experienced did show considerable interpersonal and intercase variation, but there was no significant difference between practice and exam cases. These results highlight several questions about how to optimally design future training, including how best to balance low and high CL training cases and which cases may require further training.


TAF1B depletion leads to apoptotic cell death by inducing nucleolar stress and activating p53-miR-101 circuit in hepatocellular carcinoma.
TAF1B (TATA Box Binding Protein (TBP)-Associated Factor) is an RNA polymerase regulating rDNA activity, stress response, and cell cycle. However, the function of TAF1B in the progression of hepatocellular carcinoma (HCC) is unknown. In this study, we intended to characterize the crucial role and molecular mechanisms of TAF1B in modulating nucleolar stress in HCC. We analyzed the differential expression and prognostic value of TAF1B in hepatocellular carcinoma based on The Cancer Genome Atlas (TCGA) database, tumor and paraneoplastic tissue samples from clinical hepatocellular carcinoma patients, and typical hepatocellular carcinoma. We detected cell proliferation and apoptosis by lentiviral knockdown of TAF1B expression levels in HepG2 and SMMC-7721 cells using clone formation, apoptosis, and Western blotting (WB) detection of apoptosis marker proteins. Simultaneously, we investigated the influence of TAF1B knockdown on the function of the pre-initiation complex (PIC) by WB, and co-immunoprecipitation (Co-IP) and chromatin immunoprecipitation (ChIP) assays verified the interaction between the complexes and the effect on rDNA activity. Immunofluorescence assays measured the expression of marker proteins of nucleolus stress, fluorescence in situ hybridization (FISH) assays checked the rDNA activity, and qRT-PCR assays tested the pre-rRNA levels. Regarding molecular mechanisms, we investigated the role of p53 and miR-101 in modulating nucleolar stress and apoptosis. Finally, the impact of TAF1B knockdown on tumor growth, apoptosis, and p53 expression was observed in xenograft tumors. We identified that TAF1B was highly expressed in hepatocellular carcinoma and associated with poor prognosis in HCC patients. TAF1B depletion modulated nucleolar stress and apoptosis in hepatocellular carcinoma cells through positive and negative feedback from p53-miR-101. RNA polymerase I transcription repression triggered post-transcriptional activation of miR-101 in a p53-dependent manner. In turn, miR-101 negatively feeds back through direct inhibition of the p53-mediated PARP pathway. These findings broaden our comprehension of the function of TAF1B-mediated nucleolar stress in hepatocellular carcinoma and may offer new biomarkers for exploring prospective therapeutic targets in HCC.


Sodium-dependent glucose transport by cultured proximal tubule cells.
The cotransport of sodium ion and alpha-methyl glucose, a non-metabolized hexose, was studied in rabbit proximal tubule cells cultured in defined medium. The rate of uptake of alpha-methyl glucose shows saturation kinetics, in which Km, but not Vmax, is dependent upon the Na+ concentration in the medium. The transport system was found to be of the high-affinity type, characteristic of the straight portion of the proximal tubule. Analysis of the rates of initial uptake within the context of a generalized cotransport model, suggests that two Na+ ions are bound in the activation of the hexose transport. The steady-state level of accumulation of alpha-methyl glucose also depends upon sodium concentration, consistent with the initial rate findings. The uptake of alpha-methyl glucose is inhibited by other sugars with the relative potencies of D-glucose greater than alpha-methyl glucose greater than D-galactose = 3-O methylglucose. L-Glucose, D-fructose, and D-mannose show no inhibition. Phlorizin inhibits the alpha-methyl glucose uptake with a Ki of 9 X 10(-6) M. Ouabain (10(-3) M) decreases the steady-state alpha-methyl glucose accumulation by 60%. In the absence of sodium, the accumulation of alpha-methyl glucose is 7-fold less than at 142 mM Na+, reaching a level comparable to the sodium-independent accumulation of 3-O-methyl-D-glucose. These findings are similar to those observed in the proximal tubule of the intact kidney.


Prognostic implications of cyclin B1, p34cdc2, p27(Kip1) and p53 expression in gastric cancer.
Cell cycle progression is regulated by interactions of specific cyclins and cyclin dependent kinases (CDKs) at the G1-S and G2-M checkpoints and cell cycle deregulation plays a major role in carcinogenesis of human cancers. To investigate the role of cell cycle regulators in the pathogenesis and progression of human gastric cancers, 23 cases of gastric carcinomas were examined for the expression of cyclin B1, p34cdc2, p27(Kip1) and p53 by immunohistochemical methods, and gene expression was correlated with various clinicopathologic findings. Out of 23 cases studied, cyclin B1 was diffusely expressed in 20 cases (87.0%), p34cdc2 in 14 cases (60.9%) and p53 in 12 cases (52.2%), whereas in normal gastric tissues, cyclin B1 and p34cdc2 were weakly expressed and p53 was not expressed. In contrast, p27(Kip1) was expressed in only 8.7% of gastric carcinomas compared with 78.3% of normal gastric tissues. There was correlation between the expression of cyclin B1 and expression of p34cdc2 (p=0.002), between the expression of cyclin B1 and loss of p27(Kip1) (p=0.025), and between the expression of p34cdc2 and loss of p27(Kip1) (p=0.065). In addition, expression of cyclin B1 was correlated with regional lymph node metastasis (p=0.032). Our results indicate that cyclin B1 and p34cdc2 are involved in the genesis or progression of gastric cancers. Furthermore, overexpression of cyclin B1 may play an important role in lymph node metastatic potential of gastric cancer. Thus, abnormal expression of cyclin B1 and CDKs, overexpression of p53 and loss of p27(Kip1) expression may play important roles in human gastric carcinogenesis.


Management of the hard posterior polar cataract.
I describe a technique for managing the posterior polar cataract with a dense nucleus by initially sculpting a central trough to the depth of the polar opacity. The nucleus is partially cracked in the periphery, avoiding the polar opacity, and then chopped into quadrants without rotating. Next, the phaco tip engages the core of the nuclear quadrant while a cleavage plane is manually dissected along the lenticular lamellae using a Nagahara phaco chopper, with the tip rotated slightly so it points somewhat toward the equator of the lens, to a depth estimated to leave a nuclear shell, sparing the polar cataract. The nucleus is peeled away from the outer nucleus shell, which is kept in place by the phaco chopper. The intact nuclear plate is removed last and any capsular defect managed accordingly. This technique minimizes the risk for losing a nuclear fragment into the vitreous cavity.


Vena cava filter scaffold to prevent migration of embolic materials in the treatment of a massive renal arteriovenous malformation.
Transcatheter occlusion of high-flow arteriovenous malformations (AVMs) or arteriovenous fistulas may be complicated by migration of embolic materials intended to impede flow. Reducing antegrade flow during catheter embolization via balloon occlusion or inhibiting migration with anchoring devices may improve procedural safety in high-flow settings. We describe the use of an inferior vena cava filter as a scaffold to support complete vascular occlusion of a large renal AVM.


Plasma catecholamines in man are not influenced by the inhibition of prostaglandin synthesis.
Indomethacin has been reported to potentiate the release of noradrenaline from sympathetic nerve endings in vitro and to increase urinary noradrenaline excretion in rats. We have studied the influence of indomethacin on plasma catecholamine levels in 10 normal men, using measurement of plasma renin activity (PRA) as an index of the pharmacodynamic effect of indomethacin. Both in the supine and standing positions indomethacin failed to alter the plasma concentrations of noradrenaline, adrenaline or dopamine, while PRA was markedly suppressed. It is concluded that in the intact human indomethacin does not influence catecholamine concentrations.


Beliefs and communication practices regarding cognitive functioning among consumers and primary care providers in the United States, 2009.
Limited research has examined primary care providers' communication with patients about maintaining cognitive functioning. Our study's objective was to compare the perceptions of consumers and primary care providers related to beliefs and communication practices about lifestyle behaviors beneficial for overall health and for maintaining cognitive functioning. In 2009, we submitted 10 questions to Porter Novelli's HealthStyles survey and 6 questions to their DocStyles survey. We compared consumers' (n = 4,728) and providers' (n = 1,250) beliefs, practices, and information sources related to maintaining health and cognitive functioning. We made comparisons using nonparametric statistics. Approximately 76% of consumers considered their health to be good or very good; 73.4% were concerned or very concerned about the possibility that their memory may worsen with age. Women were significantly more concerned than men, and white consumers were more concerned than black and Hispanic consumers. Consumers reported they believed that intellectual stimulation (86.6%), physical activity (82.6%), and healthful diet (82.5%) prevented or delayed cognitive impairment. Providers reported advising patients to reduce cognitive impairment risk through physical activity (85.9%), intellectual stimulation (80.3%), and social involvement (67.4%). Few consumers (7.8%) reported receiving this information from providers but reported learning about strategies to maintain memory, primarily from television (50.1%), magazines (44.1%), and newspapers (33.7%). Providers reported advising patients about how to reduce risks of cognitive impairment. Consumers reported receiving this information from other sources. Findings suggest a need to examine and assess media messages and to better understand patient-provider communication about cognitive functioning.


