Simulation of fracture healing in the tibia: mechanoregulation of cell activity using a lattice modeling approach.
In this study, a three-dimensional (3D) computational simulation of bone regeneration was performed in a human tibia under realistic muscle loading. The simulation was achieved using a discrete lattice modeling approach combined with a mechanoregulation algorithm to describe the cellular processes involved in the healing process-namely proliferation, migration, apoptosis, and differentiation of cells. The main phases of fracture healing were predicted by the simulation, including the bone resorption phase, and there was a qualitative agreement between the temporal changes in interfragmentary strain and bending stiffness by comparison to experimental data and clinical results. Bone healing was simulated beyond the reparative phase by modeling the transition of woven bone into lamellar bone. Because the simulation has been shown to work with realistic anatomical 3D geometry and muscle loading, it demonstrates the potential of simulation tools for patient-specific pre-operative treatment planning.


The prevalence of Escherichia coli O157 and O157:H7 in ground beef and raw meatball by immunomagnetic separation and the detection of virulence genes using multiplex PCR.
The present study was conducted to investigate the presence of Escherichia coli O157 and O157:H7 strains and to detect the presence of the stx1, stx2, and eaeA genes in isolates derived from 200 samples (100 samples from fresh ground beef and 100 samples from raw meatball). The samples were purchased from the Samsun Province in Turkey, over a period of 1 year. Enrichment-based immunomagnetic separation and multiplex polymerase chain reaction were applied for these analyses. E. coli O157 was detected in five of the 200 (2.5%) samples tested (one isolated from ground beef and four from meatball samples), whereas E. coli O157: H7 was not detected in any sample. During the analysis, eight strains of E. coli O157 were obtained. The genes stx1, stx2, and eaeA were detected in two E. coli O157 isolates obtained from two meatball samples, whereas only the eaeA and the stx2 genes were detected in four E. coli O157 strains that were isolated from one meatball sample. None of the stx1, stx2, and eaeA was detected in the E. coli O157 isolates obtained from the ground beef and the one meatball samples.


Does ligation of the intersphincteric fistula tract raise the bar in fistula surgery?
The ligation of the intersphincteric fistula tract procedure has been reported to have high cure rates, with minimal impairment of continence. The aim of this study was to evaluate the success rates and functional outcome after the ligation of the intersphincteric fistula tract procedure. This study was performed as a retrospective review. The study was conducted at the Division of Colon and Rectal Surgery, University of Minnesota and at affiliated hospitals in Minneapolis and St. Paul, Minnesota, between March 2007 and September 2011. Ninety-three patients with transsphincteric cryptoglandular anal fistula were included. Ligation of the intersphincteric fistula tract procedure was performed. Failure was defined as persistent or recurrent drainage, air leakage from a patent external opening, or intersphincteric incision or reoperation for recurrent fistula. Success was defined as healing of the external fistula opening and intersphincteric incision. Patients were followed up with a questionnaire to assess the recurrence rate and the Wexner incontinence score. The median follow-up time for was 19 months (range, 4-55). Thirty patients (32%) had a history of previous surgery for their fistula. The success rate of fistula healing was 40% after the first ligation of the intersphincteric fistula tract procedure. When including patients with repeat ligation of the intersphincteric fistula tract and subsequent intersphincteric fistulotomy after ligation of the intersphincteric fistula tract repair, the success rates were 47% and 57%. Patients with successful fistula closure reported a mean Cleveland Clinic Florida Fecal Incontinence score of 1.0 (SD 2.3). No predictor for successful fistula closure was found. Retrospective design, limited accuracy of diagnosing fistula failure, and lack of baseline continence were limitations of this study. The present study indicates that the ligation of the intersphincteric fistula tract procedure for transsphincteric fistulas has a significant risk for failure but good functional outcome in patients with no recurrence.


Consumption of potatoes and incidence rate of coronary artery disease: The Million Veteran Program.
Potato consumption is highly prevalent around the world. Previous studies have reported a positive association of potato intake with hypertension and type 2 diabetes. However, data are scarce on potato consumption and risk of coronary artery disease (CAD). We hypothesized that potato consumption is positively associated with the incidence of CAD among US veterans. We prospectively studied 148,671 participants from Million Veteran Program (MVP). We used a semi-quantitative food frequency questionnaire to assess consumption of baked, boiled, and mashed potatoes. The incidence of CAD was assessed through electronic health record. We used Cox Proportional hazard model to compute hazard ratios (HR) and 95% confidence intervals (95% CI) for CAD events across categories of potato intake. The average age of participants was 64 years at the time of potato assessment. A total of 6309 new cases of CAD occurred during a mean follow up of 2.7 ± 1.4 y. Median potato consumption was 1 cup/week. The crude incidence of CAD from lowest to highest category of potato consumption was 14.5, 15.0, 15.2, 16.1, and 18.9 per 1000 person-years, respectively. Hazard ratios (95% CI) of CAD were 1.00 (reference), 1.02 (0.93-1.11), 1.02 (0.93-1.12), 1.04 (0.95-1.15), and 1.21 (1.07-1.37) for potato intake of <1 cup/month, 1-3 cups/month, 1 cup/week, 2-4 cups/week, and 5+ cups/week respectively, adjusting for age, gender, race, body mass index (BMI), alcohol consumption, exercise, smoking, DASH (Dietary Approaches to Stop Hypertension) score, and education. The observed relation of potato consumption with CAD was not modified by age, BMI, gender, and ethnicity in a secondary analysis. In a sensitivity analysis, exclusion of CAD events occurred during the first year of follow up did not alter the findings. Frequent (5+ cups/week) but not infrequent potato consumption was associated with a higher risk of CAD among MVP participants.


Metabolic Syndrome and Physical Inactivity May Be Shared Etiological Agents of Prostate Cancer and Coronary Heart Diseases.
As metabolic syndrome (MetS) and a sedentary lifestyle have been associated with an increased risk of developing both prostate cancer (PCa) and cardiovascular diseases (CVDs), the 2 conditions may share a common etiology. We aimed at investigating the association between CVDs and PCa. A retrospective analysis was performed. Our dataset on patients undergone systematic prostate biopsy was searched for histopathologic and clinical data. The physical activity (PA) scale for the elderly (PASE) was collected. Coronary heart diseases (CHDs) were recorded. Prognostic Grade Group ≥3 tumors were defined as high-grade (HG). The association between MetS, PA, CHDs and PCa was assessed using logistic regression analyses. Data on 955 patients were collected; 209 (22%) presented with MetS, 79 (8%) with CHDs. PCa was diagnosed in 395 (41.3%) men and 60% (n = 238) presented with an high-grade tumor. CHDs were more common among PCa-patients (9.4% vs. 7.5%; p = 0.302) but the difference was not statistically significant. No difference was observed between low- and high-grade subgroups (9.5% vs. 9.2%; p = 0.874). PASE independently predicted PCa diagnosis (OR: 0.287; p = 0.001) and HG-PCa (OR: 0.165; p = 0.001). MetS was an independent predictor of HG-PCa only (OR: 1.50; 95% CI: 1.100-2.560; p = 0.023). CHDs were not associated with tumor diagnosis and aggressiveness.


Serum concentrations of organochlorine pesticides, biomarkers of oxidative stress, and risk of breast cancer.
Studies have documented that exposure to organochlorine pesticides (OCPs) is linked with breast cancer, but the underlying biological mechanisms are still unknown. This study included 313 women diagnosed with breast cancer and 313 controls in Wuhan, China, and measured 18 OCPs in serum and 3 oxidative stress biomarkers in urine. Multivariable adjusted regression models were used to evaluate the associations among OCPs, oxidative stress biomarkers, and breast cancer. The mediating effect of oxidative stress was assessed by mediation analysis. We observed that most OCPs were positively associated with risk of breast cancer (all FDR-P values < 0.05 or 0.10). Moreover, we found that p,p'-DDT, p,p'-DDD, dieldrin, heptachlor, and heptachlor epoxide were positively associated with 4-hydroxy-2-nonenal-mercapturic acid (HNE-MA) and 8-iso-prostaglandin F2α (8-isoPGF2α), which in turn were positively associated with risk of breast cancer. Mediation analysis indicated that HNE-MA and 8-isoPGF2ɑ mediated the positive associations between these OCPs and risk of breast cancer, with mediating proportion ranging from 6.23% to 19.9%. Our results suggest that lipid peroxidation may mediate the positive associations between OCP exposures and risk of breast cancer.


Use of induction therapy in pediatric heart transplant recipients in Switzerland - Analysis of the Swiss national database.
Data on individualized immunosuppressive protocols for the pediatric heart recipients are missing in Europe. To contribute to this very small but specialized field, we describe the use of induction therapy (IT) in pediatric heart transplant patients in Switzerland and the retrospective outcomes. This is a retrospective national database analysis of children <19 years of age at time of heart transplantation (HT) from 05/2008-01/2018. Use of IT or no IT, use of steroids, calculated panel reactive antibodies (cPRA) and outcomes (Mortality, post-transplant lymphoproliferative disease (PTLD), rejection rates) were studied within a mean follow-up period of 2.9 years (0.2-8.1 years). All 32 patients (12♂, 20♀), median age at HT of 6.4 years (24 days - 18 years) received IT using either polyclonal antibodies (ATG; 72%) or interleukin-2 receptor antagonist (anti-IL-2R mAb; 28%). Length of treatment was median of 4 (1-63) days. At time of HT all patients received steroids, while at discharge 32% and one year after HT 19%. Kaplan-Meier analysis of survival revealed a one-year survival of 86%. Three out of 7 patients with elevated cPRA (43%) died. Median time to first treated rejection was 19.4 months (±60.5 SD) without significant difference if treated with anti-IL-2R mAb or ATG (p:0.5). No development of PTLD, chronic renal failure needing ongoing renal replacement therapy or diabetes mellitus were recorded. This is the first report of the national practice use of IT within Switzerland. It reveals a high use of IT, no development of PTLD and a low use of steroids at one-year post HT.


Fms-like tyrosine kinase 3-internal tandem duplications epigenetically activates checkpoint kinase 1 in acute myeloid leukemia cells.
It is not clear how Fms-like tyrosine kinase 3-internal tandem duplications (FLT3-ITD) regulates checkpoint kinase 1 (CHK1) in acute myeloid leukemia (AML). In this study, we investigated the regulatory effect of FLT3-ITD on CHK1. Our results showed that CHK1 was highly expressed in FLT3-ITD positive AML. The overall survival rate and disease-free survival rate of AML patients with high CHK1 level were lower than those of patients with low CHK1 level. Mechanistically, FLT3-ITD recruited p300 to the CHK1 promoter and subsequently acetylated H3K27, thereby enhancing the transcription of CHK1. Interfering with the expression of CHK1 significantly inhibited the cell proliferation and induced cell apoptosis in FLT3-ITD positive MV4-11 cells. In addition, CHK1 knockdown promoted the sensitivity of MV4-11 cells to the epigenetic inhibitors JQ1 and C646. This study discovers a new therapeutic target for FLT3-ITD + AML and provided evidence for the combination of epigenetic inhibitors for AML treatment.


