The Somogyi phenomenon in insulin-dependent diabetics.
Insulin-dependent diabetics at times of changing stress levels require alterations in their insulin dosage. Determination of the required insulin dosage is not a precise science. At these times, the diabetic may experience insulin overdosage with a resulting rebounding hyperglycemia (Somogyi phenomenon). Adequate data collection and a systematic approach to documentation of the data by either the nurse or the patient will provide both the physician and the nurse with the means to recognize the occurrence of the Somogyi phenomenon. The author recommends a tool that can be used to enhance recognition of the occurrence and provide clinical criteria to evaluate its presence.


Reflection interference contrast microscopy combined with scanning force microscopy verifies the nature of protein-ligand interaction force measurements.
The integration of a stand-alone scanning force microscope (SFM) scanner with a reflection interference contrast microscope (RICM) makes it possible to measure directly the separation distance between the SFM probe and the sample surface. The SFM-RICM combination, when applied to the force measurements between ligand-derivatized SFM probe and a protein receptor-derivatized surface, showed that the anomalous force discontinuities often observed for such interacting pairs were indeed a real behavior characteristic of a particular experimental configuration. Apart from small discrepancies due to transient damping, commercially available cantilevers did behave in an ideal mechanical fashion, thus indicating that protein-ligand unbinding events were occurring at distances much larger than their maximum extended length. This external verification of separation distance requires a closer examination of the physical events occurring upon detachment of the surfaces. An alternative interpretation of such force measurements is proposed here in which the protein and/or ligand immobilization chemistry is called into question.


Surface model of the collagen fibril as determined by two-stage plastic replica.
Two-stage plastic replicas of fresh and vacuum dried rat tail collagen were examined by electron microscopy. Microphotometer tracings made from the replica micrographs showed that the surface of the collagen has a periodic undulating topography which includes an approximately 20--40 A deep depression located within the major band distance.


Use of a quantitative gene expression assay based on micro-array techniques and a mathematical model for the investigation of chlamydial generation time.
Chlamydia is an important pathogen which possesses a unique developmental cycle. We used real-time PCR technology to measure gene transcript levels in Chlamydia trachomatis strain L2. By measuring 16S rRNA transcript levels, and developing a mathematical model of the chlamydial developmental cycle fitting the data, we predict an average generation time of approximately 2.6 h. Additionally, potentially this modelling also provides the foundation for the application of emerging micro-array technology in which identification of the gene signals that trigger a chlamydial body to start replicating or transform to its infectious form can be made possible.


Glucose-based versus fat-based total parenteral nutrition (TPN): effects on hepatic function in septic patients complicated with cholestatic jaundice.
A prospective study of two types of total parenteral nutrition (TPN) was carried out in 34 patients suffering from sepsis and complicated liver dysfunction. Group 1 (18 patients) received non-protein energy as glucose plus fat emulsion in a caloric ratio of 19:1, while group 2 (16 patients) received the same energy intake but with a ratio of 1:1. Group 1 exhibited higher levels of bilirubin and alkaline phosphatase with values of 93.5 +/- 25.5 mumol/l and 160 +/- 30 IU/l respectively compared to Group 2, in which the corresponding values were 81.6 +/- 32.3 mumol/l and 120 +/- 10 IU/l (p < 0.05). On the other hand, group 1 had lower levels of serum albumin and serum transferrin with values 25 +/- 1.3 g/l and 40 +/- 20% of normal, compared to group 2 in whom the corresponding values were 28 +/- 8 g/l and 48 +/- 30% of normal (p < 0.05). There were no differences between the two groups, in the absolute number of T-lymphocytes and in transaminase levels. In sepsis complicated by liver dysfunction a 50:50 glucose: fat regimen caused less disturbance of liver function than one consisting almost entirely of glucose.


Sonographic findings in women with complications after hysterectomy.
To study the role of abdominal ultrasound in the management of complications after emergency or elective hysterectomy. The study compared 102 women who had complications after emergency peripartum hysterectomy (study group) with 102 women who had complications after elective hysterectomy for fibroids (control group). The study group had a lower mean age (31.1+/-2.1 vs 40.1+/-3.4 years) but higher parity (3.0+/-1.3 vs 2.0+/-1.2) compared with the control group. The primary complication in the study group was abdominal mass or distension (55.9%) compared with chronic abdominal pain (49.0%) in the control group. The leading sonographic abnormality in the study group was pelvic abscess (9.8%) while hydrosalpinx (6.9%) was more common in the control group. There was no statistical difference in the findings between the groups. Management of complications included ovarian cystectomy and drainage of pelvic abscesses. Ultrasound facilitates accurate diagnosis of complications after hysterectomy and enhances management.


Health care utilization among beneficiaries with diabetes from federally qualified health centers: Analysis from a Medicaid managed care organization.
We examined health care utilization among federally qualified health center (FQHCs) users from a Medicaid managed care organization based on 2016 administrative claims data (n = 8,402). FQHC users had fewer primary care visits (Adjusted Incidence Rate Ratios (aIRR): 0.82; 95% CI: 0.76-0.88) compared with non-FQHC users. Statistically significant differences were not observed in emergency department visits (aIRR: 1.19; 95% CI: 0.98-1.46) and hospitalizations (aIRR: 1.03; 95% CI: 0.80-1.34). FQHCs provide comprehensive primary care to Medicaid managed care beneficiaries with diabetes in fewer PCP visits. Results provide evidence to health policy experts and MCOs to increase provider network contracting with FQHCs.


Modulation of the Naked-Eye and Fluorescence Color of a Protonated Boron-Doped Thiazolothiazole by Anion-Dependent Hydrogen Bonding.
The reaction of a cyclic alkyl(amino)carbene (CAAC)-stabilized thiazaborolo[5,4-d]thiazaborole (TzbTzb) with strong Brønsted acids, such as HCl, HOTf (Tf=O2 SCF3 ) and [H(OEt2 )2 ][BArF 4 ] (ArF =3,5-(CF3 )2 C6 H3 ), results in the protonation of both TzbTzb nitrogen atoms. In each case X-ray crystallographic data show coordination of the counteranions (Cl- , OTf- , BArF 4 - ) or solvent molecules (OEt2 ) to the doubly protonated fused heterocycle via hydrogen-bonding interactions, the strength of which strongly influences the 1 H NMR shift of the NH protons, enabling tuning of both the visible (yellow to red) and fluorescence (green to red) colors of these salts. DFT calculations reveal that the hydrogen bonding of the counteranion or solvent to the protonated nitrogen centers affects the intramolecular TzbTzb-to-CAAC charge transfer character involved in the S0 →S1 transition, ultimately enabling fine-tuning of their absorption and emission spectral features.


Effect of several androgens, cyproterone acetate or estrogen-progesterone on the prolactin-releasing activity of arginine vasotocin in castrated male rats.
Intravenous (iv) administration of 5 microgram of arginine vasotocin (AVT) into urethane-anesthetized, castrated male rats had no effect on plasma prolactin titers as compared to the rise in prolactin levels observed in intact AVT-treated rats. However, when castrated rats were first treated for two days with 2.5 mg/day of testosterone propionate and then challenged with a 5-microgram dose of AVT, the prolactin surge values obtained were comparable to those seen in intact AVT-treated rats. Conversely, treatment of intact rats for two days with 25 mg/day of the anti-androgen, cyproterone acetate, blocked the prolactin-releasing activity of AVT. In a separate experiment, treatment of castrated rats for two days with 2.5 mg/day of testosterone, androsterone or 50 microgram of estradiol benzoate and 25 mg progesterone, completely restored the prolactin-releasing activity of AVT. Similar treatment with 2.5 mg/day of androstenedione or dihydrotestosterone was without effect in restoring this response to AVT. It is concluded that the presence of gonadal steroids is essential to the action of AVT in provoking the release of prolactin in urethane-anesthetized male rats.


Proton binding sites and conformational analysis of H+K(+)-ATPase.
It is proposed that the hydronium ion, H3O+, binds to the E1 conformation of the alpha-subunit of gastric proton pump. The H3O+ binding cavities are characterized parametrically based on valence, sequence, geometry, and size considerations from comparative modeling. The cavities have scope for accommodating monovalent cations of different ionic radii. The H3O+ transport is proposed to be aided by arenes which are arranged regularly along the pump starting from N-domain through the transmembrane region. Step-by-step structural changes accompanying H3O+ occlusion are studied in detail. The observations corroborate well with earlier experimental studies.


The atlas of cytoophidia in Drosophila larvae.
In 2010, cytidine 5'-triphosphate synthase (CTPS) was reported to form the filamentous or serpentine structure in Drosophila, which we termed the cytoophidium. In the last decade, CTPS filaments/cytoophidia have been found in bacteria, budding yeast, human cells, mice, fission yeast, plants, and archaea, indicating that this mechanism is highly conserved in evolution. In addition to CTPS, other metabolic enzymes have been identified to have the characteristics of forming cytoophidia or similar advanced structures, demonstrating that this is a basic strategy of cells. Nevertheless, our understanding of the physiological function of the cytoophidium remains incomplete and elusive. Here, we took the larva of Drosophila melanogaster as a model to systematically describe the localization and distribution of cytoophidia in different tissues during larval development. We found that the distribution pattern of CTPS cytoophidia is dynamic and heterogenic in larval tissues. Our study provides a road map for further understanding of the function and regulatory mechanism of cytoophidia.


Results of a survey on the treatment of adolescents and young adult (AYA) sarcoma patients in Spain at present time.
We aimed to analyse health care services for adolescents and young adults (AYA) with sarcomas in Spain. A survey was sent to all Spanish cancer centres, including questions about demographic, facilities, and treatment strategies for AYAs with sarcomas in the last 2 years. Thirty-five units participated in the survey, 17 paediatric and 15 adult units. There were three specialized AYA units. First line regimen varied depending on whether the treating unit was paediatric or not, for osteosarcomas, rhabdomyosarcomas, and non-rhabdomyosarcomas. By contrast, 91.4% of Ewing sarcomas were treated according to EE2012. In the relapse setting, differences between units were higher in all tumours. Additionally, 48% of the units reported not having trials for this population. There are major differences in the treatment of AYAs with sarcomas between adult and paediatric units. Enormous efforts are needed to homogenize treatments and increase the access to innovation.


