Neck as mediastinal extension: Diagnosis of sarcoidosis by core biopsy of cervical lymph nodes.
Sarcoidosis is a multi-system granulomatous disease. The diagnostic procedures for histological confirmation are invasive and a less invasive approach to diagnostic pathway is warranted. The utility of diagnostic value of neck ultrasound was retrospectively evaluated. A histological diagnosis was made by ultrasound-guided head and neck core biopsy to confirm clinically and radiologically suspected sarcoidosis. Twenty-five patients were referred for sonographic evaluation of the head and neck after CT scan in an attempt to avoid the use of more invasive tests. These patients had mediastinal adenopathy, but not clinically apparent neck nodes. Where no cervical lymph node suitable for biopsy was seen, parotid glands were biopsied if deemed abnormal. A diagnosis of sarcoidosis was made in all cases where a core biopsy of cervical lymph nodes was attempted. The cervical lymph nodes in this cohort were not particularly enlarged, short-axis dimensions being less than 10mm in majority, and they did not have any sonographic appearances to mark them as pathological. Nevertheless histological examination revealed non-caseating granulomas in all cases. In further two cases, where no neck nodes were seen, a histological diagnosis of sarcoidosis was made from biopsy of diffusely abnormal parotid glands. Given the clear advantages of cervical diagnosis in terms of invasiveness and economy compared to mediastinal alternatives, it is suggested that where the expertise for core biopsy of normal-sized cervical lymph nodes is readily available, the technique may be considered as a first-line investigation for the diagnosis of sarcoidosis.


Photobiomodulation at Multiple Wavelengths Differentially Modulates Oxidative Stress In Vitro and In Vivo.
Photobiomodulation (PBM) is emerging as an effective strategy for the management of multiple inflammatory conditions, including oral mucositis (OM) in cancer patients who receive chemotherapy or radiotherapy. Still, the poor understanding of the mechanisms by which the light interacts with biological tissues and the heterogeneity of light sources and protocols employed worldwide significantly limits its applicability. Reactive oxygen species (ROS) are massively generated during the early phases of OM and play a major role in the pathogenesis of inflammation in general. Here, we report the results of a clinical and experimental study, aimed at evaluating the effect of laser light at different wavelengths on oxidative stress in vivo in oncologic patients suffering from OM and in vitro in two cell types abundantly present within the inflamed oral mucosa, neutrophil polymorphonuclear (PMN) granulocytes, and keratinocytes. In addition to standard ROS detection methods, we exploited a roGFP2-Orp1 genetically encoded sensor, allowing specific, quantitative, and dynamic imaging of redox events in living cells in response to oxidative stress and PBM. We found that the various wavelengths differentially modulate ROS production. In particular, the 660 nm laser light increases ROS production when applied either before or after an oxidative stimulus. In contrast, the 970 nm laser light exerted a moderate antioxidant activity both in the saliva of OM patients and in both cell types. The most marked reduction in the levels of ROS was detected in cells exposed either to the 800 nm laser light or to the combination of the three wavelengths. Overall, our study demonstrates that PBM exerts different effects on the redox state of both PMNs and keratinocytes depending on the used wavelength and prompts the validation of a multiwavelength protocol in the clinical settings.


Feasibility of localized immunosuppression: 2. PLA microspheres for the sustained local delivery of a soft immunosuppressant.
While biohybrid therapy shows promise, their further development into an "artificial pancreatic" system in diabetics also requires the management of the related immuneresponse triggered by such cellular therapies. Ideally this should be on a local level within the biohybrid device. This study relates to the design of sustained release formulations of the glucocorticoid soft drug loteprednol etabonate (LE) that are intended to locally suppress the immune response within the biohybrid devices, thereby warranting high local activity and reduced systemic side effects. Poly(D,L-lactic) acid (PLA) and poly(D,L-lactic glycolic acid (PLGA) microspheres of the soft corticosteroid loteprednol etabonate (LE) were prepared by solvent evaporation. A range of particles differing in particle size, nature of the polymer, emulsification method, and emulsifier were prepared and characterized. These results showed that the approach is able to customize slow release particles with predictable release characteristics over a period of days to month. Preliminary studies were performed with particles of a drug loading of 3.9 (+/- 0.2) %, and a mean particle diameter of 5 microm. In-vitro release studies indicated that these particles released drug over a period of three months. In vitro cell toxicity studies suggested that at higher concentrations (> 1 microM), unencapsulated LE showed some effect on the viability of the MIN-6 insuloma cell line, while the sustained release microspheres showed no cytotoxicity. The ability of these microspheres to provide localized immunosuppression has been evaluated in a set of early exploratory experiments with diabetic rats receiving islet transplantation. Animals treated using a biohybrid device loaded with microspheres showed improved results compared to those treated by delivery in solution form with an osmotic mini-pump. These results show the promise of localized glucocorticoid treatment by sustained release microspheres as a possible form of localized immunosuppression regimen. However, further confirmation is required before use in cell or organ transplantation.


The effect of cognitive-behavioral therapy and haptonomy on fear of childbirth in primigravida women: a randomized clinical trial.
Considering the role of fear of childbirth (FOC) in the enhancement of unnecessary cesarean sections (CS), the present study aimed at evaluating the effect of Cognitive-Behavioral Therapy (CBT) and haptonomy on the FOC (as primary outcome) and intended birth method and final birth method (as secondary outcomes) among primigravida women. This randomized clinical trial was conducted on 99 primigravida women in Tabriz, Iran 2022. Participants were assigned to three groups with a ratio of 1:1:1 using stratified block randomization based on the fear intensity. One of the intervention groups (n = 33) received eight group sessions of CBT from 24 to 28 weeks of gestation and the other intervention group (n = 33) received haptonomy during seven sessions once a week. The control group (n = 33) received routine prenatal care. The Wijma questionnaire was completed by the participants before the intervention, after the intervention at 35-37 weeks of gestation, and after birth. The intended birth method was investigated before and after the intervention at 35-37 weeks of gestation. The final birth method and the reasons for CS were recorded based on the mother's medical profile. The one-way ANOVA was used before the intervention and RMANOVA after the intervention to compare the mean scores of FOC among the three groups. Further, chi-square test was applied to compare the intended and final birth method. The mean (standard deviation: SD) of FOC in the CBT group changed from 74.09 (11.35) at 24-28 weeks of gestation to 46.50 (18.28) at 35-37 weeks and 48.78 (20.64) after birth (P < 0.001). The means (SDs) of FOC in the haptonomy group were 76.81 (13.09), 46.59 (15.81), and 45.09 (20.11), respectively (P < 0.001). The mean (SD) of FOC in the control group decreased from 70.31 (6.71) to 66.56 (18.92) and then, increased to 71.00 (21.14) after birth (P = 0.878). After the intervention, there was no statistically significant difference among the three groups in terms of the intended birth method (P = 0.278), and final birth method (P = 0.107). The findings of the present study revealed that both CBT and haptonomy interventions reduce FOC. Although the desire for vaginal birth and final vaginal birth in the haptonomy group was more than that in the other two groups, there was no statistically significant difference among the three groups. Iranian Registry of Clinical Trials: IRCT20170506033834N9. Date of registration: 02.01.2022. URL: http://en.irct.ir .


Direct observed procedural skills assessment in the undergraduate setting.
Medical students are required to undertake procedural clinical skills training before they qualify as doctors, and an assessment of these skills is a critical element of their fitness to practice. Challenges facing educators include the logistics of observing: i.e. who is best placed to assess their competence? Evidence appears to be inconclusive about the competence of students in the workplace to adhere to standards of practice, and the time required for an effective assessment. In this article the aim is to discuss who is best placed to assess final-year medical students in the workplace. We explore the use of direct observed procedural skills (DOPS) to assess students undertaking procedural skills in a simulated workplace setting by tutor-, peer- and self-assessment. The DOPS tool has been used to assess foundation doctors, but can it be used to effectively assess undergraduate medical students? The main purpose of formative assessment in the simulated setting is to support student learning through the provision of feedback and debriefing. The use of the DOPS tool in this way can provide an insightful perspective of a students' assessment of procedural clinical skills. Tutors can use the DOPS tool to guide their teaching practice by tailoring their lessons towards areas in which students require more guidance. The DOPS assessment tool presents an opportunity to provide immediate and relevant feedback.


Understanding Technology Preferences and Requirements for Health Information Technologies Designed to Improve and Maintain the Mental Health and Well-Being of Older Adults: Participatory Design Study.
Worldwide, the population is aging rapidly; therefore, there is a growing interest in strategies to support and maintain health and well-being in later life. Although familiarity with technology and digital literacy are increasing among this group, some older adults still lack confidence in their ability to use web-based technologies. In addition, age-related changes in cognition, vision, hearing, and perception may be barriers to adoption and highlight the need for digital tools developed specifically to meet the unique needs of older adults. The aim of this study is to understand the use of technology by older adults in general and identify the potential barriers to and facilitators of the adoption of health information technologies (HITs) to support the health and well-being of older adults to facilitate implementation and promote user uptake. In addition, this study aims to co-design and configure the InnoWell Platform, a digital tool designed to facilitate better outcomes for people seeking mental health services, to meet the needs of adults 50 years and older and their supportive others (eg, family members, caregivers) to ensure the accessibility, engagement, and appropriateness of the technology. Participants were adults 50 years and older and those who self-identified as a supportive other (eg, family member, caregiver). Participants were invited to participate in a 3-hour participatory design workshop using a variety of methods, including prompted discussion, creation of descriptive artifacts, and group-based development of user journeys. Four participatory design workshops were conducted, including a total of 21 participants, each attending a single workshop. Technology use was prevalent, with a preference indicated for smartphones and computers. Factors facilitating the adoption of HITs included personalization of content and functionality to meet and be responsive to a consumer's needs, access to up-to-date information from reputable sources, and integration with standard care practices to support the relationship with health professionals. Concerns regarding data privacy and security were the primary barriers to the use of technology to support mental health and well-being. Although HITs have the potential to improve access to cost-effective and low-intensity interventions at scale for improving and maintaining mental health and well-being, several strategies may improve the uptake and efficacy of technologies by the older adult community, including the use of co-design methodologies to ensure usability, acceptability, and appropriateness of the technology; support in using and understanding the clinical applications of the technology by a digital navigator; and ready availability of education and training materials.