Feedforward and feedback pathways of nociceptive and tactile processing in human somatosensory system: A study of dynamic causal modeling of fMRI data.
Nociceptive and tactile information is processed in the somatosensory system via reciprocal (i.e., feedforward and feedback) projections between the thalamus, the primary (S1) and secondary (S2) somatosensory cortices. The exact hierarchy of nociceptive and tactile information processing within this 'thalamus-S1-S2' network and whether the processing hierarchy differs between the two somatosensory submodalities remains unclear. In particular, two questions related to the ascending and descending pathways have not been addressed. For the ascending pathways, whether tactile or nociceptive information is processed in parallel (i.e., 'thalamus-S1' and 'thalamus-S2') or in serial (i.e., 'thalamus-S1-S2') remains controversial. For the descending pathways, how corticothalamic feedback regulates nociceptive and tactile processing also remains elusive. Here, we aimed to investigate the hierarchical organization for the processing of nociceptive and tactile information in the 'thalamus-S1-S2' network using dynamic causal modeling (DCM) combined with high-temporal-resolution fMRI. We found that, for both nociceptive and tactile information processing, both S1 and S2 received inputs from thalamus, indicating a parallel structure of ascending pathways for nociceptive and tactile information processing. Furthermore, we observed distinct corticothalamic feedback regulations from S1 and S2, showing that S1 generally exerts inhibitory feedback regulation independent of external stimulation whereas S2 provides additional inhibition to the thalamic activity during nociceptive and tactile information processing in humans. These findings revealed that nociceptive and tactile information processing have similar hierarchical organization within the somatosensory system in the human brain.


Outcomes following the delayed management of open tibial fractures.
National guidelines set standards for the definitive management of open fractures within 72 h. This study aims to investigate our outcomes where this timeline was unachievable for most cases due to a split-site orthoplastic service. 116 consecutive Gustilo-Anderson grade IIIB & IIIC open tibial fractures presenting to our major trauma centre (MTC) between September 2012 and April 2018 were reviewed. The mean follow up was 46 months (17 to 88). 110 (95%) were grade IIIB and 6 (5%) grade IIIC. The most common injury mechanism included road traffic accidents (59%) and falls (28%). Primary outcomes were recorded according to; timing of initial debridement and definitive cover, rates of superficial and deep infection, non-union and amputation. Subgroups were statistically analysed according to time to initial debridement, definitive soft-tissue cover and injury severity score (ISS). The mean time to initial debridement was 11.3 h (2.9 to 38.9) and definitive soft-tissue cover 9.9 days (0 to 37). We recorded rates of: superficial infection; 42 cases (36%), deep infection; 14 cases (12%) and non-union requiring revision; 19 cases (16%). There were 20 amputations (17%) with 9 (8.6%) performed early and 11 (9.5%) delayed. Subgroup analysis showed higher rates of superficial infection (50%, p = 0.002) and amputation (26.6%, p = 0.01) for those debrided <12 h. A greater presenting ISS related to a delay to definitive cover >7 days (p = 0.05). Primary outcomes trended worse for those covered >7 days but did not reach significance. Major trauma patients are particularly vulnerable to poor outcomes resulting from the delay in definitive management of open fractures. MTC's need resources and a co-located orthoplastic service to achieve national standards and better outcomes. Current guidelines do not advise for the management of patients where a delay in definitive surgery is anticipated.


Restoration of normal function in genetically defective myotubes by spontaneous fusion with fibroblasts.
Muscular dysgenesis in mice is a genetic disease of skeletal muscle caused by the recessive mutation mdg. Muscle fibres in affected mice are paralysed because of the failure of excitation-contraction coupling. Unlike normal myotubes in primary culture, dysgenic myotubes do not contract, either spontaneously or in response to electrical stimulation. The deficiency results from mutation of the gene for the skeletal muscle dihydropyridine receptor, an essential sarcolemmal component both of excitation-contraction coupling and of the slow calcium-ion channel. It has recently been shown that the addition of fibroblasts from normal (but not dysgenic) mice to cultures of dysgenic myotubes can restore spontaneous contractions in a small fraction of these myotubes, but the mechanism for this 'rescue' was not determined. In principle, if fibroblast nuclei were able to incorporate into myotubes, such nuclei could then supply the missing muscle-specific gene product. We have now investigated this possibility using nuclear, cytoplasmic and plasmalemmal markers. We report that the rescue to contractile ability in genetically paralysed dysgenic muscle is mediated by the previously unrecognized ability of fibroblasts to fuse spontaneously with developing myotubes.


Development of individuals with thanatophoric dysplasia surviving beyond infancy.
This study aimed to analyze the physical and psychosocial development of long-term survivors (age >1 year) of thanatophoric dysplasia (TD). The participants were 20 long-term survivors recruited from a cohort obtained through a nationwide survey for TD conducted across 147 pediatric departments in Japan between 2012 and 2016. Their guardians consented to participate in this study. Medical and psychosocial information was collected through questionnaires and interviews with primary physicians and guardians. The participants were 1.2-27.8 years old, and all showed marked growth deficiency. The mean length at birth was 36 cm (-3.4 SD to -7.9 SD). The adult height (age >16 years) was <-15.2 SD. All individuals showed severely delayed psychomotor development. The highest level of psychosocial development was equivalent to that at 2 years of age. Skin disorders (acanthosis nigricans and seborrheic keratoses) were common. Eleven subjects had been hospitalized or institutionalized consistently after birth, and nine had been moved to home care, and four were exclusively orally fed. All individuals required assisted ventilation. Long-term survival of TD individuals is common. Some individuals enjoy home-based lives; however, they are severely psychosocially and physically disabled and require meticulous respiratory and nutritional support.


The relationship between primary splenic malignant lymphoma and chronic liver disease associated with hepatitis C virus infection.
An etiologically important role has been suggested for hepatitis C virus (HCV) infection in the development of B-cell non-Hodgkin's lymphoma (NHL). HCV has been recognized as the major cause of non-A, non-B chronic hepatitis throughout the world. Moreover, the occurrence of primary splenic malignant lymphoma (PSML) has been demonstrated in patients with chronic liver disease. In this study, the authors describe three patients with PSML. The clinical, histologic, and immunohistochemical features of the lymphomas were studied. Clonal immunoglobulin heavy chain gene rearrangement was investigated by polymerase chain reaction. All three cases of PSML were detected by imaging studies performed in routine follow-up of cases of chronic liver disease associated with HCV infection. Macronodular lesions were found in the three spleens; two of them were of normal weight and another was moderately enlarged. The former two were the smallest PSMLs reported to date. The histology was B-cell NHL in all cases. All 3 patients were alive after splenectomy with an average follow-up of 51.7 months (range, 35-74 months). HCV infection may play an etiologic role in the development of splenic B-cell lymphoma. The long survival of the patients in this study may have been due to early splenectomy.


The role of endogenous hydrogen sulfide in pathogenesis of chronotropic dysfunction in rats with cirrhosis.
Endogenous hydrogen sulfide is produced by cystathionine-γ-lyase and cystathionine-β-synthase in a variety of tissues and has recently been implicated in the regulation of cardiac functions. Acceleration of the heart rate in response to catecholamines is impaired in patients with cirrhosis. The present study was aimed to examine the role of endogenous hydrogen sulfide in the pathogenesis of chronotropic dysfunction in rats with cirrhosis. Cirrhosis was induced by surgical ligation of bile duct in rats. There was no significant difference in atrial cystathionine-γ-lyase and cystathionine-β-synthase mRNA levels in control and cirrhotic rats as assessed by quantitative RT-PCR. Four weeks after bile duct ligation or sham surgery the atria were isolated and chronotropic responsiveness to adrenergic stimulation was assessed using standard organ bath. Incubation of the atria with propargylglycine (PAG, a cystathionine-γ-lyase inhibitor) and amino-oxyacetic acid (AOAA, a cystathionine-β-synthase inhibitor) was associated with a significant desensitization of chronotropic response to adrenergic stimulation in controls rats. This indicates that endogenous hydrogen sulfide might be involved in modulation of adrenergic signaling in the atrium. Bile duct ligation was associated with impaired chronotropic responsiveness to adrenergic stimulation in comparison with sham-operated rats. In contrast to control group, incubation of the atria with PAG and AOAA was able to partially improve the chronotropic responsiveness to adrenergic stimulation in cirrhotic rats. Our data shows that local inhibition of endogenous hydrogen sulfide in atria has opposite effect in cirrhotic versus control rats and may play a role in physiological modulation of adrenergic signaling in the atrium.


Ni single atoms on carbon nitride for visible-light-promoted full heterogeneous dual catalysis.
Visible-light-driven organic transformations are of great interest in synthesizing valuable fine chemicals under mild conditions. The merger of heterogeneous photocatalysts and transition metal catalysts has recently drawn much attention due to its versatility for organic transformations. However, these semi-heterogenous systems suffered several drawbacks, such as transition metal agglomeration on the heterogeneous surface, hindering further applications. Here, we introduce heterogeneous single Ni atoms supported on carbon nitride (NiSAC/CN) for visible-light-driven C-N functionalization with a broad substrate scope. Compared to a semi-heterogeneous system, high activity and stability were observed due to metal-support interactions. Furthermore, through systematic experimental mechanistic studies, we demonstrate that the stabilized single Ni atoms on CN effectively change their redox states, leading to a complete photoredox cycle for C-N coupling.


The morphology and electrochemical properties of porous Fe2O3@C and FeS@C nanofibers as stable and high-capacity anodes for lithium and sodium storage.
Among an enormous variety of electrode materials for lithium and sodium storage, transition metal-oxides/sulfides stand out on account of their widespread availability and high theoretical charge capacity. However, these anodes still undergo poor capacity retention and limited cycle life. Herein, we present a simple approach to synthesize one-dimensional (1D) porous Fe2O3@C and FeS@C nanofibers in which ultra-small active nanoparticles are first distributed in the internal porous carbon matrix and further encapsulated in the external nano-carbon walls. The 1D porous nano-architecture effectively alleviates the pulverization or aggregation induced by huge volume changes during cycling as well as provides a short ion/electron diffusion path in the crystal. Furthermore, the internal porous carbon matrix and the external nano-carbon layers keep the structural and mechanical stability of the entire electrode. The as-synthesized Fe2O3@C and FeS@C nanofibers show high specific capacities, robust cycling stability as well as desirable rate capability for LIBs and SIBs. Simultaneously, the FeS@C nanofibers achieve better lithium and sodium storage properties due to good electrical property and fast ion diffusion kinetics compared with Fe2O3@C nanofibers. This novel architecture design may open an avenue to seeking out high performance electrodes for advanced energy storage.