Foley Catheter Balloon Rupture and Risk of Free Fragment Formation.
To assess the volume of foley catheter balloon rupture and the incidence of free fragment formation. Forty Latex and 30 Silicone catheters of common sizes (14-20 French) were inflated with sterile water in an aqueous environment without positive pressure. Catheters producing fragments, fragment size, and maximum instilled volume prior to rupture ("burst volume") were recorded. A subset of catheters was inflated to recommended balloon volume and the balloon was needle punctured. Of the 40 latex catheters, 32 (80%) produced a free fragment during spontaneous rupture. Fragments ranged from 2.1 to 3.2 cm with an average size of 2.74 ± 0.33 cm. Latex catheters had average burst volume of 83 mL, 90 mL, 112 mL,120 mL, and 422 mL for 14 Fr, 16 Fr, 18 Fr, 16 Fr 3-way, and 20 Fr 30 cc, respectively. Of the 30 silicone catheters, spontaneous rupture produced no fragments. Average burst volume for silicone catheters was 57 mL, 45 mL, and 55 mL for 14 Fr, 16 Fr, and 18 Fr, respectively. No catheter balloons produced fragments when needle punctures at recommended balloon volumes. Latex catheter balloons tolerate higher fill volumes but have a high likelihood of releasing fragments. Silicone catheters burst at lower volumes, but do not release fragments. At normal fill volumes, needle puncture is safe, and cystoscopy is likely unnecessary.


Single-incision versus conventional laparoscopic colectomy for colonic neoplasm: a randomized, controlled trial.
Single-incision laparoscopic colectomy (SILC) is a newly developed procedure with the benefit of better cosmetic outcome and potentially reduced wound pain compared with conventionally laparoscopic colectomy (CLC). However, the application of SILC requires careful evaluation to prove its benefit and safety. This randomized, controlled study compared the operative outcome of patients who underwent SILC and CLC. Patients who had small cancer (<4 cm) or adenomatous polyp requiring colectomy were randomized to have SILC or CLC. The patients were blinded to the procedures and the postoperative pain was used as the primary outcome measure. All patients had patient-controlled analgesia with intravenous morphine after the operation and the nominal rating score on days 1-3 and day 14 were recorded by research staff, who did not known the types of operations. Other operative outcomes of the two groups of patients also were recorded prospectively and compared. There were 25 patients in each group. The patients' demographics, tumor characteristics, operating time, blood loss, complication rate, number of lymph nodes harvested, and resection margin have no statistically significant difference between the two groups. There was no operative mortality in both groups. The SILC group had consistently lower median pain score than CLC group in the whole postoperative course and the difference was statistically significant on day 1 (0 (0-5) vs. day 3 (0-6) respectively; p = 0.002) and day 2 (0 (0-3) vs. 2 (0-8) respectively; p = 0.014). The median hospital stay in the SILC group also was shorter the CLC group. In a selected group of patients with small tumor and good operative risk, SILC is a safe alternative to CLC. Single-port laparoscopic colectomy also is associated with the benefits of less postoperative pain and shorter hospital stay than CLC.


Smoking desistance and personality change in emerging and young adulthood.
We attempted to replicate and extend (Welch, P., & Poulton, R. [2009]. Personality influences on change in smoking behavior. Health Psychology, 28, 292-299. doi:10.1037/a0013471) findings regarding the relationship between smoking involvement and personality change. Two time frames (18-25, 18-35) in a cohort of college students (N = 489 at baseline) followed for 17 years were used. Impulsivity, neuroticism, and 4 measures of smoking involvement were assessed at each time point. Individuals who desisted in smoking involvement tended to exhibit the largest decreases in neuroticism and impulsivity from ages 18 to 25. From ages 18 to 35, changes in smoking involvement were only significantly associated with changes in impulsivity. These findings add to the literature demonstrating decreasing neuroticism, and impulsivity correlates with decreased substance involvement during emerging adulthood.


Mechanism of growth inhibition of melanoma cells by 4-S-cysteaminylphenol and its analogues.
Our previous studies have shown that 4-S-cysteaminylphenol (4-S-CAP) causes a significant inhibition of in vivo melanoma growth and a marked depigmentation of black skin and hair follicles. These studies have suggested a role of tyrosinase in the manifestation of these in vivo effects. In this study 4-S-CAP and its analogues were examined for their effects on the growth of human melanoma cells in vitro. 4-S-CAP and 4-S-HomoCAP exhibited strong cytotoxicity with effects much greater than those of alpha-methyl-4-S-CAP and N,N-dimethyl-4-S-CAP. The cytotoxicity of the former two amines was completely prevented by semicarbazide, an inhibitor of plasma monoamine oxidase, while that of the latter two was not prevented by semicarbazide, catalase, and phenylthiourea, a tyrosinase inhibitor. In culture medium 4-S-CAP was rapidly converted by the action of monoamine oxidase present in fetal bovine serum to the aldehyde which was then metabolized to the alcohol and the carboxylic acid when cells were present. alpha-Methyl-4-S-CAP was found to exert higher cytotoxicity to cells with higher tyrosinase activity and melanin content. These results suggest that the in vitro cytotoxicity of 4-S-CAP and 4-S-HomoCAP is mediated through conversion to the aldehydes while that of alpha-methyl-4-S-CAP appears to be dependent on tyrosinase activity to some extent.


HPLC determination of a chemically modified nonantimicrobial tetracycline: biological implications.
Chemically modified tetracycline (4-de-dimethylamino tetracycline), like commercially available tetracyclines, is known to inhibit experimentally induced pathologic collagen breakdown. A method for measurement of chemically modified tetracycline in small volumes (50 microliters) of rat serum was developed using reversed-phase HPLC; this was necessary because this tetracycline analog lacks antimicrobial activity and, therefore, cannot be measured with standard bioassays. This method uses the same solution for extraction and elution thus providing a simple and rapid assay for both drugs. Using this technique, the concentration of chemically modified tetracycline and tetracycline were determined in rat serum at different times after oral administration. The serum concentration of chemically modified tetracycline was much higher than that for tetracycline, and its serum half-life was greater. The IC50 of chemically modified tetracycline and tetracycline, as inhibitors of collagenase from rat polymorphonuclear leukocytes, was determined and found to be 4.1 x 10(-8) M (0.02 micrograms/ml) and 2.4 x 10(-4) M (120 micrograms/ml), respectively. Based on the serum levels of these drugs after oral administration, and their IC50 values, chemically modified tetracycline is potentially a far more potent inhibitor of excess collagenase activity than tetracycline, during pathologic conditions, and may have the added advantage of not producing some of the typical complications of long-term antibiotic therapy.


Discovery of novel DNA-damaging agents through phenotypic screening for DNA double-strand break.
DNA double-strand breaks (DSBs) seriously damage DNA and promote genomic instability that can lead to cell death. They are the source of conditions such as carcinogenesis and aging, but also have important applications in cancer therapy. Therefore, rapid detection and quantification of DSBs in cells are necessary for identifying carcinogenic and anticancer factors. In this study, we detected DSBs using a flow cytometry-based high-throughput method to analyze γH2AX intensity. We screened a chemical library containing 9600 compounds and detected multiple DNA-damaging compounds, although we could not identify mechanisms of action through this procedure. Thus, we also profiled a representative compound with the highest DSB potential, DNA-damaging agent-1 (DDA-1), using a bioinformatics-based method we termed "molecular profiling." Prediction and verification analysis revealed DDA-1 as a potential inhibitor of topoisomerase IIα, different from known inhibitors such as etoposide and doxorubicin. Additional investigation of DDA-1 analogs and xenograft models suggested that DDA-1 is a potential anticancer drug. In conclusion, our findings established that combining high-throughput DSB detection and molecular profiling to undertake phenotypic analysis is a viable method for efficient identification of novel DNA-damaging compounds for clinical applications.


In vivo interactions of acrylonitrile with macromolecules in rats.
The irreversible binding of [2,3-14C]acrylonitrile (VCN) to proteins, RNA and DNA of various tissues of male Sprague-Dawley rats after a single oral dose of 46.5 mg/kg (0.5 LD50) has been studied. Proteins were isolated by chloroform-isoamyl alcohol-phenol extraction. RNA and DNA were separated by hydroxyapatite chromatography. Binding of VCN to proteins was extensive and was time dependent. Radioactivity in nucleic acids was registered in the liver and the target organs, stomach and brain. DNA alkylation, which increased by time, was significantly higher in the target organs, brain and stomach (119 and 81 pmol/mg, respectively, at 24 h) than that in the liver. The covalent binding indices for the liver, stomach and brain at 24 h after dosing were, 5.9, 51.9 and 65.3, respectively. These results suggest that VCN is able to act as a multipotent carcinogen by alkylation of DNA in the extrahepatic target tissues, stomach and brain.


"Exposing force": the COVID-19 pandemic and women's sport in Ireland during 2020-2021.
An analysis of how the pandemic served to highlight neglected weaknesses and inequalities with regard to the structures and supports available to facilitate women's sport in Ireland. A survey conducted in the summer of 2021 with 194 female athletes across the island of Ireland. These athletes were engaged with the sports of camogie, Ladies Gaelic football, hockey, and rugby, and each responded to a 28-question survey. Our findings indicate that the experiences of female athletes during the COVID-19 pandemic raise serious questions regarding equality in sport across gender lines. Concerns expressed by the surveyed athletes, especially in relation to access to facilities, inadequate sponsorship, and funding reveal salient aspects of the experiences of Irish female athletes during the pandemic, and its role as an "exposing force" of inequalities within Irish sport. The COVID-19 pandemic in Ireland has been a challenging and revealing period with regard to sport in Ireland, acting as an "exposing force" of the existing inequitable structures for both the support and coverage of women's sport. Such challenging periods can also offer opportunities to learn, progress and improve and already in the past year there is evidence of positive developments for women's sport in Ireland, developments that we contend were expedited due to the impact of the COVID-19 pandemic in Ireland.