A simple epidemic model for semi-closed community reveals the hidden outbreak risk in nursing homes, prisons, and residential universities.
We develop a general SIS model to study the epidemic transmission in such semi-closed communities. The community population is divided into susceptible and infected in terms of the infection state, and concerning the physical structure of the crowd, they are classified into mobile and fixed individuals. The mobile individuals can be inside or outside the community, while the fixed individuals can be only inside the community. There are fixed infection sources outside the community, measuring the epidemic severity in society. We attribute the spreading to two reasons: (i) clustered infection among the community population and (ii) the epidemic in society spreading to the community population. We discuss the model in two cases. In the first case, the epidemic spreads in society, such that reasons (i) and (ii) work together. The results show that concerning fixed individuals (e.g. the elderly in nursing homes), a more closed community always promotes the infection. In the second case, there is no epidemic spreading in society, such that only reason (i) works. The results show that restricting all individuals to the community produces equivalent consequences as allowing them going outside the community. We should evenly distribute individuals inside and outside to form isolation. A counterexample is residential universities implementing closed management, where only students are restricted to campus. The model shows such management may lead to severe epidemics, and to prevent the epidemic outbreaks, students should have free access to being on or off campus.


Synthesis, anticonvulsant activity, and neuropathic pain-attenuating activity of N-benzyl 2-amino-2-(hetero)aromatic acetamides.
N-Benzyl 2-acetamido-2-substituted acetamides, where the 2-substituent is a (hetero)aromatic moiety, are potent anticonvulsants. We report the synthesis and whole animal pharmacological evaluation of 16 analogues where the terminal 2-acetyl group was removed to give the corresponding primary amino acid derivatives (PAADs). Conversion to the PAAD structure led to a substantial drop in seizure protection in animal tests, demonstrating the importance of the N-acetyl moiety for anticonvulsant activity. However, several of the PAADs displayed notable pain-attenuating activities in a mouse model.


Survival analysis for a large-scale forest health issue: Missouri oak decline.
Survival analysis methodologies provide novel approaches for forest mortality analysis that may aid in detecting, monitoring, and mitigating of large-scale forest health issues. This study examined survivor analysis for evaluating a regional forest health issue - Missouri oak decline. With a statewide Missouri forest inventory, log-rank tests of the effects of covariates on the survivor function and equality of the survivor function among classes were conducted for selected oak species. Additionally, hazard functions were determined for diameter classes for damaged and undamaged oaks. Results indicate that mortality appears to vary significantly among some inventory classes such as oak species, but not among other classes such as ownership class. Indicators of individual tree vigor (i.e., crown class and ratio) were more significant predictors of oak tree mortality than site/stand attributes (i.e., density and aspect). Finally, results indicate that even fast-growing oak trees are at high risk of mortality if damaged by disease. Survival analyses, such as those applied in this study, may enable testing of forest health hypotheses using large-scale inventories. In the context of Missouri's oak forest decline, study results suggest management efforts should focus on limiting the spread of disease damage, increasing the vigor of residual trees, and emphasizing small trees when developing stand prescriptions.


Peripheral blood minimal residual disease may replace bone marrow minimal residual disease as an immunophenotypic biomarker for impending relapse in acute myeloid leukemia.
As relapses are common in acute myeloid leukemia (AML), early relapse prediction is of high importance. Although conventional minimal residual disease (MRD) measurement is carried out in bone marrow (BM), peripheral blood (PB) would be an advantageous alternative source. This study aims to investigate the specificity of leukemia-associated immunophenotypes used for MRD detection in blood samples. Consistency of PB MRD as compared with BM MRD was determined in flow cytometric data of 205 paired BM and PB samples of 114 AML patients. A significant correlation was found between PB and BM MRD (r=0.67, P<0.001), while median PB MRD percentage was factor 4-5 lower compared with BM MRD. Primitive blast (CD34+/CD117+/CD133+) frequency was significantly lower in PB (median factor 23.7), indicating that PB MRD detection is more specific than BM. Cumulative incidence of relapse 1 year after induction therapy was 29% for PB MRD-negative and 89% for PB MRD-positive patients (P<0.001). Three-year OS was 52% for MRD-negative and 15% for MRD-positive patients (P=0.034). Similar differences were found after consolidation therapy. As PB MRD appeared to be an independent predictor for response duration, the highly specific PB MRD assay may have a prominent role in future MRD assessment in AML.


A systematic review of hemorrhage and vascular injuries in civilian public mass shootings.
Civilian public mass shootings (CPMSs) are a major public health issue and in recent years several events have occurred worldwide. The aim of this systematic review was to characterize injuries and mortality after CPMSs focusing on in-hospital management of hemorrhage and vascular injuries. A systematic review of all published literature was undertaken in Medline, Embase and Web of Science January 1st, 1968, to February 22nd, 2021, according to the PRISMA guidelines. Literature was eligible for inclusion if the CPMS included three or more people shot, injured or killed, had vascular injuries or hemorrhage. The search identified 2884 studies; 34 were eligible for inclusion in the analysis. There were 2039 wounded in 45 CPMS events. The dominating anatomic injury location per event was the extremity followed by abdomen and chest. The median number of operations and operated patients per event was 22 (5-101) and 10.5 (4-138), respectively. A total of 899 deaths were reported with a median mortality rate of 36.1% per event (15.9-71.4%) Thirty-eight percent (13/34) of all studies reported on vascular injuries. Vascular injuries ranged from 8 to 29%; extremity vascular injury the most frequent. Specific vascular injuries included thoracic aorta 18% (42/232), carotid arteries 6% (14/232), and abdominal aorta 5% (12/232). Vascular injuries were involved in 8.3%-10% of all deaths. This systematic review showed an overall high mortality after CPMS with injuries mainly located to the extremities, thorax and abdomen. About one quarter of deaths was related to hemorrhage involving central large vessel injuries. Further understanding of these injuries, and structured and uniform reporting of injuries and treatment protocols may help improve evaluation and management in the future. Level of Evidence Systematic review and meta-analysis, level III.


Comparative solvent perturbation of horse heart cytochrome c and Rhodospirillum rubrum cytochrome c2.
The extent of exposure of heme to solvent in horse heart cytochrome c and Rhodospirillum rubrum c2 was investigated to determine whether a correlation exists between the properties of these oxidation-reduction proteins and their heme environments. Solvent perturbation absorption difference spectra were measured using ethylene glycol, glycerol, and sucrose at concentrations between 0 and 30%. Cytochrome c appears to exhibit a somewhat greater extent of heme exposure than cytochrome c2 for both the oxidized and reduced states. These results suggest that the lower oxidation-reduction potential of cytochrome c may in part be due to a greater extent of exposure of the heme. The oxidized state of both proteins appears to exhibit a greater exposure than that of the reduced state which is consistent with a more favorable environment for the charge on the ferric heme coordination center.


Calcitonin gene-related peptide (CGRP)-immunoreactive sensory and motor nerves of the mammalian palate.
The distribution of calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibres in the palate of rat, cat and monkey was studied using immunocytochemistry and radioimmunoassay. CGRP-containing nerve fibres were found, in all species studied, to form a rich plexus in the subepithelial and submucous layers, around excretory ducts and blood vessels. A small number of CGRP-containing nerve fibres penetrated the epithelium of the hard and soft palate, and terminated as free endings. Some CGRP-containing nerve fibres were found in the vicinity of the mucous glands. CGRP-immunoreactive motor end plates were seen in the striated muscle (tensor veli palatini) of the soft palate. Following capsaicin treatment a small depletion in CGRP-immunoreactive nerve fibres in the rat palate epithelium was noted. In contrast, CGRP immunoreactive fibres forming rich plexuses in other layers of the palate, including motor end plates, were not affected. The extractable CGRP showed no significant depletion (normal animals [n = 10] 21.7 +/- 2.4 pmol/g compared with capsaicin-treated animals [n = 10] 17.5 +/- 1.8 pmol CGRP/g wet weight). The reduction in the number of visible immunoreactive nerves following capsaicin application tends to confirm the sensory character of the CGRP-containing nerve fibres terminating in the epithelium of the hard and soft palate. The capsaicin insensitive CGRP-immunoreactive nerve fibres may thus have a predominantly motor function.(ABSTRACT TRUNCATED AT 250 WORDS)


Associations of cigarette smoking with memory decline and neurodegeneration among cognitively normal older individuals.
Cigarette smoking is associated with a higher risk of Alzheimer's disease (AD), but the underlying mechanisms remain to be clarified. In this study, we aimed to examine the effects of cigarette smoking on multiple AD biomarkers among older individuals with normal cognition (NC). Among 415 older individuals with NC from the Alzheimer's disease Neuroimaging Initiative (ADNI) cohort, we examined the associations between smoking status (non-smokers vs smokers) and global cognition, verbal memory, hippocampal volumes, cerebral glucose metabolism and CSF AD pathologies. The primary findings of this study were: (1) In NC, smokers showed worse performance on verbal memory tests [Rey Auditory Verbal Learning Test (RAVLT) total learning score and delayed recall] than non-smokers; (2) Compared with non-smokers, smokers had significantly lower HpVR; (3) Smokers, relative to non-smokers, demonstrated lower levels of cerebral glucose metabolism as measured by FDG-PET; and (4) there were no significant differences in CSF AD pathologies (CSF Aβ42, t-tau or p-tau) between non-smokers and smokers. Longitudinal studies are needed to investigate the relationship between cigarettes smoking and changes in AD-related markers over time. Further, ADNI participants were highly educated and predominantly white. This may limit the generalizability of our results. In summary, among individuals with NC, cigarette smoking was associated with memory impairment, hippocampal atrophy and cerebral glucose hypometabolism, but not CSF AD pathologies.


Conservative treatment of severe limited mouth opening after transtemporal craniotomy.
Limited mouth opening (LMO) is a common complication of the transtemporal craniotomy that is often involved in the temporalis and temporomandibular joint. The occurrence of prolonged LMO was rare. Risk factors for such limitation after these neurosurgical procedures are defined, and the strategies of treatment and prevention are further discussed.This article presents 12 patients with severe LMO after transtemporal neurosurgical procedures. Four patients received myofunctional training combined with the neuromuscular blocking in the temporalis, which methods may be more effective for LMO within 3 months. Five patients received myofunctional training combined with joint injection of hyaluronic acid, which methods may be more effective for LMO beyond 3 months. In addition, 2 patients who experienced LMO for more than 1 year had to receive coronoidectomy.Therefore, these conservative treatments have proved a useful method to restore LMO at an early stage. If intervention and diagnosis was delayed, the efficiency of these conservative therapies would decline, and coronoidectomy has to be considered. The early physiotherapy cannot be overemphasized to minimize this complication after these transtemporal neurosurgical procedures.