Evaluating scenarios for carbon reduction using different tableware in China.
Global consumption of disposable plastic tableware (DPT) is massive because it is durable, light and inexpensive. Using the life cycle assessment method, we found that DPT for per person per meal emitted 597 g of CO2 and was far more than that of reusable plastic tableware (RPT, 7.00 g), ceramic tableware (9.55 g) and straw tableware (14.6 g). If the demand growth for DPT continues, 416 MT of CO2 will be emitted due to DPT consumption by 2050 globally. We further explored strategies to reduce CO2 emissions by examining the life cycles of four types of tableware according to sensitivity analysis. According to our results, if the recycling rate of DPT reaches 60% at the end-of-life stage, 50% of CO2 emissions can be cut; if dishwashing instead of hand washing is used to clean RPT, ceramic, and straw tableware, approximately 64%, 71%, and 23% of CO2 emissions can be reduced, respectively. If 60% of DPT is replaced by RPT, this plastic tableware will halve carbon emissions. If the rate reaches 100%, carbon emissions will be reduced by 92%. Although the CO2 emissions of the three types of tableware other than DPT are relatively small, RPT will bring other environmental burdens and human health risks, ceramic tableware is bulky and its additives are toxic. Straw tableware combines practical and safety performance. The results show that the choice of straw tableware plays a significant role in curbing the greenhouse effect without compromising consumer safety.


Discovery and biosynthesis of phosphonate and phosphinate natural products.
The P-C bonds in phosphonate and phosphinate natural products endow them with a high level of stability and the ability to mimic phosphate esters and carboxylates. As such, they have a diverse range of enzyme targets that act on substrates containing such functionalities. Recent years have seen a renewed interest in discovery efforts focused on this class of compounds as well as in understanding their biosynthetic pathways. This chapter focuses on current knowledge of these biosynthetic pathways as well as tools for phosphonate discovery.


Chaperones and transport proteins regulate TLR4 trafficking and activation.
TLR4/MD-2 plays an important role in the inflammatory responses against lipopolysaccharide (LPS), a principal membrane component in Gram-negative bacteria. LPS recognition by TLR4/MD-2 is followed by the homotypic interaction of TLR4 (TLR4 dimerization) on the cell surface, leading to the activation of the downstream signaling pathways. The activated TLR4 is transported from plasma membrane to lysosomes, where TLR4 is degraded as a mechanism terminating LPS responses. TLR4 endocytosis is dependent on clathrin-coated vesicles. On the other hand, chaperones play an important role in TLR4 trafficking from endoplasmic reticulum (ER) to cell surface, where TLR4 recognizes LPS and activates. Chaperone gp96 and newly identified chaperone PRAT4A differentially regulate TLR4 translocation to the cell surface. In this review, we discuss TLR4 localization from endoplasmic reticulum to the cell surface and from the cell surface to endosome/lysosome that regulate TLR4 activation.


Sandhopper Talitrus saltator (Montagu) as a bioindicator of contamination by polycyclic aromatic hydrocarbons.
This study evaluates the use of the sandhopper Talitrus saltator as a bioindicator of PAHs contamination of the supralittoral zone of sandy beaches. Adult amphipods were collected at eight localities on the Tuscany shore in summer. Concentrations of 16 PAHs indicated as priority pollutants by the Environmental Protection Agency were determined. T. saltator is able to accumulate PAHs (total PAHs ranging from 0.75 to 62.1 ng g(-1)), with concentrations generally greater than in the sand (total PAHs ranging from 0.04 to 38 ng g(-1)). In particular, it accumulates Phe, Cry, and BbF. Preliminary laboratory tests indicate food as a possible route of PAHs intake by sandhoppers. Despite the need of further studies to clarify the variability of the PAH concentrations found in the substrata and in sandhoppers, T. saltator could be used as a bioindicator of PAHs contamination of the supralittoral zone of sandy shores.


Comparative study of raw and processed Vladimiriae Radix on pharmacokinetic and anti-acute gastritis effect through anti-oxidation and anti-inflammation.
Vladimiriae Radix (VR) is the dry root of Vladimiria souliei (Franch.) Ling or Vladimiria souliei (Franch.) Ling var. cinerea Ling. Costunolide (CO) and dehydrocostus lactone (DE) are the two most effective active ingredients of VR. Raw Vladimiriae radix (rVR) and processed Vladimiriae radix (pVR) are the two most common forms. They have been used for hundreds of years to treat gastritis, gastric ulcer and gastrointestinal pain, but their protective effects on gastric mucosa have been widely considered to be different, and the mechanism is not clear. A comparative study of in vivo process and efficacy difference of raw and processed Vladimiriae Radix was carried out to explore the treatment mechanism and to provide reference for the rationality of clinical usage. In this study, multi-batch rVR and pVR were used to establish the characteristic chromatograms through high performance liquid chromatography (HPLC) to control the qualities of their extracts. A rapid and accurate ultra-high performance liquid chromatography - mass spectrometry (UPLC-MS) method was established and verified, and the concentrations of CO and DE in plasma of rats after oral administration were determined to analyze the pharmacokinetics. The anti-inflammatory and antioxidant activities of ethanol-induced acute gastric mucosa injury (AGMI) in rats were quantitatively analyzed by ELISA and Westernblot methods. Characteristic chromatograms study showed that there were 9 common characteristic peaks between the chromatograms of rVR and pVR, and there was a high level (> 0.90) of the similarity between batches (only one batch less than 0.90). The increased levels of Tmax, T1/2 and MRT were found in rats treated with the pVR. Animal model studies indicated that both the two forms of VR could relieve AGMI, but pVR could more effectively reduce the content of ethanol in blood and lower the levels of TNF-α, IL-6, IL-1β, NO, iNOS and MDA, and increase the level of SOD. Results of Westernblot proved that pVR also could inhibit the expression of NF-κB p65, IκBα and up-regulate the expression of HO-1 and NRF2 more operatively to protect gastric mucosa through anti-inflammatory and antioxidant stress mechanisms. Compared with rVR, pVR has an accelerated absorption in vivo and its effect time was prolonged, and the observed improvement of anti-AGMI effect was achieved through anti-oxidation and anti-inflammation regulation.


Polyarteritis Nodosum of the Breast in a Patient with History of Bilateral Augmentation Mammoplasty.
Polyarteritis nodosum belongs to a group of inflammatory disorders characterized by necrotizing vasculitis of small and medium-sized blood vessels. To date, there are 14 publications that document involvement of the breast. Our publication is the second study documenting polyarteritis nodosum of the breast in a patient who had previously had augmentation mammoplasty. Level of Evidence V This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Spatio-temporal prediction of atmospheric benzene (part I).
Benzene is a carcinogenic and genotoxic pollutant which mainly affects the people health through the inhalation. Nevertheless, this pollutant is not frequently measured by air-quality networks. To solve this problem, some models have been published to estimate benzene concentrations in the atmosphere. However, the lack of measures makes difficult the application of complex models in order to get a detailed spatio-temporal analysis, namely in urban areas. In this work was developed a simple semi-empirical model to predict benzene concentrations based on the ratio of benzene and carbon monoxide concentrations in order to predict the concentrations of this pollutant in large areas and periods with lack of benzene measurements but with higher impact in the human health. The model was applied to an urban area, the Metropolitan Area of Oporto, for a period of 12 years (1995-2006). Monthly correlations between benzene and carbon monoxide concentrations at Custóias air-quality station are significant (p = 0.01) and higher in winter (r(s) > 0.7) than in summer (0.3 > r(s) > 0.7). Estimate of the monthly ratio of the concentration of these two pollutants range between 199 and 305. The methodology validation shows good results (r(s) = 0.81) which allow, assuming the availability of carbon monoxide data, the use of this tool for areas with low benzene recorded data. The application of this methodology in the study area shows an annual average trend decrease of benzene concentrations during the study period, which may be linked to a general trend decrease of benzene emissions in European urban areas, including the study domain.


Purification and functional characterization of endo-beta-mannanase MAN5 and its application in oligosaccharide production from konjac flour.
MAN5, the main extracellular saccharide hydrolase from Bacillus sp. MSJ-5, is an endo-beta-mannanase with a demand of at least five sugar moieties for effective cleavage. It has a pH optimum of 5.5 and a temperature optimum of 50 degrees C and is stable at pH 5-9 or below 65 degrees C. MAN5 has a very high ability to hydrolyze konjac flour, 10 U/mg of which could completely liquefy konjac flour gum in 10 min at 50 degrees C. HPLC analysis showed that most glucomannan in the konjac flour was hydrolyzed into a large amount of oligosaccharides with DP of 2-6 and a very small amount of monosaccharide. With the culture supernatant as enzyme source, the optimum condition to prepare oligosaccharides from konjac flour was obtained as 10 mg/ml konjac flour incubated with 10 U/mg enzyme at 50 degrees C for 24 h. With this condition, more than 90% polysaccharides in the konjac flour solution were hydrolyzed into oligosaccharides and a little monosaccharide (2.98% of the oligosaccharides). Konjac flour is an underutilized agricultural material with low commercial value in China. With MAN5, konjac flour can be utilized to generate high value-added oligosaccharides. The high effectiveness and cheapness of this technique indicates its potential in industry.


Activity of cholinephosphotransferase, lysolecithin: lysolecithin acyltransferase and lysolecithin acyltransferase in the developing mouse lung.
1. The present study presents the activity profiles of cholinephosphotransferase, lysolecithin:lysolecithin acyltransferase and lysolecithin acyltransferase at different stages of development of the mouse lung. 2. The specific activity of cholinephosphotransferase, a key enzyme in the de novo synthesis of phosphatidylcholine, increases during the later stages of fetal development until it reaches a maximal value at a gestational age of 17 days, i.e. 2 days before term. Thereafter, the activity of the enzyme declines again until around term. 2. The specific activity of lysolecithin:lysolecithin acyltransferase which catalyzes the transesterification between two molecules of 1-acyl-sn-glycero-3-phosphocholine, appears to be much lower than that of cholinephosphotransferase at gestational ages below 18 days. However, around day 18, the specific activity of lysolecithin:lysolecithin acyltransferase increases dramatically until it almost equals the maximal activity of cholinephosphotransferase measured on day 17. 4. The specific activity of lysolecithin acyltransferase, which catalyzes the direct acylation of 1-acyl-sn-glycero-3-phosphocholine, does not change significantly during the prenatal development and is lower than that of either lysolecithin:lysolecithin acyltransferase or cholinephosphotransferase at all stages of development. 5. These results are discussed in view of the possible role of these enzymes in the biosynthesis of pulmonary 1,2-dipalmitoyl-sn-glycero-3-phosphocholine.


