Validation of the Pictorial Fit-Frail Scale in a memory clinic setting.
To assess the feasibility, reliability, and validity of the Pictorial Fit-Frail Scale (PFFS) among patients, caregivers, nurses, and geriatricians in an outpatient memory clinic. Observational study. A Canadian referral-based outpatient memory clinic. Fifty-one consecutive patients and/or their caregivers, as well as attending nurses and geriatricians. Participants (patients, caregivers, nurses, and geriatricians) were asked to complete the PFFS based on the patient's current level of functioning. Time-to-complete and level of assistance required was recorded. Participants also completed a demographic survey and patients' medical history (including the Mini-Mental State Examination [MMSE], and Comprehensive Geriatric Assessment [CGA]) was obtained via chart review. Patient participants had a mean age of 77.3±10.1 years, and average MMSE of 22.0±7.0, and 53% were female. Participants were able to complete the PFFS with minimal assistance, and their average times to completion were 4:38±2:09, 3:11±1:16, 1:05±0:19, and 0:57±0:30 (mins:sec) for patients, caregivers, nurses, and geriatricians, respectively. Mean PFFS scores as rated by patients, caregivers, nurses, and geriatricians were 9.0±5.7, 13.1±6.6, 11.2±4.5, 11.9±5.9, respectively. Patients with low MMSE scores (0-24) took significantly longer to complete the scale and had higher PFFS scores. Inter-rater reliability between nurses and geriatricians was 0.74, but it was lower when assessments were done for patients with low MMSE scores (0.47, p<0.05). The correlation between PFFS and a Frailty Index based on the CGA was moderately high and statistically significant for caregivers, nurses, and geriatricians (r=0.66, r=0.59, r=0.64, respectively), but not patients. The PFFS is feasible, even among people with some slight cognitive impairment, though it may be less useful when patients with severe dementia administer it to themselves. Further, the PFFS may help inform clinicians about areas of concern as identified by patients, enabling them to contribute more to diagnostic and treatment decisions or helping with health tracking and care planning.


Absence of Tumor Necrosis Factor Receptor 1 Inhibits Osteoclast Activity in Apical Dental Resorption Caused by Endodontic Infection in Mice.
The aim of this study was to evaluate osteoclastogenesis and dental resorption resulting from endodontic infection in wild-type (WT) and tumor necrosis factor receptor 1 genetically deficient (TNFR1 KO) mice. After approval by the ethics committee on the use of animals, 40 mice were distributed into 2 experimental groups based on time periods: 14 days (n = 10 WT mice and n = 10 TNFR1 KO mice) and 42 days (n = 10 WT mice and n = 10 TNFR1 KO mice). After these periods, morphometric analysis was performed using bright field and fluorescence microscopy and tartrate-resistant acid phosphatase histoenzymology to identify osteoclasts. One-way analysis of variance followed by the Tukey post hoc test was used for the statistical analysis (α = 0.05). WT mice in the 42-day period had a greater apical dental resorption in the distal root of the first molar than TNFR1 KO mice (P < .05). On the other hand, TNFR1 KO mice showed a smaller number of osteoclasts on the dental surface than WT mice (P < .05). WT mice with apical periodontitis had more extensive apical dental resorptions and a larger number of osteoclasts on the tooth surface than TNFR1 KO mice.


Modern psychophysical tests to assess olfactory function.
The sense of smell significantly contributes to quality of life. In recent years much progress has been made in understanding the biochemistry, physiology and pathology of the human olfactory system. Olfactory disorders may arise not only from upper airway phlogosis but also from neurodegenerative disease. Hyposmia may precede motor signs in Parkinson's disease and cognitive deficit in Alzheimer's disease. These findings suggest the complementary role of olfactory tests in the diagnosis and management of neurodegenerative diseases. In this report we present a review of modern olfactory tests and their clinical applications. Although rarely employed in routine clinical practice, the olfactory test evaluates the ability of odour identification and is a useful diagnostic tool for olfaction evaluation. Olfactory screening tests are also available. In this work we strongly recommend the importance of an ENT evaluation before the test administration and dissuade from a self-administration of an olfactory test.


Epigenome-wide association study of plasma lipids in West Africans: the RODAM study.
DNA-methylation has been associated with plasma lipid concentration in populations of diverse ethnic backgrounds, but epigenome-wide association studies (EWAS) in West-Africans are lacking. The aim of this study was to identify DNA-methylation loci associated with plasma lipids in Ghanaians. We conducted an EWAS using Illumina 450k DNA-methylation array profiles of extracted DNA from 663 Ghanaian participants. Differentially methylated positions (DMPs) were examined for association with plasma total cholesterol (TC), LDL-cholesterol, HDL-cholesterol, and triglycerides concentrations using linear regression models adjusted for age, sex, body mass index, diabetes mellitus, and technical covariates. Findings were replicated in independent cohorts of different ethnicities. We identified one significantly associated DMP with triglycerides (cg19693031 annotated to TXNIP, regression coefficient beta -0.26, false discovery rate adjusted p-value 0.001), which replicated in-silico in South African Batswana, African American, and European populations. From the top five DMPs with the lowest nominal p-values, two additional DMPs for triglycerides (CPT1A, ABCG1), two DMPs for LDL-cholesterol (EPSTI1, cg13781819), and one for TC (TXNIP) replicated. With the exception of EPSTI1, these loci are involved in lipid transport/metabolism or are known GWAS-associated loci. The top 5 DMPs per lipid trait explained 9.5% in the variance of TC, 8.3% in LDL-cholesterol, 6.1% in HDL-cholesterol, and 11.0% in triglycerides. The top DMPs identified in this study are in loci that play a role in lipid metabolism across populations, including West-Africans. Future studies including larger sample size, longitudinal study design and translational research is needed to increase our understanding on the epigenetic regulation of lipid metabolism among West-African populations. European Commission under the Framework Programme (grant number: 278901).


Nurses' Finnegan Scoring of Newborns with Neonatal Abstinence Syndrome Not Affected by Time or Day of the Week.
Newborns exposed to drugs in utero are at risk of developing neonatal abstinence syndrome (NAS), characterized by behavioral changes and physiologic instability. Finnegan scoring tool quantifies severity of symptoms and guides treatment. This article evaluates whether time of day and the number of shift hours affects modified Finnegan scores, and the subjective component of these scores. Institutional review board-approved, retrospective chart review of newborns admitted to neonatal intensive care or transitional nursery from 2011 to 2014. > 35 weeks' gestation, known maternal substance use, positive maternal or newborn urine, or meconium drug screen, NAS treatment. A total of 101 charts were evaluated. Mean treatment duration was 31.8 days (standard deviation ±18.3). There was no significant relationship between observer shift hour and high scores (> 8) (p = 0.83). Highest scores occurred in the afternoon, decreased at night (p = 0.03), and throughout admission (p < 0.0001). Weekend and weekday scores were similar (p = 0.4). The objective component of the scores remained similar throughout the day (p = 0.91) and week (p = 0.52). Finnegan scores given by nurses were not influenced by shift hour. Time of day did not influence overall high scores or the proportion of objective to total Finnegan score. Inter-rater reliability was maintained regardless of time of day or day of the week.