Intestinal absorptive and digestive function in pernicious anemia.
We examined the digestive and absorptive function of the small intestinal mucosa in three patients with pernicious anemia to determine the functional correlates of the morphologic changes previously described. Digestive brush border enzymes (disaccharidases and leucyl-naphthylamidase) in jejunal biopsy specimens from the patients followed a normal distribution compared with those in the control group. With the exception of one patient with mild steatorrhea, the rest of the absorption test results were within the normal range. Jejunal perfusion studies, however, with glucose, sodium and water showed intestinal secretion of sodium and water, i.e., net movement of sodium and water from plasma to lumen, in the presence of normal glucose absorption. Follow-up studies in two patients after treatment with vitamin B12 did not reveal any significant improvement in the absorption rates from the pretreatment period. This abnormality of sodium and water transport in pernicious anemia represents another intestinal defect of a systemic disease which is not corrected by vitamin B12 replacement therapy.


Adhesion dynamics, morphology, and organization of 3T3 fibroblast on chitosan and its derivative: the effect of O-carboxymethylation.
Chitosan and O-carboxymethylchitosan (OCMCS) have been proved to have biocompatibility and have been extensively researched in the field of biomaterials. In this study, Confocal-reflectance interference contrast microscopy (C-RICM) in conjunction with phase contrast imaging was used to investigate the adhesion contact dynamics of 3T3 fibroblasts on chitoan and OCMCS surface-modified silica coverslips. The C-RICM results demonstrate that the weak cell contact forms on OCMCS surface while a much stronger contact area forms on the chitosan surface. 3T3 fibroblasts are found to spread randomly with spindlelike morphology on the chitosan surface, while they exhibit elongated morphology and align on the OCMCS surface. It is believed that fibroblast behaviors such as migration, spreading with an elongated morphology, and alignment on the OCMCS surface are correlated with the weak cell contact. The mechanisms to form cell adhesion contact on chitosan and OCMCS were discussed.


Factors affecting relapse in node-negative breast cancer. A multivariate analysis including the labeling index.
Between 1975 and 1982, 167 patients with carcinoma of the breast without axillary lymph node metastases were studied. The thymidine labeling index (LI), representing the percentage of cells in the DNA synthesis phase, was measured in all these patients. High LI values were more frequently encountered in young patients (P = 0.05), in low estrogen receptor (ER) tumor content (P = 0.007) and in high grade tumors (P = 0.0002). The overall 8-year relapse-free survival (RFS) was 68%. Univariate analysis demonstrated that RFS was influenced by histological grading (P = 0.03), ER (P = 0.03), PR (P = 0.02) and LI (P = 0.01). Multivariate analysis using the Cox regression model selected the LI as the single significant prognostic factor with regard to RFS (P = 0.037). These results emphasize the important role of cell proliferation kinetics in defining node-negative breast cancer patients with a high risk of relapse.


The changing scene in graduate medical education: implications for pathology.
During the past several years a number of unrelated events have occurred that will have a marked impact upon residency training in pathology in the United States. These factors include: (1) a twofold increase in the annual number of graduates from American medical schools, (2) Public Law 94-484, which will reduce the number of foreign medical graduates eligible for postgraduate training in this country by 70 to 80 per cent, and (3) increased emphasis on permitting Americans studying abroad to transfer to United States medical schools. Barring unforeseen changes, the culmination of these events is expected circa 1984 when the number of applicants for first year house office positions will equal or exceed the pool of available slots. Therefore, a significant proportion of graduates either will not be matched at all or will not be matched with the specialty(ies) of their choice. The implications of these trends with respect to residency training in pathology, with particular reference to the motives and motivation of persons seeking a first year slot, are discussed and alternative strategies for maximizing benefits discussed.


Development and validation of prediction model for incident overactive bladder: The Nagahama study.
We aimed to develop models to predict new-onset overactive bladder in 5 years using a large prospective cohort of the general population. This is a secondary analysis of a longitudinal cohort study in Japan. The baseline characteristics were measured between 2008 and 2010, with follow-ups every 5 years. We included subjects without overactive bladder at baseline and with follow-up data 5 years later. Overactive bladder was assessed using the overactive bladder symptom score. Baseline characteristics (demographics, health behaviors, comorbidities, and overactive bladder symptom scores) and blood test data were included as predictors. We developed two competing prediction models for each sex based on logistic regression with penalized likelihood (LASSO). We chose the best model separately for men and women after evaluating models' performance in terms of discrimination and calibration using an internal validation via 200 bootstrap resamples and a temporal validation. We analyzed 7218 participants (male: 2238, female: 4980). The median age was 60 and 55 years, and the number of new-onset overactive bladder was 223 (10.0%) and 288 (5.8%) per 5 years in males and females, respectively. The in-sample estimates for C-statistic, calibration intercept, and slope for the best performing models were 0.77 (95% confidence interval 0.74-0.80), 0.28 and 1.15 for males, and 0.77 (95% confidence interval 0.74-0.80), 0.20 and 1.08 for females. Internal and temporal validation gave broadly similar estimates of performance, indicating low optimism. We developed risk prediction models for new-onset overactive bladder among men and women with good predictive ability.


Community-based perinatal mental health peer support: a realist review.
Peer support has been suggested as an alternative or complement to professional support for mothers with perinatal mental health difficulties. The aim of this realist review was to synthesise the evidence on perinatal mental health peer support programmes outside mental health services, to understand what is it about community-based perinatal mental health peer support that works, for whom, in what circumstances, in what respects, and why. Applying realist methodology, an initial theoretical model was tested against evidence from empirical studies. 29 empirical studies were included, covering 22 antenatal and postnatal mental health interventions that offered one-to-one or group peer support, in person or by telephone. Data extraction identified the configurations of contexts (C), mechanisms (M) and outcomes (O) relevant to mothers' use of peer support and to the positive and negative effects of using peer support. 13 C-M-O configurations explained take-up of peer support. These were based on mothers' perceptions that peer support would offer empathetic understanding and non-judgemental acceptance outside their social circle; their relationships with primary health professionals; their cultural background and perspectives on mental health; their desire for professional support; overcoming practical barriers; the format of the support; and the use of volunteers. A further 13 C-M-O configurations explained positive impact on mothers. These were based on receiving empathetic listening, acceptance, affirmation and normalisation; peers sharing ideas about self-care, coping, and services; peers using therapeutic techniques; the opportunity to give support to others; meaningful social relationships with volunteers and other mothers; and other benefits of attending a group. There were 8 C-M-O configurations explaining negative impact. These were based on lack of validation; self-criticism from downward and upward social comparison; a culture of negativity; peers being judgemental or directive; not feeling heard; peer support as a stressful social relationship; and distress at endings. Peer support works in complex ways that are affected by personal and social contexts. Providers, commissioners and evaluators can use this review to understand and maximise the valuable benefits of peer support, to minimise potential risks, and to devise ways of reaching mothers who do not currently engage with it.


Thermal adaptations in lizard muscle function.
This study was undertaken to investigate thermal adaptations in muscle contractile properties in closely-related lizards with different preferred body temperatures (PBT). The species examined all belong to the Sphenomorphus group of Australian skinks (Scincidae: Lygosominae). Preferred body temperatures are conservative at the generic level as follows: Ctenotus, 35 degrees C; Sphenomorphus, 30 degrees C; Eremiascincus, 25 degrees C. Contractile properties of the fast glycolytic portion of the iliofibularis muscle were measured. Translational adaptations are evident in several isometric factors, including tetanic tension (Po), twitch tension (Pt), twitch time to peak tension (TPT), and twitch half-relaxation time (1/2 RT). Capacity adaptations are not evident in rates of tetanic tension development (dPo/dt) or in maximal velocities of isotonic shortening (Vmax). Rotational adaptations are not evident in any contractile properties. Thermal limits on upper response temperatures are about 5 degrees C warmer in Ctenotus than in the more cryophilic species, indicative of resistance adaptation in muscle performance. Despite these adaptive shifts, there is little indication that muscle functional capacities are optimized or equalized at PBT in these lizards.


Dexamethasone induces p21cip1/waf1 expression via FoxO3a independently of the Lamin A/C-HDAC2 interaction in Ataxia Telangiectasia.
Ataxia-Telangiectasia (A-T) is a very rare autosomal recessive multisystemic disorder which to date is still uncurable. The use of glucocorticoid analogs, such as dexamethasone (dex), can improve neurological symptoms in patients, but the molecular mechanism of action of these analogs remains unclear. Here, we report the effects of dex in regulating the interaction between Lamin A/C and HDAC2 in WT and A-T cells. Upon administration of dex to A-T cells, we first observed that the accumulation of HDAC2 on the CDKN1A promoter did not exert a repressive role on p21cip1/waf1 expression, and second, we established that HDAC2 accumulation was not dependent on Lamin A/C. Both of these results are contrary to previous reported outcomes in other cellular models. Furthermore, large amounts of LAP2α and FoxO3a were found to occupy the CDKN1A promoter with matched p21cip1/waf1 overexpression. Hence, in A-T cells p21 could be activated as a result of a dex-induced rearrangement of a multicomponent complex, composed of Lamin A/C, HDAC2, LAP2α, pRb, E2F1, and FoxO3a, at the CDKN1A gene promoter.


Tolerance of adjuvant letrozole outside of clinical trials.
Recently aromatase inhibitors have become a standard care as an adjuvant treatment for many postmenopausal patients with hormone receptor positive early breast cancer. Adjuvant letrozole was made available either immediately postoperative, after 2-3 years of tamoxifen, or as an extended treatment after 5 years of tamoxifen. Between October 2003 and October 2005, we analyzed the subjective tolerance in 185 postoperative early breast cancer patients receiving letrozole outside of a clinical trial. The most prominent toxicity was musculoskeletal pain. In addition hot flushes, increased fatigue, nausea, vomiting, anorexia, mood disturbances, vaginal dryness, hair loss and rash were also recorded. In contrast to the prospective randomized clinical trials, a high drop-out rate of 20% was documented, mainly due to aromatase inhibitor-associated arthralgia syndrome interfering significantly with the daily life of our patients. Although adjuvant aromatase inhibitors have proven to be generally superior to tamoxifen in the adjuvant setting, it is important to focus attention on the tolerance during the adjuvant therapy and to balance this against the potential benefit in individual patients. Alternative options including switching to tamoxifen remain available.