Results of a model analysis to estimate cost utility and value of information for intravenous immunoglobulin in Canadian adults with chronic immune thrombocytopenic purpura.
The aim of this work was to estimate the cost-effectiveness of intravenous immunoglobulin (IVIg) compared with oral prednisone as a treatment for Canadian adults with persistent chronic immune thrombocytopenic purpura (ITP). The lifetime costs and effectiveness of IVIg and prednisone were estimated from the perspective of a publicly funded health care system in Canada, using a Markov model that was developed based on a systematic clinical and economic review and recommendations of clinical experts in Canada. Transition probabilities (ie, point estimates and 95% CIs) were estimated from the studies identified in a systematic literature review using a random-effect meta-analysis; point estimates were weighted-mean values from the meta-analysis. No published studies directly estimate the utility weight for patients with relapsed or refractory ITP; therefore, a value of 0.76 was used, based on the mean of the utilities for thrombocytopenia without major bleeding or hemorrhagic stroke. Costs and incremental cost-effectiveness ratios were reported as year-2007 Can $. The incremental costs and quality-adjusted life-years (QALYs) of IVIg versus prednisone were Can $8080 and 0.0071, respectively, resulting in an incremental cost-effectiveness ratio of Can $1.13 million/ QALY in the base-case analysis. The probability of IVIg being cost-effective was 0 if the maximum willingness-to-pay (WTP) value for an additional QALY was below Can $40,000. The probability that IVIg would be cost-effective was only 20%, even if the WTP increased to Can $100,000. The expected value of perfect information (EVPI) and expected value of partial perfect information (EVPPI) were 0 if the WTP was less than Can $30,000. If WTP increased to Can $100,000, the EVPI was Can $1700, and the EVPPI was Can $1010 for utility weights of relapse/refractory states, Can $136 for initial response rates of the treatments, and Can $6 for first-year relapse rates for the treatments. Based on the current available clinical evidence, this model analysis of hypothetical patients suggests that IVIg may not be a cost-effective option for adults with persistent chronic ITP in Canada.


Outcomes with ventricular assist device versus extracorporeal membrane oxygenation as a bridge to pediatric heart transplantation.
Extracorporeal membrane oxygenation (ECMO) has long been the sole means of mechanical support for pediatric patients with end-stage cardiac failure, but has a high waitlist mortality and a reported survival to hospital discharge of less than 50%. The purpose of this study was to compare waitlist mortality and survival for ECMO versus ventricular assist device (VAD) support. A review was conducted of all patients listed for heart transplantation (HTx) since 2002 and requiring mechanical support. VAD support has been available from 2004 (Berlin Heart Excor Pediatrics). Competing risks analysis was used to model survival to one of four outcomes (HTx, death on waitlist, delisting, improvement). Thirty-six patients were on mechanical support while awaiting HTx (21 ECMO, 12 VAD, three both). Median age at listing was 1.2 years (birth-16.6 years) for ECMO and 11.3 years (0.3-14.6 years) for VAD. Diagnosis was cardiomyopathy in 33% for ECMO and 93% for VAD. Median time to HTx was 37 days (1-930) overall, 20 days (1-85) for ECMO, and 39 days (5-108) for VAD. Mechanical support was associated with increased odds of HTx (hazard ratio [HR] 2.4 [1.7-3.3], P < 0.0001) but also delisting or death waiting (HR 3.0 [1.1-7.8], P = 0.03). Waitlist mortality of 38% on ECMO was reduced to 13% with VAD use. Survival post-HTx to hospital discharge was better in the group on VAD support (92 vs. 80%). Pediatric patients requiring mechanical support as a bridge to HTx have short wait times but high waitlist mortality. Those patients who survived to be put on the Berlin Heart Excor Pediatric device based on individualized clinical decision making then had a lower waitlist mortality, a longer duration of support, and a higher survival to transplantation and hospital discharge.


FDA Benchmark Medical Device Flow Models for CFD Validation.
Computational fluid dynamics (CFD) is increasingly being used to develop blood-contacting medical devices. However, the lack of standardized methods for validating CFD simulations and blood damage predictions limits its use in the safety evaluation of devices. Through a U.S. Food and Drug Administration (FDA) initiative, two benchmark models of typical device flow geometries (nozzle and centrifugal blood pump) were tested in multiple laboratories to provide experimental velocities, pressures, and hemolysis data to support CFD validation. In addition, computational simulations were performed by more than 20 independent groups to assess current CFD techniques. The primary goal of this article is to summarize the FDA initiative and to report recent findings from the benchmark blood pump model study. Discrepancies between CFD predicted velocities and those measured using particle image velocimetry most often occurred in regions of flow separation (e.g., downstream of the nozzle throat, and in the pump exit diffuser). For the six pump test conditions, 57% of the CFD predictions of pressure head were within one standard deviation of the mean measured values. Notably, only 37% of all CFD submissions contained hemolysis predictions. This project aided in the development of an FDA Guidance Document on factors to consider when reporting computational studies in medical device regulatory submissions. There is an accompanying podcast available for this article. Please visit the journal's Web site (www.asaiojournal.com) to listen.


Pharmacokinetics of mycophenolic acid and its effect on CD4+ and CD8+ T cells after oral administration of mycophenolate mofetil to healthy cats.
Mycophenolate mofetil (MMF) is an immunosuppressant used in human and veterinary medicine. Little pharmacokinetic and pharmacodynamic information on MMF is available in cats. To evaluate the plasma disposition of mycophenolic acid (MPA) and assess its effect on total peripheral blood mononuclear cells and CD4+ /CD8+ ratios after PO administration of MMF. Healthy cats (n = 10). Mycophenolate mofetil was administered at a dosage of 10 mg/kg q12h (n = 3), 15 mg/kg q12h (n = 3), and 15 mg/kg q8h (n = 4) for 7 days. Concentrations of MPA and derivatives were determined using ultra-high-performance liquid chromatography. Flow cytometry was used to assess CD4+ /CD8+ T-cell ratios. All cats biotransformed MMF into MPA. Half of the cats (5/10) had adverse effects within 1 week of MMF administration. Area under the curve limit of quantification (AUC0-LOQh ) of MPA ranged from 1.27 to 2.03 hours·μg/mL and from 1.77 to 8.54 hours·μg/mL after the first and last PO dose of 10 mg/kg. The AUC0-loqh of MPA ranged from 2.18 to 31 hours·μg/mL after the first dose of 15 mg/kg of MMF. Before the first dose of MMF, the average total number of PBMC ranged from 1.2 to 9.3 million/mL. At the last dose of MMF, the average total number of PBMC ranged from 3 to 5 million/mL. Mycophenolic acid was detected in all cats. The dose 10 mg/kg given q12h for 1 week was tolerated (n = 3). The efficacy of MMF as an immunosuppressant and long-term safety in cats of this dosage regimen is unknown.


Axial heterogeneity of intracellular pH in rat proximal convoluted tubule.
In the proximal convoluted tubule (PT), the HCO3- reabsorptive rate is higher in early (EPS) compared with late proximal segments (LPS). To examine the mechanism of this HCO3- reabsorption profile, intracellular pH (pHi) was measured along the superficial PT of the rat under free-flow and stationary microperfusion using the pH-sensitive fluorescence of 4-methylumbelliferone (4MU). With 4MU superfusion, pHi was found to decline along the PT. Observation with 365-nm excitation revealed that EPS were brightly fluorescent and always emerged away from their star vessel. Midproximal segments were darker and closer to the star vessel which was surrounded by the darkest LPS. Decreasing luminal HCO3- from 15 to 0 mM lowered pHi in both EPS and LPS, but pHi remained more alkaline in EPS with both perfusates. Thus the axial decline in pHi along the PT is due to both luminal factors and intrinsic differences in luminal H+ extrusion in PT cells.


Determinants of RNA-dependent RNA polymerase (in)fidelity revealed by kinetic analysis of the polymerase encoded by a foot-and-mouth disease virus mutant with reduced sensitivity to ribavirin.
A mutant poliovirus (PV) encoding a change in its polymerase (3Dpol) at a site remote from the catalytic center (G64S) confers reduced sensitivity to ribavirin and forms a restricted quasispecies, because G64S 3Dpol is a high-fidelity enzyme. A foot-and-mouth disease virus (FMDV) mutant that encodes a change in the polymerase catalytic site (M296I) exhibits reduced sensitivity to ribavirin without restricting the viral quasispecies. In order to resolve this apparent paradox, we have established a minimal kinetic mechanism for nucleotide addition by wild-type (WT) FMDV 3Dpol that permits a direct comparison to PV 3Dpol as well as to FMDV 3Dpol derivatives. Rate constants for correct nucleotide addition were on par with those of PV 3Dpol, but apparent binding constants for correct nucleotides were higher than those observed for PV 3Dpol. The A-to-G transition frequency was calculated to be 1/20,000, which is quite similar to that calculated for PV 3Dpol. The analysis of FMDV M296I 3Dpol revealed a decrease in the calculated ribavirin incorporation frequency (1/8,000) relative to that (1/4,000) observed for the WT enzyme. Unexpectedly, the A-to-G transition frequency was higher (1/8,000) than that observed for the WT enzyme. Therefore, FMDV selected a polymerase that increases the frequency of the misincorporation of natural nucleotides while specifically decreasing the frequency of the incorporation of ribavirin nucleotide. These studies provide a mechanistic framework for understanding FMDV 3Dpol structure-function relationships, provide the first direct analysis of the fidelity of FMDV 3Dpol in vitro, identify the beta9-alpha11 loop as a (in)fidelity determinant, and demonstrate that not all ribavirin-resistant mutants will encode high-fidelity polymerases.


CD39 is highly involved in mediating the suppression activity of tumor-infiltrating CD8+ T regulatory lymphocytes.
CD39 is an ectoenzyme, present on different immune cell subsets, which mediates immunosuppressive functions catalyzing ATP degradation. It is not known whether CD39 is expressed and implicated in the activity of CD8+ regulatory T lymphocytes (Treg). In this study, CD39 expression and function was analyzed in both CD8+ and CD4+CD25(hi) Treg from the peripheral blood of healthy donors as well as from tumor specimens. CD39 was found expressed by both CD8+ (from the majority of healthy donors and tumor patients) and CD4+CD25(hi) Treg, and CD39 expression correlated with suppression activity mediated by CD8+ Treg. Importantly, CD39 counteraction remarkably inhibited the suppression activity of CD8+ Treg (both from peripheral blood and tumor microenvironment) suggesting that CD39-mediated inhibition constitutes a prevalent hallmark of their function. Collectively, these findings, unveiling a new mechanism of action for CD8+ Treg, provide new knowledge on intratumoral molecular pathways related to tumor immune escape, which could be exploited in the future for designing new biological tools for anticancer immune intervention.