Decolorization and biodegradation of melanoidin contained in beet molasses by an anamorphic strain of Bjerkandera adusta CCBAS930 and its mutants.
We used a ligninolytic strain of the white-rot fungus B. adusta CCBAS 930 and its mutants with modified ligninolytic activity to assess their potential to remove of molasses. The analyzed strains have been shown to be able to decolorize 1% or 2% molasses solutions containing brown-colored toxic melanoidins. It was found that the decolorization process was determined by the transition to the stage of production of sporulating aerial mycelium (liquid and agar cultures) coupled with an increase in peroxidase activity, which was accompanied by a decrease in the level of melanoidin, free radicals, and phenolic compounds. Four different peroxidase activities were detected in post-culture liquids, i.e. horseradish-like (HRP-like), manganese-dependent (MnP), lignin (LiP), and versatile (VP) peroxidase activities. The HRP-like peroxidase was characterized by the highest activity. The efficiency of removal of melanoidins from a 1% molasses solution by the parental strain and the mutants was dependent on the culture method. The highest efficiency was noted in immobilized cultures (threefold higher than in the mycelium-free cultures), which was accompanied by stimulation of HRP-like peroxidase activity. Mutant 930-5 was found to be the most effective in the decolorization and decomposition of melanoidin. The HRP-like activity in the immobilized cultures of B. adusta 930-5 was 640-fold higher than in the mycelium-free cultures of the fungus. Moreover, decolorization and biodegradation of melanoidin by B. adusta CCBAS 930 and 930-5 was coupled with detoxification.


Erratum: Changes in B-type Natriuretic Peptide and Related Hemodynamic Parameters following a Fluid Challenge in Critically ill Patients with Severe Sepsis or Septic Shock.
[This corrects the article on p. 117 in vol. 21, PMID: 28400680.].


A novel approach to diagnose reversible cerebral vasoconstriction syndrome: a case series.
Reversible cerebral vasoconstriction syndrome (RCVS) is classically diagnosed based on the presence of severe thunderclap headache, focal neurologic symptoms, and the radiographic findings of reversible diffuse segmental cerebral vasoconstriction. We present a diagnostic test that may assist in the clinical diagnosis and facilitate treatment. From October 1, 2010, to August 1, 2013, we identified consecutive patients who presented with a presumptive diagnosis of RCVS and underwent cerebral diagnostic angiography with intra-arterial (IA) vasodilator therapy. Medical records including clinical presentation, radiographic, and angiographic images were all reviewed. We identified a total of 7 patients (4 females; age range, 22-56; mean, 45 years) who met our inclusion criteria. Four patients received a combination of milrinone and nicardipine infusion either in the internal carotid arteries or in the left vertebral artery; the remaining patients received IA therapy solely with either nicardipine or milrinone. Five patients had a positive angiographic response, defined as significant improvement or resolution of the blood vessels irregularities. All 5 patients had a definite discharge diagnosis of RCVS. The remaining 2 patients had a negative angiographic response and based on their clinical and radiographic course had a final diagnosis of intracranial atherosclerotic disease. Our small case series suggest that IA administration of vasodilators is safe and may aid in distinguishing vasodilator responsive syndromes such as RCVS from other causes. Further study is required with long-term clinical outcome to determine the utility of this diagnostic test.


The suicidal desire of Tolstoy.
To explore whether a healthy, successful individual may experience suicidal desires. Examination of "A Confession" by Leo Tolstoy. Confirmation that a physically and mentally healthy, well resourced individual may experience suicidal desires. To reduce suicide rates, a broader understanding of the factors which contribute to suicidal desires is required.


Development of microwave-assisted extraction procedure for organic impurity profiling of seized 3,4-methylenedioxymethamphetamine (MDMA).
Organic impurity profiling of seized 3,4-methylenedioxymethamphetamine (MDMA) tablets aims to link tablets to common production sources. Conventionally, organic impurities are extracted from tablets using a liquid-liquid extraction (LLE) procedure prior to analysis by gas chromatography-mass spectrometry (GC-MS). In this research, the development of an alternative microwave-assisted extraction/headspace solid-phase microextraction (MAE/HS-SPME) procedure is described. The optimal procedure used phosphate buffer (1 M, pH 8), with an HS-SPME extraction temperature of 70 °C for 40 min, using a divinylbenzene/Carboxen™/polydimethylsiloxane (DVB/CAR/PDMS) fiber. Impurities were extracted from seized MDMA exhibits using the MAE/HS-SPME procedure, as well as HS-SPME alone, and a conventional LLE procedure. The HS-SPME procedure was deemed to be the most practical because of the affordability and need for less analyst involvement. Although the LLE was limited in the number of impurities extracted, the procedure is still useful for the extraction of less volatile impurities that are not extracted by HS-SPME.


Primary Ciliary Dyskinesia: An Update on New Diagnostic Modalities and Review of the Literature.
Primary ciliary dyskinesia (PCD) is a genetic condition affecting approximately 1 in 15,000-20,000 individuals, and the majority of cases exhibit an autosomal recessive inheritance pattern. However, genetic heterogenicity is seen in PCD and reflects the complexity of ciliary structure and biogenesis. There have been many recent advances in the diagnosis and management of PCD in the last few years, including advanced genetic sequencing, nasal nitric oxide assay, and ciliary motility tests. This article focuses on the ultrastructure and pathophysiology of ciliary dyskinesias, along with a review of clinical features, screening, and diagnostic tests. It also reflects upon the diagnostic challenge caused by the diverse clinical presentation, which will be of great value to pediatricians for considering PCD in their differential list, henceforth leading to early recognition and management, along with awareness of the recent advances in the field of genetics and other techniques for diagnosis of this condition.


Randomized comparison between fluconazole and itraconazole for the treatment of candidemia in a pediatric intensive care unit: a preliminary study.
Candida bloodstream infections have shown an increase in hospitalized patients, especially those receiving intensive care. The effectiveness of various azoles, especially itraconazole, in treatment of candidemia has not been fully evaluated. Our objective was to compare the efficacy and safety of enterally administered itraconazole vs. fluconazole in treatment of candidemia. Randomized, double-blind, controlled trial. Pediatric intensive care unit of a referral and teaching hospital. Forty-three pediatric patients with candidemia, Patients received either fluconazole (n = 22) or itraconazole (n = 21), about 10 mg/kg orally or through a gastric tube, and were monitored for clinical and mycological cure (sterile fungal blood culture), blood counts, and liver and renal functions. The clinical characteristics of two groups were comparable. The cure rate was similar in both the groups: itraconazole 17 of 21 (81%) and fluconazole 18 of 22 (82%). Crude mortality rate (itraconazole 9.5% and fluconazole 13.6%) was also comparable in two groups of patients. The frequency of electrolyte disturbance was very low and similar in both the groups. Blood urea, creatinine, liver enzymes, and serum bilirubin were not adversely affected. Itraconazole was as effective as fluconazole in nosocomial candidiasis in children receiving intensive care and was devoid of serious side effects.