Direct determination of impurities in high purity silicon carbide by inductively coupled plasma optical emission spectrometry using slurry nebulization technique.
A novel method for the determination of Al, Ca, Cr, Cu, Fe, Mg, Mn, Ni and Ti in high purity silicon carbide (SiC) using slurry introduction axial viewed inductively coupled plasma optical emission spectrometry (ICP-OES) was described. The various sizes of SiC slurry were dispersed by adding dispersant polyethylene imine (PEI). The stability of slurry was characterized by zeta potential measurement, SEM observation and signal stability testing. The optimal concentration of PEI was found to be 0.5 wt% for the SiC slurry. Analytical results of sub-mum size SiC by the slurry introduction were in good accordance with those by the alkaline fusion method which verified that determination could be calibrated by aqueous standards. For mum size SiC, results of most elements have a negative deviation and should be calibrated by the Certified Reference Material slurry. Owing to a rather low contamination in the sample preparation and stability of the slurry, the limits of detection (LODs), which are in the range of 40-2000 ng g(-1), superior to those of the conventional nebulization technique by ICP-OES or ICP-MS.


Does resveratrol improve insulin signaling in chronically ischemic myocardium?
Resveratrol is a naturally occurring polyphenol believed to be cardioprotective. We previously demonstrated that resveratrol improves insulin signaling and glucose metabolism in liver and skeletal muscle of swine with metabolic syndrome. Although resveratrol has metabolic benefits in peripheral tissues, its effect on insulin signaling in ischemic myocardium (IM) is unclear. Therefore, we developed a clinically relevant swine model of metabolic syndrome and chronic myocardial ischemia to investigate the effects of resveratrol on insulin signaling in cardiac tissue. Thirteen male Yorkshire swine were fed a high-cholesterol diet for 4 wk then underwent surgical placement of an ameroid constrictor to their circumflex artery to induce chronic myocardial ischemia. The high-cholesterol control group was given no drug (n = 7). The experimental group was provided the same diet and received supplemental resveratrol (100 mg/kg/d) (n = 6). Tissue was harvested 7 wk after ameroid placement for western blot and histological analyses. In IM, there was no significant difference between the two groups in the insulin signaling markers studied. In nonischemic myocardium, there was a significant decrease in phosphorylated AMP-activated protein kinase α (P = 0.021) in the group treated with resveratrol; otherwise, there were no significant differences between the groups. Immunostaining for glucose transporter 4 and Periodic acid-Schiff staining for myocardial glycogen stores was similar between the groups. Resveratrol has complex effects on glucose metabolism. Although prior studies demonstrated that resveratrol supplementation improves insulin sensitivity in peripheral tissues, in chronically IM, there are no significant alterations.


Relation of thrombogenesis in systemic hypertension to angiogenesis and endothelial damage/dysfunction (a substudy of the Anglo-Scandinavian Cardiac Outcomes Trial [ASCOT]).
Increasing evidence points toward a prothrombotic state in hypertension and atherosclerosis, conditions associated with thrombosis-related complications, such as myocardial infarction and stroke. We hypothesized that this increased risk of thrombogenesis may be related to endothelial damage/dysfunction and abnormal angiogenesis, and thus, an increased risk of future cardiovascular disease. Thrombogenesis, endothelial damage/dysfunction, and angiogenesis can be assessed by measurement of tissue factor (TF), von Willebrand Factor (vWF), flow-mediated dilatation (FMD), and vascular endothelial growth factor (VEGF), respectively. To test this hypothesis, we measured TF, vWF, FMD, and VEGF in 76 patients with systemic hypertension (71 men; mean age 64; mean blood pressure 167/72 mm Hg), considered additional risk factors such as diabetes, and related them to the patient's 10-year cardiovascular and cerebrovascular risk score using the Framingham equation. Patients were compared with 48 healthy normotensive controls. In these patients, the effects of 6 months of intensified blood pressure and (where appropriate) lipid-lowering treatment were investigated. In our patients, TF, VEGF, and vWF levels were higher, but FMD was lower (all p <0.001) compared with the controls. All markers correlated with each other and with both cardiovascular and cerebrovascular risk scores (all p <0.001). After intensified blood pressure and hypercholesterolemia treatment, total cholesterol, blood pressure, TF, VEGF, and vWF levels all decreased, whereas FMD increased (all p <0.001). Thus, in subjects with hypertension and other risk factors, endothelial damage/dysfunction (and thus, atherogenesis), thrombogenesis, and angiogenesis are abnormal, correlate with overall cardiovascular risk, and importantly, can be related to each other in a "Birmingham Vascular Triangle." Furthermore, these processes are beneficially affected by intensive blood pressure and lipid treatment.


The comparative cariogenicity and plaque-forming ability in vivo of four species of the bacterium Actinomyces in gnotobiotic rats.
Gnotobiotic WAG/RIJ rats on a high sucrose diet were monoinfected with 12 strains of Actinomyces spp. Moderate levels of caries were induced by a single strain of Actinomyces naeslundii and low levels by two strains of Actinomyces viscosus, three strains of A. naeslundii and one strain of Actinomyces israelii. No caries was induced by single strains of A. viscosus and A. israelii or by three Actinomyces odontolyticus strains. Only fissure caries was observed. Scanning electron microscopy showed that all strains colonized the fissures and most colonized the lingual surface of the teeth, but to a limited extent. Production of abundant and dense plaque was not always accompanied by caries.


Exosome nanovesicles as potential biomarkers and immune checkpoint signaling modulators in lung cancer microenvironment: recent advances and emerging concepts.
Lung cancer remains the leading cause of cancer-related deaths globally, and the survival rate remains low despite advances in diagnosis and treatment. The progression of lung cancer is a multifaceted and dynamic phenomenon that encompasses interplays among cancerous cells and their microenvironment, which incorporates immune cells. Exosomes, which are small membrane-bound vesicles, are released by numerous cell types in normal and stressful situations to allow communication between cells. Tumor-derived exosomes (TEXs) possess diverse neo-antigens and cargoes such as proteins, RNA, and DNA and have a unique molecular makeup reflecting tumor genetic complexity. TEXs contain both immunosuppressive and immunostimulatory factors and may play a role in immunomodulation by influencing innate and adaptive immune components. Moreover, they transmit signals that contribute to the progression of lung cancer by promoting metastasis, epithelial-mesenchymal transition (EMT), angiogenesis, and immunosuppression. This makes them a valuable resource for investigating the immune environment of tumors, which could pave the way for the development of non-invasive biomarkers that could aid in the prognosis, diagnosis, and immunotherapy of lung cancer. While immune checkpoint inhibitor (ICI) immunotherapy has shown promising results in treating initial-stage cancers, most patients eventually develop adaptive resistance over time. Emerging evidence demonstrates that TEXs could serve as a prognostic biomarker for immunotherapeutic response and have a significant impact on both systemic immune suppression and tumor advancement. Therefore, understanding TEXs and their role in lung cancer tumorigenesis and their response to immunotherapies is an exciting research area and needs further investigation. This review highlights the role of TEXs as key contributors to the advancement of lung cancer and their clinical significance in lung immune-oncology, including their possible use as biomarkers for monitoring disease progression and prognosis, as well as emerging shreds of evidence regarding the possibility of using exosomes as targets to improve lung cancer therapy.


Different Trajectories of Apathy and Depression Among Subjective Cognitive Impairment Individuals with or without Conversion to Dementia: Results from the Memento Cohort in France.
Apathy and depression are two early behavioral symptoms in Alzheimer's disease (AD) and related disorders that often occur prior to the onset of cognitive decline and memory disturbances. Both have been associated with an increased risk of conversion to dementia, with a distinct neuropathology. The assessment of the trajectories of apathy and depression and their independent impact on dementia conversion. Apathy and Depression were measured using the Neuropsychiatric Inventory for caregiver (NPI) and clinician (NPI-C), among the nondemented individuals reporting subjective cognitive decline (SCD) at baseline. They were followed up over a 60-month period. Some converted to dementia, according to the methodology carried out by the French Memento Cohort. Among individuals with SCD (n = 2,323), the levels of apathy and depression were low and did not evolve significantly over the 60-month period, despite a trend in apathy increasing as of month 24. Regarding SCD individuals who converted to dementia within the 60-month period (n = 27), the prevalence of depression remained globally steady, while the levels of apathy increased over time. Apathy and depression have different trajectories among individuals with SCD and apathy alone is more likely-compared to depression-to be associated with conversion to dementia.


A brief intervention for preparing ICU families to be proxies: A phase I study.
Family members of critically ill patients report high levels of conflict with clinicians, have poor understanding of prognosis, struggle to make decisions, and experience substantial symptoms of anxiety, depression, and post-traumatic stress regardless of patient survival status. Efficient interventions are needed to prepare these families to act as patient proxies. To assess a brief "patient activation" intervention designed to set expectations and prepare families of adult intensive care unit (ICU) patients to communicate effectively with the clinical team. Phase I study of acceptability and immediate side effects. 122 healthcare proxies of 111 consecutive patients with a stay of ≥24 hours in the Johns Hopkins Hospital Medical ICU (MICU), in Baltimore, Maryland. Reading aloud to proxies from a booklet (Flesch-Kincard reading grade level 3.8) designed with multidisciplinary input including from former MICU proxies. Enrolled proxies had a median age of 51 years old with 83 (68%) female, and 55 (45%) African-American. MICU mortality was 18%, and 37 patients (33%) died in hospital or were discharged to hospice. Among proxies 98% (95% CI: 94% - 100%) agreed or strongly agreed that the intervention was appropriate, 98% (95% CI: 92% - 99%) agreed or strongly agreed that it is important for families to know the information in the booklet, and 54 (44%, 95% CI 35%- 54%) agreed or strongly agreed that parts of the booklet are upsetting. Upset vs. non-upset proxies were not statistically or substantially different in terms of age, sex, education level, race, relation to the patient, or perceived decision-making authority. This patient activation intervention was acceptable and important to nearly all proxies. Frequently, the intervention was simultaneously rated as both acceptable/important and upsetting. Proxies who rated the intervention as upsetting were not identifiable based on readily available proxy or patient characteristics.