Nutritional aspects of malabsorption. Short gut adaptation.
Adaptation of remaining small bowel after extensive intestinal resection plays an important part in maintaining adequate nutrition. The metabolic consequences of short bowel depend upon the anatomic extent of bowel removed, the functional condition of remaining digestive organs and the degree to which adaptive changes occur. The pathophysiology of short bowel results primarily from the abnormal absorption of nutrients and selective metabolites leading to malnutrition and other organ complications. Nutrients in the lumen of the small intestine are required to stimulate the process of intestinal adaptation (mucosal hyperplasia). This stimulation may be mediated via direct mucosal absorption or metabolism of nutrients, the presence of pancreaticobiliary secretions in the intestine, trophic effects of circulating enteric hormones, or neurovascular influences. Understanding the mechanisms by which mucosal growth may be stimulated could lead to important clinical applications.


Predictors of venous thromboembolism in hospitalized patients with inflammatory bowel disease and colon cancer: A retrospective cohort study.
Venous thromboembolism (VTE) in patients with inflammatory bowel disease (IBD) and colon cancer (CC) increases morbidity and mortality. Risk of thrombosis in IBD and CC is well established. Still, it remains unclear how interaction of thrombotic properties in patients with both diseases predict development of VTE. The Nationwide Inpatient Sample was sourced (2005-2014) for data on patients admitted with IBD-CC who developed VTE. The main outcome was predictors of VTE. Secondary outcomes were length of stay and total charge of admission. 7625 adults were admitted from 2005 to 2014 with a co-diagnosis of IBD and CC. 197 (2.6%) were coded to have VTE as a top three diagnosis. Multivariate logistic regression showed that black patients (11.9% vs 6.0%; aOR 2.04, 95% CI = 1.26-3.31, P < 0.004) and patients with metastatic disease (27.9% vs 16.7%; aOR 1.77, 95% CI = 1.27-2.47, P = 0.001) had higher odds of having VTE. Patients with uncomplicated diabetes (8.1% vs 15.5%; aOR 0.48, 95% CI = 0.28-0.84, P = 0.010) had lower odds. Obesity and anemia were significantly associated with VTE in univariate logistic regression, but lost significance after multivariate regression. Additionally, VTE was associated with increased length of stay (8.41 vs 6.87 days, P = 0.006) and admission cost ($64,388 vs $50,874, P = 0.010). Patients with IBD and CC likely have unique procoagulant properties that differ from patients with IBD or CC alone. Knowledge of these predictors can assist efforts to risk stratify IBC-CC patients, and can aid development of an individualized approach to DVT prophylaxis in this population.


Paratesticular liposarcoma: a report of two cases and review of the literature.
Paratesticular liposarcoma is extremely rare. Only 41 cases have been reported in the literature. This article reports two cases, reviews the literature, discusses the pathology, and proposes the use of combined surgery and radiation therapy in selected cases.


Human complement component C3: characterization of active C3 S and C3 F, the two common genetic variants.
The two common genetic variants of human C3, C3 S and C3 F, were purified and characterized by SDS-PAGE, agarose gel electrophoresis, isoelectric focusing and amino acid analysis. The difference in electrophoretic mobility between the two variants was conserved after purification, and by isoelectric focusing of the hemolytically active proteins, pI values of 5.86 and 5.81 were determined for C3 S and C3 F, respectively. Any difference in amino acid composition was too small to be detected by amino acid analysis, and the two proteins had the same molecular weight as determined by SDS-PAGE.


Time-lapse analysis of aggregate formation in an inducible PC12 cell model of Huntington's disease reveals time-dependent aggregate formation that transiently delays cell death.
Huntington's disease (HD) is a progressive neurodegenerative disease caused by a CAG repeat expansion in the coding region of the HD gene. The translated polyglutamine expansion causes the formation of insoluble aggregates in the brains of HD patients and transgenic mouse models. However, the relationship between aggregate formation and neuropathology remains unknown. We used fluorescent protein tagging and live-cell time-lapse microscopy to study visible aggregate formation and its relationship to cell death in transgenic PC12 cells. We used cell lines expressing a fragment of huntingtin exon 1 with either 23 (wild type) or 74 (mutant) glutamines fused to enhanced green fluorescent protein under the control of an inducible promoter. Live cells were observed in real time after transgene induction for up to 96 h. We found that aggregate formation was time-dependent and predominantly nuclear in these cells. We followed inclusion formation in individual cells, examining the cells every 10 min for up to 48 h. This revealed new details of inclusion formation. Initial aggregate formation was rapid (often <1 h), but many (18->48) h were needed to establish a final aggregate phenotype. Aggregates formed in a dynamic manner and were in constant motion within cell nuclei throughout their maturation. The formation of large aggregates occurred more frequently in cells that survived longer. However, aggregate size was not a good predictor of cell death, since cells could die with either large (>2 microm), small (<0.5 microm) or no visible aggregates. Cells that formed large aggregates survived longer than cells that formed small aggregates or no aggregates at all. However, the time taken for a cell to die decreased as a function of increasing size of final aggregate formed. Further, cells that formed aggregates earlier tended to die earlier. Together our data are compatible with a toxic role for aggregates/aggregation and support the 'toxic precursor' hypothesis. However, they also suggest that at some stages, the process of aggregate formation is cytoprotective.


Waste disposal-An attractive strategy for cancer therapy.
Targeted therapies for cancer are typically small molecules or monoclonal antibodies that act by inhibiting the activity of specific proteins that drive tumor growth. Although many of these drugs are effective in cancer patients, the response is often not durable because tumor cells develop resistance to the drugs. Another limitation of this strategy is that not all oncogenic driver proteins are "druggable" enzymes or receptors with activities that can be inhibited. Here we describe an alternative approach to targeted therapy that is based on co-opting the cellular quality-control machinery-the ubiquitin-proteasome system-to remove specific cancer-causing proteins from the cell. We first discuss examples of existing cancer drugs that work by degrading specific proteins and then review recent progress in the rational design and preclinical testing of small molecules that induce selective degradation of specific target proteins.


HIV-2 Vpx neutralizes host restriction factor SAMHD1 to promote viral pathogenesis.
SAMHD1, a human host factor found in myeloid cells which restricts HIV-1 replication. It depletes the dNTPs pool for viral cDNA syntheses, thus preventing the viral replication in the cells. The viral accessory protein, Vpx, exists only in SIVmac/HIV-2 particles. Vpx in SIVmac can induce proteosomal degradation of SAMHD1, which then leads to a decrease in the cytoplasmic dNTP pool. The protein-protein interaction between Vpx and SAMHD1 and its consequences are still unclear. Methods: In this study, we cloned, for the first time, Vpx gene from a HIV-2 infected patient and found up to 30% sequence variation compared to known HIV-2 strains. We then analyzed the role of SAMHD1 protein expression in transfected THP-1 and U937 cells by transfecting with the Vpx gene derived from SIVmac, HIV-2 from the NIH sample as well as HIV-2 from a Saudi patient. We found that Vpx gene expression led to reduced levels of intracellular SAMHD1. When the supernatants of the transfected cell lines were examined for secreted cytokines, chemokines and growth factors, Vpx expression seemed to be suppressive of pro-inflammatory response, and skewed the immune response towards an anti-inflammatory response. These results suggest that Vpx can act at two levels: clearance of intracellular restriction factor and suppression of cytokine storm: both aimed at long-term latency and host-pathogen stand-off, suggesting that Vpx is likely to be a potential therapeutic target.


FebriDx for rapid screening of patients with suspected COVID-19 upon hospital admission: systematic literature review and meta-analysis.
A systematic literature review and meta-analysis was undertaken of the lateral flow-based FebriDx immunoassay for triaging patients with suspected coronavirus disease 2019 (COVID-19) upon admission to healthcare facilities. An electronic search was conducted in Scopus and Medline using the keywords 'FebriDx' AND 'COVID-19' OR 'SARS-CoV-2', with no language or date (i.e. up to 4th February 2022) limits, selecting studies where FebriDx was used for triaging patients with suspected COVID-19 in acute care settings, and reporting sufficient data to construct a 2×2 table. Five studies were included in the final analysis, totalling 2309 patients. The pooled diagnostic sensitivity and specificity were 0.91 [95% confidence interval (CI) 0.88-0.93] and 0.92 (95% CI 0.90-0.93), whilst the area under the curve, accuracy and kappa statistics were 0.971 (95% CI 0.962-0.980), 91.4% (95% CI 90.2-92.5%) and 0.762 (95% CI 0.731-0.793), respectively, thus reflecting substantial agreement with reference molecular testing techniques. Negative and positive predictive values were 0.974 (95% CI 0.966-0.981) and 0.742 (95% CI 0.711-0.770), respectively. This pooled analysis demonstrated that FebriDx has clinical value for rapid screening of patients with suspected COVID-19 in acute care settings, especially in regions with high viral circulation in which the pre-test probability is high, and enables prioritization for confirmatory laboratory testing.


Single step purification procedure for the rapid separation of equine leucocytes.
Percoll gradients have been used to separate relatively pure populations of viable equine polymorphonuclear (PMN) and mononuclear (MN) cells. In preliminary studies, a continuous density gradient of 70% Percoll solution was used to separate two distinct leucocyte-rich bands. After measurement of the density of each band on the continuous gradient, discontinuous Percoll gradients, using 60% and 75% Percoll solutions, were used to provide a rapid means of separating PMN and MN cells. The yield of viable cells per ml of blood was 3.0 X 10(6) and 3.2 X 10(6) for MN and PMN cells, respectively. Corresponding values for recovery were 45% and 72%. The purity was 94% for PMNs and 99% for MNs.