Portal hypertension hemorrhage secondary to a stomal varicose vein in a patient with two consecutive trombosis of transjugular intrahepatic portosystemic shunt (TIPS).
Less than 5% of patients with liver cirrhosis (LC) with portal hypertension (PH) develop atypical shunt (in regions other than the esophagus or the stomach). Within this group are varices associated with a stoma, for example the ones associated with an uretero-ileostomy which are infrequent. They are a diagnostic and therapeutic challenge, as they can cause hemorrhages due to PH. We present a clinical case about stoma varicose bleeding as the latest guidelines for the management of PH do not mention them or their treatment due to their low incidence.


Plasmalogens Inhibit Endocytosis of Toll-like Receptor 4 to Attenuate the Inflammatory Signal in Microglial Cells.
Microglial activation is a pathological feature of many neurodegenerative diseases and the role of cellular lipids in these diseases is mostly unknown. It was known that the special ether lipid plasmalogens (Pls) were reduced in the brain and blood samples of Alzheimer's disease (AD) patients. It has recently been reported that the oral ingestion of scallop-derived Pls (sPls) improved cognition among mild AD patients, which led us to investigate the role of sPls in the microglial activation. We used the lipopolysaccharides (LPS)-induced microglial activation model and found that sPls inhibit the LPS-mediated TLR4 endocytosis and the downstream caspases activation. By using the specific inhibitors, we also confirmed that the TLR4 endocytosis and the caspases activation strictly controlled the pro-inflammatory cytokine expression. In addition, the reduction of cellular Pls by sh-RNA-mediated knockdown of GNPAT (glyceronephosphate O-acyltransferase), a Pls synthesizing enzyme, enhanced the endocytosis of TLR4 and activation of caspase-3 which resulted in the enhanced pro-inflammatory cytokine expression. We also report for the first time that the TLR4 endocytosis was significantly higher in the cortex of aged mice and AD model mice brains, proposing a significant link between the age-related reduction of Pls and microglial activation. Interestingly, the sPls drinking in AD model mice significantly reduced the TLR4 endocytosis. Our cumulative data indicates that the cellular Pls attenuate the microglial activation by maintaining the endocytosis of TLR4, suggesting a possible mechanism of the cognition improvement effect of sPls among mild AD patients.


Polysubstituted pyridazinones from sequential nucleophilic substitution reactions of tetrafluoropyridazine.
4,5,6-trifluoropyridazin-3(2H)-one can be used as a scaffold for the synthesis of various 4,5- and 4,6-disubstituted and ring-fused pyridazinone systems by sequential nucleophilic aromatic substitution processes. Although the regioselectivity of nucleophilic substitution can be affected by the nature of the nucleophile and the substituent attached to the pyridazinone ring, a variety of polyfunctional systems can be readily accessed by sequential nucleophilic substitution methodology which may have applications in the drug discovery arena. For example, reaction of 4,5,6-trifluoropyridazin-3(2H)-one with nitrogen nucleophiles leads to a mixture of aminated products arising from substitution of fluorine located at the 4- and 5-positions. The ratio of isomers obtained depends on the nucleophile where the 4-isomer is the major product for reaction with primary and secondary amines such as butylamine, morpholine, and aniline derivatives. Subsequent reaction of representative 4-aminated products gave 4,5-disubstituted systems and ring fused derivatives may be formed by reaction of 4,5,6-trifluoropyridazin-3(2H)-one or 4-substituted systems with N,N'-dimethylethylenediamine.


Defining peripherally inserted central catheter tip position and an evaluation of insertions in one unit.
Peripherally inserted central catheters are increasingly used to provide access to the central venous circulation. They are commonly positioned 'blind' using a variety of anthropometric techniques and operator experience to direct insertion length. Malposition rates are poorly defined because of differing insertion techniques, difficulties defining anatomical tip position on chest radiographs, controversy over what constitutes an adequate catheter position and possible differences between patient groups. We have developed a reproducible method to define catheter positions on chest radiograph and have applied this in a retrospective analysis of 256 ICU and 243 non-ICU catheter insertions over a 6-month period. Two different definitions were used for adequate position. 'Blind' positioning of peripherally inserted central catheters was associated with a definition-dependent malposition rate of 42-76%. Malposition rates were significantly higher in ICU patients. Emerging technologies may assist in reducing these high rates.


Colloid osmotic parameterization and measurement of subcellular crowding.
Crowding of the subcellular environment by macromolecules is thought to promote protein aggregation and phase separation. A challenge is how to parameterize the degree of crowding of the cell interior or artificial solutions that is relevant to these reactions. Here I review colloid osmotic pressure as a crowding metric. This pressure is generated by solutions of macromolecules in contact with pores that are permeable to water and ions but not macromolecules. It generates depletion forces that push macromolecules together in crowded solutions and thus promotes aggregation and phase separation. I discuss measurements of colloid osmotic pressure inside cells using the nucleus, the cytoplasmic gel, and fluorescence resonant energy transfer (FRET) biosensors as osmometers, which return a range of values from 1 to 20 kPa. I argue for a low value, 1-2 kPa, in frog eggs and perhaps more generally. This value is close to the linear range on concentration-pressure curves and is thus not crowded from an osmotic perspective. I discuss the implications of a low crowding pressure inside cells for phase separation biology, buffer design, and proteome evolution. I also discuss a pressure-tension model for nuclear shape, where colloid osmotic pressure generated by nuclear protein import inflates the nucleus.


Use of a genome-wide haploid genetic screen to identify treatment predicting factors: a proof-of-principle study in pancreatic cancer.
The ability to develop a comprehensive panel of treatment predicting factors would significantly improve our ability to stratify patients for cytotoxic or targeted therapies, and prevent patients receiving ineffective treatments. We have investigated if a recently developed genome-wide haploid genetic screen can be used to reveal the critical mediators of response to anticancer therapy. Pancreatic cancer is known to be highly resistant to systemic therapy. Recently epigenetic changes have been shown to be a key determinant in the maintenance of subpopulations of cancer cells with high-level resistance to cytotoxic therapy. We show that in human pancreatic cancer cell lines, treatment with the potent class I histone deacetylase inhibitor, entinostat, synergistically enhances gemcitabine-induced inhibition of cell proliferation and apoptosis. Using a genome-wide haploid genetic screen, we identified deoxycytidine kinase (DCK) as one of the genes with the highest degree of insertional enrichment following treatment with gemcitabine and entinostat; DCK is already known to be the rate-limiting activating enzyme for gemcitabine. Immunoblotting confirmed loss of DCK protein expression in the resistant KBM7 cells. CRISPR/Cas-9 inactivation of DCK in pancreatic cancer cell lines resulted in resistance to gemcitabine alone and in combination with entinostat. We have identified gemcitabine and entinostat as a potential new combination therapy in pancreatic cancer, and in this proof-of-principle study we have demonstrated that a recently developed haploid genetic screen can be used as a novel approach to identify the critical genes that determine treatment response.


Squamous cell carcinoma of the esophagus and multiple primary tumors of the upper aerodigestive tract.
Squamous cell carcinoma of the esophagus is frequently associated with other, synchronous or metachronous tumors, in the upper aerodigestive tract. All 264 patients with squamous cell carcinoma of the esophagus, treated in the Gastrointestinal Surgery, Esophagus section, of the "Hospital das Clínicas" (São Paulo University Medical School, Brazil), between 1979 and 1989 were analyzed retrospectively with regards to the occurrence of multiple primary tumors in the upper aerodigestive tract. Multiple primary tumors were encountered in 10 (3.8%) patients. All patients were male and the mean age at the time of the first primary was 52.2 years. Tobacco smoke and alcohol were the principal carcinogens in these patients (n = 10). The sites of the tumors were: larynx (n = 4), tongue (n = 4), lung (n = 2), and oral cavity (n = 1). Two simultaneous, three synchronous and five metachronous multiple primary carcinomas were detected. The esophagus was the second primary tumor in nine patients. The mean overall survival after the diagnosis of the second primary was 2.8 months (SD = 0.89). Inquiry regarding other malignancies, associated with panendoscopy should be carry out prior to the treatment of the first primary to diagnose simultaneous or synchronous primary tumors, and careful follow-up should be performed after treatment of the first primary to detect new tumors in these high-risk patients.


Differential DNA methylation and expression of inflammatory and zinc transporter genes defines subgroups of osteoarthritic hip patients.
We have previously shown that the cartilage DNA methylome delineates two clusters of osteoarthritic (OA) hip patients, characterised by differential methylation of inflammatory genes, while others have demonstrated a link between zinc homeostasis and inflammation in OA. We aimed to investigate these effects at the methylation and gene expression level. We used our previously generated methylation data while quantitative PCR was used to measure gene expression using RNA from the hip cartilage of members of both clusters and from control individuals without hip OA. One of the OA clusters is characterised by the promoter hypomethylation and increased expression of inflammation-associated genes including IL1A and TNF. Furthermore, we show that the increase in expression of these genes is accompanied by increased expression of several zinc transporter genes. In addition, the zinc responsive transcription factor MTF1 is also upregulated, which is accompanied by an increase in the expression of its targets the metalloproteinases MMP13 and ADAMTS5. We have identified a subgroup of OA hip patients that are epigenetically and transcriptiomically characterised by a cartilage inflammatory phenotype with concurrent differential regulation of zinc regulators. The identification of subgroups enhances stratified phenotyping of OA patients and has important implications for future therapeutic applications.


Physician Factors Associated with Polypharmacy and Potentially Inappropriate Medication Use.
Despite accumulating evidence about the harm of polypharmacy in family medicine, few studies have investigated factors related to polypharmacy. The objective of this study was to explore factors related to physicians' prescribing behavior. We conducted a survey of physicians at 5 family medicine residency practices and a linked health record review of their patients ≥65 years old. The determinants of physicians' mean number of prescriptions and potentially inappropriate medications (PIMs) were examined using a generalized linear model. A total of 61 physicians (38 residents, 23 fellows/faculty) completed the survey, and 2103 visits by 932 patients seen by these physicians were analyzed. The mean numbers of prescriptions and PIMs per visit per physician were 9.50 and 0.46, respectively. After controlling for patient race and age, low prescribers were more likely to consider the number of medications (P = .007) and benefit/risk information for deprescribing (P = .017) when making prescribing decisions. Use of the Beers List was marginally significant in lower PIM prescribing (P = .05). Physicians' sex, duration of experience, and perceived confidence were not associated with prescribing patterns. Conscious consideration concerning the number of medications and benefit/risk information, as well as using the Beers List, were associated with less polypharmacy and fewer PIMs.