Bis[4-amino-N-(pyrimidin-2-yl)benzene-sulfonamidato]-κN,N';κN-aqua-bis-(dimethyl-formamide-κO)cadmium(II) monohydrate.
In the title compound, [Cd(C(10)H(9)N(4)O(2)S)(2)(C(3)H(7)NO)(2)(H(2)O)]·H(2)O, the Cd(II) ion displays a grossly distorted octa-hedral (or irregular) mer-CdN(3)O(3) coodination polyhdron arising from its coordination by one N,N'-bidentate sulfadiazinate anion, one monodentate sulfadiazinate anion, two O-bonded dimethyl-formamide molecules and one water mol-ecule. A short Cd⋯N contact [2.890 (3) Å] to the monodentate sulfadiazinate ion also occurs. The dihedral angles between the planes of the aromatic rings of the anions are 86.81 (14) and 68.65 (14)°. The crystal structure features inter-molecular N-H⋯O, O-H⋯O and O-H⋯N hydrogen bonds.


Maudsley reactive and non-reactive rat strains: differential performance in a conflict task.
Based on open field defecation rates, distinct strains of Maudsley rats have been identified. The Maudsley Reactive strains exhibit high levels of open field defecation, while the Maudsley Non-Reactive strains exhibit low levels of open field defecation. This difference in open field behavior has been interpreted as an indicator of "emotionality." The present studies were designed to compare the performance of two Maudsley strains, MR/Har and MNRA/Har, in the conditioned suppression of drinking (CSD) paradigm, another "model" behavior for the study of emotionality and/or anxiety in animals. In daily 10-minute sessions, water-deprived rats were trained to drink from a tube that was occasionally electrified (0.5 mA), electrification being signalled by a tone. In this procedure, MNRA/Har rats accepted significantly more shocks than did MR/Har rats. This difference between MR/Har and MNRA/Har strains, initially somewhat slight, became quite impressive over weeks of repeated testing. Although present in both male and female subjects, this strain difference was more prominent in the males. Thus, the MR/Har and MNRA/Har rat strains, bred originally for their differences in open field behavior, also differ markedly in their performance in the CSD paradigm. These findings suggest that the Maudsley rat strains may represent a useful genetically-based "animal model" for the study of emotionality and/or anxiety.


[Quantitative growth analysis of limbic nuclei areas fresh volume in diencephalon and mesencephalon of an albino mouse ontogenic series. I. Nucleus habenulare].
The fresh volumes of the Nucl. mediales et laterales habenulae of 52 male white mice aged between 17 and 60 days of ontogenesis have been determined. The data of both nuclei have been fitted by a sum of logistic functions. There is an increasing component and a decreasing component. The degree of maturity and its differential quotient have been discussed. The Nucl. medialis habenulae belongs to those structures of the central nervous system developing extremely early, the Nucl. lateralis habenulae belongs to those structures of the central nervous system developing extremely early, the Nucl. lateralis habenulae belongs to those structures developing early. It has been pointed out, that there is a complex heterochrony of regional development in the brain. There have to be more quantitative analyses.


Factors related to pregnancy status and unwanted pregnancy among lebanese women during the COVID-19 lockdown: a cross-sectional study.
Home confinement and lockdowns have created challenges and vulnerabilities, causing relevant changes in sexual health and couple stability, particularly in women. The objective of this study was to evaluate the socio-economic and psychological factors related to current pregnancy status and unwanted pregnancy among Lebanese women during the COVID-19 lockdown. A cross-sectional online study conducted between June 8 and August 1, 2020, enrolled 369 Lebanese women using the snowball technique for data collection. All married women between 18 and 51, with access to the internet and currently living with their partners, were eligible to participate. Current pregnancy status and unwanted pregnancy were assessed using binary questions. The SPSS software 25 was used for data analysis, and multivariable analysis was performed, taking the pregnancy status and unwanted pregnancy as the dependent variables. The statistical significance was set at a p-value < 0.05. Our results showed that 11.1% of women were pregnant, of whom 22.0% reported unwanted pregnancies. Having children (ORa = 0.183) and taking contraceptives (ORa = 0.231) were significantly associated with a reduced chance of becoming pregnant. Higher psychological violence would negatively affect pregnancy, but the association was not significant (p = 0.065). Regular visits to the physician for routine checks were also linked to a decreased risk of unwanted pregnancy (ORa = 0.053). Higher psychological violence would affect unwanted pregnancy; however, the association was insignificant (p = 0.056). The study findings showed that having children and taking contraceptives are associated with a reduced pregnancy. Additionally, psychological violence was found to be related to current pregnancy status and unplanned pregnancy. During a pandemic, vulnerable women should be identified and given adequate care, knowledge, and awareness regarding their reproductive health.


Pyoderma gangrenosum: an Indian perspective.
Pyoderma gangrenosum (PG) is an uncommon, idiopathic, neutrophilic skin disease sometimes associated with systemic diseases. To our knowledge, there have been no case series of PG reported from India. To study the clinical features, associated systemic diseases and treatment methods in our case series. In total, 18 cases of PG diagnosed at our institution in Mangalore were evaluated in a prospective study. Ulcerative PG was the most common variant, seen in all except one case. Paediatric patients constituted less than a quarter (27.77%) of the patients. Systemic diseases were associated with the disease in over half of the patients (55.55%). The pathergy test was positive in seven patients (38.88%), and all but one had associated systemic disease. Corticosteroids were mainly used for management. PG is a rare disease in India, but paediatric PG cases were relatively common. Ulcerative PG was the commonest type, and over half of the patients had associated systemic diseases. There were a significant number of patients with associated systemic disease who had a positive pathergy test.


Healthcare needs of HIV-infected persons in hospital, outpatient, home, and long-term care settings.
In this descriptive study, healthcare needs reported by 386 HIV-infected persons from all risk categories and their primary nurses from four care settings (hospital, outpatient, long-term care, and home) are identified. Healthcare needs were classified into four domains of nursing practice using a conceptual framework from community health nursing--physiologic, psychosocial, health behaviors, environmental--and were assessed using the Healthcare Needs Scale for Patients with HIV/AIDS. Nurses' perceptions of major healthcare needs across all settings centered around decreased physical endurance, limited physical mobility, and sensory deficits. Patients' needs focused on financial issues. The results reflect the multidimensional nature of this infection and provide direction for clinical practice.


Survival outcomes of elderly patients with pathological stages II and III gastric cancer following curative gastrectomy.
Survival outcomes in elderly patients with pathological stages (pStages) II and III gastric cancer remain inadequately elucidated. We retrospectively analyzed outcomes of elderly and nonelderly patients who underwent curative gastrectomy for this cancer and considered clinical results of the Estimation of Physiologic Ability and Surgical Stress (E-PASS) scoring system for prediction. Among 1041 patients who underwent gastrectomy for pStages II and III gastric cancer between 2008 and 2013 consecutively, 898 patients were enrolled. Of these, 158 patients (17.6%) were elderly and 740 patients (82.4%) were nonelderly. Disease-specific survival (DSS) in the elderly group with pStage III cancer was significantly worse than that in the same stage nonelderly group (P = .001), while there was no difference in DSS for pStage II cancer between the groups (P = .45). Overall survival (OS) was significantly worse in elderly patients for both pStages II and III. Elderly patients with pStage II cancer had larger survival gaps between OS and DSS compared with those with pStage III cancer. OS for elderly patients with comprehensive risk score (CRS) > 0.159 was significantly worse than that for elderly patients with CRS ≤ 0.159 in pStage II cancer. Compared with nonelderly patients, different characteristics were observed in the survival outcomes of elderly patients between pStages II and III gastric cancer. The survival gap between OS and DSS of elderly patients was larger in pStage II cancer than in pStage III cancer. The E-PASS scoring system could be a relatively useful predictor in elderly patients.