End-stage renal disease among silica-exposed gold miners. A new method for assessing incidence among epidemiologic cohorts.
To examine the association between silica exposure and end-stage renal disease (ESRD). Retrospective cohort study. A cohort of 2412 white male gold miners was studied. Eligible gold miners worked underground for at least 1 year between 1940 and 1965 in a South Dakota gold mine and were alive on January 1,1977. Of primary interest was exposure to silica. The ESRD Program Management and Medical Information System (PMMIS) was used to identify members of the gold mine cohort who had treated ESRD and to create a US rate file for treated ESRD. The ESRD incidence among the gold miners was compared with that in the US population. Based on the 11 cohort members identified with treated ESRD, the risk for ESRD in the cohort was elevated (standardized incidence ratio [SIR], 1.37; 95% confidence interval [CI], 0.68-2.46). The risk was greatest for nonsystemic ESRD (ESRD caused by glomerulonephritis or interstitial nephritis) for which the SIR was 4.22 (95% CI, 1.54-9.19), increasing to 7.70 (95% CI, 1.59-22.48) among workers with 10 or more years of employment underground. To our knowledge this is the first epidemiologic study to examine ESRD incidence in an occupational cohort. This study provides evidence that silica exposure is associated with an increased risk for ESRD, especially ESRD caused by glomerulonephritis. This study also demonstrates the usefulness of the ESRD PMMIS to assess ESRD risk among cohorts exposed to potential nephrotoxins.


[The use of recombinant human interleukin-1 alpha and -1 beta as agents in the early treatment of acute radiation sickness in an experiment].
Human recombinant interleukins 1 alpha and 1 beta (IL-1 alpha and IL-1 beta) have a good dose-dependent therapeutic effect for an acute radiation damage. For mice, the optimum therapy dose of IL-1 is 100.0 micrograms/kg-1 (survival rates: CD70--50%, CD100--17%; p < 0.05). For dogs, the dose is 1.0 microgram/kg-1 (survival rate: CD100 in combination with antibacterial therapy-up to 80%; p < 0.05).


Personal constructs in anorexia.
A repertory grid of 12 elements and 12 constructs was presented to 15 female subjects suffering from primary anorexia nervosa and 15 matched control subjects. Cluster analysis, as used by Makhlouf-Norris and Norris (1973) revealed that the anorexic group had a prevalence of segmented and monolithic structures and the normal group a prevalence of articulated structures. The relationships between the elements was examined and the anorexic group showed significantly more "ideal self" isolation, "social" and "present self" alienation than the control group. The average distance between the elements was significantly greater in the anorexic group than in the control group. Analysis of the first two components showed that the first component accounted for more variance than the second when compared to the normals. It is concluded that the repertory grid is a valid method of investigating the conceptual disturbance in anorexia nervosa.


Structure-based design of oxygen-linked macrocyclic kinase inhibitors: discovery of SB1518 and SB1578, potent inhibitors of Janus kinase 2 (JAK2) and Fms-like tyrosine kinase-3 (FLT3).
Macrocycles from our Aurora project were screened in a kinase panel and were found to be active on other kinase targets, mainly JAKs, FLT3 and CDKs. Subsequently these compounds became leads in our JAK2 project. Macrocycles with a basic nitrogen in the linker form a salt bridge with Asp86 in CDK2 and Asp698 in FLT3. This residue is conserved in most CDKs resulting in potent pan CDK inhibition. One of the main project objectives was to achieve JAK2 potency with 100-fold selectivity against CDKs. Macrocycles with an ether linker have potent JAK2 activity with the ether oxygen forming a hydrogen bond to Ser936. A hydrogen bond to the equivalent residues of JAK3 and most CDKs cannot be formed resulting in good selectivity for JAK2 over JAK3 and CDKs. Further optimization of the macrocyclic linker and side chain increased JAK2 and FLT3 activity as well as improving DMPK properties. The selective JAK2/FLT3 inhibitor 11 (Pacritinib, SB1518) has successfully finished phase 2 clinical trials for myelofibrosis and lymphoma. Another selective JAK2/FLT3 inhibitor, 33 (SB1578), has entered phase 1 clinical development for the non-oncology indication rheumatoid arthritis.


New steroidal saponin from Hosta sieboldiana.
An extract of the edible parts of Hosta sieboldiana suppressed the growth of P388 mouse leukemic cells. A new steroidal saponin was isolated from Hosta sieboldiana in this study, and the structure was determined by a spectroscopic analysis to be (25R)-3beta-(alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranosyl)-5alpha-spirostan-2alpha-ol.


Efficacy and safety of pasireotide for Cushing's disease: A protocol for systematic review and meta-analysis.
Cushing's disease (CD) is associated with increased risk of mortality, myocardial infarction, stroke, peptic ulcers, fractures and infections. The prevalence of CD is nearly 40 per million and higher in women than in men. When surgery has failed, is not feasible, or has been refused, pharmacotherapy can be considered a valuable option. Pasireotide is the first medical therapy officially approved for adult patients with CD. We will conduct a comprehensive systematic review and meta-analysis to systematically evaluate the efficacy and safety of pasireotide for CD. Five English databases (PubMed, Web of Science, Embase, Cochrane Library, and OVID) and 3 Chinese databases (China National Knowledge Infrastructure, China Science and Technology Journal Database, and Chinese Biomedical Literature Database) will be searched from their respective inception of databases to December 2020. Two reviewers will select articles, extract data and assess the risk of bias independently. Any disagreement will be resolved by discussion with the third reviewer. Review Manager 5.3 software will be used for data synthesis. The Cochrane risk of bias assessment tool will be used to evaluate the bias risk. This systematic review and meta-analysis will conduct a comprehensive literature search and provide a systematic synthesis of current published data to explore the efficacy and safety of pasireotide for CD. This systematic review and meta-analysis will provide clinical evidence for the efficacy and safety of pasireotide for CD, and inform our understanding of the value of pasireotide in improving CD clinical signs and symptoms. The conclusions drawn from this study may be beneficial to patients, clinicians, and health-related policy makers. INPLASY2020110070.


[Results of the neurosurgical treatment of patients with drug-resistant epilepsy: psychosocial aspects].
Evaluation of psychological and social functioning of patients after neurosurgical treatment for pharmacologically intractable epilepsy. Descriptive study. University Hospital Utrecht in collaboration with the Institute for Epilepsy in Heemstede, the Netherlands. After two years' follow-up, social functioning, vocational adjustment and emotional stability were assessed in a group of 13 adult patients treated surgically between 1985 and 1992 for intractable seizures. The majority of the patients improved or at least maintained their social and employment status. Emotional functioning showed minor changes for anxiety and emotional stability. The outlook for improvement in psychosocial functioning for epileptic patients treated neurosurgically is promising. Minor changes were found in emotional stability.


DRG ripple and the shifting burden of care to home health.
It is no secret to any nurse in community health care that the DRG system has caused major shifts in the nature of the patient load, types of patients served, and level of care technology required. Phillips et al. detail the mechanism by which the care demands of the community have changed.


Neurotrophin stimulation of human melanoma cell invasion: selected enhancement of heparanase activity and heparanase degradation of specific heparan sulfate subpopulations.
Heparanase is an endo-beta-D-glucuronidase whose enzymatic targets are the glycosaminoglycan chains of heparan sulfate proteoglycans (50). Elevated levels of heparanase are associated with the metastatic potential of melanoma cells, and treatment of murine and human melanoma cells with the prototypic neurotrophin nerve growth factor (NGF) increases the production of heparanase by melanoma cells. We previously reported that physiological concentrations of NGF increased invasion of early passage human brain-metastatic 70W melanoma cells but not melanoma cells metastatic to other sites or nonmetastatic melanoma cells as measured in Matrigel invasion assays. Here we found that treatment of 70W melanoma cells with neurotrophin-3 (NT-3) increased Matrigel invasion, whereas treatment with neurotrophins other than NGF or NT-3 did not influence invasion. Mutants of NGF that do not bind to the neurotrophin receptor p75NTR or other nonneuronal growth factors were not able to enhance the invasion of 70W melanoma cells. When 70W cells were exposed to antisense oligonucleotides directed against p75NTR mRNA, there was a reduction in NGF and NT-3 binding, and the neurotrophins failed to enhance Matrigel invasion. To study the properties of heparanase in neurotrophin-regulated malignant melanoma invasive processes, we developed a sensitive heparanase assay consisting of purified [35S]HS subpopulations separated by agarose gel electrophoresis. Incubation of 70W cells with NGF or NT-3 but not brain-derived neurotrophic factor, neurotrophin-4/5 or mutant NGF resulted in increased release of heparanase activity that was capable of degrading a subpopulation of heparan sulfate molecules.


A comparative analysis of nuclear DNA content and N-myc gene amplification in neuroblastoma.
Nuclear DNA content and N-myc amplification have been found to have prognostic significance in neuroblastoma. To investigate the interrelationships between these two parameters, tumor samples from 18 patients with neuroblastoma were analyzed for both total DNA and N-myc gene content. Quantitative DNA analysis was performed by image analysis. Quantitative Southern blot hybridization was used to determine N-myc gene copy number and to distinguish between low level gene amplification or excess gene copies secondary to aneuploidy. Six of the 18 patients have died. Five of the six had nonaneuploid tumors, but only two of the six exhibited major N-myc amplification. Low-level amplification was detected in one Stage II patient. The authors' results suggest that neuroblastomas with N-myc amplification are a subset of nonaneuploid tumors.


Theory of mind and irony comprehension in children with cerebral palsy.
The main goal of the present study was to characterise the pragmatic abilities of French children with cerebral palsy through their understanding of irony and other people's mental states. We predicted that children with cerebral palsy would have difficulty understanding false-belief and ironic remarks, due to the executive dysfunction that accompanies the motor disorders of cerebral palsy. We conducted an experiment in which children with cerebral palsy and typically developing matched controls performed theory-of-mind and executive function tasks. They then listened to ironic stories and answered questions about the speakers' beliefs and attitudes. The groups differed significantly on second-order theory of mind, irony comprehension and working memory, indicating pragmatic difficulties in children with cerebral palsy.


Reduced Nm23/Awd protein in tumour metastasis and aberrant Drosophila development.
Tumour metastasis is the principal cause of death for cancer patients. We have identified the nm23 gene, for which RNA levels are reduced in tumour cells of high metastatic potential. In this report we identify the cytoplasmic and nuclear Nm23 protein, and show that it also is differentially expressed in metastatic tumour cells. We also find that the human Nm23 protein has sequence homology over the entire translated region with a recently described developmentally regulated protein in Drosophila, encoded by the abnormal wing discs (awd) gene. Mutations in awd cause abnormal tissue morphology and necrosis and widespread aberrant differentiation in Drosophila, analogous to changes in malignant progression. The metastatic state may therefore be determined by the loss of genes such as nm23/awd which normally regulate development.