Hereditary factor VII deficiency: heterogeneity defined by combined functional and immunochemical analysis.
Twenty-six patients with hereditary factor VII deficiency (VII:C less than 10%) were evaluated using a panel of three thromboplastins of varying species and tissue origin in both coagulant and chromogenic assay systems. Normal values for the coagulation and chromogenic assays were 104% +/- 7% and 108% +/- 21%, respectively. Factor VII antigen was measured by a specific radioimmunoassay (normal, 470 +/- 112 ng/mL). The patients were divided into two groups based on the factor VII:Ag assay results. Group 1, 18 patients, had decreased levels of factor VII:Ag and group 2, eight patients, had normal levels of factor VII:Ag. The two groups were further subdivided on the basis of discrepant factor VII:C levels in the chromogenic and coagulant assays. The number of observed patterns of functional factor VII:C activity suggests a high degree of complexity of factor VII and thromboplastin interaction. Whereas no clinical bleeding was reported in any of the nine black patients evaluated, all Caucasians (16) and one Hispanic presented with mild to severe bleeding. Patients with factor VII:C greater than 10% using a human thromboplastin had a negative bleeding history, regardless of the activity measured with other thromboplastins. Factor VII activity measured with a human thromboplastin appears to correlate best with the clinical picture.


Systematic Review and Meta-analysis of Stepping Stones Triple P for Parents of Children with Disabilities.
The aim of this systematic review and meta-analysis was to examine the effectiveness of Stepping Stones Triple P (SSTP) parent training programs on child behavior problems and parenting outcomes in families of children with developmental disabilities. Sixteen suitable studies including data from over 900 families were identified in a search for English language published and unpublished controlled outcome studies. SSTP has five levels on a graded continuum of increasing intensity targeting families with differing degrees of treatment need from low intensity media-based parenting information campaigns at level 1, through brief interventions at levels 2 and 3, to more intensive parent training and family therapy interventions at levels 4 and 5. Analyses were conducted on the combination of all levels of SSTP and separately for each level. For combined levels, significant overall effect sizes were found for parent-reported child problems (d = 0.46), researcher observed child behavior (d = 0.51), parenting style (d = 0.70), parenting satisfaction/self-efficacy (d = 0.44), parental adjustment (d = 0.27), and coparental relationship (d = 0.26), but not researcher-observed parent behavior. Strong support was found for level 4 SSTP as an effective intervention for improving child and parent outcomes in families of children with disabilities who have clinically significant problems. Less intensive SSTP interventions for cases with circumscribed difficulties yielded fewer significant treatment effects, and there were relatively few studies of such interventions.


Physiological and structural anorectal abnormalities in patients with systemic sclerosis and fecal incontinence.
Fecal incontinence is common in systemic sclerosis (SSc), but the underlying mechanisms are not fully understood. The objectives of this study were to characterize anorectal physiological and morphological defects in SSc patients and to correlate the results with incontinence symptoms. Twenty-five SSc patients underwent anorectal neurophysiological investigations, anal manometry, and ultrasound. Eleven patients (44%) reported incontinence to solid or liquid feces, but no patient reported diarrhea. Increased fiber density (FD) was recorded in 78% of patients with and in 86% of patients without fecal incontinence not significant (NS). Incontinent patients had lower squeeze pressure (SP; median 49.5 mm Hg) in the high-pressure zone (HPZ) than continent patients (median 72 mm Hg; p = 0.01). In two of the incontinent patients, sonographic abnormalities of the internal anal sphincter (IAS) and the external anal sphincter (EAS) were present, whereas in another two patients isolated IAS abnormalities were seen. These four individuals had lower resting pressure at 1 cm and in the HPZ, and lower SP at 2 cm than patients with normal anorectal sonographic findings (p < 0.05). Lower voluntary SP in incontinent patients and EAS sonographic abnormalities only in patients with incontinence suggest that the EAS is more important in maintaining fecal continence in SSc patients than has previously been reported. The finding of increased FD in most patients further supports involvement of the EAS function in SSc and could indicate previous nerve injury with consequent incomplete reinnervation.


Age-based computer-aided diagnosis approach for pancreatic cancer on endoscopic ultrasound images.
The aim was to develop a high-performance computer-aided diagnosis (CAD) system with image processing and pattern recognition in diagnosing pancreatic cancer by using endosonography images. On the images, regions of interest (ROI) of three groups of patients (<40, 40-60 and >60) were extracted by experts; features were obtained from images using three different techniques and were trained separately for each age group with an Artificial Neural Network (ANN) to diagnose cancer. The study was conducted on endosonography images of 202 patients with pancreatic cancer and 130 noncancer patients. 122 features were identified from the 332 endosonography images obtained in the study, and the 20 most appropriate features were selected by using the relief method. Images classified under three age groups (in years; <40, 40-60 and >60) were tested via 200 random tests and the following ratios were obtained in the classification: accuracy: 92%, 88.5%, and 91.7%, respectively; sensitivity: 87.5%, 85.7%, and 93.3%, respectively; and specificity: 94.1%, 91.7%, and 88.9%, respectively. When all the age groups were assessed together, the following values were obtained: accuracy: 87.5%, sensitivity: 83.3%, and specificity: 93.3%. It was observed that the CAD system developed in the study performed better in diagnosing pancreatic cancer images based on classification by patient age compared to diagnosis without classification. Therefore, it is imperative to take patient age into consideration to ensure higher performance.


Guard-band influence on orthogonal-band-multiplexed coherent optical OFDM.
Coherent optical orthogonal frequency-division multiplexing (OFDM) has recently been demonstrated beyond 100 Gb/s using orthogonal band multiplexing (OBM). OBM-OFDM gives a multitude of benefits in implementing high-speed optical OFDM transmission. We investigate the system sensitivity to the guard band of OBM-OFDM. The result suggests that when the guard band equals a multiple of the subcarrier spacing, the Q penalty practically vanishes. We also show that two OFDM bands can be demodulated simultaneously with one fast Fourier transform, owing to interband orthogonality, and its performance is almost the same as one-band detection.


DOSIMETRIC CHARACTERISATION OF A SUB-NATURAL BACKGROUND RADIATION ENVIRONMENT FOR RADIOBIOLOGY INVESTIGATIONS.
Living systems have evolved in the presence of naturally occurring ionising radiation. REPAIR is a research project investigating the biological effects of sub-natural background radiation exposure in SNOLAB, a deep-underground laboratory. Biological systems are being cultured within a sub-background environment as well as two control locations (underground and surface). A comprehensive dosimetric analysis was performed. GEANT4 simulation was used to characterise the contribution from gamma, muons and neutrons. Additionally, dose rates from radon, 40K and 14C were calculated based on measured activity concentrations. The total absorbed dose rate in the sub-background environment was 27 times lower than the surface control, at 2.48 ± 0.20 nGy hr-1, including a >400-fold reduction in the high linear energy transfer components. This modelling quantitatively confirms that the environment within SNOLAB provides a substantially reduced background radiation dose rate, thereby setting the stage for future sub-background biological studies using a variety of model organisms.


COVID-19 in Long-Term Care Facilities: A Rapid Review of Infection Correlates and Impacts on Mental Health and Behaviors.
Long-term care facilities (LTCFs) with compact, group-living arrangements have become COVID-19 hot spots during the pandemic. Systematic research is needed to understand factors associated with COVID-19 infections in LTCFs and the inadvertent effects of preventive measures adopted by LTCFs. This rapid review identifies factors associated with LTCF residents' COVID-19 infections and the impacts of the pandemic and the corresponding preventive measures on residents' mental health and behavioral problems. Following the preferred reporting items for systematic reviews and meta-analyses guidelines, we identified and reviewed relevant literature in Medline, PsycINFO, and AgeLine. Thirty-seven articles were identified and reviewed, including 30 reporting factors associated with COVID-19 infections in LTCFs and seven reporting the impact of the pandemic and corresponding prevention measures on LTCF residents. Results revealed four domains of factors associated with COVID-19 infections: facility physical environments, resident characteristics, facility management and testing, and community factors. The pandemic and infection control measures increased residents' depression, anxiety, loneliness, and behavioral problems (e.g., agitation, hallucinations). Residents without cognitive impairments were more vulnerable to these adverse effects. LTCF managers/policymakers and healthcare designers can help mitigate COVID-19 infections by (1) providing additional resources to vulnerable LTCFs; (2) enhancing the training of personal protective equipment use and guideline compliance; and (3) investing in amenities, such as sinks, quarantine rooms, and outdoor spaces. Digital activities and accessible green spaces can mitigate mental health and behavior issues. Future LTCF design can benefit from flexible spaces, natural ventilation, and reducing crowding.


Dietary Methionine Restriction Regulates Liver Protein Synthesis and Gene Expression Independently of Eukaryotic Initiation Factor 2 Phosphorylation in Mice.
Background: The phosphorylation of eukaryotic initiation factor 2 (p-eIF2) during dietary amino acid insufficiency reduces protein synthesis and alters gene expression via the integrated stress response (ISR).Objective: We explored whether a Met-restricted (MR) diet activates the ISR to reduce body fat and regulate protein balance.Methods: Male and female mice aged 3-6 mo with either whole-body deletion of general control nonderepressible 2 (Gcn2) or liver-specific deletion of protein kinase R-like endoplasmic reticulum kinase (Perk) alongside wild-type or floxed control mice were fed an obesogenic diet sufficient in Met (0.86%) or an MR (0.12% Met) diet for ≤5 wk. Ala enrichment with deuterium was measured to calculate protein synthesis rates. The guanine nucleotide exchange factor activity of eIF2B was measured alongside p-eIF2 and hepatic mRNA expression levels at 2 d and 5 wk. Metabolic phenotyping was conducted at 4 wk, and body composition was measured throughout. Results were evaluated with the use of ANOVA (P < 0.05).Results: Feeding an MR diet for 2 d did not increase hepatic p-eIF2 or reduce eIF2B activity in wild-type or Gcn2-/- mice, yet many genes transcriptionally regulated by the ISR were altered in both strains in the same direction and amplitude. Feeding an MR diet for 5 wk increased p-eIF2 and reduced eIF2B activity in wild-type but not Gcn2-/- mice, yet ISR-regulated genes altered in both strains similarly. Furthermore, the MR diet reduced mixed and cytosolic but not mitochondrial protein synthesis in both the liver and skeletal muscle regardless of Gcn2 status. Despite the similarities between strains, the MR diet did not increase energy expenditure or reduce body fat in Gcn2-/- mice. Finally, feeding the MR diet to mice with Perk deleted in the liver increased hepatic p-eIF2 and altered body composition similar to floxed controls.Conclusions: Hepatic activation of the ISR resulting from an MR diet does not require p-eIF2. Gcn2 status influences body fat loss but not protein balance when Met is restricted.