Extended access to methamphetamine self-administration affects sensorimotor gating in rats.
Disturbed information processing observed in neuropsychiatric disorders is reflected by deficient sensorimotor gating, measured as prepulse inhibition (PPI) of the acoustic startle response (ASR). Long-term, higher dose methamphetamine (METH) abuse patterns are associated with cognitive impairments, mania and/or schizophrenia-like psychosis. The present study investigated in rats METH-induced impairment of sensorimotor gating using an intravenous self-administration (IVSA) escalating dose procedure. In this procedure, rats escalated drug intake during weekly extended access periods to METH IVSA (1, 3, and 6h), where PPI was assessed after each access period and thus at various times of drug exposure. Despite increased drug intake over the course of extended access to METH, disruption of sensorimotor gating was only seen after the access period of 6h. The data suggest that METH-induced impairment of sensorimotor gating in IVSA-tasks is rather attributed to continuous and higher dose exposure than to actual amounts of drug present at the time of testing. IVSA procedures, comprising stepwise stimulant escalation may serve as a useful translational model in rats that approximate important aspects of human abuse pattern in the context of stimulant-induced cognitive and behavioral deficits.


Temporal order in yeast chromosome replication.
Previous work with bacteria has shown that a gene is maximally sensitive to mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine (NG) at the time it is being replicated. NG was used to test for temporal order in the replication of the genome of the unicellular eucaryote, Saccaromyces cerevisiae. Yeast cells growing exponentially were more sensitive to mutagenesis by NG than cells in which DNA synthesis had been inhibited. Further, in a synchronized population of cells, individual genetic markers exhibited maximum sensitivity to muta-genesis at distinct limited intervals within the DNA synthesis period. The peaks of sensitivity are interpreted as reflecting the times of replication of different genes. Since markers for five genes on four different chromosomes showed discrete periods of maximum sensitivity, it is likely that temporal ordering of replication exists for most genes in the yeast genome. These results imply that sites for initiation of DNA replication occur at fairly specific regions along yeast chromosomal DNA moleucles, and are activated at predetermined times in the DNA synthesis period.


IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity.
Tumours evade immune control by creating hostile microenvironments that perturb T cell metabolism and effector function1-4. However, it remains unclear how intra-tumoral T cells integrate and interpret metabolic stress signals. Here we report that ovarian cancer-an aggressive malignancy that is refractory to standard treatments and current immunotherapies5-8-induces endoplasmic reticulum stress and activates the IRE1α-XBP1 arm of the unfolded protein response9,10 in T cells to control their mitochondrial respiration and anti-tumour function. In T cells isolated from specimens collected from patients with ovarian cancer, upregulation of XBP1 was associated with decreased infiltration of T cells into tumours and with reduced IFNG mRNA expression. Malignant ascites fluid obtained from patients with ovarian cancer inhibited glucose uptake and caused N-linked protein glycosylation defects in T cells, which triggered IRE1α-XBP1 activation that suppressed mitochondrial activity and IFNγ production. Mechanistically, induction of XBP1 regulated the abundance of glutamine carriers and thus limited the influx of glutamine that is necessary to sustain mitochondrial respiration in T cells under glucose-deprived conditions. Restoring N-linked protein glycosylation, abrogating IRE1α-XBP1 activation or enforcing expression of glutamine transporters enhanced mitochondrial respiration in human T cells exposed to ovarian cancer ascites. XBP1-deficient T cells in the metastatic ovarian cancer milieu exhibited global transcriptional reprogramming and improved effector capacity. Accordingly, mice that bear ovarian cancer and lack XBP1 selectively in T cells demonstrate superior anti-tumour immunity, delayed malignant progression and increased overall survival. Controlling endoplasmic reticulum stress or targeting IRE1α-XBP1 signalling may help to restore the metabolic fitness and anti-tumour capacity of T cells in cancer hosts.


Comparisons of accelerometer and pedometer determined steps in free living samples.
The purpose of this study was to compare steps·d-1 between an accelerometer and pedometer in 2 free-living samples. Data from 2 separate studies were used for this secondary analysis (Sample 1: N = 99, Male: n = 28, 20.9 ± 1.4 yrs, BMI = 27.2 ± 5.0 kg·m-2, Female: n = 71, 20.9 ± 1.7 yrs, BMI = 22.7 ± 3.0 kg·m-2; Sample 2: N = 74, Male: n = 27, 38.0 ± 9.5 yrs, BMI = 25.7 ± 4.5 kg·m-2, Female: n = 47, 38.7 ± 10.1 yrs, BMI = 24.6 ± 4.0 kg·m-2). Both studies used identical procedures and analytical strategies. The mean difference in steps·d-1 for the week was 1643.4 steps·d-1 in Study 1 and 2199.4 steps·d-1 in Study 2. There were strong correlations between accelerometer- and pedometer-determined steps·d-1 in Study 1 (r = .85, P < .01) and Study 2 (r = 0.87, P < .01). Bland-Altman plots indicated agreement without bias between steps recorded from the devices in Study 1 (r = -0.14, P < .17) and Study 2 (r = -0.09, P < .40). Correlations examining the difference between accelerometer-pedometer steps·d-1 and MVPA resulted in small, inverse correlations (range: r = -0.03 to -0.28). These results indicate agreement between accelerometer- and pedometer-determined steps·d-1; however, measurement bias may still exist because of known sensitivity thresholds between devices.


Cancer-Related Activity Limitations Among Head and Neck Cancer Survivors.
To characterize self-reported cancer-related activity limitations among a broad population of head and neck (HNC) survivors and identify sociodemographic factors associated with these limitations. Cross-sectional analysis of data from the National Health Interview Survey. The study population included individuals who completed the National Health Interview Survey (NHIS) from 1997 to 2018 and self-reported a cancer diagnosis. Data regarding activity limitations, cancer history, mental health, and demographics were extracted from the NHIS. Poisson regression was used to evaluate associations between demographics and cancer-related limitations, and a descriptive analysis was performed to identify the most common types of cancer-related limitations experienced by HNC survivors. Individuals with HNC were more than twice as likely to report having a disability caused by cancer when compared to individuals with other cancers (24% vs. 11%, P < .001). Cancer-related disability was highest among HNC survivors who were Black (adjusted prevalence ratio (aPR) = 1.57, 95% CI = 1.13-2.18), were aged 50 to 64 (aPR = 1.74, 95% CI = 1.1-2.74), had high school or lower education (aPR = 2.40, 95% CI = 1.07-5.37), and had Medicaid insurance (aPR = 2.58, 95% CI = 1.62-4.10). Among HNC patients who reported a cancer-related limitation, the most common limitations included difficulty working (78%), going out (51%), and socializing (42%). Cancer-related activity limitations are more common among HNC survivors compared to survivors of other cancers, and disproportionately affect socioeconomically disadvantaged HNC survivors. Clinicians should be aware of the limitations experienced by HNC survivors to provide counseling and resources to help patients cope with these limitations. 3 Laryngoscope, 132:593-599, 2022.