Determination of plasmid copy number by fluorescence densitometry.
A simple and reliable method for the determination of plasmid copy numbers by direct fluorescence densitometry of ethidium bromide-stained electrophoretic gels was developed. In developing the method, the following parameters were evaluated and controlled: plasmid DNA trapping in the linear chromosomal DNA, staining-destaining kinetics for ethidium bromide, linearity of the fluorescence response, and the effect of the molecular topology of DNA on ethidium bromide binding to DNA in agarose.


A structure and evolutionary-based classification of solute carriers.
Solute carriers are an operationally defined diverse family of membrane proteins involved in the transport of nutrients, metabolites, xenobiotics, and drugs. Here, we provide an integrative classification of solute carriers by combining evolutionary information with proteome-wide structure models recently made available through the AlphaFold resource. Analyses of orthologous relations among 455 protein-coding genes currently classified as human solute carriers, over the fully sequenced genomes of 2,100 species, suggest no more than approximately 180 independent evolutionary origins. Structural comparative analyses provided further insight revealing a total of 24 structurally distinct transmembrane folds, increasing by approximately 40% the number of previously described SLC structural folds. In addition, a structural comparative analysis identified a new human solute carrier member and revealed details of noncanonical ones. Our analyses uncover new ancestral relations between solute carrier genes, provide insights into the evolution of remote homologs and a platform to test hypotheses of functional deorphanization.


A potential role for thalamocingulate circuitry in human maternal behavior.
Little is known about the regional brain basis of human maternal behavior. To understand this better, we have been examining brain activity in mothers listening to infant cries. We measured functional Magnetic Resonance Imaging brain activity in healthy, breastfeeding first-time mothers with young infants while they listened to infant cries, white noise control sounds, and a rest condition. Based on the thalamocingulate theory of maternal behavior and pilot work, we hypothesized that the cingulate, medial thalamus, medial prefrontal cortex, and right orbitofrontal cortex would display more activity with infant cries than with white noise (comparison 1) and would uniquely activate with the cries, meaning that these regions would display activity with cry minus rest but not with white noise minus rest (comparison 2). In hypothesized regions, the group displayed more activity in the medial thalamus, medial prefrontal and right orbitofrontal cortices with both comparisons. The anterior and posterior cingulate cortex displayed more activity only with comparison 1. In non-hypothesized brain regions, several other structures thought important in rodent maternal behavior displayed activity with both comparisons including the midbrain, hypothalamus, dorsal and ventral striatum, and vicinity of the lateral septal region. Our results partially support our hypotheses and are generally consistent with neuroanatomical studies of rodent maternal behavior.


Inhibition of neuropathic hyperalgesia by intrathecal bone marrow stromal cells is associated with alteration of multiple soluble factors in cerebrospinal fluid.
Injury-induced neuropathic pain remains a serious clinical problem. Recent studies indicate that bone marrow stromal cells (BMSCs) effectively attenuate chronic neuropathic pain in animal models. Here, we examined the therapeutic effect of intrathecal administration of BMSCs isolated from young (1-month-old) rats on pain hypersensitivity induced by tibial nerve injury. Cerebrospinal fluid (CSF) was collected and analyzed to examine the effect of BMSC administration on the expression of 67 soluble factors in CSF. A sustained remission in injury-induced mechanical hyperalgesia was observed in BMSC-treated rats but not in control animals. Engrafted BMSCs were observed in spinal cords and dorsal root ganglia at 5 weeks after cell injection. Injury significantly decreased the levels of six soluble factors in CSF: intercellular adhesion molecule 1 (ICAM-1), interleukin-1β (IL-1β), IL-10, hepatocyte growth factor (HGF), Nope protein, and neurogenic locus notch homolog protein 1 (Notch-1). Intrathecal BMSCs significantly attenuated the injury-induced reduction of ICAM-1, IL-1β, HGF, IL-10, and Nope. This study adds to evidence supporting the use of intrathecal BMSCs in pain control and shows that this effect is accompanied by the reversal of injury-induced reduction of multiple CSF soluble factors. Our findings suggest that these soluble factors may be potential targets for treating chronic pain.


Concurrent androgen deprivation therapy during salvage prostate radiotherapy improves treatment outcomes in high-risk patients.
To determine whether concurrent androgen deprivation therapy (ADT) during salvage radiotherapy (RT) improves prostate cancer treatment outcomes. A total of 630 postprostatectomy patients were retrospectively identified who were treated with three-dimensional conformal RT. Of these, 441 were found to be treated for salvage indications. Biochemical failure was defined as prostate-specific antigen (PSA) of 0.2 ng/mL or greater above nadir with another PSA increase or the initiation of salvage ADT. Progression-free survival (PFS) was defined as the absence of biochemical failure, continued PSA rise despite salvage therapy, initiation of systemic therapy, clinical progression, or distant failure. Multivariate-adjusted Cox proportional hazards modeling was performed to determine which factors predict PFS. Low-, intermediate-, and high-risk patients made up 10%, 24%, and 66% of patients, respectively. The mean RT dose was 68 Gy. Twenty-four percent of patients received concurrent ADT (cADT). Regional pelvic nodes were treated in 16% of patients. With a median follow-up of 3 years, the 3-year PFS was 4.0 years for cADT vs. 3.4 years for cADT patients (p = 0.22). Multivariate analysis showed that concurrent ADT (p = 0.05), Gleason score (p < 0.001), and pre-RT PSA (p = 0.03) were independent predictors of PFS. When patients were stratified by risk group, the benefits of cADT (hazard ratio, 0.65; p = 0.046) were significant only for high-risk patients. This retrospective study showed a PFS benefit of concurrent ADT during salvage prostate RT. This benefit was observed only in high-risk patients.


Enhancement of Edwardsiella piscicida infection, biofilm formation, and motility caused by N-acetylneuraminate lyase.
Sialic acid and its catabolism are involved in bacterial pathogenicity. N-acetylneuraminate lyase (NAL), which catalyzes the reversible aldol cleavage of sialic acid to form N-acetyl-D-mannosamine in the first step of sialic acid degradation, has been recently investigated to elucidate whether NAL enhances bacterial virulence; however, the role of NAL in bacterial pathogenicity remains unclear. In the present study, we demonstrated that the existence of two enzymes in Edwardsiella piscicida, referred to as dihydrodipicolinate synthase (DHDPS) and NAL, induced the cleavage/condensation activity toward sialic acids such as N-acetylneuraminic acid, N-glycolylneuraminic acid and 3-deoxy-D-glycero-D-galacto-non-2-ulopyranosonic acid. NAL enhanced cellular infection in vitro and suppressed the survival rate in zebrafish larvae in bath-infection in vivo, whereas DHDPS did not. Furthermore, NAL strongly activated the expression of E. piscicida phenotypes such as biofilm formation and motility, whereas DHDPS did not. Besides, the gene expression level of nanK, nanE, and glmU were up-regulated in the NAL-overexpressing strain, along with an increase in the total amount of N-acetylglucosamine.


[Behavior of vasomotor tonus after lung autotransplantation].
In eight lung autotransplants and six normal lungs the vasomotor activity was tested with norepinephrine and acetylcholine. In autotransplants and normal lungs both substances elicit the same effects. The long-term interruption of the central innervation does not provoke a vasoconstriction in lung autotransplants.


[Indications and results of colostomy for anorectal malformations in girls].
On the basis of an analysis of 122 cases the authors present options on the strategy and methods of operations depending on the anatomical patterns, the degree of anorectal malformations and the type of coexisting pathology, the technique of placing unnatural anus. Preference is given to a one-stage surgical method, the indications to the graded treatment with the formation of colostomy are described in detail.


Impact of Serum Vancomycin Trough Levels in the Treatment of Central Nervous System Shunt Infections Caused by Coagulase-Negative Staphylococci.
Coagulase-negative staphylococci (CoNS) are a common cause of pediatric ventricular shunt infections. The Infectious Diseases Society of America recommends vancomycin serum troughs of 15-20 µg/mL when treating CoNS shunt infections in adult patients. We report a series of pediatric cases of CoNS shunt infections in which clinical cure was obtained with troughs < 15 µg/mL. These findings question the relevance of this recommendation in pediatric patients.


Biomarkers from circulating neutrophil transcriptomes have potential to detect unruptured intracranial aneurysms.
Intracranial aneurysms (IAs) are dangerous because of their potential to rupture and cause deadly subarachnoid hemorrhages. Previously, we found significant RNA expression differences in circulating neutrophils between patients with unruptured IAs and aneurysm-free controls. Searching for circulating biomarkers for unruptured IAs, we tested the feasibility of developing classification algorithms that use neutrophil RNA expression levels from blood samples to predict the presence of an IA. Neutrophil RNA extracted from blood samples from 40 patients (20 with angiography-confirmed unruptured IA, 20 angiography-confirmed IA-free controls) was subjected to next-generation RNA sequencing to obtain neutrophil transcriptomes. In a randomly-selected training cohort of 30 of the 40 samples (15 with IA, 15 controls), we performed differential expression analysis. Significantly differentially expressed transcripts (false discovery rate < 0.05, fold change ≥ 1.5) were used to construct prediction models for IA using four well-known supervised machine-learning approaches (diagonal linear discriminant analysis, cosine nearest neighbors, nearest shrunken centroids, and support vector machines). These models were tested in a testing cohort of the remaining 10 neutrophil samples from the 40 patients (5 with IA, 5 controls), and model performance was assessed by receiver-operating-characteristic (ROC) curves. Real-time quantitative polymerase chain reaction (PCR) was used to corroborate expression differences of a subset of model transcripts in neutrophil samples from a new, separate validation cohort of 10 patients (5 with IA, 5 controls). The training cohort yielded 26 highly significantly differentially expressed neutrophil transcripts. Models using these transcripts identified IA patients in the testing cohort with accuracy ranging from 0.60 to 0.90. The best performing model was the diagonal linear discriminant analysis classifier (area under the ROC curve = 0.80 and accuracy = 0.90). Six of seven differentially expressed genes we tested were confirmed by quantitative PCR using isolated neutrophils from the separate validation cohort. Our findings demonstrate the potential of machine-learning methods to classify IA cases and create predictive models for unruptured IAs using circulating neutrophil transcriptome data. Future studies are needed to replicate these findings in larger cohorts.