Carotid endarterectomy under local anaesthesia does not increase plasma homocysteine concentration.
The purpose of this cohort study was to evaluate the effect of carotid endarterectomy under local anaesthesia on homocysteine (Hcy) concentrations. Of 100 patients with internal carotid artery (ICA) stenosis >70%, the complete data set was available for 91 patients (39 asymptomatic and 52 symptomatic). All patients underwent eversion endarterectomy of the ICA under regional anaesthesia. Thirty-two percent of the examined patients had a total Hcy above 15 micromol/l. The mean Hcy levels preoperatively were 13.9+/-4.8 micromol/l. The Hcy levels on day 5 were 13.1+/-5.0 micromol/l and after 6 months 14.0+/-5.8 micromol/l. There was no significant change during follow-up. No intraoperative strokes and deaths were observed and during the 6 months follow-up no recurrent strokes, TIAs or deaths occurred. Patients undergoing carotid endarterectomy under regional anaesthesia do not have an increase in total Hcy postoperatively. This finding is in contrast to results from cardiac surgery and carotid endarterectomy in a recently published animal study, both performing surgery under general anaesthesia.


Degradation of nitroesters by plant tissue cultures.
Nitrate esters are widely used as effective explosives, important components of explosive ranges, and energetic plasticizers. The environmental problem arising from the production and use of these compounds can be solved using biotechnology. Phytoremediation appears as an efficient technology for this purpose. The uptake and transformation of nitroglycerine (NG) and ethylene glycol dinitrate (EGDN) from wastewater by plants using in vitro regenerants of Juncus inflexus and Phragmites australis were investigated. The plants were exposed to the NG, (600 mg l(-1)), the parent compound disappeared during 20 days and degradation products as dinitroglycerine (DNG) and mononitroglycerine (MNG) were identified in the medium. During 20 days the starting concentration of 100 mg l(-1) EGDN disappeared in the case of J. inflexus or decreased to 5% in the case of P. australis. Ethylene glycol mononitrate as the degradation product was identified. Using this approach directly to the wastewater from production of explosives, the starting concentration of nitroesters mixture (total concentration 270 mg l(-1)) was decreased by in vitro regenerants of reed (P. australis) during 6 weeks to the water contained only MNG (48 mg l(-1)).


Angiotensin II and its 3-7 fragment improve recognition but not spatial memory in rats.
The effects of angiotensin II (AII), its 3-7 fragment [AII(3-7)] and the substituted 3-7 fragment [Leu-5,AII(3-7)] given intracerebroventricularly (ICV) at the dose of 1 nmole each, on spatial memory and recognition were tested. AII(3-7) increased while Leu-5,AII(3-7) slightly decreased session to session foot shock reinforced runtime to the goal in a complex 6 chamber maze. The animals treated with AII performed in the maze similarly to saline injected controls. Overall number of errors was unchanged in all peptide treated groups in comparison with the control group. Object recognition was significantly improved in all the peptide treated groups except for the Leu-5,AII(3-7) group. The results point to the facilitation of recognition and lack of influence on, or even attenuation of, spatial memory by AII and its 3-7 fragment. Leu-5,AII(3-7) caused similar though less pronounced effects.


Surveillance of Dialysis Events: one-year experience at 33 outpatient hemodialysis centers in China.
A multicenter prospective surveillance on dialysis events was carried in 33 dialysis centers in China. Maintenance hemodialysis (HD) outpatients who were dialyzed on the first two days of each month during 2014 were monitored for dialysis events and other infections. During the one-year period, 52,680 patient-months were monitored. Fistula and tunneled or non-tunneled central line were used for 73.70%, 15.70% and 8.85% of vascular access, respectively. There were 773 dialysis events occurred in 671 patients including 589 IV antimicrobial starts, 74 positive blood cultures and 110 local access site infections (LASI). The incidence of dialysis events was 1.47 per 100 patient-months. Among the 74 cases with bloodstream infection (BSI), 38 were access-related BSI (ARB) and there were therefore 148 cases with vascular-related infection (VAI; 38 ARB and 110 LASI). There were 740 cases (1.40 per 100 patient-months) with infections other than BSI and LASI, most (79.19%) of which were respiratory tract infections. For those with dialysis events, there were 425 cases (425/671, 63.34%) admitted to hospital and 12 cases of death (12/671, 1.79%). In conclusion, the surveillance revealed a relatively low incidence of dialysis events and the surveillance may be tailored to target those using central lines in resource-limited settings.


MS-DIAL: data-independent MS/MS deconvolution for comprehensive metabolome analysis.
Data-independent acquisition (DIA) in liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS) provides comprehensive untargeted acquisition of molecular data. We provide an open-source software pipeline, which we call MS-DIAL, for DIA-based identification and quantification of small molecules by mass spectral deconvolution. For a reversed-phase LC-MS/MS analysis of nine algal strains, MS-DIAL using an enriched LipidBlast library identified 1,023 lipid compounds, highlighting the chemotaxonomic relationships between the algal strains.


Pseudomonas infection of the catheter exit site successfully managed with topical acetic acid.
A case of recurrent catheter exit site infection with Pseudomonas aeruginosa is presented in a patient receiving home parenteral nutrition. The past episodes were managed by elective catheter replacement following extrusion of the catheter cuff. We describe the successful use of acetic acid to the exit site which resulted in the eradication of the organism and complete resolution of all signs.


Renal Toxicity.
With the increasing use of immunotherapy, there has been an associated increased survival in many cancers but has also resulted in unregulated organ-specific toxicities. In this review, we will discuss the renal toxicities associated with a checkpoint inhibitor (CPI) from the typical acute tubulointerstitial nephritis to glomerulonephritis and their proposed mechanisms and treatments. We also discuss the use of CPI and reactivation of preexisting autoimmune disease with a focus on renal cell cancer in setting of chronic kidney disease (CKD). Transplant rejection in setting of CPI use has been further evaluated with single-center and multicenter retrospective studies, and available data will be presented in this chapter.


Intensity-modulated radiotherapy for locally advanced cancers of the larynx and hypopharynx.
Limited data evaluate intensity-modulated radiotherapy (IMRT) for cancers of the hypopharynx and larynx. We report clinical outcomes and failure patterns for these patients. Between September 2001 and December 2007, 42 patients with squamous cell carcinoma (SCC) of the hypopharynx (n = 23) and larynx (n = 19) underwent IMRT, 11 postoperatively and 31 definitively. Thirty-six received systemic therapy. Median follow-up was 30 months among surviving patients. Three local failures occurred within the high-dose region and 3 occurred in regional nodes. Seven patients developed distant metastasis as the initial failure. Three-year actuarial estimates of locoregional control, freedom from distant metastasis, and overall survival rates were, respectively, 80%, 72%, and 46%. IMRT provides good locoregional control for SCC of the hypopharynx and larynx compared with historical controls. Locoregional relapses occurred in the high-dose volumes, suggesting adequate target volume delineation. Hypopharyngeal tumors, which fare worse than laryngeal tumors, warrant investigation of more aggressive treatment.


Antibacterial capacity of buccal epithelial cells from healthy donors and children with recurrent urinary tract infections.
The effect of buccal epithelial cells (BEC) on bacterial growth was investigated in healthy subjects as well as in patients with recurrent urinary tract infections (UTI) and compared to the antibacterial capacity of uroepithelial cells (UEC) of the same individuals. Epithelial cells were obtained from the following groups: healthy female controls; females without further UTI after reflux operation; females with asymptomatic bacteriuria (ABU); females with further UTI despite successful reflux operation; and patients with meningomyelocele (MMC) and recurrent UTI due to significant residual urinary volume. Cocultivation of Escherichia coli with BEC as well as UEC from healthy females or patients with MMC resulted in significant suppression of bacterial growth. However, neither type of epithelial cell showed an antibacterial effect when they were obtained from patients with recurrent UTI in the absence of urological abnormalities (ABU patients; reflux-corrected patients with further UTI). From these results it is concluded that a generalised epithelial defence defect is one important pathogeneic factor for recurrent idiopathic UTI.


Could BRAF mutations in melanocytic lesions arise from DNA damage induced by ultraviolet radiation?
Although BRAF V600 mutations in melanocytic tumors are not UV-signature mutations, it is plausible that they could still arise from error-prone replication of UV-damaged DNA. We propose a mechanism for their origin, taking into consideration melanocytic-specific BRAF tandem mutations, nearby potential pyrimidine dimer sites, the properties of specialized DNA polymerases, and biological selection.


A ferritin gene in the marine copepod Acartia tonsa as a highly sensitive biomonitor for nano-contamination.
Toxicology is not only for eco-risk assessments, but also for the real-time environmental monitoring based on the quick response of specific biomarkers. Ferritin gene (ftn) is a potential biomarker involving in crucial protective responses in biota. However, little information is available concerning the ftn in marine copepod Acartia tonsa (A. tonsa), a model organism widely applied in toxicology assessments. Our study for the first time identified and characterized the ftn in A. tonsa, along with its time-dependent transcriptional response to the reproductive toxicity of two newly emerged nanomaterials. The full-length cDNA of ftn contains a 114-bp 5'-untranslated region (UTR), a 236-bp 3'-untranslated region, and a 510-bp open reading frame which encodes an 18.51 kDa polypeptide composed of 169 amino acids. The ftn sequence has an iron binding signature and a potential phosphorylation site, which is closely-related to the ftn of Calanus sinicus and Pseudodiaptomus annandalei genes at the phylogenetical level. The ftn showed a quick and highly sensitive response to nanomaterial exposures, even at no observed effect concentrations. In detail, after exposure to nickel nanomaterials (up to 17.0 mg/L), the ftn was significantly upregulated immediately at 0.5 h and peaked at 9.5-fold in adults within 48 h, along with a significant reduction of egg hatching rate. When exposed to CdSe/ZnS quantum dots (up to 135 mg/L), no significant change in egg productions or hatching rates was observed, while the expression of ftn still significantly increased to over 3.0-fold in the initial 48 h. After that, the upregulation of ftn induced by CdSe/ZnS quantum dots or nickel nanoparticles both gradually returned back within 96 h. These findings demonstrate the highly sensitive response of this new cloned ftn to nanomaterial exposures, and highlight the suitability of ftn in A. tonsa as a promising biomonitor for nano-contamination in marine environments.


Termination of second trimester pregnancy with intraamniotic administration of 16-phenoxy-omega-tetranor-PgE2-methylsulfonamide (SHB 286) alone and combined with oxytocin and calcium gluconate.
Midtrimester abortion was succesfully induced within 30 hours in 62 out of 90 patients by intraamniotic administration of 16-phenoxy-omega-17,18,19, 20-tetranor-prostaglandine-E2-methylsulfonamide. The Pg-analogue was tested in 5 groups of patients which received 1,2 or 3 mg. In group IV, 2 mg of the analogue was combined with oxytocin. In group V, 3 mg of the analogue was combined with 2.75 grams of calcium gluconate. The success rate was significantly influenced by the amount of the drug administered. Side effects were minimal. Multiparous women respond better to the therapy than nulliparous patients. The combination with oxytocin was more effective than the Pg-analogue alone and the combination of sulprostone with calcium gluconate was 87% succesful within 30 hours and 100% succesful within 36 hours.