Cyclin D1 expression, cell proliferation, and clonal persistence characterize primary cutaneous CD4+ small or medium T-cell lymphoproliferative disorder.
The aim was to gain insight into the biology of primary cutaneous CD4+ small/medium T-cell lymphoproliferative disorder (PCSM-LPD). We describe the histopathological and clinical characteristics of 177 PCSM-LPD diagnosed at our consultation centre. We performed immunohistochemical multistaining in a subset of cases (n = 46) including PD1, Cyclin D1, and multiple markers of proliferation. We evaluated clonal T-cell-receptor-(TCR) rearrangements and used tissue microdissection to analyse TCR-clonality of PD1(+) cells. The cohort of n = 177 PCSM-LPD included 84 males and 93 females (median age 57, range 13-85). Clinical presentation was as a solitary nodule or plaque (head and neck > trunk > extremities). Most patients were treated by local excision or steroids (96%, 69/72); relapses occurred in 12/65 (18%) of patients with follow up. Histopathology revealed the predominance of a nodular pattern (75%, 134/177) and frequent clustering of PD1(+) large cells (70%, 103/147). We detected Cyclin D1 and PD1 coexpression (>10% of PD1(+)-cells) in 26/46 (57%), which was not associated with CCND1 breaks or amplifications. PD1(+)-cells in PCSM-LPDs showed a significantly higher expression of proliferation-associated proteins compared to PD1(-)-cells. A clonal TCR-rearrangement was present in 176/177 (99%), with a clonal persistence in 7/8 patients at relapse including distant sites. Tissue-microdissection revealed PD1(+)-cells as the source of clonality, whilst PD1(-)-cells remained polyclonal. PCSM-LPD is a clinically indolent, albeit neoplastic, disease driven by clonal expansion of PD1(+)-cells. We demonstrate Cyclin D1-expression associated with accelerated proliferation as a surprising new biological feature of the disease.


New Assay System Elecsys Anti-p53 to Detect Serum Anti-p53 Antibodies in Esophageal Cancer Patients and Colorectal Cancer Patients: Multi-institutional Study.
Several recent studies suggest that serum anti-p53 antibodies (s-p53-Abs) may be combined with other markers to detect esophageal and colorectal cancer. In this study, we assessed the sensitivity and specificity of s-p53-Abs detection of a new electrochemiluminescence immunoassay (ECLIA; Elecsys anti-p53). Elecsys anti-p53 assay was used to analyze the level of s-p53-Abs in blood sera from patients with esophageal or colorectal cancer taken before treatment. Control blood sera from healthy volunteers, patients with benign diseases, and patients with autoimmune diseases served as a reference. In addition, squamous cell carcinoma antigen (SCC-Ag) and cytokeratin 19 fragments (CYFRA21-1) were assessed in patients with esophageal cancer, and carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9 were assessed in patients with colorectal cancer. Samples from 281 patients with esophageal cancer, 232 patients with colorectal cancer, and 532 controls were included in the study. The median value of s-p53-Abs in control samples was < 0.02 μg/mL (range < 0.02-29.2 μg/mL). Assuming 98% specificity, the cut-off value was determined as 0.05 μg/mL. s-p53-Abs were detected in 20% (57/281) of patients with esophageal cancer and 18% (42/232) of patients with colorectal cancer. In combination with SCC-Ag and CEA, respectively, s-p53-Abs detected 51% (144/281) of patients with esophageal and 53% (124/232) of patients with colorectal cancer. The new s-p53-Abs assay Elecsys anti-p53 was useful in detecting esophageal and colorectal cancers with high specificity. Adding s-p53-Abs to conventional markers significantly improved the overall detection rates.


The utility of remdesivir in SARS-CoV-2: A single tertiary care center experience from a developing country.
Remdesivir is a monophosphoramidate prodrug of an adenosine analog, and it has a broad-spectrum antiviral activity against paramyxoviruses, flaviviruses, and coronaviruses. Remdesivir is associated with decreased hospital stay and improved outcomes in coronavirus- disease 2019 (COVID-19). Of 846 suspected COVID-19 patients admitted to the hospital, 612 SARS-CoV-2 nasopharyngeal RT-PCR positive patients were evaluated for enrollment in this prospective cohort study. 159 RT-PCR positive patients were given remdesivir. Their clinical, biochemical parameters, hospital stay, and outcomes related to morbidity and mortality were followed. Out of the 159 patients, 141 recovered after remdesivir use. The Chi-square test for independence examined the relation between the day of the first dose, dose of remdesivir, and clinical outcome. The standardized case fatality ratio (CFR) in the 453 hospitalized patients who did not receive remdesivir was 32.89% (N = 149) as compared to 11.32% (N = 18) in the patients who received remdesivir. These findings are in keeping with the therapeutic value of remdesivir in symptomatic SARS-CoV-2 infection of varying severity. The use of remdesivir is associated with a decrease in the severity of the SARS-CoV-2 infection. Its use is also associated with a decreased length of hospital stay and lower mortality than the patients who did not receive remdesivir.


Prevalence and predictors of long-delayed (> 120 h) chemotherapy-induced nausea and vomiting (CINV)-a systematic review and individual patient data meta-analysis.
Although there have been reports of chemotherapy-induced nausea and vomiting (CINV) beyond 120 h, its overall prevalence has not been systematically examined. The aim of this review and meta-analysis was to report on the prevalence of this long-delayed CINV. This review was registered on PROSPERO (CRD42022346963). PubMed (Medline), Embase, and Cochrane Central were searched from inception until August 2022. Articles were included if they reported on CINV > 120 h after initiation of the chemotherapy regimen and patients received a single-agent highly emetogenic (HEC) or moderately emetogenic (MEC) antineoplastic agent for 1 day alone or in combination with low/minimal emetogenic chemotherapy. For all eligible articles, individual study authors were contacted and requested to provide individual patient-level data of demographics, emetogenicity of chemotherapy regimens, and daily incidence of nausea and vomiting. Forward stepwise logistic regression identified predictors for the incident day's CINV based on prior day's CINV episodes, controlling for patient demographics, and stratified by regimen emetogenicity. A total of 2048 patients from 2 studies were included in this individual patient data meta-analysis: 1333 patients (65%) received HEC and 715 (35%) received MEC. Among those receiving HEC, 325 (24%) experienced acute, 652 (49%) delayed, and 393 (31%) long-delayed nausea; 107 (8%) experienced acute, 179 (14%) delayed, and 79 (6%) long-delayed vomiting. Among those receiving MEC, 48 (7%) experienced acute, 272 (38%) delayed, and 167 (24%) long-delayed nausea; 12 (2%) experienced acute, 97 (14%) delayed, and 42 (6%) long-delayed vomiting. Nausea in the long-delayed phase was as severe as in the delayed phase. Patients experiencing nausea and vomiting on days 4 and 5 were at significant risk of experiencing long-delayed CINV. While not as prevalent as delayed nausea and vomiting, long-delayed CINV affects a significant proportion of patients and severity is similar. Patients with delayed CINV, specifically on days 4-5, are at risk of experiencing long-delayed CINV.


Akt blockade downregulates collagen and upregulates MMP1 in human dermal fibroblasts.
Acutely transforming retrovirus AKT8 in rodent T-cell lymphoma (Akt) is a serine/threonine kinase that plays important roles in survival, cell-cycle progression, and cell proliferation, and has recently been implicated in collagen regulation. The aim of this study was to determine the role of Akt in collagen deposition by normal dermal fibroblasts, and to determine the sensitivity of cultured systemic sclerosis (SSc) fibroblasts to Akt inhibition. We show that blockade of Akt using pharmacological inhibitors, small interfering RNA (siRNA), and a dominant-negative Akt mutant led to inhibition of the basal type I collagen production. Furthermore, inhibition of Akt upregulated basal matrix metalloproteinase 1 (MMP1) production and reversed the inhibitory effect of transforming growth factor-beta (TGF-beta) on MMP1 gene expression. In addition, SSc fibroblasts were more sensitive to Akt inhibition, with respect to collagen and MMP1 production. These findings suggest that in human dermal fibroblasts, Akt has dual profibrotic effects, increasing collagen synthesis and decreasing its degradation via downregulation of MMP1. Akt could directly contribute to elevated collagen in SSc fibroblasts and it may represent an attractive target for therapy of SSc fibrosis.


Scoring and identifying organism-specific functional patterns and putative phosphorylation sites in protein sequences using mutual information.
The co-variance of amino acid positions within a multiple alignment of 294 protein kinases from mammals, plants, and bacteria was studied. Applying mutual information (MI), characteristic amino acid sites have been identified markedly discriminating the different organisms. The relation of surface accessibility of these sites in the 3D structure of a kinase and their MI content is studied. We extended the method to score a predicted phosphorylation site of this highly conserved catalytic protein kinase region. Based on this score mammalian and plant protein kinases were grouped together apart from the bacterial kinases. Thus, the presented method allows us to analyse putative phosphorylation sites in the context of their organism-specific origin.


Endoscopic Resection of Lateral Synovial Cyst of the Knee.
Synovial cysts on the lateral side of the knee can cause pain, limitation of joint mobility, compression of the neighboring neurovascular structures, rupture leading to compartment syndrome, infection, erosion of adjacent bone, and iliotibial band friction syndrome. Open resection is commonly performed for symptomatic cysts. We describe an alternative, endoscopic approach to cyst resection. This has the advantages of minimally invasive surgery of smaller wounds, better cosmesis, and less surgical trauma.


Name and shame: can taxonomists agree on systematic reforms?
The Linnean nomenclatural system dominates in taxonomy, despite various ethical issues being raised regarding the rules applied to this approach. Some recent proposals to adapt the system have been dismissed as destabilizing by many taxonomists. However, simple rule changes and amendments to address these issues could be implemented without excessive disruption.


Protein-Losing Enteropathy Associated with Thrombotic Microangiopathic Features Revealing Hirschsprung's Disease in a Child.
We report the case of a 4-year-old boy with protein-losing enteropathy, leading to severe malnutrition. Associated thrombotic microangiopathic features made diagnosis difficult and challenging.