Nucleotide sequence of the Erwinia chrysanthemi NCPPB 1066 L-asparaginase gene.
The complete nucleotide sequence of the Erwinia chrysanthemi NCPPB 1066 gene coding for the chemotherapeutic enzyme L-asparaginase has been determined. The structural gene consists of an open reading frame commencing with an ATG start codon of 1044 bp followed by a TGA stop codon. Confirmation of the nucleotide sequence was obtained by comparing the predicted amino acid (aa) sequence with that derived by N-terminal aa sequencing of the purified protein. The gene has been shown to code for a 21-aa signal peptide at its N terminus which closely resembles the signal peptides of other secreted proteins. In common with highly expressed Escherichia coli genes, little use is made of modulator codons. The predicted aa sequence of the enzyme exhibits 46% identity with the determined primary sequence of the E. coli L-asparaginase, although the predicted secondary structure of both proteins indicates more extensive homology. Downstream of the TGA stop codon is a G + C-rich region of dyad symmetry (delta G = -25.4 kcal) characteristic of E. coli Rho-independent transcription terminators. Upstream of the structural gene there are no sequences which bear a strong resemblance to the consensus -35 and -10 regions of E. coli promoters. A sequence is present (CTGGCTCTCCTCTTGAT), however, which exhibits strong homology to the nif promoter consensus sequence (CTGGCACN5TTGCA). Upstream of this region is a sequence which strongly resembles the consensus sequence for promoter regions which are subject to catabolite repression.


Identification of new demands regarding prehospital care based on 35,188 missions in 2018.
Population ageing and increased prevalence of chronic diseases result in the emergence of new demands in prehospital care. The prehospital system is facing an increase of cases without acute threat to life (so-called "non-urgent"), which generates tension due to a higher number of admissions to emergency departments and a greater use of prehospital resources. Our aim is to understand this transition in prehospital activities and to delineate the primary missions performed by paramedics in 2018 with a focus on the population concerned, the severity of cases encountered and the typology of health issues. The study is retrospective, and descriptive, using a statistical description of 35,188 primary missions realized in 2018 in the State of Vaud (Switzerland). The characteristics taken into consideration are the age and gender of patients, as well as the health issue, the severity of cases based on National Advisory Committee for Aeronautics score (NACA score), and the time and place of intervention. The results describe the primary missions in the State of Vaud in 2018 and show that 87% of missions concern "non-urgent" situations (without acute threat to life). Over half of patients are 65 or older, the highest proportion of health issues, 49%, are medical and only 23% of missions are for traumas. Mission related to mental health issues reach 7% and those for intoxication 6%. Most missions take place between 7:00 am and 6:00 pm (67%), and around 12% of missions lead to the non-transport of the patient. The prehospital sector is confronted with a major transition in terms of patient care. An increase of non-urgent cases is observed, associated with the care of persons aged 65 or more. Our results question the adequacy between the needs in terms of prehospital care and the paramedic profession as it is currently defined, as well as the place of this profession within the health network. Reflecting upon the role of paramedics with respect to the socio-demographic evolution of populations appears necessary, to analyse the adequacy of the paramedics' skills to respond to the current needs.


Protocatechudehyde improves mitochondrial energy metabolism through the HIF1α/PDK1 signaling pathway to mitigate ischemic stroke-elicited internal capsule injury.
The internal capsule is vulnerable to ischemia, and mild ischemic stroke often results in lesion of the internal capsule, manifested as contralateral hemiplegia. Protocatechudehyde (PCA), a potential neuroprotective agent, has shown therapeutic effects in the study of a variety of nervous system diseases, including ischemic stroke. The aim of this study was to evaluate the effects of PCA on cerebral ischemia reperfusion (CI/R)-elicited internal capsule injury and to elucidate the role of mitochondrial energy metabolism in the underlying mechanism of neuroprotective effects on ischemic stroke. A rat tMCAO model was established to investigate the therapeutic effects of intravenous PCA (20, 40, and 80 mg/kg, once per day, continued for 7 days) on CI/R-induced internal capsule injury and the regulation of PCA on molecules related to mitochondrial energy metabolism. In vitro, an OGD/R model of PC12 cells was established to further verify the therapeutic mechanism of PCA. Results showed that PCA dose-dependently attenuated neurological deficit, reduced cerebral infarction, alleviated histopathological damage, and improved mitochondrial ultrastructure of the internal capsule after CI/R. Moreover, PCA reversed the upregulation of HIF1α, PDK1 and pPDHA1 expression induced by CI/R and significantly increased the content of acetyl-CoA, ATP, and the activity of ATP synthase. In vitro, PCA treatment promoted cell survival, inhibited apoptosis, attenuated the dissipation of mitochondrial membrane potential in OGD/R-treated PC12 cells, and these therapeutic effects were reversed by the combination of cobalt chloride (CoCl2), a specific pharmacological inducer of HIF1a expression. These results indicate that PCA exerts a protective effect against CI/R-induced internal capsule injury and improves mitochondrial energy metabolism in the internal capsule, and the mechanism is associated with the inhibition of HIF1α/PDK1 signaling pathway.


Perinatal outcomes after selective feticide via umbilical cord occlusion in complicated monochorionic pregnancies: A systematic review and meta-analysis.
We aimed to compare perinatal outcomes between umbilical cord occlusion techniques in monochorionic pregnancies, including umbilical cord ligation, fetoscopic laser coagulation, interstitial laser coagulation, bipolar cord occlusion, radiofrequency ablation, and microwave ablation. This study was registered with PROSPERO (CRD42020158861). PubMed, Web of Science, Cochrane Library, and Embase were searched for studies published up to May 2020. The DerSimonian-Laird random-effects model was used for the meta-analysis. Subgroup analyses were performed to compare the outcomes among the six techniques. Meta-regression was used to adjust for confounders. Thirty-four studies with 1646 participants were included. The overall survival was 76.5% after umbilical cord ligation, 78.8% after fetoscopic laser coagulation, 60.3% after interstitial laser coagulation, 79.2% after bipolar cord occlusion, 77.5% after radiofrequency ablation, and 75.0% after microwave ablation. Subgroup comparison showed the rates of overall survival and preterm premature rupture of membranes were not significant different among six techniques. However, rates of fetal loss, premature birth, live birth, and neonatal death differed significantly among the six groups. The choice of umbilical cord occlusion techniques will affect perinatal outcomes. We suggested that the choice of umbilical cord occlusion techniques should fully consider these differences among techniques.


COVID-19 Infections and Predictors of Sickness Related Absences Among Healthcare Workers: The Experience of a Tertiary Care Center With the COVID-19 Pandemic.
Little has been published on predictors of prolonged sick leaves during the COVID-19 pandemic. This study aims to determine the rate of COVID-19 infections among healthcare workers (HCWs) and to identify the predictors of longer sick leave days. We identified predictors of longer sick leave using linear regression analysis in a cross-sectional study design. Thirty-three percent of the total workforce contracted COVID-19. On average, HCWs took 12.5 sick leave days after COVID-19 infection. The regression analysis revealed that older employees, nurses, and those who caught COVID-19 earlier in the pandemic were more likely to take longer sick leave. Age, job position, and month of infection predicted sick leave duration among HCWs in our sample. Results imply that transmission was most likely community-based. Public health interventions should consider these factors when planning for future pandemics.