Acute pyridostigmine overdose: a report of nine cases.
Pyridostigmine is known as a pre-treatment drug against intoxication with organophosphorus nerve agents. During the Persian Gulf war, we encountered a cluster of nine cases of pyridostigmine self-poisoning, of which three presented with mixed drug poisoning. The clinical and laboratory features of pyridostigmine toxicity are presented. Doses ranged between 390 and 900 mg. Pyridostigmine ingestion resulted in mild to moderate cholinergic symptoms such as abdominal cramps, diarrhea, emesis, nausea, hypersalivation, urinary incontinence, fasciculations, muscle weakness and blurred vision. No central nervous system manifestations were evident. The symptoms developed within several minutes and lasted up to 24 h. All patients underwent gastric emptying followed by administration of activated charcoal. Atropine (1-8 mg) was required in only three patients. Measurement of serum cholinesterase inhibition was found to be a reliable and sensitive diagnostic tool in pyridostigmine poisoning. No clear correlation was found between the extent of cholinesterase inhibition and the incidence or severity of the cholinergic signs. The clinical recovery was faster than the spontaneous recovery of the enzyme. Pyridostigmine intoxication is self-limited and well tolerated by young healthy adults.


Concentration and occupancy of dopamine transporters in cocaine abusers with [11C]cocaine and PET.
The concentration (Bmax) of the dopamine transporter (DAT) and the maximum and effective occupancies by cocaine doses of 0.1 mg/kg or 0.05 mg/kg were measured in the striatum of cocaine abusers (n = 12) by using [11C]cocaine as a radiotracer for the DAT and positron emission tomography (PET). Two methods based on a three-compartment model with one binding site (the nonlinear least squares (NLSQ) and the Farde pseudoequilibrium method) were used to estimate Bmax. Effective occupancies and maximum occupancies were calculated from the distribution volume ratios (DVR) and a three-compartment model, respectively. The NLSQ and Farde methods gave similar values of Bmax (average, 650 +/- 350 pmol/ml and 776 +/- 400 pmol/ml, respectively), but the individual estimates of Bmax were found to be very sensitive to small variations in other model parameters and were not correlated with the parameter Bmax/Kd (r = .07). The average maximum (and effective) occupancies were found to be 67% (50%) and 52% (39%) for the 0.1-mg/kg and the 0.05-mg/kg studies, respectively. The ED50 based on the effective occupancy corresponds to 0.1 mg/kg, which is significantly smaller than the ED50 of 3 mg/kg calculated from studies in which [123]beta-CIT is displaced by cocaine. The effect on the Bmax estimate of two binding sites with different Kd's is also considered by simulation. We conclude (1) that the lack of robustness in the Bmax estimate limits the usefulness of any one subject's Bmax and suggests that the combination parameter Bmax/Kd (or the DVR), which has been used extensively, is a more stable measure of free receptor/transporter concentration. The average Bmax may, however, provide an estimate of the expected concentration in humans. (2) The DVR can be used as a measure of DAT occupancy without applying an explicit model.


Cardiac and renal outcomes of direct oral anticoagulants in patients with atrial fibrillation.
Oral anticoagulation therapy with warfarin or direct oral anticoagulants (DOACs) is the mainstay for stroke prevention in patients with non-valvular atrial fibrillation (AF). The DOACs might have a lower risk of declining renal function than warfarin. This study aimed to compare renal outcomes among rivaroxaban, edoxaban, dabigatran, and warfarin. This cohort study identified 2203 adults with AF who started anticoagulation therapy between 1 July 2013 and 31 December 2020, in a clinical database at a single centre. Inverse probability of treatment weighting was adopted to balance baseline characteristics among four anticoagulants treatment groups. The primary outcome was a composite of cardiac and renal outcomes, involving a ≥30% decline in estimated glomerular filtration rate (eGFR), renal failure and cardiovascular death. After propensity score weighting, dabigatran was associated with significantly lower risks of a ≥30% decline in eGFR (hazard ratio [HR]: .69, 95% confidence interval [CI]: .497-.951, p = .0237), doubling of the serum creatinine level (HR: .49, 95% CI: .259-.927, p = .0282) and the cardiac and renal outcome composite (HR: .67, 95% CI: .485-.913, p = .0115) than warfarin. Rivaroxaban and edoxaban did not show significant protective effects on renal outcomes compared to warfarin. In this study, patients treated with dabigatran had significantly reduced risks of declining renal function and composite cardiac and renal events than those treated with warfarin. However, rivaroxaban and edoxaban were not associated with lower risks of any renal outcomes than warfarin. More studies are warranted to investigate and compare the impact of renal function between different DOACs in patients with AF.


Retrospective cohort study of neonatal blood transfusion in China.
Blood transfusion therapy is extremely important for certain neonatal diseases, but the threshold for neonatal blood transfusion is not the same in different countries. Until now, clinical studies to determine the suitable threshold for newborns in China are lacking. Therefore, it is of high importance to establish a multi-center cohort study to explore appropriate transfusion thresholds for newborns in China. This retrospective cohort study investigated neonatal blood transfusion therapy administered from January 1, 2017 to June 30, 2018, with the aim of evaluating the effect of restricted and nonrestricted blood transfusion on neonatal health. The subjects were enrolled in 46 hospitals in China. A total of 5669 neonatal cases were included in the study. Clinical diagnosis and transfusion treatment of these neonates were collected and the data were retrospectively analyzed. The neonates were followed up 1 week and 1 month after leaving the hospital. The newborns' and their mothers' data were collected containing 280 variables in the database. The primary outcome of the study was mortality, and the secondary outcomes were complications, hospital stays, NICU hospital stays and hospital costs. Results from the < 1500 g group showed that there was a higher mortality rate in the restricted transfusion group (11.41%) when compared with the non-restricted transfusion group (5.12%) (P = 0.000). Among the secondary outcomes, the restricted transfusion group had fewer costs. Results from the 1500-2500 g group showed that the mortality rates of the restricted and non-restricted transfusion groups were 3.53% and 4.71%, respectively, however there was no statistical significance between the two groups (P = 0.345). Among the secondary outcomes, the restricted transfusion group had fewer hospital stays, NICU hospital stays and hospital costs. The incidence of necrotizing enterocolitis was lower in the restricted transfusion group (OR, 2.626; 95% confidence interval [CI], 1.445 to 4.773; P = 0.003). The results from the ≥ 2500 g restricted transfusion group suggested that the mortality rate of (3.02%) was significantly lower than that of non-restricted transfusion group (9.55%) (P = 0.000). Among the secondary outcomes, the restricted transfusion group had fewer hospital stays and hospital costs. The incidence of retinopathy of prematurity was lower in the restricted transfusion group (OR, 4.624; 95% confidence interval [CI], 2.32 to 9.216; P = 0.000). Current transfusion protocols for newborns weighing less than 1500 g may be inappropriate and lead to higher mortality. The current transfusion threshold performed better for the other two weight groups.


Lactational transfer of PCBs and chlorinated pesticides in pups of southern elephant seals (Mirounga leonina) from Antarctica.
Seven pairs of southern elephant seals (Mirounga leonina) pups and their dams were sampled during the late weaning season among a breeding population of seals on Elephant Island in Antarctica. The blubber of the pups and the milk and blubber of their dams were analyzed for lipid-normalized concentrations of PCBs and organochlorines compounds in order to evaluate the lactational transfer of these contaminants. The lipid-normalized concentrations in these tissues were in the ppb range (i.e., ngg(-1) lipid). The levels of contaminants in southern elephant seals were low in comparison with residues that have been reported in pinnipeds from the northern hemisphere. The relative tissue concentrations of the analytes measured followed the pattern: SigmaDDT>mirex>SigmaPCB>Sigmachlordane>HCB>heptachlor epoxide>dieldrin>methoxychlor>SigmaHCH>other organochlorines. The very high DDE/SigmaDDT ratio (0.91) in the blubber of dams and pups was an indicative of long-term, extremely distant pollution. On the other hand, the relatively high levels of some other organochlorine pesticides (e.g. mirex, heptachlor epoxide, dieldrin, methoxychor) may reflect the continued use of these insecticides in developing countries located in the southern hemisphere. For most of the analytes measured, the lipid-normalized concentrations were lower in pup blubber and in the milk than in the maternal blubber. Lactational transfer rates were dependent on the logK(ow) (octanol/water partition coefficient) values of the analytes measured, less lipophilic compounds being more readily transferred to the pups by the lactational route.


Quality of Beverage Intake and Cardiometabolic and Kidney Outcomes: Insights From the STANISLAS Cohort.
Background and Aims: Beverages are an important aspect of diet, and their quality can possibly affect health. The Healthy Beverage Index (HBI) has been developed to take into account these effects. This study aimed to highlight the relationships between health and beverage quality by assessing the association of the HBI and its components with kidney and cardiometabolic (CM) outcomes in an initially healthy population-based familial cohort. Methods: This study included 1,271 participants from the STANISLAS cohort. The HBI, which includes 10 components of habitual beverage consumption, was calculated. Associations of the HBI and its components with estimated glomerular filtration rate (eGFR), albuminuria, hypertriglyceridemic waist (HTG waist), metabolic syndrome (MetS), carotid-femoral pulse wave velocity (cfPWV), carotid intima-media thickness (cIMT), and left ventricular mass (LV mass) were analyzed using multivariable linear or logistic regression models. Results: The median HBI score was 89.7 (78.6-95) out of 100 points. While the overall HBI score was not significantly associated with any of the studied outcomes, individual HBI components were found differently associated with the outcomes. cfPWV and cIMT were lower in participants who did not meet the full-fat milk criteria (p = 0.03 and 0.001, respectively). In men, higher cfPWV was observed for the "low Fat milk" (p = 0.06) and "alcohol" (p = 0.03) non-adherence criteria. Odds of HTG waist were higher with the non-adherence to sugar-sweetened beverages criteria (p < 0.001). eGFR was marginally higher with non-adherence to the coffee/tea criteria (p = 0.047). Conclusions: In this initially healthy population, HBI components were differently associated with kidney and cardiometabolic outcomes, despite a good overall HBI score. Our results highlight specific impacts of different beverage types and suggest that beverages could have an impact on kidney and cardiometabolic health.