Development of a transferrin receptor-targeting HVJ-E vector.
The development of more effective cancer treatments is anticipated. Tumor-targeted drug delivery is an important strategy in cancer therapy. We have developed an HVJ (hemagglutinating virus of Japan; Sendai virus) envelope (HVJ-E) vector using inactivated Sendai virus. The HVJ-E vector has been observed to target a number of cell lines since its hemagglutinin-neuraminidase (HN) protein recognizes the sialic acids of host cells. Thus, to reduce non-specific binding of the HVJ-E vector, we eliminated HN protein using HN-specific short interfering RNA (siRNA). Then, to further increase its tumor-targeting ability, we constructed HN-depleted HVJ containing the F-transferrin chimeric protein. The modified vectors containing Q-dots demonstrated 32-fold greater tumor-targeting efficiency than wild-type HVJ-E.


Pharmacokinetics and side-effects of clonidine.
A single oral dose of clonidine 300 microgram was administered to 8 healthy, normotensive subjects and the time course of its plasma concentrations was followed for 24 h. The plasma concentration of clonidine rose to a peak of 1.17 +/- 0.12 ng/ml at about 2 h: the absorption half-life was 0.6 +/- 0.2 h. Elimination followed first order kinetics with a half-life of 7.7 +/- 2.0 h. The correlation between the two most common side-effects of clonidine, sedation and dryness of the mouth, with the time course of its plasma concentrations was highly significant, p less than 0.01. All the subjects complained of severe sedation. During continuous administration of clonidine (75 microgram t.i.d.) for one week a steady state serum level of 0.30-0.35 ng/ml was achieved. One 75 microgram tablet of clonidine raised the serum level to about 0.69 +/- 0.13 ng/ml in two hours. After cessation of dosing, the serum level declined with a half-life of 7.5 +/- 1.5 h. The urinary excretion of unchanged clonidine was found to be about one third of the administered dose in 24 h during continuous administration and in the first 24 h after the single oral dose.


A longitudinal study of digit ratio (2D:4D) and other finger ratios in Jamaican children.
It has been hypothesised that the ratio between the length of the 2nd and 4th digits (2D:4D) is a correlate of prenatal sex steroids, and this relationship is strongest for the right hand. Furthermore, it has been suggested that 2D:4D is sexually dimorphic, the dimorphism is determined early, and 2D:4D among children is stable with growth. Here, we present the first longitudinal study of right and left hand 2D:4D. Our sample was 108 (54 males) Jamaican children. The first measurements were made in 1998 when mean age was 9.68 +/- 1.39 years, and a second set of measurements were made in 2002. We found that: (i) there was a small increase in 2D:4D with age which was lowest in the right hand; (ii) 2D:4D was sexually dimorphic, the means for males and females differed in the same direction in the 1998 and 2002 samples, and the sex difference was significant in the 1998 but not in the 2002 sample; (iii) the correlation between the 1998 and 2002 measurements of 2D:4D was high, indicating that rank order of the ratio was stable across year groups; and (iv) the rate of change in 2D:4D did not differ significantly across year groups. We conclude that 2D:4D increases slightly with age in children with the effect less marked for the right hand (i.e. the hand which is likely to show the strongest association with prenatal steroids), 2D:4D is sexually dimorphic from an early age, and the rank order of 2D:4D is stable in children. We discuss the implications of our findings for the status of 2D:4D as a correlate of prenatal sex steroids. The patterns of change in other finger ratios are also considered.


Environmental contaminants in Canadian shorebirds.
Canadian shorebirds are exposed to environmental contaminants throughout their annual cycle. Contaminant exposure among species varies with diet, foraging behaviour and migration patterns. We sampled twelve species of shorebirds from four locations across Canada to assess their exposure to PCBs, organochlorine pesticides, as well as four trace elements (Hg, Se, Cd, As). SigmaPCB and SigmaDDT followed by SigmaCHL were most frequently found above trace level in the shorebird carcasses. In general, the plover species (American golden, semipalmated, black-bellied) appear to be the most contaminated with organochlorines, whereas Hudsonian and marbled godwits appear to be the least contaminated. Among adult birds, the greater and lesser yellowlegs had the highest hepatic Hg concentrations (2.4-2.7 microg g(-1) dw), whereas American golden plovers as well as Hudsonian and marbled godwits contained relatively low levels of Hg (<1 microg g(-1) dw). Renal Se concentrations varied from 3.2 to 16.7 microg g(-1) dw and exhibited little interspecific or seasonal variation. Renal Cd levels in adult birds were highest in Hudsonian godwits from Quill Lakes (43 microg g(-1) dw) and Cape Churchill (12 microg g(-1) dw), and lowest (0.8-1.5 microg g(-1) dw) in greater and lesser yellowlegs from Cape Churchill and Bay of Fundy. Renal As concentrations varied from 0.06 microg g(-1) dw in golden plovers from Cape Churchill to 4.6 and 5.1 microg g(-1) dw in dunlin samples from the Pacific coast. There is no evidence that contaminants were adversely affecting the shorebirds sampled from the Canadian locations in this study.


Variation in amobarbital metabolism: evaluation of a simplified population study.
Kinetic constants of amobarbital metabolism were established for 52 subjects on the basis of urinary analysis extending over several days, usually 96 hr. There was no evidence of effect of age or sex on any of the constants. C-Hydroxylation was induced by cigarette smoking as much as 100%, but glucosidation was not affected. A factor influencing the constants was ethnicity of subjects (Caucasian or Oriental). This study confirms ethnic differences in amobarbital metabolism that were reported after measuring the concentration of metabolites in single samples of urine, that is, urine specimens voided during the postdistributive phase after oral drug intake. It appears that extreme simplification of sampling methods may be contemplated in the design of metabolic investigations of populations.


Direct aerobic epoxidation of alkenes catalyzed by metal nanoparticles stabilized by the H5PV2Mo10O40 polyoxometalate.
Ag and Ru nanoparticles stabilized by H5PV2Mo10O40, prepared by a sequence of redox reactions and supported on alpha-alumina, were effective catalysts for the direct aerobic epoxidation of alkenes in the liquid phase.


Aligned electrospun fiber film loaded with multi-enzyme mimetic iridium nanozymes for wound healing.
A film with elaborate microstructures that offers biomimetic properties and multi functionalities is highly desired in wound healing. Here, we develop an aligned hydrogel fiber film integrated with multi-active constituents to promote wound healing. Such fiber films are designed and constructed by photo-crosslinking the methacrylate gelatin (GelMA) doped with silver nanoparticles (Ag NPs) and iridium nanoparticles coated with polyvinylpyrrolidone (PVP-Ir NPs) in the precursor solution using electrospinning. The nature of GelMA hydrogel and the aligned arrangement of nanofibers endow the film with high-water content, self-degradability, improved bionic characteristics, oriented cell growth, and improved cell proliferation and migration. Moreover, the encapsulated nanozymes and Ag NPs offer the fiber film with superior reactive oxygen species (ROS) scavenging and antibacterial capability. The infected wound model shows that the multi-active hydrogel fiber film can reduce inflammation by killing bacteria and decomposing ROS, which accelerates the growth of new blood vessels and granulation tissue. Benefitting from these features, the versatile aligned GelMA fiber film demonstrates the clinically translational potential for wound healing.


Chelating bis(thiophosphinic amidate)s as versatile supporting ligands for the group 3 metals. An application to the synthesis of highly active catalysts for intramolecular alkene hydroamination.
Bis(thiophosphinic amidate) complexes (i.e., 1) of representative group 3 and lanthanide metals have been quantiatively prepared in situ from the corresponding thiophosphinic amides and Ln[N(TMS)2]3. These unusual pentacoordinate complexes exhibit very high activity as catalysts for intramolecular alkene hydroamination.


The relationship between spermine content of human milk during the first postnatal month and allergy in children.
Qualitative case study and mathematical model. Belgium. To evaluate the correlation between the polyamine mean concentration of the milk drunk during the first postnatal month and the appearance of allergy in children who drank this milk. A model that describes the dependence of the allergy appearance with the spermine mean concentration of milk drunk during the first postnatal month was established. This model shows that 5.02 nmol ml(-1) of spermine is a critical value to prevent the appearance of allergy.


Interleukin 6 modulates PPARα and PGC-1α and is involved in high-fat diet induced cardiac lipotoxicity in mouse.
Interleukin 6 (IL-6) may be involved in regulation of cardiac lipid metabolism and mitochondrial function through its influence on peroxisome proliferator-activated receptors (PPARs). In this study we evaluated the impact of the physiological level of IL-6 on the expression of PPARα and PGC-1α in the heart and the effect of lack of this cytokine on high-fat diet (HFD) induced lipotoxicity. Male C57BL6/J wild type (WT) and IL-6 knock-out (IL-6KO) mice were used. 20 animals of each genotype were fed with HFD for 15-18weeks. Cardiac function was assessed using echocardiography and cardiomyocyte ultrastructure was examined using electron microscopy. QT-PCR and Western blotting were applied to estimate the expression of PPARα and PGC-1α at the transcriptional and protein levels. At baseline WT and IL-6KO mice had similar size and function of the left ventricle. HFD induced similar left ventricular hypertrophic response in both groups without causing heart failure, but only WT animals had increased resting ejection fraction of the LV. Ultrastructure of HFD groups showed markers of lipotoxicity, that were more pronounced in IL-6KO group. In basal conditions IL-6KO animals had lower PPARα and similar PGC-1α expression as compared to WT. HFD induced downregulation of both PPARα and PGC-1α in WT animals, while in IL-6KO mice this effect was constrained. IL-6 is involved in basal regulation of PPARα and PGC-1α expression in cardiomyocytes. The lack of this cytokine promotes high-fat diet induced lipotoxicity but without overt manifestations of cardiac failure.