Lateral thoracic perforator flap: additional perforator flap option from the lateral thoracic region.
Perforator flaps from the lateral thoracic region have not been as popular as other donor sites because of the misconception that the vascular anatomy in this region is less than predictable. However, the skin over the lateral thoracic region is vascularised by three rows of perforators of varied vascular dominance. Two perforator flaps from this region based on the middle and the posterior row of perforators from the thoracodorsal artery have been described. The lateral thoracic perforator flap based on the anterior row of perforators is another useful option. Nine patients underwent reconstructions using the lateral thoracic perforator flap for various defects in the head and neck region and lower limbs as a result of tumour extirpation, crush injury and chronic wound with osteomyelitis. All flaps were raised in the supine position. Three flaps were raised in a chimaeric fashion. The largest flap was 20×12 cm and the mean size was 106 cm2. All flaps survived without major complication. The lateral thoracic perforator flap is a reliable reconstructive option. It can be readily configured in terms of size, thickness and tissue composition. However, it is not the first-choice flap from this region because the resultant donor scar tends to extend visibly beyond the anterior axillary fold and the arterial and venous pedicles frequently have separate courses. The lateral thoracic region has become a versatile and universal donor site for free-style flap harvest with this additional flap option.


Attenuation of defined Brucella melitensis wboA mutants.
Rough Brucella mutants have been sought as vaccine candidates because they do not induce seroconversion. In this study, two defined nonreverting rough mutants were derived from virulent Brucella melitensis strain 16M: a wboA deletion mutant designated WRR51 and a wboA purEK dual deletion mutant designated WRRP1. Strain WRRP1 exhibited reduced survival in human monocyte-derived macrophages (hMDMs) compared with parent strain WRR51 or with ΔpurEK strain WR201. Strain WRRP1 persisted for 1 week or less in BALB/c mice after intraperitoneal infection, while less severe attenuation was exhibited by the two single mutants in this model. Trans complementation of wboA restored the survival of WRR51 in hMDMs comparable to strain 16M and the survival of WRRP1 comparable to strain WR201.


Associations Between Baseline Total PTSD Symptom Severity, Specific PTSD Symptoms, and 3-Month Quality of Life in Neurologically Intact Neurocritical Care Patients and Informal Caregivers.
The objective is to pilot test the feasibility of assessing severity of posttraumatic stress disorder (PTSD) symptoms, as well as specific lower-order PTSD symptoms, experienced during neurological intensive care unit (neuro-ICU) admission and their relationship with 3-month quality of life (QoL) scores in multiple domains (i.e., physical, psychological, social, and environmental) in both patients and caregivers. Between 2015 and 2016, we enrolled neurologically intact patients and informal caregivers of patients who reported demographics and PTSD symptoms (PTSD Checklist-Specific; PCL-S) during neuro-ICU admission and completed a QoL assessment (World Health Organization Quality of Life; WHOQOL-BREF) 3 months later. Clinical data were extracted from medical records. We ran two bivariate correlation matrices among PTSD symptom dimensions in patients and caregivers. Then, we used multiple linear regression to examine the prospective association of total PCL-S scores with each QoL domain in both patients and caregivers after adjusting for clinically important variables. Next, we explored differential associations between the 4 PTSD symptom dimensions and each QoL domain in both patients and caregivers, adjusting for sex and age. A total of 70 patients (45.7% women, 84.3% white, mean age 52.08 years) and 64 caregivers (64.1% women, 90.6% white, mean age 53.12 years) were included. PTSD symptom dimensions were moderately strongly correlated among patients (r = 0.65-0.79, p < 0.001) and caregivers (r = 0.55-0.78, p < 0.001). For both patients and caregivers, greater PTSD symptom severity was prospectively associated with lower QoL in all domains (β = - 0.289 to - 0.622; p < 0.05). Our exploratory analysis revealed that greater numbing symptoms were associated with lower psychological QoL in patients (β: - 0.397, p = 0.038), and lower physical (β: - 0.409, p = 0.014), psychological (β: - 0.519, p = 0.001), and social QoL (β: - 0.704, p < 0.001) in caregivers. Greater re-experiencing symptoms were associated with lower physical QoL in both patients (β: - 0.422, p = 0.047) and caregivers (β: - 0.4, p = 0.041). Our preliminary results indicated that greater severity of PTSD symptoms, and specifically numbing and re-experiencing symptoms, experienced by patients and caregivers during neuro-ICU admission was predictive of worse 3-month QoL. Continued study is needed to identify treatment targets for PTSD and QoL in this population.


On-grid and in-flow mixing for time-resolved cryo-EM.
Time-resolved cryo-electron microscopy (TrEM) allows the study of proteins under non-equilibrium conditions on the millisecond timescale, permitting the analysis of large-scale conformational changes or assembly and disassembly processes. However, the technique is developing and there have been few comparisons with other biochemical kinetic studies. Using current methods, the shortest time delay is on the millisecond timescale (∼5-10 ms), given by the delay between sample application and vitrification, and generating longer time points requires additional approaches such as using a longer delay line between the mixing element and nozzle, or an incubation step on the grid. To compare approaches, the reaction of ATP with the skeletal actomyosin S1 complex was followed on grids prepared with a 7-700 ms delay between mixing and vitrification. Classification of the cryo-EM data allows kinetic information to be derived which agrees with previous biochemical measurements, showing fast dissociation, low occupancy during steady-state hydrolysis and rebinding once ATP has been hydrolysed. However, this rebinding effect is much less pronounced when on-grid mixing is used and may be influenced by interactions with the air-water interface. Moreover, in-flow mixing results in a broader distribution of reaction times due to the range of velocities in a laminar flow profile (temporal spread), especially for longer time delays. This work shows the potential of TrEM, but also highlights challenges and opportunities for further development.


Effects of intermittent very-low calorie diet on glycemic control and cardiovascular risk factors in obese patients with type 2 diabetes mellitus: A randomized controlled trial.
Very few studies assess the effectiveness of different protocols of intermittent very-low calorie diet (VLCD) in patients with diabetes. This study was designed to compare the effects of 2 days/week and 4 days/week of intermittent VLCD on glycemic control, diabetes remission, metabolic parameters and quality of life in patients with type 2 diabetes and obesity. Participants with obesity and type 2 diabetes were recruited and randomly assigned to three groups, consisting of control, 2 days/week and 4 days/week of intermittent VLCD. In the intermittent VLCD groups, participants received a 600-kcal diet per day on restricted days and ad libitum food consumption on non-restricted days. Glycemic control, rate of diabetes remission, metabolic parameters and quality of life were evaluated at baseline, weeks 2, 10 and 20. A total of 40 participants were enrolled. The mean body mass index was 30.1 ± 5.9 kg/m2 , and the mean glycated hemoglobin was 7.4 ± 1.2%. At week 20, there was an improvement in glycemic control in both intermittent VLCD groups with significant decreases in glycated hemoglobin levels and insulin resistance index throughout the study periods. Diabetes remission without the need for medications was equally found in 29% of participants in both intermittent VLCD groups. Serum triglyceride, bodyweight, body mass index and fat mass were also significantly decreased in both VLCD groups. No serious adverse events were encountered. Intermittent VLCD was highly effective in achieving optimal glycemic control. The effects of 2 days/week and 4 days/week of intermittent VLCD on diabetes remission were relatively similar.