Surveillance for gonorrhea and primary and secondary syphilis among adolescents, United States--1981-1991.
During the 1980s, an increasing proportion of adolescent women reported having had premarital sexual intercourse, thus potentially placing an increasing number of young persons at higher risk of acquiring a sexually transmitted infection. To determine rates and examine trends of sexually transmitted infections among adolescents, we analyzed data for reported cases of gonorrhea and primary and secondary syphilis among 10- to 19-year-olds for 1981 through 1991. Summary data for cases of gonorrhea and primary and secondary syphilis that were identified and reported to state health departments were sent annually to CDC. These data included total number of cases by disease (gonorrhea, primary and secondary syphilis), sex, racial/ethnic group (white, not of Hispanic origin; black, not of Hispanic origin; Hispanic; Asian/Pacific Islander; or American Indian/Alaskan Native), 5-year age group, and source of report (public, private). From 1981 through 1991, 24%-30% of the reported morbidity from gonorrhea and 10%-12% of the reported morbidity from primary and secondary syphilis in the United States affected the adolescent age groups. Some of the highest rates of gonorrhea during that time period were among 15- to 19-year-olds. Gonorrhea rates among adolescents increased or remained unchanged from 1981 through 1991, while the rates among older age groups decreased. Although primary and secondary syphilis rates were lower among adolescents than older age groups, adolescents contributed to the epidemic of syphilis that occurred from 1987 through 1990. Differences in reported rates of both syphilis and gonorrhea among white, black, and Hispanic adolescents increased during the latter half of the 1980s. Reporting biases could account for some the differences among rates for white, black, and Hispanic adolescents. However, if gonorrhea has been underreported for any racial group, the high rates of gonorrhea among 15- to 19-year-olds represented an underestimate of the true infection rate. Increases in sexual activity among adolescents and a lack of clinical services in settings convenient to adolescents could have contributed to the increasing rates of gonorrhea and syphilis among these young persons during this time period. If gonorrhea and other sexually transmitted infections are cofactors for facilitating the transmission of human immunodeficiency virus (HIV), the high incidence of gonorrhea in some locales among some populations of adolescents could result in dramatic increases in HIV acquisition, a situation that demands attention from public health organizations.


Safety and stable survival of stem-cell-derived retinal organoid for 2 years in patients with retinitis pigmentosa.
Transplantation of induced pluripotent stem cell (iPSC)-derived retinal organoids into retinal disease animal models has yielded promising results, and several clinical trials on iPSC-derived retinal pigment epithelial cell transplantation have confirmed its safety. In this study, we performed allogeneic iPSC-derived retinal organoid sheet transplantation in two subjects with advanced retinitis pigmentosa (jRCTa050200027). The primary endpoint was the survival and safety of the transplanted retinal organoid sheets in the first year post-transplantation. The secondary endpoints were the safety of the transplantation procedure and visual function evaluation. The grafts survived in a stable condition for 2 years, and the retinal thickness increased at the transplant site without serious adverse events in both subjects. Changes in visual function were less progressive than those of the untreated eye during the follow-up. Allogeneic iPSC-derived retinal organoid sheet transplantation is a potential therapeutic approach, and the treatment's safety and efficacy for visual function should be investigated further.


Lingonberry (Vaccinium vitis-idaea) and European cranberry (Vaccinium microcarpon) proanthocyanidins: isolation, identification, and bioactivities.
European, small-fruited cranberries (Vaccinium microcarpon) and lingonberries (Vaccinium vitis-idaea) were characterized for their phenolic compounds and tested for antioxidant, antimicrobial, antiadhesive, and antiinflammatory effects. The main phenolic compounds in both lingonberries and cranberries were proanthocyanidins comprising 63-71% of the total phenolic content, but anthocyanins, hydroxycinnamic acids, hydroxybenzoic acids, and flavonols were also found. Proanthocyanidins are polymeric phenolic compounds consisting mainly of catechin, epicatechin, gallocatechin, and epigallocatechin units. In the present study, proanthocyanidins were divided into three groups: dimers and trimers, oligomers (mDP 4-10), and polymers (mDP > 10). Catechin, epicatechin, A-type dimers and trimers were found to be the terminal units of isolated proanthocyanidin fractions. Inhibitions of lipid oxidation in liposomes were over 70% and in emulsions over 85%, and in most cases the oligomeric or polymeric fraction was the most effective. Polymeric proanthocyanidin extracts of lingonberries and cranberries were strongly antimicrobial against Staphylococcus aureus, whereas they had no effect on other bacterial strains such as Salmonella enterica sv. Typhimurium, Lactobacillus rhamnosus and Escherichia coli. Polymeric fraction of cranberries and oligomeric fractions of both lingonberries and cranberries showed an inhibitory effect on hemagglutination of E. coli, which expresses the M hemagglutin. Cranberry phenolic extract inhibited LPS-induced NO production in a dose-dependent manner, but it had no major effect on iNOS of COX-2 expression. At a concentration of 100 μg/mL cranberry phenolic extract inhibited LPS-induced IL-6, IL-1β and TNF-α production. Lingonberry phenolics had no significant effect on IL-1β production but inhibited IL-6 and TNF-α production at a concentration of 100 μg/mL similarly to cranberry phenolic extract. In conclusion the phenolics, notably proanthocyanidins (oligomers and polymers), in both lingonberries and cranberries exert multiple bioactivities that may be exploited in food development.


Multiple domains of MASP-2, an initiating complement protease, are required for interaction with its substrate C4.
The complement system is fundamental to both innate and adaptive immunity and can be initiated via the classical, lectin or alternative pathways. Cleavage of C4 by MASP-2, the initiating protease of the lectin pathway, is a crucial event in the activation of this pathway, preceding the eventual formation of the C3 convertase (C4bC2a) complex on the pathogen surface. Interactions required for the cleavage of C4 by MASP-2 are likely to be facilitated by the initial binding of C4 to an exosite on the protease. We have shown that both proteolytically active and catalytically inactive CCP1-CCP2-serine protease (CCP1-CCP2-SP) forms bind C4 with similar affinity. Interestingly, proteins containing the CCP1-CCP2 domains or the SP domain alone bound C4 with much lower affinity than the CCP1-CCP2-SP protein, suggesting that the CCP domains cooperate positively with the active site to mediate efficient binding and cleavage of C4. In addition, mutation of residue K342 to alanine in the CCP1 domain abolished binding to both C4 and C4b in its CCP1-CCP2 form, suggesting a key electrostatic role for this amino acid. The presented data indicates that all of the domains are required in order to mediate high affinity interaction with C4.


Health insurance in India: need for managed care expertise.
Health insurers in India currently face many challenges, including poor consumer awareness, strict regulations, and inefficient business practices. They operate under a combination of stifling administrative costs and high medical expense ratios which have ensured that insurers operate under steep losses. External factors (eg, onerous regulations, lack of standards, high claims payouts) and internal factors (eg, high administrative costs, dependence on indemnity models that cover inpatient treatment costs only) have forced the health insurance industry into a regressive spiral. To overcome these challenges, health insurers need to innovate in their product offerings and tighten their existing processes and cost structures. But as a long-term strategy, it is imperative that health insurers deploy managed care concepts, which will go a long way toward addressing the systemic issues in the current operational models of health plans.


Cell cycle control and cancer.
This review consists of two parts. In the first part normal mechanisms regulating the progression of cells through the cell cycle are briefly reviewed. Besides mitogenic stimulation, cyclin kinase inhibition, the G1 restriction point and the prb pathway, accuracy of DNA replication and DNA repair, the G2 to M transition, apoptosis and the p 53 pathway, proteolytic, in particular ubiquitin-dependent mechanisms involved in the initiation of DNA synthesis in the separation of sister chromatids and in the telophase to GO/G1 transition, are discussed. In the second part oncogene and tumor suppressor gene products are briefly characterized. Aberrations of cell cycle control mechanisms associated with cancer are grouped as follows: deregulation of protooncogenes by translocations juxtaposing protooncogenes to immunoglobulin--or T cell receptor genes; translocations producing chimeric proteins unique to cancer cells; inversions and amplifications resulting in over expression of regulator genes; and deletions and mutations of tumor suppressor genes. It is emphasized that cancer is the result of a multistep process and that uncontrolled cell production and other alterations are, as a rule, late phenomena.


Diagnosing disruptive mood dysregulation disorder: Integrating semi-structured and unstructured interviews.
The newest iteration of the Diagnostic and Statistical Manual-fifth edition (DSM-5), is the first to include the diagnosis of disruptive mood dysregulation disorder (DMDD). The assessment and diagnosis of psychopathology in children are complicated, particularly for mood disorders. Practice can be guided by the use of well-validated instruments. However, as this is a new diagnosis existing instruments have not yet been evaluated for the diagnosis of DMDD. This study seeks to provide a method for using existing structured interview instruments to assess for this contemporary diagnosis. The Children's Interview for Psychiatric Syndromes (ChIPS) and the Mini-International Neuropsychiatric Interview for Children and Adolescents (MINI-KID) are reviewed and existing items consistent with a diagnosis of DMDD are identified. Finally, a case is presented using both measures and applying the theoretical items identified to illustrate how one might use these measures to assess DMDD. Limitations and future directions are discussed.


Languages evolve in punctuational bursts.
Linguists speculate that human languages often evolve in rapid or punctuational bursts, sometimes associated with their emergence from other languages, but this phenomenon has never been demonstrated. We used vocabulary data from three of the world's major language groups-Bantu, Indo-European, and Austronesian-to show that 10 to 33% of the overall vocabulary differences among these languages arose from rapid bursts of change associated with language-splitting events. Our findings identify a general tendency for increased rates of linguistic evolution in fledgling languages, perhaps arising from a linguistic founder effect or a desire to establish a distinct social identity.


[Poisoning with spotted and red mushrooms--pathogenesis, symptoms, treatment].
Amanita pantherina and Amanita muscaria are commonly occurring mushrooms in Polish forests. They contain ibotenic acid and muscimol: the substances reacting with neurotransmitter receptors in central nervous system. The ingestion of these mushrooms produces a distinctive syndrome consisting of alternating phases of drowsiness and agitation with hallucinations, and sometimes with convulsions. The diagnosis of Amanita pantherina or Amanita muscaria poisoning is established by means of mycologic investigation of gastric lavage. The treatment is only symptomatic, and the prognosis is usually good.