Antagonists of substance P from emphasis on position 11.
Ten analogs of substance P (SP) were designed and synthesized. The agonist and antagonist activities against SP were assayed on the isolated guinea pig ileum. The prime designs were changes of the important Met11 to include Leu11, Thr11, D-Leu11 and D-Ala11. Step-wise designs of changing D-Arg1 and D-Pro2 to the corresponding L-configurations resulted in decreasing antagonist activity. Changing Leu11 to D-Leu11 and D-Ala11 reduced antagonist activity. [D-Arg1,D-Pro2,D-Trp7,D-Trp9,Leu11]-SP is the most potent antagonist of this group of analogs, and required a 100-fold increase in the concentration of SP to give 50% of the maximal response caused by SP.


Systemic minor histocompatibility antigen expression in blood endothelial cells prevents T cell-mediated vascular immunopathology.
Attenuation of T cell-mediated damage of blood endothelial cells (BECs) in transplanted organs is important to prevent transplant vasculopathy (TV) and chronic rejection. Here, we assessed the importance of minor histocompatibility antigen (mHA) distribution and different coinhibitory molecules for T cell-BEC interaction. A transgenic mHA was directed specifically to BECs using the Tie2 promoter and cellular interactions were assessed in graft-versus-host disease-like and heterotopic heart transplantation settings. We found that cognate CD4(+) T-cell help was critical for the activation of BEC-specific CD8(+) T cells. However, systemic mHA expression on BECs efficiently attenuated adoptively transferred, BEC-specific CD4(+) and CD8(+) T cells and hence prevented tissue damage, whereas restriction of mHA expression to heart BECs precipitated the development of TV. Importantly, the lack of the coinhibitory molecules programmed death-1 (PD-1) and B and T lymphocyte attenuator fostered the initial activation of BEC-specific CD4(+) T cells, but did not affect development of TV. In contrast, TV was significantly augmented in the absence of PD-1 on BEC-specific CD8(+) T cells. Taken together, these results indicate that antigen distribution in the vascular bed determines the impact of coinhibition and, as a consequence, critically impinges on T cell-mediated vascular immunopathology.


Experimental and Theoretical Investigation on the Extractive Mass Transfer of Eu3+ Ions Using Novel Amide Ligands in 1-Hexyl-3-methylimidazolium Bis(trifluoromethylsulfonyl)imide.
Novel amide ligands in the ionic liquid (1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide) were utilized for the liquid-liquid biphasic mass transfer of Eu3+ ions from aqueous acidic waste solution. The cation exchange mechanism was found to be operative with the formation of [Eu(NO3)2L3]+ species (L = 4-chloro-N-(1-methyl-1H-pyrazol-3-yl)picolinamide). However, the presence of an inner-sphere water molecule was revealed by density functional theory (DFT) calculations. The viscosity-induced slower kinetics was evidenced during mass transfer, which was improved by increasing temperature. The process was exothermic in nature. The improvement in the kinetics of extractive mass transfer at higher temperatures is evinced by a reduction in the distribution ratio value. The spontaneity of the reaction was evidenced through the negative Gibbs free energy value, whereas the process enhances the entropy of the system, probably by releasing water molecules at least partially during complexation. The structures of bare ligands and complexes have been optimized by using DFT calculations. A high value of complexation energy, solvation energy, and associated enthalpy and free energy change reveal the efficacy in binding Eu with O and N donor atoms. In addition, natural population analysis, atoms-in-molecules analysis, and energy decomposition analysis have been employed to explore the nature of bonding existing in Eu-O and Eu-N bonds.


Reply to Jones et al. Comment on "Suresh et al. The Short-Term Effects and Tolerability of Low-Viscosity Soluble Fibre on Gastroparesis Patients: A Pilot Clinical Intervention Study. Nutrients 2021, 13, 4298".
We would like to thank Jones and colleagues for taking the time to comment [...].


[Topical beclomethasone dipropionate (BDP) in intestinal inflammatory diseases: the results of a multicentre trial].
The authors assessed the efficacy and tolerability of BDP in an open protocol using rectal enemas and suppositories and in a double-blind protocol vs mesalazine using rectal enemas. A total of 47 patients suffering from ulcerous rectocolitis were enrolled in the study and treated for 42 days while undergoing endoscopic, histologic and clinical controls. In conclusion, the authors affirm that BDP may represent a useful new therapeutic instrument in the treatment of slight to moderately severe forms of inflammatory intestinal disease.


Repressive effect of Rhus coriaria L. fruit extracts on microglial cells-mediated inflammatory and oxidative stress responses.
Rhus coriaria L. represents a herbal shrub that is used widely in traditional medicine in the Middle East region to treat different diseases including inflammation-related disorders. R. coriaria extracts have been well characterized in terms of their biological activities, pharmacological potential and phytochemical components. However, the effect of R. coriaria on neuro-inflammation has not been studied previously in detail. In the present study, we performed a qualitative phytochemical analysis and investigated the antioxidant and anti-neuro-inflammatory potential of R. coriaria extracts on BV-2 microglial cells. R. coriaria extracts were prepared using two different solvents: distilled water and ethanol. Phytochemical screening was performed to determine the principal bioactive components. The radical scavenging activity was assessed by DPPH method (2,2-diphenyl-1-picrylhydrazyl). The effect of R. coriaria on neuro-inflammation was studied upon measuring the production of oxidative stress and inflammatory factors using DCF (2',7'-dichlorofluorescein) and Nitric oxide (NO) assays respectively, and by analyzing the mRNA (TNFα, IL-10, iNOS and COX-2) and protein (NFκβ) levels of genes involved BV-2 microglia cells-mediated inflammation using quantitative Real Time PCR and Western blot, respectively. We found that R. coriaria extracts contain high phenolic and flavonoid contents. Interestingly, the ethanolic extract exerted a potent anti-inflammatory potential on insulted BV-2 cells manifested by: i) inhibition of Reactive Oxygen species (ROS) production and nitric oxide (NO) release; ii) suppressing TNFα, iNOS and COX-2 mRNA levels; iii) reducing NFκβ activation; and iiii) enhancing IL-10 transcription levels. Our results indicate that the neuro-inflammation inhibitory activity of R. coriaria extracts involves the inhibition of NF-κB signaling pathway. These findings suggest that R. coriaria might carry therapeutic potential against neurodegenerative diseases.


Expression patterns of class I histone deacetylases in osteosarcoma: a novel prognostic marker with potential therapeutic implications.
Epigenetic aberrations are recognized as having pivotal roles in cancer etiology and progression. Histone deacetylases are among the most studied epigenetic modulators in various cancer types. The expression levels of class I histone deacetylase isoforms 1, 2, and 3 in patient-derived primary osteosarcoma cells (6 cases) was investigated, comparing them to normal bone graft-derived osteoblasts (6 cases) using the immunoblotting technique. Expression profiles of histone deacetylases in high-grade osteosarcoma tissue of 89 patients were examined and their association with clinicopathologic parameters and the patient survival was evaluated. Histone deacetylases were immunohistochemically stained on formalin-fixed paraffin-embedded biopsied tissue. Primary osteosarcoma cells expressed higher levels of histone deacetylase 1 and histone deacetylase 2, but lower levels of histone deacetylase 3 compared to benign osteoblasts. Overall, 82, 99, and 93% of 89 osteosarcomas showed nuclear expression of the histone deacetylase isoforms 1, 2, and 3, respectively. Low levels of histone deacetylase 1 were significantly associated with a high Enneking stage (P=0.014) and the presence of initial metastasis (P=0.040), while low levels of histone deacetylase 3 were significantly correlated with age >15 years (P=0.026). Univariate survival analysis found significantly shorter survival in the patients with a high Enneking stage (P<0.001), axial location (P=0.009), presence of initial metastasis (P<0.001), low-histone deacetylase 1 expression (P=0.038), and low-all-histone deacetylases expression (P=0.016). Multivariate survival analysis showed that only axial location (P=0.011) and low-all-histone deacetylases expression (P=0.039) were independent prognostic factors. In subgroup analysis of stage IIB patients (n=45), only axial location and low-all-histone deacetylases expression were associated with shorter survival in both univariate and multivariate analysis (axial location, P=0.008 and 0.010; low-all-HDACs, P=0.013 and 0.038, respectively). Low levels of all-histone deacetylases expression were significantly associated with advanced disease status and short survival. These findings may be a guide to future use of histone deacetylase inhibitors in osteosarcoma patients.


Evidence for association between vitamin D receptor BsmI polymorphism and type 1 diabetes in Japanese.
Type 1 diabetes is considered to be T-helper 1 (Th1) type autoimmune disease. Because the vitamin D receptor is expressed on CD4+T cells and is known to affect cytokine responses, several groups have investigated the association between the vitamin D receptor gene BsmI polymorphism and type 1 diabetes. However, this issue is still controversial; therefore, we examined this gene polymorphism in a large number of type 1 diabetic patients as a multi-center collaborative study in Japan. A total of 1,373 subjects, including 774 cases and 599 control subjects of Japanese origin, were studied. The frequency of carriers of the BB genotype in type 1 diabetic patients was significantly higher than that in controls (p<0.01, odds ratio 3.65). Moreover, IFN-gamma production upon anti-CD3 stimulation in the BB genotype group was significantly higher than that in the Bb and bb genotype groups (p<0.05), suggesting that the polyclonal T cell response in BB genotype patients is Th1 dominant. Based upon these results, we propose that it may be worthwhile to focus on subjects with the BB genotype of this gene polymorphism as having high risk for type 1 diabetes.