Prognosis of extremity osteosarcoma in patients aged 40-60 years: a cohort/case controlled study at a single institute.
The outcome of older osteosarcoma patients with multi-disciplinary management has not been clearly defined. We conducted a cohort (n=375) and a case-control (n=78) study on 26 older age patients (40-60 years) with localized osteosarcoma of extremity. In the case-control study, controls were matched for location and initial tumor volume. Compared to 349 younger patients, older age patients showed an osteolytic pattern on plain radiographs (P=0.05), fibroblastic subtype (P<0.01), and poor histologic response (P=0.03). Multivariate analysis revealed that a large absolute tumor volume (P<0.01), a tumor location in the proximal humerus (P=0.02), and a poor histologic response to preoperative chemotherapy (P<0.01) independently predicted poorer metastasis-free survival. However, an older age showed marginal significance (P=0.09). A case-control study showed a higher proportion of the fibroblastic subtype and poor histologic response in the case group. Five-year metastasis-free survival rates for the 26 cases and 52 controls were 40.1+/-10.1% and 61.5+/-6.8%, respectively (P=0.02). Older age osteosarcoma patients showed an unfavorable histologic response to chemotherapy and lower survival than younger patients. Nevertheless, a further larger-scale study is required to confirm our observations.


Carbon nanotube-mediated DNA delivery without transgene integration in intact plants.
Exogenous biomolecule delivery into plants is difficult because the plant cell wall poses a dominant transport barrier, thereby limiting the efficiency of plant genetic engineering. Traditional DNA delivery methods for plants suffer from host-species limitations, low transformation efficiencies, tissue damage, or unavoidable and uncontrolled DNA integration into the host genome. We have demonstrated efficient plasmid DNA delivery into intact plants of several species with functionalized high-aspect-ratio carbon nanotube (CNT) nanoparticles (NPs), enabling efficient DNA delivery into a variety of non-model plant species (arugula, wheat, and cotton) and resulting in high protein expression levels without transgene integration. Herein, we provide a protocol that can be implemented by plant biologists and adapted to produce functionalized single-walled CNTs (SWNTs) with surface chemistries optimized for delivery of plasmid DNA in a plant species-independent manner. This protocol describes how to prepare, construct, and optimize polyethylenimine (PEI)-functionalized SWNTs and perform plasmid DNA loading. The authors also provide guidance on material characterization, gene expression evaluation, and storage conditions. The entire protocol, from the covalent functionalization of SWNTs to expression quantification, can be completed in 5 d.


CXCR3+ T cells in multiple sclerosis correlate with reduced diversity of the gut microbiome.
Multiple sclerosis (MS) is a genetically mediated autoimmune disease characterized by inflammation in the central nervous system (CNS). Disease onset is thought to occur when autoreactive T cells orchestrate a cascade of events in the CNS resulting in white and grey matter inflammation and axonal degeneration. It is unclear what triggers the activation of CNS-reactive T cells and their polarization into inflammatory subsets. Mounting evidence from animal and human studies supports the hypothesis that the gut microbiome affects MS pathogenesis. We investigated the association between the gut microbiome and inflammatory T cell subsets in relapsing-remitting MS patients and healthy controls. Gut microbiome composition was characterized by sequencing the V4 region of the 16S rRNA gene from fecal DNA, and inflammatory T cell subsets were characterized by flow cytometry. We identified an altered gut microbiome in MS patients, including decreased abundance of Coprococcus, Clostridium, and an unidentified Ruminococcaceae genus. Among circulating immune cells, patients had increased expression of CXCR3 in both CD4 and CD8 T cells, and both CD4+CXCR3+ and CD8+CXCR3+ populations expressing the gut-homing α4β7 integrin receptor were increased. Finally, we show that alpha diversity inversely correlated with a CXCR3+ Th1 phenotype in MS. These findings indicate the presence of an aberrant gut-immune axis in patients with MS.


Simple bone cysts. A review of 59 cases with special reference to their treatment.
In a retrospective study, 21 simple bone cysts (SBC) treated by curettage (with or without bone grafting) are compared to 20 SBC treated by intralesional injections of methylprednisolone. Curettage led to 43% favourable results and 29% recurrences. Cortisone injections led to 90% favourable results and 5% recurrences. Combined therapy (curettage and injections) led to results comparable to injections only. In our experience, curettage and hydroxyapatite grafting led to 100% complete healing (only 2 cases). We recommend intralesional methylprednisolone injections because the method is easy, effective and safe.


Filling space with hypercubes of two sizes - The pythagorean tiling in higher dimensions.
We construct a unilateral lattice tiling of R n into hypercubes of two differnet side lengths p or q. This generalizes the Pythagorean tiling in R 2 . We also show that this tiling is unique up to symmetries, which proves a variation of a conjecture by Bölcskei from 2001. For positive integers p and q, this tiling also provides a tiling of ( Z / ( p n + q n ) Z ) n .


Non-reciprocal control of rhythmic activity in respiratory-modulated XII motoneurons.
To elucidate the nature of phasic respiratory inputs to hypoglossal (XII) motoneurons, we assessed the phasic changes in membrane resistance (RN) in inspiratory- and expiratory-modulated motoneurons. In expiratory motoneurons, RN was consistently lower in inspiration than in expiration and intracellular injection of negative current reversed the phasic hyperpolarizing membrane potential wave in inspiration. In inspiratory motoneurons, RN changed less but also was minimal in inspiration. In these neurons, injection of a negative current sufficient to reverse a lingual nerve-evoked IPSP did not reverse the phasic expiratory hyperpolarization. Thus, respiratory-related inhibitory inputs are non-reciprocal in inspiratory and expiratory XII motoneurons, with inspiratory motoneurons probably lacking a somatic, amino acid-mediated inhibitory input in expiration.


MicroRNAs and lung cancers: from pathogenesis to clinical implications.
Lung cancer is the leading cause of cancer-related deaths in the US and worldwide. Better understanding of the disease is warranted for improvement in clinical management. Here we summarize the functions of small-RNA-based, posttranscriptional gene regulators, i.e. microRNAs, in the pathogenesis of lung cancers. We discuss the microRNAs that play oncogenic as well as tumor suppressive roles. We also touch on the value of microRNAs as markers for diagnosis, prognosis and the promising field of microRNA-based novel therapies for lung cancers.


The COL5A1 gene is associated with increased risk of carpal tunnel syndrome.
The direct causes of idiopathic carpal tunnel syndrome (CTS), a common upper limb entrapment neuropathy, remain unknown. It is however generally accepted that an increase in pressure within the carpal tunnel structure, which contains nine flexor tendons, causes compression of the median nerve. The involvement of these tendons in the aetiology of CTS cannot be excluded. Variants within the collagen, type V, alpha 1 (COL5A1) gene, which encodes for the α1 chain of type V collagen, an important regulator of fibril assembly in tendons, have previously been associated with Achilles tendinopathy. The aim of this study was to determine whether these COL5A1 variants are also associated with CTS. One hundred and three self-reported coloured participants, with a history of carpal tunnel release surgery (CTS) and 150 matched control (CON) participants without any reported history of CTS symptoms were genotyped for the COL5A1 rs13946 (C/T), rs14774622 (C/T)/rs55748801 (G/A) (W/M where W = CG), rs12722 (C/T) and rs71746744 (-/AGGG) variants. The TT genotype of COL5A1 rs13946 was significantly over-represented (p = 0.007) in the CON (69.3 %) compared to that in the CTS (50.6 %) group. When the combined rs14774622/rs55748801 and rs12722 genotypes were analysed, the WW + CC (41.7 %, p = 0.008) and WW + CT (40.3 %, p = 0.009) genotypes were significantly over- and under-represented in the CON group, respectively, when compared to the CTS group (24.5 % WW + CC, 59.2 % WW + CT). Furthermore, the T-W-C (51.2 %, p < 0.001) and C-W-C (15.9 %, p = 0.005) inferred haplotypes were significantly over- and under-represented in the CON compared to the CTS (34.9 % T-W-C, 26.8 % C-W-C). In conclusion, this is the first study to report that variants within the functional COL5A1 3'-untranslated region are associated with the CTS. Further studies are required to replicate these findings.


HIV Pre-exposure Prophylaxis Use and Persistence among Black Ciswomen: "Women Need to Protect Themselves, Period".
Disparities in HIV incidence and PrEP use among Black ciswomen remain. We examine factors associated with PrEP persistence using mixed methods. Black ciswomen in Chicago, IL, prescribed PrEP at a federally qualified health center (FQHC). We used electronic health data to determine PrEP persistence (Proportion of Days Covered ≥86% at 6 months) and tested demographic and clinical factors in logistic regressions. We interviewed eight Black ciswomen, purposefully selected by PrEP persistence. Among 112 Black ciswomen, 18% were persistent. In adjusted models, neighborhood, visit reason (at initiation), and initiation year were significantly associated with persistence. Qualitatively, we found little evidence of cost or adherence as barriers; participants reported low community awareness, importance of providers, and concerns around stigma, side effects, and pregnancy while using PrEP. While persistence among Black ciswomen was low, patients were often making decisions based on perceived HIV risk. We identified real-world barriers to address in future interventions.


Mechanism of synchronized Ca2+ oscillations in cortical neurons.
Dissociated rat cortical neurons reassociate in vitro to form synaptically connected networks. Removal of Mg2+ from the extracellular medium then induces neurons in the network to undergo synchronized oscillations of cytoplasmic calcium. Previous studies have shown that these calcium oscillations involve the activation of NMDA receptors and that the rising phase of each calcium spike is coincident with a brief burst of action potentials (Robinson et al., Jpn. J. Physiol. 43 (Suppl. 1) (1993) S125-130; Robinson et al., J. Neurophysiol. 70 (1993) 1606-1616; Murphy et al., J. Neurosci. 12 (1992) 4834-4845). We have found that these calcium oscillations are dependent on an influx of extracellular calcium but are independent of mobilization of calcium from intracellular stores. The influx of extracellular Ca2+ occurs primarily through L-type voltage-gated calcium channels (VGCCs), since diltiazem inhibits calcium oscillations under all conditions. On the other hand, N-, P/Q-, and T-type VGCCs are not required for calcium oscillations, although inhibitors of these channels may act as partial antagonists. In addition to removal of Mg2+, oscillations can also be induced by the inhibition of voltage-gated K+ channels with 4-aminopyridine (4-AP), a treatment known to increase neurotransmitter release. In the presence of 4-AP, synchronized calcium oscillations become independent of NMDA receptor activation, although they continue to require activation of AMPA/KA receptors. A model for the mechanism of neuronal calcium oscillations and the reason for their synchrony is presented.