The comparative biology of pulmonary intravascular macrophages.
Pulmonary intravascular macrophages are an important part of the mononuclear phagocyte system in some species of mammals, mainly sheep and other ruminants, pigs, and horses. These cells phagocytize foreign particles, cell debris and pathogens that pass through the pulmonary circulation. Species with intravascular macrophages localize intravenously injected tracer particles and bacteria predominantly in the lung rather than the liver, and exhibit pulmonary hypertension when these cells are activated. Both in vivo and in vitro studies show that pulmonary intravascular macrophages have distinct secretory and immune capabilities. Consequently, the pulmonary intravascular macrophages play an important role in pulmonary inflammation in species that have them


[Malfunctions of pacemakers].
No pacemaker is absolutely reliable : more or less threatening failures can be observed after implantation. Pacemaker runaway has completely disappeared. Sudden battery failure, intermittent or permanent, remains too frequent. Other incidents, less frequent or severe, are also reviewed. Failures are often isolated and sporadic, but can sometimes involve a complete manufacture series. It is a medical duty to follow-up carefully the patients and to announce the failures with a view to permit early recognition of imperfect series. Pacing failures do not play a major role in the mortality in the group of patients with permanently implanted pacemakers. Nevertheless, anticipated replacement of pacemakers increases the cost of the technique.


Drug discovery and regulatory considerations for improving in silico and in vitro predictions that use Caco-2 as a surrogate for human intestinal permeability measurements.
There is a growing need for highly accurate in silico and in vitro predictive models to facilitate drug discovery and development. Results from in vitro permeation studies across the Caco-2 cell monolayer are commonly used for drug permeability screening in industry and are also accepted as a surrogate for human intestinal permeability measurements by the US FDA to support new drug applications. Countless studies carried out in this cell line with published permeability measurements have enabled the development of many in silico prediction models. We identify several common cases that illustrate how using Caco-2 permeability measurements in these in silico and in vitro predictive models will not correlate with human intestinal permeability and will further lead to inaccuracies in these models. We provide guidelines and recommendations for improving these models to more accurately predict clinically relevant information, thereby enhancing the drug discovery, development, and regulatory approval processes.


The circular dichroism of ribosomal ribonucleic acids.
1. The c.d. (circular dichroism) of Drosophila melanogaster rRNA (42% G+C) and of G+C-rich fragments (78% G+C) obtained by partial hydrolysis of rabbit L-rRNA (the largest RNA species isolated from the large subribosomal particle) were measured and found to differ substantially. 2. To interpret these spectra a relation between c.d. of bihelical RNA and % G+C was derived, namely delta epsilonfG = AFG2+bfG+c, where deltaepsilonfG is the c.d. of RNA characterized by a mole fraction, fG, of guanine nucleotides and a, b and c are constants. 3. A frame of reference was established by studying the c.d. of a range of rRNA species, including S-rRNA (the RNA species isolated from the smaller subribosomal particle) and L-rRNA of Escherichia coli. 4. It was found for the rRNA species studied that 0.60+/-0.05 of residues appear to form bihelical secondary structure. 5. A higher helical content, 0.66+/-0.05, was found for the G+C-rich fragment of L-rRNA. The difference in the c.d. of rabbit L-rRNA and of D. melanogaster rRNA is attributable to the dependence of c.d. of the bihelical parts on %G+C. 6. The minimum in c.d. at 295 nm increases with increasing %G+C. The c.d. of rRNA was compared with that of the parent subparticle in this region of the spectrum, where high precision may be attained.


Vesting of private pension benefits in 1979 and change from 1972.
This article examines the prevalence of vested private pension benefits in 1979 as reported by full-time private wage and salary workers actively participating in pension plans. It also analyzes the effect of selected demographic, economic, labor-force, and plan characteristics on vested status. Years of participation in a plan was by far the most important predictor of vested status. Age was positively related to vested status even after accounting for the longer plan participation of older workers. Evidence pertaining to occupation, size of firm, and type of employer plan suggests that multi-employer plans and those covering professional and managerial workers and the employees of the smallest firms have more rapid vesting schedules than plans covering other workers. Sex, race, union representation, and earnings had little meaningful effect on vested status. This article also examines change in the prevalence of vesting after passage of the Employee Retirement Income Security Act (ERISA) by comparing reported vested rates for 1972 and 1979. The proportion of plan participants reporting vested benefits rose substantially during this period, a finding suggestive of ERISA's substantial impact. The magnitude of the change is examined by years of service with employer, age, sex, earnings, occupation, and industry.


Therapeutic effect of transplantation of human bone marrow‑derived mesenchymal stem cells on neuron regeneration in a rat model of middle cerebral artery occlusion.
Human bone marrow‑derived mesenchymal stromal cells (hBMSCs) have been revealed to be beneficial for the regeneration of tissues and cells in several diseases. The present study aimed to elucidate the mechanisms underlying the effect of hBMSC transplantation on neuron regeneration in a rat model of middle cerebral artery occlusion (MCAO). The hBMSCs were isolated, cultured and identified. A rat model of MCAO was induced via the modified Longa method. Neurological severity scores (NSS) were adopted for the evaluation of neuronal function in the model rats after cell transplantation. Next, the expression levels of nestin, β‑III‑tubulin (β‑III‑Tub), glial fibrillary acidic protein (GFAP), HNA and neuronal nuclear antigen (NeuN) were examined, as well as the positive expression rates of human neutrophil alloantigen (HNA), nestin, NeuN, β‑III‑Tub and GFAP. The NSS, as well as the mRNA and protein expression of nestin, decreased at the 1st, 2nd, 4 and 8th weeks, while the mRNA and protein expression of NeuN, β‑III‑Tub and GFAP increased with time. In addition, after treatment, the MCAO rats showed decreased NSS and mRNA and protein expression of nestin, but elevated mRNA and protein expression of NeuN, β‑III‑Tub and GFAP at the 2nd, 4 and 8th weeks, and decreased positive expression of HNA and nestin with enhanced expression of NeuN, β‑III‑Tub and GFAP. Therefore, the present findings demonstrated that hBMSC transplantation triggered the formation of nerve cells and enhanced neuronal function in a rat model of MCAO.


Mitochondrial network morphology: building an integrative, geometrical view.
The morphology of mitochondrial networks is complex and highly varied, yet vital to cell function. The first step toward an integrative understanding of how mitochondrial morphology is generated and regulated is to define the interdependent geometrical features and their dynamics that together generate the morphology of a mitochondrial network within a cell. Distinct aspects of the size, shape, position, and dynamics of mitochondrial networks are described and examples of how these features depend on one another discussed.


Rheumatic fever and streptococci: the Wairoa College study.
The decline in incidence of rheumatic fever seen in many countries is not evident in New Zealand. Wairoa county has the highest hospital admission rate within New Zealand, 232 per 100 000 per year, five times the national average, 43 per 100 000. Studies in Wairoa included a cross-sectional survey of the local secondary school. Throat swabbing yielded 13.6% of subjects positive for beta-haemolytic streptococcus group A. Boys had a higher rate than girls, and girls aged 14 and over had a lower rate than younger girls. Discriminant analysis failed to identify ethnic, social or geographic factors as contributing to positivity. Sibling concordance was not significant. ASO titres were high in comparison with another New Zealand study (Rotorua Lakes). There was an association between high ASO titre and throat swab positive for group A and group C streptococci. The prevalence of definite rheumatic fever and rheumatic heart disease was established as 11/715 (1.5%). A wide range of residential geographic, climatic and socioeconomic factors failed to discriminate between cases and noncases. Ethnicity alone was a significant discriminator, the cases being more Maori than noncases.


Limiting loss to follow-up in a multicenter randomized trial in orthopedic surgery.
Even the best-designed, randomized controlled trials suffer when patients are lost to follow-up. Incomplete follow-up biases the results of a trial when patients who drop out are different from those who complete follow-up. This is exaggerated further when there are differential dropout rates between study groups. Previous randomized controlled trials in orthopedic trauma have reported up to 28% loss to follow-up. Only by striving to achieve a 0% loss to follow-up rate can we be certain that this type of bias does not affect our results. In our ongoing multicenter, randomized controlled trial comparing reamed and nonreamed intramedullary nailing of tibial shaft fractures, we have implemented several innovative strategies to minimize loss to follow-up. The exclusion criteria and consent process are designed to minimize losses. Study staff are carefully trained in communication and negotiation with patients. Additionally, a central methods center monitors all patient follow-up and aids in finding lost patients. Through these primary, secondary, and tertiary interventions, we have achieved 94% complete 1-year follow-up for the first 440 patients enrolled in the trial. Eleven patients withdrew consent, and we are unable to locate 17 patients. We have successfully minimized the loss to follow-up rate in our trial by incorporating innovative prevention and retention strategies into its design and conduct. Through planning, organization, and committing time and resources to minimizing loss to follow-up, other orthopedic trauma trials can hope to achieve the same high rates of follow-up.


The Importance of Religiosity to the Well-Being of African American Older Adults Living With Dementia.
This study was a post hoc analysis of a larger qualitative descriptive study exploring family involvement in health promotion activities for African Americans living with dementia where participants identified religious practices as meaningful health promotion activities. The purpose of this study was to explore ways in which religiosity may influence the well-being of older adults living with dementia. Semi-structured interviews were conducted among a sample of 22 family caregivers and 15 older adults living with dementia (N = 37). Three themes emerged: Engagement, Promotion of Faith and Spiritual Connectedness, and Maintenance of Religious Practices. It is imperative for family caregivers to understand the important contributions of religious activities and beliefs to the well-being of their family member. This information might be of use for faith communities, policy makers, and health care providers in the provision of optimal person-centered care and the promotion of quality of life for persons living with dementia.


Sperm aggregation by water extracts from two Bursera species.
Some plants have more than the common utility value, as is the case of some members of the Bursera species such as the Mexican copal, a plant used for worship. Water extracts of several plants have vaginal contraceptive properties. The authors evaluated the sperm agglutinating activity of two Bursera species on human and boar sperm. Extracts from stems and leaves were obtained. Capacitated sperm samples were used in all cases. There were different agglutinating capacities, which were not observed in the vehicle-only samples. The most frequent sperm agglutination response was that involving the heads. Agglutinating activity was higher from stem- than leaf-derived extracts. The results indicate that proteins present in the extracts are responsible for the aggregation of sperm heads.