Cutting-edge mass spectrometry strategy based on imaged capillary isoelectric focusing (icIEF) technology for characterizing charge heterogeneity of monoclonal antibody.
Imaging capillary isoelectric focusing (icIEF) technology has been becoming the gold criteria of monitoring monoclonal antibody (mAb) charge heterogeneity that is one of the major product-related variants in recombinant biopharmaceuticals, since the first commercial instrument developed twenty years ago. However, the protein identification in icIEF separation is just based on isoelectric point (pI) measurement of protein. Although high resolution mass spectrometry (HRMS) is currently the most powerful means of qualitative protein analysis, traditional icIEF cannot compatibly be used in conjunction with MS due to the use of less volatile reagents. In addition, protein heterogeneity characterization in depth such as peptide mapping by high performance liquid chromatography (HPLC) requires the focused protein bands to be collected as fractions after the icIEF separation, which is a great challenge in biopharmaceutical discovery. In this work, pembrolizumab was employed as targeting mAb (a highly selective anti-PD-1 humanized mAb), an integrated icIEF platform was developed including analytical profiling, MS coupling and fraction collections for charged variant preparation. Multiple operation modes can be rapidly and flexibly switched just by changing customized capillary separation cartridges without more configurations. Main component, four acidic variants (A1-A4) and three basic variants (B1-B3) were baseline separated then directly detected by icIEF-HRMS online coupling for rapid screening of intact protein heterogeneity where reliable and accurate molecular weight of protein charged variants were obtained. Next, by installing preparative capillary separation cartridge, fractions of major charge variants (A2-3 and B1-2) and main component were collected for following LC-MS peptide mapping characterization. The whole workflow of icIEF-based MS strategy for protein heterogeneity is straight forward, reliable and accurate, which provides a comprehensive and revolutionary technology for protein drug quality control (QC) monitoring, MS coupling for fingerprinting intact protein and HPLC-MS peptide mapping in depth.


Breast volume replacement using the latissimus dorsi miniflap.
Mastectomy is often recommended to women with early breast cancer who have large tumours or where the breast volume requiring resection to achieve adequate tumour clearance is too great to allow for a satisfactory cosmetic result after breast conservation surgery. The use of a latissimus dorsi muscular flap (latissimus dorsi miniflap (LDMF)) to replace the volume loss after major breast sector resection is an option where the tumour to breast volume ratio is large. The present study describes the technique and evaluates the experience of the LDMF at Royal Adelaide Hospital, Adelaide, Australia. Between August 1997 and April 2002, 18 women aged 37-64 years underwent wide local excision for primary breast cancer with LDMF reconstruction. Tumour characteristics, breast specimen weight and postoperative sequelae were assessed. Quality of life measurements and objective assessments of aesthetic outcome were evaluated. Tumour diameter ranged from 13 to 80 mm (median 30 mm). Nine patients had multifocal or extensive intraductal component positive tumours. The weight of the resected specimens ranged from 75 to 395 g (median 130 g). There were no major postoperative complications, with a range of inpatient stay of 3-10 days. Seromas were aspirated in 14 patients but did not delay adjuvant treatment. Quality of Life results showed high patient satisfaction in all but one patient. A satisfactory cosmetic result was achieved in all but one patient who subsequently required mastectomy. The LDMF procedure allows breast conservation to be achieved in women with large tumour to breast volume ratios, with satisfactory resection margins and good cosmetic and functional results. In the present experience standard oncological adjuvant treatment is not compromised.


Spectrophotometric determination of hyoscine butylbromide and famciclovir in pure form and in pharmaceutical formulations.
A simple, rapid, and extractive spectrophotometric method was developed for the determination of hyoscine butylbromide (HBB) and famciclovir (FCV) in pure and pharmaceutical formulations. These methods are based on the formation of yellow ion-pair complexes between the basic nitrogen of the drug and four sulphonphthalein acid dyes, namely; bromocresol green (BCG), bromothymol blue (BTB), bromocresol purple (BCP) and bromophenol blue (BPB) in phthalate buffer of pH range (3.0-3.5). The formed complexes were extracted with chloroform and measured at 420, 412, 409 and 415nm for HBB and at 418, 412, 407 and 414nm for FCV using BCG, BTB, BCP and BPB, respectively. The analytical parameters and their effects on the reported systems are investigated. Beer's law was obeyed in the range 1.0-20microgmL(-1) with correlation coefficient (n=6)> or =0.9997. The molar absorptivity, Sandell sensitivity, detection and quantification limits were also calculated. The composition of the ion pairs was found 1:1 by Job's method in all cases and the conditional stability constant (Kf) of the complexes have been calculated. The free energy changes (DeltaG) were determined for all complexes formed. The proposed methods have been applied successfully for the analysis of the studied drugs in pure and pharmaceutical formulations with percentage recoveries ranges from 99.84 to 100.26. The results were in good agreement with those obtained by the official methods.


Lysolecithin acyltransferase and lysolecithin: lysolecithin acyltransferase in adult rat lung alveolar type II epithelial cells.
1. The specific activity of lysolecithin acyltransferase (EC 184.108.40.206) in sonicated adult rat lung alveolar type II epithelial cells, measured either alone or in combination with acyl-CoA synthetase (EC 18.104.22.168), was found to be an order of magnitude greater than that of lysolecithin:lysolecithin acyltransferase. 2. Lysolecithin acyltransferase in type II cells was found to prefer palmitoyl-CoA over oleoyl-CoA as substrate. The combination of lysolecithin acyltransferase and acyl-CoA synthetase was found to prefer palmitate over oleate for incorporation into phosphatidylcholine. 3. Compared to whole lung homogenate, sonicated adult rat type II cells are highly enriched in lysolecithin acyltransferase but not in lysolecithin:lysolecithin acyltransferase. 4. These observations indicate that in normal adult rat type II cells the deacylation-reacylation cycle is more important for the formation of dipalmitoyl phosphatidylcholine than the deacylation-transacylation process.


Contribution of odd-numbered fatty acid oxidation to propionate production in neonates with methylmalonic and propionic acidaemias.
Newborns with disturbed propionate metabolism (propionic and methylmalonic acidaemias) accumulate during fetal life large amounts of odd-numbered long-chain fatty acids in adipose tissue (8% to 10% of total fatty acids vs 1% in normal controls) and other body lipids. During periods of acute catabolism, such as in the early days of life, mobilization and oxidation of odd-chain fatty acids from adipose tissue yield extensive amounts of toxic propionyl-CoA in the mitochondria and thus might contribute to the severe illness of the patients. Our data suggest that not only catabolism of protein but also that of adipose tissue has to be avoided in order to alleviate the neonatal illness of patients with propionic and methylmalonic acidaemias. It seems prudent to avoid prolonged fasting during the 1st year of life.


DNA damage: a trigger of innate immunity but a requirement for adaptive immune homeostasis.
Chromosome breakage is frequently associated with viral infection and cellular transformation, but it is also required for two processes that are crucial for the development and function of adaptive immunity: V(D)J recombination and class-switch recombination. The cellular responses that result from this type of DNA damage, which are mostly activated by the protein kinase ataxia-telangiectasia mutated (ATM), lead to cell-cycle arrest at several checkpoints and efficient DNA repair. This Review focuses on the important roles of these DNA-damage responses in the activation of innate immunity and the targeting of the innate immune response to infected or transformed cells, as well as in the development and function of adaptive immunity.


Effect of cyclosporin A on daunorubicin accumulation in multidrug-resistant P388 leukemia cells measured by real-time flow cytometry.
We investigated the mode of action of cyclosporin A (Cy-A) as a modifier of multidrug resistance in P388 mouse leukemia cells. A fluorescence-activated flow cytometer (FCM) was modified with a flow-through cuvette to allow continuous on-line monitoring of daunorubicin uptake in vitro. The addition of Cy-A to multidrug-resistant P388/R cells at steady-state daunorubicin uptake, led to a dose-dependent increase in cellular daunorubicin accumulation, as measured by FCM and high-performance liquid chromatography (HPLC). A linear relationship was found between the daunorubicin concentration in the incubation medium and the Cy-A concentration required for optimal stimulation of cellular anthracycline accumulation. The results of a cytotoxicity assay indicated that Cy-A completely restored the chemosensitivity of the P388/R cells. Intracellular Cy-A measurements in P388/S and P388/R cells showed that P388/R cells accumulated significantly less Cy-A than P388/S cells. Relatively high daunorubicin concentrations could not restore that accumulation defect. These results suggest that Cy-A promotes cellular anthracycline accumulation by competing for an outward drug-transport system that operates in multidrug-resistant cells.


Palliative surgery for unresectable carcinoma of the head of the pancreas, ampulla, and distal end of the common bile duct in Japan. A statistical review.
To elucidate the approximate present status of palliative surgery for unresectable carcinoma of the head of the pancreas, ampulla, and distal end of the common bile duct in Japan, surgical methods in 151 institutions were collected and analyzed. Biliary drainage alone was performed in 144 institutions (95 percent) and in 80 of them, cholecystectomy and choledochojejunostomy with a Roux-Y procedure was adopted. Gastric bypass with biliary drainage was routinely performed in 13 institutions (8.7 percent), whereas in 62 institution (41.3 percent), combined biliary and gastric drainage operations were performed as the primary procedures in selected cases. In over half the institutions, neither gastrectomy nor vagotomy was provided. Some means to alleviate the pain caused by tumor invasion were performed in about a third of the institutions (49), and about 80 percent of them added some manipulations of the celiac plexus.