Immunological consequences of dioctyltin dichloride (DOTC)-induced thymic injury.
Inbred rats fed diets containing 75 ppm dioctyltin dichloride (DOTC) for 8 or 12 weeks demonstrated marked reduction in thymic weight. This reduction was accompanied by a decrease in the number of W3/25 positive phenotypic circulating lymphocytes and an impaired ability to respond to suboptimal mitogenic stimulation. Responsiveness to alloantigenic stimulation, as measured by mixed leucocyte reactions, was depressed in DOTC-fed animals whereas no effect on natural cytotoxicity in nonimmunized treated or control rats was evident. No difference was found in the ability of treated compared to untreated animals to mount specific antibody responses against sheep red blood cells, and no evidence was found to indicate toxicity to lymphocytes in vivo. The investigation showed that DOTC administration resulted in a selective immunodeficiency.


Interspecific competitive interactions between pairs of fungal species in natural substrates.
The role of interspecific competition in fungal communities in natural substrates is poorly understood because fungi do not form easily definable populations. A new approach to investigating fungal competition, using natural substrates containing a range of known biomass concentrations of each of two species, is described. Relative competitive success of each species is assessed over time in terms of propagule production and substrate colonisation by each species. In an agricultural soil Mucor hiemalis usually out-competed Trichoderma harzianum. After 27 days, the success of both species in the mixtures was independent of the initial biomass concentration of either species, although the success of T. harzianum in these mixtures was substantially inhibited relative to the T. harzianum monocultures. In a forest soil, T. polysporum maintained a competitive advantage over M. hiemalis, and induced M. hiemalis to produce propagules rather than mycelia. Coexistence of both species always occurred in both experiments, and in the forest soil experiment the two-species mixtures all contained a higher total microbial biomass than the monocultures of either species by day 47, suggesting some niche differentiation.


Contrasting allocation of magnesium, calcium and manganese in leaves of tea (Camellia sinensis (L.) Kuntze) plants may explain their different extraction efficiency into tea.
During tea preparation mineral elements are extracted from the dried leaves of tea (Camellia sinensis (L.) Kuntze) plants into the solution. Micro-particle induced X-ray emission was employed to investigate the spatial distribution of magnesium (Mg), calcium (Ca) and manganese (Mn) in the young and old leaves of tea plants grown in the absence and presence of aluminium (Al) in the substrate. Results revealed that in tea leaves the largest concentrations of Mg occurred in the epidermis, of Ca in oxalate crystals and of Mn in epidermis and oxalate crystals; there was a leaf-age effect on tissue-specific concentrations of Mg, Ca and Mn with all tissues of old leaves containing larger concentrations of Mg, Ca and Mn than young leaves; supplementation of substrate with Al reduced concentrations of Mg, Ca and Mn in the old leaves, and a link between the distribution of Mg, Ca and Mn in the tea leaves with the extraction efficiencies of these elements into the tea was possible. We conclude that old leaves of tea plants cultivated under conditions of low Al availability will have the largest concentrations of Mg, Ca and Mn and may represent most acceptable ingredient for the preparation of tea.


Histologic changes in the gastroduodenal mucosa after long-term medical treatment with cimetidine or parietal cell vagotomy in patients with juxtapyloric ulcer disease.
Biopsy specimens were collected during endoscopy from pre-established sites in the corpus (n = 60), antrum (n = 53), and the duodenal bulb (n = 54) from the same patient before and 2-3 years after parietal cell vagotomy (PVC) or after a similar period of treatment with cimetidine. There was a significant increase in scores of chronic body gastritis after PCV (p less than 0.001) even in comparison with the cimetidine group (p less than 0.01), for which the scores of chronic body gastritis remained essentially unchanged. The scores of chronic antral gastritis and the incidence of intestinal metaplasia of the antrum increased significantly (p less than 0.05) in both the PCV and the cimetidine groups when the two treatment groups were analyzed together. The degree of polymorphonuclear infiltration in the body and antral mucosa, the incidence and severity of duodenitis, and the incidence of gastric metaplasia in the duodenal cap were unaffected by the treatment. In contrast to maintenance treatment with cimetidine PCV seems to accelerate the development of chronic body gastritis. The kappa statistics, as indicator of the reproducibility of histopathologic scoring, were acceptable.


[Variability of Chinese hamster clones in resistance to 8-azaguanine].
The stability of the 8-azaguanine resistance marker during the number of cell generations has been analyzed for 63 clones of Chinese hamster cells. The resistant clones analyzed were spontaneous or induced after the treatment of parent cells by 5-BUdR, EMS, or histone H1. The clones have revealed different levels of AG-resistance as well as different ability to maintain the above level, under the cultivation in the drug-free medium. Subclones, isolated from the unstable clone, were found to be different in both the resistance level and the stability of resistant state.


Control of immunity in parental and F1 hybrid mouse strains to a spontaneously arising parental tumor. I. Correlation between in vitro and in vivo anti-tumor reactivity in responsive and unresponsive animals.
This study has investigated the in vivo and in vitro cellular immunity of the AKR/J, (AKR/J x C57BL/6)F1 and (AKR/J x DBA/2)F1 mouse strains to a spontaneously arising AKR/J tumor. The study concludes that: 1) there is an excellent correlation between the growth of the tumor in vivo, and the ability of spleen cells from the respective strains to generate both primary and secondary cell-mediated cytotoxic responses in vitro; and 2) one F1 host can be considered a responder against this tumor, whereas the other F1 strains studied appears to be genetically nonresponsive. Possible mechanisms relating to the observed patterns of response are discussed.


Clinical spectrum of TB in elderly in a TB & respiratory institution.
Tuberculosis has maximum burden among young population in developing countries like India. However, children and elderly form a special group where TB may have atypical presentation. This presents with epidemiological, diagnostic and treatment challenges in these groups which may need special attention in the national programmes for TB. Due to atypical presentation, elderly population is vulnerable to frequent misdiagnosis and disease may already be in advanced stage when correct diagnosis is made. Not only this, adjustment of drug dosages and high chances of adverse drug reaction make the management of TB more complicated in elderly. Mortality due to TB is also higher in this group as compared to young patients of TB. This view point briefly highlights the epidemiological, clinical and disease outcome aspects of TB disease in elderly patients.


Publisher Correction: Damage-tolerant architected materials inspired by crystal microstructure.
In Fig. 4a of this Article, owing to an error in the production process, the scale bar inadvertently read 1 mm instead of 1 m. This error has been corrected online.


Force sense deficits in functionally unstable ankles.
The purpose of this study was to determine whether participants with functional ankle instability have deficits in force sense reproduction compared to uninjured participants. Twenty participants with no history of injury and 20 with functional ankle instability volunteered for this investigation. Participants in the instability group had a history of at least one ankle sprain and frequent episodes of giving way. Ankle force sense testing was performed at 10, 20, and 30% of eversion maximal voluntary isometric contraction. Ten trials were performed at each force. Absolute and variable errors were then calculated. We observed significant (p < 0.05) main effects for group for absolute and variable errors. Force sense absolute error was significantly greater in the functionally unstable ankles (3.7 +/- 2.2 N) compared to uninjured ankles (2.8 +/- 1.1 N). Variable error was also significantly greater in the functionally unstable ankles (3.2 +/- 1.8 N) than the uninjured ankles (2.4 +/- 0.8 N). Functional ankle instability is associated with deficits in an individual's ability to accurately reproduce a given force. This deficit may impair an individual's ability to set appropriate muscle force levels to provide stability to the joint.


Production of volatile fatty acid from fruit waste by anaerobic digestion at high organic loading rates: Performance and microbial community characteristics.
This study examined the performance and microbial community dynamics of an anaerobic volatile fatty acid (VFA) production reactor for treating fruit waste by stepwise increasing organic loading rates (OLRs) from 8 to 24 g volatile total solids (VTS)/(L·d). Results showed that higher VFA concentrations of 52.25-61.90 g chemical oxygen demand (COD)/L can be maintained at each OLR, thereby resulting to a production of 0.70-0.76 g chemical oxygen demand (COD)VFA/g VTS. Notably, an increase in OLR from 8 to 14 g VTS/(L·d) was beneficial for achieving higher VFA concentrations and yields. Moreover, an increase in OLR affected the VFA distribution significantly; acetate and butyrate became dominant in the fermentation liquid at OLRs ≥ 14 g VTS/(L·d). Microbial community dynamics analysis revealed that phyla Firmicutes and Actinobacteriota were predominant at each OLR, and the genera Lactobacillus, Clostridium_sensu_stricto_12, and Caproiciproducens were closely related to anaerobic VFA production.


Cannabinoid receptor activation potential of the next generation, generic ban evading OXIZID synthetic cannabinoid receptor agonists.
In recent years, several nations have implemented various measures to control the surge of new synthetic cannabinoid receptor agonists (SCRAs) entering the recreational drug market. In July 2021, China put into effect a new generic legislation, banning SCRAs containing one of seven general core scaffolds. However, this has driven manufacturers towards the synthesis of SCRAs with alternative core structures, exemplified by the recent emergence of "OXIZID SCRAs." Here, using in vitro β-arrestin2 recruitment assays, we report on the CB1 and CB2 potency and efficacy of five members of this new class of SCRAs: BZO-HEXOXIZID, BZO-POXIZID, 5-fluoro BZO-POXIZID, BZO-4en-POXIZID, and BZO-CHMOXIZID. All compounds behaved as full agonists at CB1 and partial agonists at CB2 . Potencies ranged from 84.6 to 721 nM at CB1 and 2.21 to 25.9 nM at CB2 . Shortening the n-hexyl tail to a pentyl tail enhanced activity at both receptors. Fluorination of this pentyl analog did not yield a higher receptor activation potential, whereas an unsaturated tail resulted in decreased potency and efficacy at CB1 . The cyclohexyl methyl analog BZO-CHMOXIZID was the most potent compound at both receptors, with EC50 values of 84.6 and 2.21 nM at CB1 and CB2 , respectively. Evaluation of the activity of a seized powder containing BZO-4en-POXIZID suggested a high purity, in line with high-performance liquid chromatography coupled to diode-array detection (HPLC-DAD), gas chromatography coupled to mass spectrometry (GC-MS), liquid chromatography coupled to time-of-flight mass spectrometry (LC-QTOF-MS), and Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) analysis. Furthermore, all tested compounds showed a preference for CB2 , except for BZO-POXIZID. Overall, these findings inform public health officials, law enforcement agencies, and clinicians on these newly emerging SCRAs.