Dihedral representations and statistical geometric optics. I. Spherocylindrical lenses.
The linear 2-dim irreducible representations of the dihedral groups (Dn) are interpreted as classical linear operators of geometrical optics. It is shown that the 2-dim irreducible representation of D4 is simply the refractive group described by Campbell [Optom. Vision Sci. 74, 381 (1997)]. The dihedral Fourier-inverse mechanism is introduced and shown to provide a systematic connection between the standard refractive data and their vector space representation, as proposed by Thibos et al. [Vision Sci. Appl. 2, 14 (1994)].


Identifying environmental reservoirs of Clostridium difficile with a scent detection dog: preliminary evaluation.
Prompted by an article describing a dog trained to detect Clostridium difficile in patients, our institution evaluated a dog's ability to detect C. difficile scent from equipment and surfaces to assist in strategic deployment of adjunctive cleaning measures. An expert in drug and explosives scent dog handling trained a canine to identify odours from pure cultures and/or faecal specimens positive for C. difficile. Methods used to assess explosive and drug detection dogs were adapted and included evaluation of (i) odour recognition, using containers positive and negative for the scent of C. difficile, and of (ii) search capability, on a simulation ward with hidden scents. After demonstration that the canine could accurately and reliably detect the scent of C. difficile, formal assessments of all clinical areas began. Odour recognition (N = 75 containers) had a sensitivity of 100% and specificity of 97%. Search capability was 80% sensitive and 92.9% specific after removal of results from one room where dog and trainer fatigue influenced performance. Both odour recognition and search capability had an overall sensitivity of 92.3% and specificity of 95.4%. The clinical unit sweeps over a period of five months revealed a sensitivity of 100% in alerting on positive quality control hides. These clinical unit sweeps also resulted in 83 alerts during 49 sweep days. A dog can be trained to accurately and reliably detect C. difficile odour from environmental sources to guide the best deployment of adjunctive cleaning measures and can be successfully integrated into a quality infection control programme.


[The effect of different doses of esmolol on hemodynamic, bispectral index and movement response during orotracheal intubation: prospective, randomized, double-blind study].
A prospective, randomized and double-blind study was planned to identify the optimum dose of esmolol infusion to suppress the increase in bispectral index values and the movement and hemodynamic responses to tracheal intubation. 120 patients were randomly allocated to one of three groups in a double-blind fashion. 2.5mgkg(-1) propofol was administered for anesthesia induction. After loss of consciousness, and before administration of 0.6mgkg(-1) rocuronium, a tourniquet was applied to one arm and inflated to 50mmHg greater than systolic pressure. The patients were divided into 3 groups; 1mgkg(-1)h(-1) esmolol was given as the loading dose and in Group Es50 50μgkg(-1)min(-1), in Group Es150 150μgkg(-1)min(-1), and in Group Es250 250μgkg(-1)min(-1) esmolol infusion was started. Five minutes after the esmolol has been begun, the trachea was intubated; gross movement within the first minute after orotracheal intubation was recorded. Incidence of movement response and the ΔBIS max values were comparable in Group Es250 and Group Es150, but these values were significantly higher in Group Es50 than in the other two groups. In all three groups in the 1st minute after tracheal intubation heart rate and mean arterial pressure were significantly higher compared to values from before intubation (p<0.05). In the study period there was no significant difference between the groups in terms of heart rate and mean arterial pressure. In clinical practise we believe that after 1mgkg(-1) loading dose, 150μgkg(-1)min(-1) iv esmolol dose is sufficient to suppress responses to tracheal intubation without increasing side effects.


Reconstruction of erythrocyte shape during modified morphological response.
Changes in erythrocyte shape during morphological response modified by benzalkonium chloride (BzA) were studied in sucrose solutions. Fixation of the cells with glutaraldehyde- and formaldehyde-containing fixatives at some time points is usually inadequate to maintain the current cell shape. Considering the reconstruction of erythrocyte shape, which takes into account the mode of fixative action, we showed that the echinocyte-forming activity of BzA depends on the concentration of this surfactant. It can induce a direct spherostomatocyte-spheroechinocyte transition without altering the near-spherical shape of the cells. On the other hand, the reverse spheroechinocyte-spherostomatocyte transition was always accompanied by some flattening of the cells, although in some instances discoidal shape was not achieved. The data point to asymmetric shape transitions of erythrocytes in sucrose solution, which contradicts the continuum and bilayer-couple models of shape regulation. It seems that the nonuniform structure of native erythrocyte membrane plays a more important role in morphological transitions of these cells than suggested earlier.


On the toxicity of kynurenic acid in vivo and in vitro.
Kynurenic acid (KYNA), a tryptophan metabolite is an antagonist of ionotropic glutamate receptors and alpha-7 nicotinic receptor. Moreover, it is an agonist of G-protein receptor GPR35. Its neuroprotective, anticonvulsant, anti-inflammatory and antioxidant activity was documented. KYNA is present in food and herbal medicines. However, the data on effects induced by a long-lasting treatment with KYNA is lacking. The aim of the study was the assessment of toxicity of a prolonged administration of KYNA in rodents. The cytotoxicity of KYNA in vitro was also examined. Adult mice and rats were used. KYNA was administered in the drinking water in concentrations of 25 or 250mg/L for 3-21 days. The following cells were cultured in an in vitro study: mouse fibroblast (NIH/3T3), green monkey kidney cells and primary chick embryo cells (CECC). Cell viability was determined with methyl thiazol tetrazolium reduction assay, neutral red uptake assay and lactate dehydrogenase leakage assay. KYNA affected neither body gain nor body composition. Blood counts were also unaffected. The viability of cells in the culture was lowered at high millimolar concentrations of KYNA. An elevated viability of GMK and CECC cells was detected in the presence of KYNA in micromolar concentrations. The obtained results showed that a long-term application of KYNA in the drinking water is well-tolerated by rodents. No evidence of a toxic response was recorded. Achieved results indicate that diets containing a high amount of KYNA or enriched with KYNA should not cause any risk to the human health.


Characterization of acetohydroxyacid synthase from Mycobacterium tuberculosis and the identification of its new inhibitor from the screening of a chemical library.
Acetohydroxyacid synthase (AHAS) is a thiamin diphosphate- (ThDP-) and FAD-dependent enzyme that catalyzes the first common step in the biosynthetic pathway of the branched-amino acids such as leucine, isoleucine, and valine. The genes of AHAS from Mycobacterium tuberculosis were cloned, and overexpressed in E. coli and purified to homogeneity. The purified AHAS from M. tuberculosis is effectively inhibited by pyrazosulfuron ethyl (PSE), an inhibitor of plant AHAS enzyme, with the IC(50) (inhibitory concentration 50%) of 0.87 microM. The kinetic parameters of M. tuberculosis AHAS were determined, and an enzyme activity assay system using 96-well microplate was designed. After screening of a chemical library composed of 5600 compounds using the assay system, a new class of AHAS inhibitor was identified with the IC(50) in the range of 1.8-2.6 microM. One of the identified compounds (KHG20612) further showed growth inhibition activity against various strains of M. tuberculosis. The correlation of the inhibitory activity of the identified compound against AHAS to the cell growth inhibition activity suggested that AHAS might be served as a target protein for the development of novel anti-tuberculosis therapeutics.