Photoperiodic influence on the innervation of the ductus epididymidis and ductus deferens of the Djungarian hamster, Phodopus sungorus: electron-microscopic and biochemical results.
The influence of long (light:dark 16:8) and short (light:dark 8:16) photoperiods on the autonomic innervation of the ductus epididymidis and the ductus deferens of Phodopus sungorus was studied using electron microscopy with morphometric analyses, and biochemical methods. At short photoperiods, only the large smooth muscle cells in the ductus deferens became atrophic, the number of mainly adrenergic varicosities in the smooth muscle layer decreased, and the mean distance between varicosity and smooth muscle cells increased. The content of noradrenaline was determined by high-performance liquid chromatography with electrochemical detection. For the ductus deferens, the noradrenaline content was reduced at LD 8:16 to less than 10% of the initial value. Short photoperiods are proposed to influence only the adrenergic innervation of the large smooth muscle cells of the ductus deferens. These cells are believed to exert a trophic influence on their nerves.


Morphological characteristics of low-threshold primary afferents in the trigeminal subnuclei interpolaris and caudalis (the medullary dorsal horn) of the golden hamster.
Intra-axonal recording and horseradish peroxidase (HRP) injection techniques were employed to define the response characteristics of low-threshold, rapidly conducting trigeminal primary afferents and the morphological features of their axon arbors in subnucleus interpolaris and subnucleus caudalis (or the medullary dorsal horn; these last two terms are used synonomously throughout the paper). A total of 61 such afferents were characterized and recovered. Of these, ten gave rapidly adapting (RA) and 17 slowly adapting (SA type I) responses to vibrissa deflection. Twenty were sensitive to guard hair deflection and 14 were responsive to indentation of the hairy skin. The vibrissa-sensitive primary afferents were all quite similar morphologically. Primary collaterals proceeded directly, in a radial fashion, to their zone of termination and gave rise to dense and compact arbors. These tended to be larger in the medullary dorsal horn (MDH) than in interpolaris and they also gave rise to more boutons in the former nucleus. Guard hair afferents generally had smaller arbors and gave rise to fewer boutons than vibrissa-sensitive axons. Like vibrissa afferents, their arbor were generally circumscribed in both interpolaris and MDH, but they were larger in the latter nucleus. Skin-sensitive afferents had arbors that tended to be somewhat larger than those of vibrissa- or guard-hair-related fibers. Unlike the other fiber types, the arbors of skin-sensitive afferents were on average larger in interpolaris than MDH. Quantitative analysis of the morphological data from well-filled examples from each of these four functional types verified our qualitative impressions regarding differences between interpolaris and MDH collaterals of a given fiber-type. Statistical comparison of data from different functional classes indicated trends that supported our qualitative impressions, but none of these was statistically significant. The topography of the trigeminal primary afferent input to interpolaris was organized such that the head was inverted and fibers with caudal receptive fields terminated in the lateral portion of the nucleus. This was true for all of the functional afferent types that we examined. Vibrissa-related fibers differed from nonvibrissa afferents in that they tended to avoid the most rostral portion of interpolaris. In the MDH, the primary afferent representation of the head was also inverted, but fibers with caudal facial receptive fields tended to terminate medially rather than laterally.(ABSTRACT TRUNCATED AT 400 WORDS)


Global longitudinal strain by two-dimensional speckle tracking imaging predicts exercise capacity in patients with chronic heart failure.
Left ventricular ejection fraction (LVEF) predicts mortality in patients with chronic heart failure (CHF). However, a weak correlation was found between LVEF and peak oxygen uptake ([Formula: see text]) in CHF patients. Global longitudinal strain measured by two-dimensional (2D) strain is regarded as a more useful predictor of cardiac events than LVEF. We investigated whether 2D strain obtained at rest could predict peak [Formula: see text] in patients with CHF. Fifty-one patients (mean age of 54.0 ± 12.0 years, 14 females, LVEF 46.0 ± 15.0%) with stable CHF underwent resting echocardiography and cardiopulmonary exercise testing. Leg muscle strength was measured for the evaluation of peripheral factors. Global longitudinal strain (GLS) in the apical 4-, 3-, and 2-chamber views and global circumferential strain (GCS) in the parasternal mid short-axis view were measured. In all patients, peak [Formula: see text] correlated with leg muscle strength (r = 0.55, p < 0.0001), LVEF (r = 0.46, p < 0.001), GLS (r = -0.45, p < 0.001), and GCS (r = -0.41, p = 0.005), respectively. No significant correlation was found between the ratio of early transmitral velocity to peak early diastolic mitral annulus velocity (E/E') and peak [Formula: see text]. In the patients with heart failure and reduced LVEF, a multiple stepwise linear regression analysis based on leg muscle strength, LVEF, E/E', GLS, and GCS was performed to identify independent predictors of peak [Formula: see text], resulting in leg muscle strength and GLS (R (2) = 0.888) as independent predictors of peak [Formula: see text]. Global longitudinal strain at rest could possibly predict exercise capacity, which appeared to be more useful than LVEF, E/E', and GCS in CHF patients with reduced LVEF.


Identification of an anti-herpetic compound isolated from Pistacia vera L. male floral buds.
Due to the numerous side effects of conventional drugs against herpetic infections and the growing phenomenon of resistance, the researchers turned to natural compounds as a source of new drugs because they are less toxic than the synthetic molecules. This study aimed to analyse the activity of Pistacia vera L. male floral bud extracts, against the replication of herpes simplex virus type 2, as well as to investigate their mode of action, isolate, and identify the active compound. Cell viability and anti-herpes virus activity were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and the plaque reduction assay, respectively. Three extracts (ethanolic, aqueous and polysaccharide extracts) were tested, only aqueous and polysaccharide extracts had anti-herpetic activity with a selectivity index of 29.12 and 20.25, respectively. Investigation about the mechanism of action indicated that the two active extracts inhibited the virus replication by direct contact with virucidal selectivity indexes of 39.15 and 32.09, respectively. An active compound was isolated from the aqueous extract using TLC bio-guided assay: it was identified as gallic acid by high-performance liquid chromatography-diode array detection coupled with electrospray ionization mass spectrometry (HPLC-DAD-ESI-MSn). The antiviral activity of Pistacia vera L. has been previously shown. The selectivity index of gallic acid is much lower than that of the active extract from which it has been isolated. Therefore, we can consider the aqueous extract prepared from Pistacia vera L. male floral buds as a promising natural product for treating herpetic diseases.


Egg yolk lipoprotein, a new supplement for the growth of mammalian cells in serum-free medium : Egg yolk lipoprotein for serum-free growth of cells.
Egg yolk lipoprotein promoted growth of a wide variety of mammalian cell lines, including plasma-cytomas and epithelial cell lines, in serum-free medium. The lipoprotein was active for cell growth when used with insulin, transferrin, ethanolamine and selenite. The most active lipoprotein fraction (YLP-pI7.5) was purified to give a single peak by chromatofocusing and gel filtration, and was homogeneous on a 0.35% agarose gel electrophoretogram. The lipoprotein was characterised as a very low density lipoprotein with a protein content of only 1.3%. This lipoprotein had an optimal concentration of 300 μg/ml (4 μg protein/ml). It was easily separable from proteinous molecules secreted into the serum-free medium by the cells, since it floated on the surface of the medium after addition of ammonium sulfate, to precipitate protein, and centrifugation. An associated structure of lipid and protein seemed to be still necessary for the lipoprotein to exhibit a growth promoting activity.


Multiplexed Analysis of Peptide Functionality Using Lanthanide-based Structural Shift Reagents.
Functionally selective lanthanide-based ion mobility shift reagents are presented as a method to elucidate protein or peptide structural information as well as relative quantitation of protein expression profiles. Sequence information and site localization of primary amines (n-terminus and lysine), phosphorylation sites, and cysteine residues can be obtained in a data dependent manner using ion mobility-mass spectrometry (IM-MS). The high mass of the incorporated lanthanide ensures a significant shift of where the signal occurs in IM-MS conformation space. Peptide sequence information provided by the use of IM-MS shift reagents allows for both a more confident identification of peptides from complex mixtures and site localization following tandem MS experiments. Stable isotopes of the lanthanide series may also be used as relative quantitation labels since several lanthanides can be utilized in differential sample analyses.


Study on the interaction between cyanobacteria FBP/SBPase and metal ions.
Fructose-1,6-/sedoheptulose-1,7-bisphosphatase (FBP/SBPase) is a potential important target enzyme for finding inhibitors to solve harmful algal bloom. In this paper, the interactions between FBP/SBPase and metal ions were studied by enzyme activity analysis, fluorescence and molecular modeling method. The enzyme activity analysis showed that FBP/SBPase can be activated by Mg2+ or Mn2+ but cannot be activated by Ca2+ or Zn2+. Spectroscopic analysis of emission quenching showed that quenching mechanism of FBP/SBPase with Mg2+ or Mn2+ was static quenching mechanism while that of Ca2+ or Zn2+ was dynamic quenching process. Hydrogen bonds and van der Waals interaction might be the predominant intermolecular forces in stabilizing FBP/SBPase-Mg2+ while hydrophobic forces were the predominant intermolecular forces in stabilizing FBP/SBPase-Mn2+. Microenvironment and conformation of FBP/SBPase were changed in binding reaction. The effect of metal ions and important amino acid residues on FBP/SBPase-metal ion complex was also discussed by molecular modeling study.