Fluorescent linear polyurea based on toluene diisocyanate: Easy preparation, broad emission and potential applications.
In contrast to conventional fluorescent polymers featured by large conjugation structures, a new class of fluorescent polymers without above conjugations are gaining constant interest owing to their significant academic importance and promising applications in diverse fields. These unconventional fluorescent polymers are in general composed of heteroatoms (e.g. N, O, P, and S) under different forms. Here we report our recent study on polyurea, prepared by a very simple one step precipitation polymerization of toluene diisocyanate in a binary solvent of water-acetone. This polyurea, basically consisting of phenyl ring and urea group, shows fluorescent emission in a broad concentration range, from very low (10-5 mg/mL) to its solubility limit (50 mg/mL), and in a wide range of emission wavelength from UV to visible regions of up to 500 nm under varied excitation wavelength. The emission behaviors were fully studied under different concentrations and excitations. It was concluded that the emission in UV region was intrinsic due to the conjugation between the phenyl and the adjacent urea unit; while the emission in visible region, strongly excitation dependent, was caused by the cluster formation of the molecular chains, in accordance with the cluster-triggered-emission (CTE) mechanism. The formation of the cluster was tested through dynamic light scattering, FTIR and UV absorbance. Tested in presence of different metal ions, Fe3+ demonstrated a quenching effect with high selectivity. Based on this study, different paper-based sensors were designed to detect Fe3+, H2O2 in bioanalysis and for data encryption. This work provides a simple way to prepare a polyurea, a novel type of unconventional fluorescent polymer, with high emission performance distinct from its known analogues.


Permanent increase in endothelin serum levels in vasovagal syncope.
Of 40 normal volunteers tilted on two separate occasions, seven subjects had vasovagal syncope only during one upright tilt, but showed increased endothelin plasma levels in the supine position and during head-up tilt on both occasions, independent of vasovagal syncope, compared to control subjects.


Medicare's 3-Day Rule: Time for a Rethink.
A pilot study was undertaken to examine the impact of Medicare's 3-day rule on length of stay (LOS). One hundred consecutive patients who underwent primary total joint arthroplasty and were discharged to extended care facilities were retrospectively reviewed. Based on readiness for discharge criteria, delaying discharge until the third postoperative day increased LOS by 1.1 days (P<0.001). 60.6% of patients were ready for discharge by the second postoperative day, none of whom required re-admission within 30 days of discharge. There were no rehabilitation gains by staying an additional hospital day beyond readiness for discharge (P=0.092). This pilot study calls into question the value of Medicare's 3-day rule and demonstrates the feasibility and need for further research to address this seemingly antiquated policy.


Botulinum toxin a in the treatment of chromhidrosis.
Chromhidrosis is an uncommon disorder characterized by secretion of colored sweat by apocrine glands, typically localized to the face or axilla. The current treatments available for chromhidrosis are time consuming and frequently ineffective. Our purpose is to demonstrate a novel approach to the treatment of apocrine chromhidrosis. We report a case of apocrine chromhidrosis successfully treated with botulinum toxin A (BTX-A; Botox). BTX-A therapy successfully controlled facial chromhidrosis, and the effects were visible at 19 weeks post-treatment. The therapeutic benefits may be attributed to its inhibitory effects on cholinergic stimulation, adrenergic stimulation, and substance P release, although further studies are necessary to elucidate the precise mechanism of action. This report demonstrates a new therapeutic approach to patients suffering from chromhidrosis.


Serum CA 125 antigen levels and disease severity in patients with endometriosis.
We wished to test whether there was a relationship between CA 125 levels and the severity of endometriosis that would allow CA 125 to be used to monitor disease progression. CA 125 levels were measured in 52 women with laparoscopically staged endometriosis and in eight women with normal pelvic findings at laparoscopy. There was a positive correlation (r = 0.63; P less than .0001) between disease severity and CA 125 levels. CA 125 was also elevated, compared with women with a normal pelvis, in patients with mild and moderate disease. There was no relationship between CA 125 levels and the day of the menstrual cycle. These results suggest that after malignancy has been excluded, CA 125 levels may offer a useful method of monitoring disease progress.


Removal of hexavalent chromium ions using CuO nanoparticles for water purification applications.
Copper(II) oxide nanoparticles were synthesized at low temperature using cold finger assisted magnetron sputtering technique and were applied as adsorbent for the rapid removal of noxious Cr(VI) ions from the solvent phase. The average size of CuO nanoparticles from TEM analysis was found to be 8nm in addition to this the BET surface area (84.327m(2)/g) was found to be significantly high in comparison to the previously CuO nanoparticles synthesized via green route. The synthesized CuO nanoparticles is crystalline in nature and exhibits monoclinic phase, which was confirmed using various analytical techniques such as SAED, XRD and Raman analysis. The impact of influential parameters including pH, adsorbent dose, contact time, stirring speed, initial Cr(VI) ions concentration, and temperature were optimized using batch adsorption method in order to obtain maximum removal of Cr(VI) ions. From the thermodynamic parameters, the positive value of enthalpy (ΔH) and negative value of Gibbs free energy (ΔG) indicate the endothermic and spontaneous nature of Cr(VI) ions adsorption, respectively. The adsorption kinetics data was well fitted and found to be in good agreement with the pseudo second order kinetic behaviour.


Brackish water improves the taste quality in meat of adult male Eriocheir sinensis during the postharvest temporary rearing.
We investigated the effect of temporary rearing in brackish water on the taste quality in meat of crab cooked. The main salinity-responsive factors included 5'-nucleotides and free amino acids (FAAs) in crab meat that were identified using tri-step infrared spectroscopy. Compared to the fresh water group, the contents of 5'-adenosine monophosphate and 5'-inosine monophosphate in the brackish water group significantly increased in the 2nd week and decreased in the 6th week, respectively. The contribution ratio of umami FAAs increased from 8.1 to 13.5% in the 4th week in the brackish water group, showing maximum value of equivalent umami concentration. Moreover, Ca2+ and Cl- contents significantly increased in the 4th and 6th weeks, respectively (P < 0.05). Infrared spectroscopy was an effective method to identify the taste components. With respect to the taste quality, four weeks were determined as the best period for temporary rearing of the crab in brackish water.


The impact of tofacitinib on fatigue, sleep, and health-related quality of life in patients with rheumatoid arthritis: a post hoc analysis of data from Phase 3 trials.
Fatigue, a common symptom of rheumatoid arthritis (RA), is detrimental to health-related quality of life (HRQoL). We evaluated the impact of tofacitinib on fatigue, sleep, and HRQoL and explored associations between fatigue, related patient-reported outcomes (PROs), and disease activity in RA patients. This post hoc analysis pooled data from three Phase 3 studies of tofacitinib (ORAL Scan; ORAL Standard; ORAL Sync) in RA patients. Patients received tofacitinib 5 or 10 mg twice daily, placebo, or adalimumab (active control; ORAL Standard only, not powered for superiority) with conventional synthetic disease-modifying antirheumatic drugs. Assessed through Month (M)12 were changes from baseline in disease activity, Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), Medical Outcomes Study Sleep scale (MOS-SS), and Short Form-36 Health Survey (SF-36) composite/domain scores, and proportions of patients reporting improvements from baseline in FACIT-F total and SF-36 domain scores ≥ minimum clinically important differences (MCIDs) or ≥ population normative values. Pearson correlations examined associations among PROs at M6. Treatment comparisons were exploratory, with p < 0.05 considered nominally significant. Generally, active treatment led to significant improvements from baseline in FACIT-F total, and MOS-SS and SF-36 composite/domain scores vs placebo, observed by M1 and maintained through M6 (last placebo-controlled time point). Through M6, more patients achieved improvements from baseline ≥ MCID and achieved scores ≥ population normative values in FACIT-F total and SF-36 domain scores with tofacitinib vs placebo. Through M12, some nominally significant improvements with tofacitinib vs adalimumab were observed. With active treatment at M6, FACIT-F scores were moderately (0.40-0.59) to highly (≥ 0.60) correlated with SF-36 composite/domain scores (particularly vitality), moderately correlated with most MOS-SS domain scores, and highly correlated with MOS-SS Sleep Problems Index I scores. Disease activity correlations were moderate with FACIT-F scores and low (0.20-0.39) to moderate with SF-36 general health domain/composite scores. Tofacitinib and adalimumab generally conferred significant, clinically meaningful improvements in fatigue, sleep, and HRQoL (including vitality) vs placebo through M6, with improvements maintained to M12. M6 correlations between FACIT-F, PROs of sleep, HRQoL, and disease activity underscore the interrelatedness of multiple PROs and disease activity in RA. ClinicalTrials.gov NCT00847613 (registered: February 19, 2009); NCT00853385 (registered: March 2, 2009); NCT00856544 (registered: March 5, 2009).


Analysis of polypeptide disposition in human erythrocyte membranes employing membrane inversion.
High resolution segregation of erythrocyte membrane polypeptides achieved by isoelectric focusing in 8 M urea was employed in conjunction with surface-restricted radioiodination to analyze the disposition of polypeptides within the human erythrocyte membrane. Several membrane polypeptides showed significant uptake of radioiodine, with the principal labeled component migrating between pH values of 3.0 and 3.5. Two approaches were taken in examining membrane polypeptide disposition on both faces of the erythrocyte membrane. Saturation labeling of the outer face of the membrane with one iodine isotope followed by cell lysis and reiodination with a second iodine isotope did not prove feasible and another procedure based on surface iodination with 125-I, formation of sealed inside-out vesicles and re-iodination with 131-I was adopted. Studies of sialic acid release from the membrane surface and trypsin cleavage of radioiodinated peptides indicated that selectively labeled, sealed inside-out vesicles had been formed. The ratio of 125-I to 131-I in membrane polypeptides separated by isoelectric focusing confirmed the existence of externally disposed, internally disposed and spanning proteins.