IL-13 R130Q, a common variant associated with allergy and asthma, enhances effector mechanisms essential for human allergic inflammation.
Genetic factors are known to strongly influence susceptibility to allergic inflammation. The Th2 cytokine IL-13 is a central mediator of allergy and asthma, and common single-nucleotide polymorphisms in IL13 are associated with allergic phenotypes in several ethnically diverse populations. In particular, IL13+2044GA is expected to result in the nonconservative replacement of arginine 130 (R130) with glutamine (Q). We examined the impact of IL13+2044GA on the functional properties of IL-13 by directly comparing the activity of WT IL-13 and IL-13 R130Q on primary human cells involved in the effector mechanisms of allergic inflammation. Our results show that IL-13 R130Q was significantly more active than WT IL-13 in inducing STAT6 phosphorylation and CD23 expression in monocytes and hydrocortisone-dependent IgE switching in B cells. Notably, IL-13 R130Q was neutralized less effectively than WT IL-13 by an IL-13R2 decoy. Decreased neutralization of the minor variant could contribute to its enhanced in vivo activity. Neither IL-13 variant was able to engage T cells, which suggests that increased allergic inflammation in carriers of IL13+2044A depends on enhanced IL-13-mediated Th2 effector functions rather than increased Th2 differentiation. Collectively, our data indicate that natural variation in the coding region of IL13 may be an important genetic determinant of susceptibility to allergy.


Trajectories of musculoskeletal pain from adolescence to middle age: the role of early depressive symptoms, a 27-year follow-up of the Northern Swedish Cohort.
Depression and musculoskeletal pain are associated, but long-term follow-up studies are rare. We aimed to examine the relationship of early depressive symptoms with developmental patterns of musculoskeletal pain from adolescence to middle age. Adolescents ending compulsory school (age 16) in Luleå, Northern Sweden, in 1981 (n = 1083) were studied and followed up in 1986, 1995, and 2008 (age 43) for musculoskeletal pain. Attrition was very low. Indicators for any and severe pain were based on pain in the neck-shoulders, low back, and the extremities. Latent class growth analyses were performed on 563 men and 503 women. Associations of a depressive symptoms score (DSS, range 0.0-2.0) at age 16 with pain trajectory membership were assessed by logistic and multinomial regression, adjusting for parental socioeconomic status, social adversities, smoking, exercise, body mass index, and alcohol consumption at age 16. For any pain, 3 trajectories emerged: high-stable (women 71%, men 61%), moderate (11%, 17%), and low-increasing (18%, 22%). With the low-increasing trajectory as reference, for each 0.1-point increase in the DSS, the odds ratio of belonging to the high-stable trajectory was 1.25 (95% confidence interval 1.11-1.41) in women and 1.23 (1.10-1.37) in men. For severe pain, 2 trajectories were found: moderate-increasing (women 19%, men 9%) and low-stable. For each 0.1-point increase in the DSS, the odds ratio of membership in the moderate-increasing trajectory was 1.14 (1.04-1.25) in women and 1.17 (1.04-1.31) in men in the fully adjusted model. Thus, depressive symptoms at baseline are strongly associated with pain trajectory membership.


[Substituted benzimidazoles--a new dimension in ulcer therapy?].
Substituted benzimidazoles represent a new class of gastric secretory inhibitors, which suppress acid secretion via direct inhibition of the parietal cell H+/K+-ATPase. The most potent derivative so far is omeprazole, which inhibits basal acid secretion and prevents the stimulatory effect of all classical acid secretory stimulants. In man, a single dose of 80 mg leads to almost complete achlorhydria over 24 hours. The results of the first duodenal ulcer trial with omeprazole are encouraging. No definite side effects have been attributed to the drug so far. Omeprazole may possibly initiate a new era in the medical treatment of peptic ulcer disease.


Inhibition of mitochondrial calcium uptake slows down relaxation in mitochondria-rich skeletal muscles.
Isolated fibres from various muscles were skinned mechanically in oil. From a Ca2+-loaded micropipette, local applications of Ca2+ were made. These produced a limited contraction which relaxed spontaneously. The time-course of sarcomere shortening and re-lengthening was recorded by microcinephotography. Application of Ruthenium Red, a potent and specific inhibitor of Ca2+ uptake by mitochondria, did not affect the contraction-relaxation cycles of typical glycolytic white fibres (frog sartorius, pigeon breast). By contrast, Ruthenium Red greatly slowed down the relaxation rate in mitochondria-rich fibres (rat soleus and rabbit masseter). In these fibres, Ca2+ uptake by mitochondria seems to play an active role in promoting relaxation.


The role of pseudolysogeny in bacteriophage-host interactions in a natural freshwater environment.
Bacteriophages occur in high numbers in environmental ecosystems and are thus significant mediators of microbial survival and activities. However, interactions between microbial populations and phages in situ have been largely ignored. Current understanding of the process relies on studies performed with well-fed, laboratory-grown host bacteria. The purpose of the experiments reported here was to determine bacteriophage-host interactions under environmentally relevant conditions of nutrient limitation. These studies have revealed the importance of a phenomenon called pseudolysogeny in the maintenance of viral genetic material for extended periods of time in natural ecosystems. Pseudolysogeny is a form of phage-host cell interaction in which the nucleic acid of the phage resides within its starved host in an unstable, inactive state. It is hypothesized that pseudolysogeny occurs due to the cell's highly starved condition. In such cells, there is insufficient energy available for the phage to initiate genetic expression leading to either a true temperate response or to the lytic response. However, upon nutrient addition, the pseudolysogenic state is resolved, resulting in either the establishment of true lysogeny or the initiation of the lytic production of progeny virions. The pseudolysogenic state may explain the long-term survival of viruses in unfavourable environments in which the infective half-life of their virions is relatively short.


Prostate specific antigen based biennial screening is sufficient to detect almost all prostate cancers while still curable.
We evaluated whether biennial screening with prostate specific antigen (PSA) only is sufficient to detect prostate cancer while still curable. In Göteborg, Sweden 9,972 men 50 to 65 years old were randomized to PSA screening. During 1995 and 1996 these men were invited for a first PSA screening and invited during 1997 and 1998 for a second screening. The screening procedure included PSA measurement in all men and in those with a PSA of 3 ng./ml. or greater also it included digital rectal examination, transrectal ultrasound and sextant biopsies. In the first screening 5,854 men participated and 145 cancers were detected. In the second screening 5,267 men participated and 111 cancers were detected. Only 9 interval cancers were diagnosed. In the second screening 102 cancers (92%) were associated with PSA less than 10 ng./ml. Of 465 men with increased PSA and who underwent biopsy with a benign outcome in the first screening 50 had cancer at the second screening. Of 241 men in whom PSA increased between screenings 1 and 2 cancer was detected in 46. None of the 2,950 men with an initial PSA of less than 1 ng./ml. had a PSA of greater than 3 ng./ml. or interval cancer. In men with a PSA of less than 2 ng./ml. it seems safe to offer repeat screening after 2 years with PSA only. Men with a PSA of 2 to 3 ng./ml. or a value of greater than 3 ng./ml. with negative biopsy may be better served by a shorter screening interval. Thus, different screening intervals are implied depending on baseline PSA.


Gray Matter Matters: A Longitudinal Magnetic Resonance Voxel-Based Morphometry Study of Primary Progressive Multiple Sclerosis.
Background: Multiple Sclerosis (MS) lesions in white matter (WM) are easily detected with conventional MRI which induce inflammation thereby generating contrast. WM lesions do not consistently explain the extent of clinical disability, cognitive impairment, or the source of an exacerbation. Gray matter (GM) structures including the cerebral cortex and various deep nuclei are known to be affected early in Primary Progressive Multiple Sclerosis (PPMS) and drive disease progression, disability, fatigue, and cognitive dysfunction. However, little is known about how rapidly GM lesions develop and accumulate over time. Objective: The purpose of this study is to analyze the degree and rate of progression in 25 patients with PPMS using voxel-based automated volumetric quantitation. Methods: This is a retrospective single-center study which includes a cohort of 25 patients with PPMS scanned utilizing NeuroQuant® 3 dimensional voxel-based morphometry (3D VBM) automated analysis and database and restudied after a period of ~1 year (11-14 months). Comparisons with normative data were acquired for whole brain, forebrain parenchyma, cortical GM, hippocampus, thalamus, superior and inferior lateral ventricles. GM volume changes were correlated with their clinical motor and cognitive scores using Extended Disability Status Scales (EDSS) and Montreal Cognitive Assessments (MoCA). Results: Steep reductions occurred in cerebral cortical GM and deep GM nuclei volumes which correlated with each patient's clinical and cognitive impairment. The median observed percentile volume losses were statistically significant compared with the 50th percentile for each GM component. Longitudinal assessments of an unselected sample of one dozen patients involved in the PPMS study showed prominent losses occurring mainly in cortical GM and hippocampus which were reflected in their EDSS and MoCA. The longitudinal results were compared with a similar sample of patients having Relapsing MS (RMS) whose GM values were largely in normal range, annualized volume GM changes were much less, while WM hyperintensities were in abnormal range in half the unselected cases. Conclusions: Knowledge of the degree and rapidity with which cortical atrophy and deep GM volume loss develops clarifies the source of progressive cognitive and clinical decline in PPMS.


Fear learning induces α7-nicotinic acetylcholine receptor-mediated astrocytic responsiveness that is required for memory persistence.
Memory persistence is a fundamental cognitive process for guiding behaviors and is considered to rely mostly on neuronal and synaptic plasticity. Whether and how astrocytes contribute to memory persistence is largely unknown. Here, by using two-photon Ca2+ imaging in head-fixed mice and fiber photometry in freely moving mice, we show that aversive sensory stimulation activates α7-nicotinic acetylcholine receptors (nAChRs) in a subpopulation of astrocytes in the auditory cortex. We demonstrate that fear learning causes the de novo induction of sound-evoked Ca2+ transients in these astrocytes. The astrocytic responsiveness persisted over days along with fear memory and disappeared in animals that underwent extinction of learned freezing behavior. Conditional genetic deletion of α7-nAChRs in astrocytes significantly impaired fear memory persistence. We conclude that learning-acquired, α7-nAChR-dependent astrocytic responsiveness is an integral part of the cellular substrate underlying memory persistence.


The Role of Biomarkers in Clinical Trials of Inflammatory Bowel Disease.
Clinical trials require valid outcome measures to assess the therapeutic benefit of investigational agents. Recently, regulatory authorities have mandated the use of patient-reported outcomes in combination with an objective measure of disease activity as primary outcome measures in inflammatory bowel disease trials. Endoscopy has commonly fulfilled the latter role; however, due to the costs and complexity of these assessments, interest has emerged in the use of noninvasive biomarkers. The role of C-reactive protein, fecal calprotectin, and fecal lactoferrin in clinical research is discussed.