Auricular Vagal Nerve Stimulation Inhibited Central Nerve Growth Factor/Tropomyosin Receptor Kinase A/Phospholipase C-Gamma Signaling Pathway in Functional Dyspepsia Model Rats With Gastric Hypersensitivity.
Functional dyspepsia (FD), which has a complicated pathophysiologic process, is a common functional gastrointestinal disease. Gastric hypersensitivity is the key pathophysiological factor in patients with FD with chronic visceral pain. Auricular vagal nerve stimulation (AVNS) has the therapeutic effect of reducing gastric hypersensitivity by regulating the activity of the vagus nerve. However, the potential molecular mechanism is still unclear. Therefore, we investigated the effects of AVNS on the brain-gut axis through the central nerve growth factor (NGF)/ tropomyosin receptor kinase A (TrkA)/phospholipase C-gamma (PLC-γ) signaling pathway in FD model rats with gastric hypersensitivity. We established the FD model rats with gastric hypersensitivity by means of colon administration of trinitrobenzenesulfonic acid on ten-day-old rat pups, whereas the control rats were given normal saline. AVNS, sham AVNS, K252a (an inhibitor of TrkA, intraperitoneally), and K252a + AVNS were performed on eight-week-old model rats for five consecutive days. The therapeutic effect of AVNS on gastric hypersensitivity was determined by the measurement of abdominal withdrawal reflex response to gastric distention. NGF in gastric fundus and NGF, TrkA, PLC-γ, and transient receptor potential vanilloid 1 (TRPV1) in the nucleus tractus solitaries (NTS) were detected separately by polymerase chain reaction, Western blot, and immunofluorescence tests. It was found that a high level of NGF in gastric fundus and an upregulation of the NGF/TrkA/PLC-γ signaling pathway in NTS were manifested in model rats. Meanwhile, both AVNS treatment and the administration of K252a not only decreased NGF messenger ribonucleic acid (mRNA) and protein expressions in gastric fundus but also reduced the mRNA expressions of NGF, TrkA, PLC-γ, and TRPV1 and inhibited the protein levels and hyperactive phosphorylation of TrkA/PLC-γ in NTS. In addition, the expressions of NGF and TrkA proteins in NTS were decreased significantly after the immunofluorescence assay. The K252a + AVNS treatment exerted a more sensitive effect on regulating the molecular expressions of the signal pathway than did the K252a treatment. AVNS can regulate the brain-gut axis effectively through the central NGF/TrkA/PLC-γ signaling pathway in the NTS, which suggests a potential molecular mechanism of AVNS in ameliorating visceral hypersensitivity in FD model rats.


Changes in risk factors for hypoxic-ischaemic seizures in term infants.
Term infants with seizures and evidence of perinatal asphyxia were prospectively identified in 1 city and 2 time periods: 1978-1981 and 1991. Infants with multiple congenital abnormalities, hypocalcaemia or infection were excluded. Although there was little change in the overall incidence of neonatal seizures between 1978-1981 (1.9 per 1,000) and 1991 (1.78 per 1,000, N.S.) there was a marked reduction in small for dates infants with seizures: 8 of 19 infants in 1978-1981 compared to none of 16 in 1991 (p < 0.005). In contrast, infants > or = 41 weeks continued to show a markedly increased risk for asphyxia (relative risk 4.48, 95% CI: 1.7-12.3). The mechanism of this improved outcome for small for gestational age infants is unknown, but speculatively may be due to improved obstetric monitoring techniques allowing early identification of compromised infants.


Implications for genetic toxicology of the chromosomal breakage syndromes.
The activation of oncogenes and our knowledge of the chromosome breakage syndromes show that both intragenic mutations and chromosomal aberrations are important in carcinogenesis. Each suggests that an agent could produce genetic changes in a tissue without producing cancer there, if the types of genetic change do not match: chromosomal aberrations may be irrelevant in the mammary epithelium but be very significant in the bone marrow, and vice versa. This has vital implications for genetic toxicology: (1) both gene mutations and chromosomal aberrations should be measured, and (2) carcinogens may be mutagenic in tissues in which they are not carcinogenic. One might therefore expect in vivo assays for mutagenicity to correlate rather well with cancer bioassays; unfortunately, the bioassays themselves seem faulty. If cancer bioassays are valid, they would be reproducible. If bioassays are reproducible, they would be internally consistent. The information supplied by Tennant et al. (1987) for their validation of in vitro assays gives data from both sexes in rats and mice for 70 chemicals. When the data are analyzed site-by-site, positive results were not replicated in the other sex or in the other species much of the time: in half the cases the other sex does not give the same result; in two-thirds of the cases the other species does not give the same result. There are 3 potential explanations for these differing results: (1) genuine sex-specific carcinogens are common, (2) genuine species-specific carcinogens are common, or (3) the bioassay does not replicate well, i.e., is erratic. The third possibility best explains the data. The apparent inability of short-term in vitro tests to discriminate well between carcinogens and non-carcinogens may be more a reflection of the cancer bioassays that were used to determine which chemicals were carcinogenic than any defect in the assays. In this situation in vivo assays can scarcely be expected to do better even if they are better.


TNF-α released from retinal Müller cells aggravates retinal pigment epithelium cell apoptosis by upregulating mitophagy during diabetic retinopathy.
Retinal pigment epithelium (RPE) cell damage, including mitophagy-associated cell apoptosis, accelerates the pathogenesis of diabetic retinopathy (DR), a common complication of diabetes that causes blindness. Müller cells interact with RPE cells via pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α). Herein, we investigated the role of the RPE cell epidermal growth factor receptor (EGFR)/p38 mitogen-activated protein kinase (p38)/nuclear factor kappa B (NF-κB) pathway in Müller cell-derived TNF-α-induced mitophagy-associated apoptosis during DR. Our results showed that TNF-α released from Müller cells activated the EGFR/p38/NF-κB/p62 pathway to increase mitophagy and apoptosis in RPE cells under high glucose (HG) conditions. Additionally, blockade of the TNF-α/EGFR axis alleviates blood-retina barrier breakdown in diabetic mice. Our data further illustrate the effects of the Müller cell inflammatory response on RPE cell survival, implying potential molecular targets for DR treatment.


Identification and functional characterization of a γ-terpinene synthase in Nigella sativa L (black cumin).
Nigella sativa (Black cumin) has many applications in food and pharmaceutical industries. Thymoquinone has been considered as a main effective compound in N. sativa seeds and attracted researchers' attention mainly due to its medicinal potential. In this study, the essential oil components of leaves, flowers and seed developmental stages including half black seeds, soft black seeds and hard black seeds were analyzed in N. sativa. Whereas no terpenes were detected in flowers and leaves, seeds showed an essential oil composition that increased in its thymoquinone content during seed maturation. To study the proposed first step of thymoquinone biosynthesis, the formation of γ-terpinene from geranyl diphosphate (GDP), we identified and functionally characterized a γ-terpinene synthase (NsTPS1) in N. sativa. This monoterpene synthase was identified in RNA sequence data derived from seeds. After heterologous expression in Escherichia coli, partially purified NsTPS1 converted GDP to γ-terpinene. NsTPS1 is the first functionally characterized terpene synthase from N. sativa and displays a higher similarity to other terpene synthases from Ranunculaceae than known γ-terpinene synthases from more distant plant species. Characterization of NsTPS1 elucidates the first dedicated step in the biosynthesis of thymoquinone in N. sativa and paves the way towards metabolic engineering for high-level thymoquinone production.


Immunohistochemical localization of tissue-type plasminogen activator in ovaries before and after induced and spontaneous ovulation in the rat.
The observation that tissue-type plasminogen activator (tPA) activity increased dramatically in preovulatory follicles has led to the hypothesis that plasminogen activation is causally related to follicle rupture. With immunohistochemistry, we have studied the appearance of tPA in ovaries of immature rats induced to ovulate and in adult cycling rats. Treatment of immature female rats with a single dose of pregnant mare serum gonadotropin (PMSG) induced follicular maturation. A subsequent human chorionic gonadotropin (hCG) injection resulted in follicle rupture 12-14 h later. PMSG treatment alone did not induce appearance of tPA-immunoreactive cells in any ovarian compartment. After hCG stimulation, however, theca cells, granulosa cells, and oocytes of pre- and postovulatory follicles displayed distinct tPA immunoreactivity. Fibroblast-like cells in the theca layers and tunica albuginea of the follicle apex also demonstrated localized cytoplasmic tPA reactivity. In addition to tPA synthesis in preovulatory follicles, hCG also induced tPA staining in the theca (but not granulosa) layers of non-ovulatory follicles. At 24 h after hCG treatment, there was a marked tPA staining in developing corpora lutea, ovulated ova, and oviductal epithelium. Ovaries from regularly cycling adult rats displayed a similar ovulation-related pattern of tPA immunostaining. The appearance of tPA in different cell types of the preovulatory follicle and in the fibroblast-like cells at the follicle apex, strengthens the hypothesis of a direct involvement of tPA in follicle rupture. Presence of tPA in postovulatory oocytes, cumulus cells, and surrounding oviductal epithelium may also indicate a role for tPA in the transfer of eggs in the oviduct.


Refinement of Draft Genome Assemblies of Pigeonpea (Cajanus cajan).
Genome assembly of short reads from large plant genomes remains a challenge in computational biology despite major developments in next generation sequencing. Of late several draft assemblies have been reported in sequenced plant genomes. The reported draft genome assemblies of Cajanus cajan have different levels of genome completeness, a large number of repeats, gaps, and segmental duplications. Draft assemblies with portions of genome missing are shorter than the referenced original genome. These assemblies come with low map accuracy affecting further functional annotation and the prediction of gene components as desired by crop researchers. Genome coverage, i.e., the number of sequenced raw reads mapped onto a certain location of the genome is an important quality indicator of completeness and assembly quality in draft assemblies. The present work aimed to improve the coverage in reported de novo sequenced draft genomes (GCA_000340665.1 and GCA_000230855.2) of pigeonpea, a legume widely cultivated in India. The two recently sequenced assemblies, A1 and A2 comprised 72% and 75% of the estimated coverage of the genome, respectively. We employed an assembly reconciliation approach to compare the draft assemblies and merge them, filling the gaps by employing an algorithm size sorting mate-pair library to generate a high quality and near complete assembly with enhanced contiguity. The majority of gaps present within scaffolds were filled with right-sized mate-pair reads. The improved assembly reduced the number of gaps than those reported in draft assemblies resulting in an improved genome coverage of 82.4%. Map accuracy of the improved assembly was evaluated using various quality metrics and for the presence of specific trait-related functional genes. Employed pair-end and mate-pair local libraries helped us to reduce gaps, repeats, and other sequence errors resulting in lengthier scaffolds compared to the two draft assemblies. We reported the prediction of putative host resistance genes against Fusarium wilt disease by their performance and evaluated them both in wet laboratory and field phenotypic conditions.