Neuro-ophthalmology Emergency Department and Inpatient Consultations at a Large Academic Referral Center.
Prompt neuro-ophthalmology consultation prevents diagnostic errors and improves patient outcomes. The scarcity of neuro-ophthalmologists means that the increasing outpatient demand cannot be met, prompting many emergency department (ED) referrals by non-neuro-ophthalmologists. We describe our quaternary care institution's ED and inpatient neuro-ophthalmology consultation patterns and patient outcomes. Prospective observational study. Consecutive neuro-ophthalmology ED and inpatient consultation requests over 1 year. We collected patient demographics, distance traveled, insurance status, referring provider details, consultation question, final diagnosis, complexity of consultation, time of consultation, and need for outpatient follow-up. Consultation patterns and diagnoses, complexity, and follow-up. Of 494 consecutive adult ED and inpatient neuro-ophthalmology consultations requested over 1 year, 241 of 494 consultations (49%) occurred at night or during weekends. Of ED consultations (322 of 494 [65%]), 127 of 322 consultations (39%) occurred during weekdays, 126 of 322 consultations (39%) occurred on weeknights, and 69 of 322 consultations (22%) occurred on weekends or holidays. Of 322 ED consultations, 225 of 322 consultations (70%) were patients who initially sought treatment in the ED with a neuro-ophthalmic chief symptom. Of the 196 patients sent to the ED by a health care professional, 148 patients (148/196 [76%]) were referred by eye care specialists (74 optometrists and 74 ophthalmologists). The most common ED referral questions were for papilledema (75 of 322 [23%]) and vision loss (72 of 322 [22%]). A total of 219 of 322 patients (68%) received a final active neuro-ophthalmic diagnosis, 222 of 322 patients (69%) were cases of high or very high complexity, and 143 of 322 patients (44%) required admission. Inpatient consultations (n = 172) were requested most frequently by hospitalists, including neurologists (71 of 172 [41%]) and oncologists (20 of 172 [12%]) for vision loss (43 of 172 [25%]) and eye movement disorders (36 of 172 [21%]) and by neurosurgeons (58 of 172 [33%]) for examination for mass or a preoperative evaluation (19 of 172 [11%]). An active neuro-ophthalmic diagnosis was confirmed in 67% of patients (116 of 172). Outpatient neuro-ophthalmology follow-up was required for 291 of 494 patients (59%). Neuro-ophthalmology consultations are critical to the diagnosis and management in the hospital setting. In the face of a critical shortage of neuro-ophthalmologists, this study highlights the need for technological and diagnostic aids for greater outpatient access. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.


Energetic coupling between native-state prolyl isomerization and conformational protein folding.
In folded proteins, prolyl peptide bonds are usually thought to be either trans or cis because only one of the isomers can be accommodated in the native folded protein. For the N-terminal domain of the gene-3 protein of the filamentous phage fd (N2 domain), Pro161 resides at the tip of a beta hairpin and was found to be cis in the crystal structure of this protein. Here we show that Pro161 exists in both the cis and the trans conformations in the folded form of the N2 domain. We investigated how conformational folding and prolyl isomerization are coupled in the unfolding and refolding of N2 domain. A combination of single-mixing and double-mixing unfolding and refolding experiments showed that, in unfolded N2 domain, 7% of the molecules contain a cis-Pro161 and 93% of the molecules contain a trans-Pro161. During refolding, the fraction of molecules with a cis-Pro161 increases to 85%. This implies that 10.3 kJ mol(-1) of the folding free energy was used to drive this 75-fold change in the Pro161 cis/trans equilibrium constant during folding. The stabilities of the forms with the cis and the trans isomers of Pro161 and their folding kinetics could be determined separately because their conformational folding is much faster than the prolyl isomerization reactions in the native and the unfolded proteins. The energetic coupling between conformational folding and Pro161 isomerization is already fully established in the transition state of folding, and the two isomeric forms are thus truly native forms. The folding kinetics are well described by a four-species box model, in which the N2 molecules with either isomer of Pro161 can fold to the native state and in which cis/trans isomerization occurs in both the unfolded and the folded proteins.


Studying pellet formation of a filamentous fungus Rhizopus oryzae to enhance organic acid production.
Using pelletized fungal biomass can effectively improve the fermentation performance for most of fugal strains. This article studied the effects of inoculum and medium compositions such as potato dextrose broth (PDB) as carbon source, soybean peptone, calcium carbonate, and metal ions on pellet formation of Rhizopus oryzae. It has been found that metal ions had significantly negative effects on pellet formation whereas soybean peptone had positive effects. In addition PDB and calcium carbonate were beneficial to R. oryzae for growing small smooth pellets during the culture. The study also demonstrated that an inoculum size of less than 1.5x10(9) spores/L had no significant influence on pellet formation. Thus, a new approach to form pellets has been developed using only PDB, soybean peptone, and calcium carbonate. Meanwhile, palletized fungal fermentation significantly enhanced organic acid production. Lactic acid concentration reached 65.0 g/L in 30 h using pelletized R. oryzae NRRL 395, and fumeric acid concentration reached 31.0 g/L in 96 h using pelletized R. oryzae ATCC 20344.


Regulation of prostaglandin E2 binding to a murine macrophage cell line, P388D1, by insulin.
Preincubation of murine macrophage-like P388D1 cells with physiological amounts of insulin resulted in an increase in prostaglandin E2 binding to these cells, by approximately 2-fold, when compared to untreated cells. Scatchard analysis of the binding of PGE2 to insulin-treated cells indicated that the enhanced binding was due to an increase in receptor number (from 0.30 +/- 0.02 to 0.63 +/- 0.03 fmol/10(6) cells for the high affinity receptor binding sites, and from 2.4 +/- 0.31 to 5.0 +/- 0.41 fmol/10(6) cells for the low affinity receptor binding sites) rather than to an increase in the affinity of the binding sites. The insulin-stimulation of PGE2 binding appeared to be associated with a lowering of the cAMP level in these cells; treatment of cells with insulin lowered the cAMP level by increasing the cAMP phosphodiesterase activity of both the membrane and cytosolic fractions. However, enhanced PGE2 binding to the cells resulted in an increase in cAMP level in the cells. This increase in cAMP level may help to enhance the immunosuppressive action of this prostanoid, as PGE2 is known to suppress many steps in the immune response, including interleukin-1 expression, by raising cAMP levels via activation of receptor-linked adenylate cyclase. Our data suggest that insulin at physiological concentrations may enhance the immunosuppressive action of PGE2.


Bacteremia due to Fusobacterium species.
Twenty-six patients were identified as having bacteremia with Fusobacterium species over a five-year period at Boston City Hospital. They represented 0.9 percent of bacteremic patients and were equally divided as to sex. Bacteremia with Fusobacterium occurred primarily in young adults and in patients over 60 years of age and was not observed in children. In 16 patients (62 percent), Fusobacterium was the only blood culture isolate. The most common primary foci of infection were the female genital tract, the upper respiratory tract, the oral cavity, and the lower respiratory tract. Five patients had primary foci of infection that were initially occult. Three of these patients were found to have unappreciated oral and pharyngeal lesions, and one had a liver abscess; no primary infection was established in the remaining patient. Shock related to bacteremia developed in six patients (23 percent), four of whom had Fusobacterium species as the only blood culture isolate. Death occurred in three patients (12 percent), all of whom were over 60 years old. Metastatic infection occurred in only one patient in whom hematogenous osteomyelitis developed. Postpartum fusobacterial bacteremia was uniformly benign. Evaluation of bacteremia with Fusobacterium species in nonpostpartum patients, without an overt focus of infection, should be directed to a search for occult abscess, especially of the upper respiratory tract and oral cavity.


Simultaneous determination of plant hormones in peach based on dispersive liquid-liquid microextraction coupled with liquid chromatography-ion trap mass spectrometry.
Fruit development is influenced greatly by endogenous hormones including salicylic acid (SA) and abscisic acid (ABA). Mass spectrometry with high sensitivity has become a routine technology to analyze hormones. However, pretreatment of plant samples remains a difficult problem. Thus, dispersive liquid-liquid microextraction (DLLME) was used to concentrate trace plant hormones before liquid chromatography-ion trap mass spectrometry (LC-ITMS) analysis. Standard curves were linear within the ranges of 0.5-50, 0.2-20ng/mL for SA and ABA, respectively. The correlation coefficients were greater than 0.9995 with recoveries above 87.5%. The limits of detection were 0.2ng/mL for SA and 0.1ng/mL for ABA in spiked water solution, respectively (injection 20μL). The successful analysis of SA and ABA in fruit samples indicated our DLLME-LC-ITMS approach was efficient, allowing reliable quantification of both two compounds from very small amounts of plant material. Moreover, this research revealed the relationship between SA and ABA content and development of peach fruit at different growth stages.


Rapid establishment of murine gastrointestinal organoids using mechanical isolation method.
Gastrointestinal (GI) diseases, including pathological dysplasia, inflammation, neoplasia and injury, suffer millions of patients globally per year. Organoids, three-dimensional cell mass structures supported by biomaterials in dishes, are currently used as a research model for diseases of the small intestine. However, the traditional enzymatic-digestion method for establishing small-intestinal organoids (EnzyOs) is time consuming and often loses the bulk of crypts, a more efficient and reliable method needs to be developed. In this study, using mouse GI organoids as a model, we formulated a rapid mechanical isolation method that could efficiently isolate and culture villi-crypts into small intestinal organoids (MechOs). Primary duodenum organoids generated by MechOs displayed three types of morphology: spheroid, semi-budding and budding, while EnzyOs produced much less budding. Moreover, primary duodenum organoids from MechOs could be subcultured and presented similar gene expression profiles of small intestine specific markers as that from EnzyOs. Importantly, the MechOs method could also be used to generate other types of organoids derived from the stomach, jejunum-ileum, sigmoid-rectum and bile cysts. Taken together, the results show that MechOs could efficiently and economically generate digestive system organoids, providing a potential basis of epithelial organoids for the clinical treatment of gastroenterological diseases.


Stereolithographic models of biopolymers.
Stereolithography (STL) has been used to make plastic models of the solvent accessible surfaces of biopolymers. Models have been made of proteins and proteins bound to DNA and RNA. The STL process uses a laser to photopolymerize a liquid resin. Using the ACES (accurate, clear, epoxy, solid) building technique, parts are made with minimum postcure shrinkage. Protein Data Bank files are converted to STL files that represent the surface topology of the biopolymer as a series of triangles and an index that describes their orientation. The models are useful in teaching biomolecular structure and the principle of docking. They are especially useful to the visually impaired.


The influence of ethanol on learned and reflexive heart rate responses of rats during classical aversive conditioning.
An unconditioned response was produced in rats by electric shock and resulted in a biphasic deceleration-acceleration of heart rate after 2.4 g of ethanol per kg. A monophasic acceleration occurred with saline and 0.8 g of ethanol per kg.