Centrally Delivered BACE1 Inhibitor Activates Microglia, and Reverses Amyloid Pathology and Cognitive Deficit in Aged Tg2576 Mice.
Multiple small-molecule inhibitors of the β-secretase enzyme (BACE1) are under preclinical or clinical investigation for Alzheimer's disease (AD). Prior work has illustrated robust lowering of central amyloid β (Aβ) after acute administration of BACE1 inhibitors. However, very few studies have assessed the overall impact of chronically administered BACE1 inhibitors on brain amyloid burden, neuropathology, and behavioral function in aged preclinical models. We investigated the effects of a potent nonbrain-penetrant BACE1 inhibitor, delivered directly to the brain using intracerebroventricular infusion in an aged transgenic mouse model. Intracerebroventricular infusion of the BACE1 inhibitor (0.3-23.5 μg/d) for 8 weeks, initiated in 17-month-old Tg2576 mice, produced dose-dependent increases in brain inhibitor concentrations (0.2-13 μm). BACE1 inhibition significantly reversed the behavioral deficit in contextual fear conditioning, and reduced brain Aβ levels, plaque burden, and associated pathology (e.g., dystrophic neurites), with maximal effects attained with ∼1 μg/d dose. Strikingly, the BACE1 inhibitor also reversed amyloid pathology below baseline levels (amyloid burden at the start of treatment), without adversely affecting cerebral amyloid angiopathy, microhemorrhages, myelination, or neuromuscular function. Inhibitor-mediated decline in brain amyloid pathology was associated with an increase in microglial ramification. This is the first demonstration of chronically administered BACE1 inhibitor to activate microglia, reverse brain amyloid pathology, and elicit functional improvement in an aged transgenic mouse model. Thus, engagement of novel glial-mediated clearance mechanisms may drive disease-modifying therapeutic benefit with BACE1 inhibition in AD.


Protective antigen in the membranes of mouse erythrocytes infected with Plasmodium chabaudi.
A malarial antigen, Pc96, in the plasma membrane of erythrocytes infected with Plasmodium chabaudi has been identified. It is synthesized by the parasite and present during most of the growth stages of the intra-erythrocytic cycle as demonstrated by immunofluorescence. The antigen has a molecular weight of approximately 96,000. Monoclonal antibodies raised against this antigen were used to isolate the protein by affinity chromatography. Mice immunized with affinity-purified Pc96 were partially protected against blood induced-P. chabaudi infection. This result indicates the existence of a protective antigen in the membranes of erythrocytes parasitized by a rodent malaria and encourages the search for analogous antigens in human malaria parasites as possible candidate molecules for malaria vaccination.


Health classification of Meibomian gland images using keratography 5M based on AlexNet model.
We aim to present effective diagnostics in the field of ophthalmology and improve eye health. The purpose of this study is to examine the capability of health classification of Meibomian gland dysfunction (MGD) using Keratography 5M and AlexNet method. A total of 4,609 meibomian gland images were collected from 2,000 patients using Keratography 5M in the hospital. Then, MGD dataset for eyelid gland health recognition was constructed through image pre-processing, labelling, cropping and augmentation. Furthermore, AlexNet network was used to identify the eyelid gland health. The effects of different optimization methods, different learning rates, Dropout methods and different batch sizes on the recognition accuracy were discussed. The results show that the effect of model recognition is the best when the optimized method is Adam, the number of iterations is 150, the learning rate is 0.001, and the batch size is 80, then, the overall test accuracy of health degree is 94.00%. Our research provides a reference to the clinical diagnosis or screening of eyelid gland dysfunction. In future implementations, ophthalmologists can implement more advanced learning algorithms to improve the accuracy of diagnosis.


A field study to evaluate runoff quality from green roofs.
Green (vegetated) roofs are emerging as practical strategies to improve the environmental quality of cities. However, the impact of green roofs on the storm water quality remains a topic of concern to city planners and environmental policy makers. This study investigated whether green roofs act as a source or a sink of various metals (Na, K, Ca, Mg, Al, Fe, Cu, Cd, Pb, Zn, Mn, Cr, Ni, Li and Co), inorganic anions (NO3-, NO2-, PO4(3-), SO4(2-), Cl-, F- and Br-) and cation (NH4+). A series of green roof assemblies were constructed. Four different real rain events and several artificial rain events were considered for the study. Results showed that concentrations of most of the chemical components in runoff were highest during the beginning of rain events and subsided in the subsequent rain events. Some of the important components present in the runoff include Na, K, Ca, Mg, Li, Fe, Al, Cu, NO3-, PO4(3-) and SO4(2-). However, the concentration of these chemical components in the roof runoff strongly depends on the nature of substrates used in the green roof and the volume of rain. Based on the USEPA standards for freshwater quality, we conclude that the green roof used in this study is reasonably effective except that the runoff contains significant amounts of NO3- and PO4(3-).


Reversal of Osseointegration as a Novel Perspective for the Removal of Failed Dental Implants: A Review of Five Patented Methods.
Osseointegration is the basis of successful dental implantology and the foundation of cementless arthroplasty and the osseointegrated percutaneous prosthetic system. Osseointegration has been considered irreversible thus far. However, controlled heating or cooling of dental implants could selectively damage the bone at the bone-implant interface, causing the reversal of osseointegration or "osseodisintegration". This review compares five methods for implant removal, published as patent documents between 2010 and 2018, which have not yet been discussed in the scientific literature. We describe these methods and evaluate their potential for reversing osseointegration. The five methods have several technical and methodological similarities: all methods include a handpiece, a connecting device for coronal access, and a controlling device, as well as the application of mechanical and/or thermal energy. The proposed method of quantifying the temperature with a sensor as the sole means for regulating the process seems inadequate. A database used in one of the methods, however, allows a more precise correlation between a selected implant and the energy needed for its removal, thus avoiding unnecessary trauma to the patient. A flapless, microinvasive, and bone-conserving approach for removing failed dental implants, facilitating successful reimplantation, would benefit dental implantology. These methods could be adapted to cementless medical implants and osseointegrated percutaneous prosthetics. However, for some of the methods discussed herein, further research may be necessary.


Change in Seasonal Beliefs Mediates the Durability Advantage of Cognitive-Behavioral Therapy Over Light Therapy for Winter Depression.
In treating an acute episode of winter depression, cognitive-behavioral therapy for seasonal affective disorder (CBT-SAD) and light therapy are comparably efficacious, with improvement in depression symptoms during CBT-SAD mediated by reduced seasonal beliefs (i.e., maladaptive thoughts about the seasons, light availability, and weather). Here, we tested whether the enduring benefit of CBT-SAD over light therapy following treatment is associated with offsetting seasonal beliefs during CBT-SAD. Currently depressed adults with Major Depression, Recurrent with Seasonal Pattern (N = 177) were randomized to 6 weeks of light therapy or group CBT-SAD and followedup one and two winters after treatment. Outcomes measured during treatment and at each follow-up included depression symptoms on the Structured Clinical Interview for the Hamilton Rating Scale for Depression-SAD Version and Beck Depression Inventory-Second Edition. Candidate mediators measured at pre-, mid-, and posttreatment were SAD-specific negative cognitions (Seasonal Beliefs Questionnaire; SBQ); general depressogenic cognitions (Dysfunctional Attitudes Scale; DAS); brooding rumination (Ruminative Response Scale-Brooding subscale; RRS-B); and chronotype (Morningness-Eveningness Questionnaire; MEQ). Latent growth curve mediation models found a significant positive path from treatment group to the slope of SBQ during treatment, with CBT-SAD showing larger improvements in seasonal beliefs with overall change in seasonal beliefs in the medium-effect range, and significant positive paths from SBQ slope to depression scores at the first and second winter follow-ups, indicating greater change towards more flexible seasonal beliefs during active treatment was associated with less severe depression symptoms following treatment. Estimated indirect effects (treatment group → SBQ change * SBQ change → outcome) were also significant at each follow-up for each outcome with βindirect ranging from .091 to .162. Models also found significant positive paths from treatment group to the slope of MEQ and RRS-B during treatment, with light therapy showing a greater increase in "morningness" and CBT-SAD showing a greater decrease in brooding during active treatment; however, neither construct emerged as a mediator of follow-up depression scores. Change in seasonal beliefs during treatment mediates both the acute antidepressant and long-term effects of CBT-SAD and explains lower depression severity following CBT-SAD relative to light therapy.


Coordinate regulation of contractile protein synthesis during myoblast differentiation.
The synthesis of contractile proteins has been studied during the differentiation of quail skeletal muscle myoblasts in culture. Myoblast differentiation was synchronized by transferring secondary cultures of rapidly dividing myoblasts into medium lacking cell division-promoting factors. Cultures at various stages of differentiation were then pulse-labeled with 35S-methionine, and cell extracts were resolved by electrophoresis on two-dimensional gels. Incorporation into specific proteins was quantitated by autoradiography and fluorography using a scanning densitometer. Contractile proteins synthesized by muscle cultures were identified by their co-electrophoresis on two-dimensional gels with contracile proteins purified from quail breast muscle. Our results show that the synthesis of myosin heavy chain, two myosin light chains, two subunits of troponin and two subunits of tropomyosin is first detected at the time of myoblast fusion and then rapidly increase at least 500 fold to maximum rates which remain constant in muscle fibers. Both the kinetics of activation and the molar rates of synthesis of these contractile proteins are virtually identical. Muscle-specific actin (alpha) synthesis also increases at the time of myoblast fusion, but this actin (alpha) is synthesized at 3 times the rate of other contractile proteins. The synthesis of 30 other muscle cell proteins was quantitated, and most of these are shown to follow different patterns of regulation. From these results, we conclude that the contractile proteins are regulated coordinately during myoblast differentiation.


Triple antibiotics for the treatment of infected joint replacements. Case report and commentary.
Combination antibiotics are frequently used in the treatment of infected joint arthroplasties to achieve synergistic activity against the infecting organisms. A case is presented in which triple antibiotic therapy was used in the treatment of an infected hip arthroplasty. The antibiotic combination proved to have an antagonistic effect despite documented sensitivities to the individual drugs. Serum antimicrobial assays are recommended in the treatment of all prosthetic joint infections.