Three -(pyridin-2-yl)-2-(pyridin-2-ylimino)thiazolidin-4-one as a novel inhibitor of cerebral MAO-B activity with antioxidant properties and low toxicity potential.
Some brain diseases are associated with oxidative stress and altered monoamine oxidase (MAO) activity. The objective of this study was to evaluate the antioxidant and neuroprotective actions through MAO inhibition of 3-(pyridin-2-yl)-2-(pyridine-2-ylimino) thiazolidin-4-one (PPIT, a synthetic molecule containing a thiazolidinone nucleus), as well as its effects on toxicity parameters in Swiss female mice. Five in vitro assays were carried out to verify the PPIT antioxidant capacity: protein carbonylation (PC), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picryl-hydrazil (DPPH), ferric ion (Fe3+ ) reducing antioxidant power (FRAP), and superoxide dismutase (SOD)-like activity. The results showed that PPIT reduced the level of PC in the homogenate of the brain. This compound did not demonstrate SOD mimetic activity, but it acted as a free radical scavenger (ABTS and DPPH) and exhibited reducing activity in the FRAP assay. In addition, the effects of PPIT on cerebral MAO activity (MAO-A and B isoforms) were investigated in vitro. Our data revealed inhibition of the MAO-B activity by PPIT with no effects on MAO-A. Lastly, an acute oral toxicity test was conducted in mice. No changes in food intake, body weight, and biochemical markers of kidney and liver damage were detected in mice treated with a high dose of PPIT (300 mg/kg). In conclusion, the present study demonstrated that PPIT exhibits antioxidant activity and selectively inhibits the MAO-B isoform without causing apparent toxicity. These findings suggest PPIT as a potential therapeutic candidate to be tested in preclinical models of brain diseases involving perturbations of MAO-B activity and redox status.


Mechanism of molybdate insertion into pterin-based molybdenum cofactors.
The molybdenum cofactor (Moco) is found in the active site of numerous important enzymes that are critical to biological processes. The bidentate ligand that chelates molybdenum in Moco is the pyranopterin dithiolene (molybdopterin, MPT). However, neither the mechanism of molybdate insertion into MPT nor the structure of Moco prior to its insertion into pyranopterin molybdenum enzymes is known. Here, we report this final maturation step, where adenylated MPT (MPT-AMP) and molybdate are the substrates. X-ray crystallography of the Arabidopsis thaliana Mo-insertase variant Cnx1E S269D D274S identified adenylated Moco (Moco-AMP) as an unexpected intermediate in this reaction sequence. X-ray absorption spectroscopy revealed the first coordination sphere geometry of Moco trapped in the Cnx1E active site. We have used this structural information to deduce a mechanism for molybdate insertion into MPT-AMP. Given their high degree of structural and sequence similarity, we suggest that this mechanism is employed by all eukaryotic Mo-insertases.


Microdialysis of GABA and glutamate: analysis, interpretation and comparison with microsensors.
GABA and glutamate sampled from the brain by microdialysis do not always fulfill the classic criteria for exocytotic release. In this regard the origin (neuronal vs. astroglial, synaptic vs. extrasynaptic) of glutamate and GABA collected by microdialysis as well as in the ECF itself, is still a matter of debate. In this overview microdialysis of GABA and glutamate and the use of microsensors to detect extracellular glutamate are compared and discussed. During basal conditions glutamate in microdialysates is mainly derived from non-synaptic sources. Indeed recently several sources of astrocytic glutamate release have been described, including glutamate derived from gliotransmission. However during conditions of (chemical, electrical or behavioral) stimulation a significant part of glutamate might be derived from neurotransmission. Interestingly accumulating evidence suggests that glutamate determined by microsensors is more likely to reflect basal synaptic events. This would mean that microdialysis and microsensors are complementary methods to study extracellular glutamate. Regarding GABA we concluded that the chromatographic conditions for the separation of this transmitter from other amino acid-derivatives are extremely critical. Optimal conditions to detect GABA in microdialysis samples--at least in our laboratory--include a retention time of approximately 60 min and a careful control of the pH of the mobile phase. Under these conditions it appears that 50-70% of GABA in dialysates is derived from neurotransmission.


Non-identical behaviour of the subunits of rabbit muscule creatine kinase.
1. The dimeric enzyme creatine kinase (ATP: creatine N-phospotransferase, EC 18.104.22.168) from rabbit muscle was reacted with three separate reagents, each of which specifically modifies one thiol group per subunit. 2. The reactions of the enzyme with these reagents (4-chloro-7-nitrobenzofurazan, 5,5'-dithiobis-(2-nitrobenzoic acid) and iodoacetate) all behave as normal second-order processes. This indicates that the thiol groups on the two subunits of the enzyme react at the same rate as each other in all three cases. 3. The effects of various ligands (Mg2+, ADP and creatine, and combinations of these) on the kinetics of the reactions were studied. In all cases the reactions behave as normal second-order processes. 4. In the presence of the ligand combination Mg2+ plus ADP plus creatine plus nitrate, which has been postulated to form a "transition state analogue" complex with the enzyme, the reactions of the thiol group show considerable deviation from second-order kinetics. This indicates that the thiol groups on the two subunits react at different rates from each other. A similar effect is also noted in the presence of the combination ADP plus creatine plus nitrate. 5. The binding of ADP to the enzyme (studied by equilibrium dialysis) is hyperbolic in the absence of other ligands or in the presence of Mg2+ or Mg2+ plus creatine. The dissociation constant is similar in all three cases. 6. In the presence of creatine plus nitrate (with or without Mg2+) the bindings of ADP to the enzyme is tightened considerably and the binding plots indicate the presence of either negative interactions between the subunits or two distinct types of binding sites. 7. Possible causes for the observed non-identical behaviour of the two subunits of the enzyme are discussed.


Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR.
To assess the extent of Listeria monocytogenes in causation of human spontaneous abortions by isolation methods and PCR analysis for the presence of virulence-associated genes. A total of 305 samples comprising blood, urine, placental bits, faecal and vaginal swabs were collected from 61 patients with spontaneous abortions. Listeria spp. were isolated from 10 samples collected from nine (14.8%) patients. Confirmation of these isolates was based on biochemical tests, haemolysis on blood agar, CAMP test, phosphatidylinositol-specific phospholipase C (PI-PLC) assay followed by in vivo pathogenicity tests and multiplex PCR to detect virulence-associated genes (prfA, plcA, hlyA, actA and iap). Three isolates were confirmed as L. monocytogenes. Of these, two isolates turned out to be pathogenic and found to posses all five genes. However, the remaining two haemolytic L. monocytogenes isolates lacking the plcA gene and activity in the PI-PLC assay were found to be nonpathogenic by in vivo tests. The occurrence of pathogenic L. monocytogenes in cases of spontaneous abortions was 3.3%. It seems that the plcA gene and its expression have an important role as essential virulence determinants in pathogenic Listeria spp. The recovery of pathogenic L. monocytogenes isolates from cases of spontaneous abortion indicates the significance of listeric infection in pregnant women.


Single-atom boosted electrochemiluminescence via phosphorus doping of Fe-N/P-C catalysts.
Single-atom catalyst (SAC), one of the most attractive catalysts in the field of energy conversion and storage, was proven as efficient accelerator for luminol-dissolved oxygen electrochemiluminescence (ECL) via the catalysis of oxygen reduction reaction (ORR). In this work, we synthesized heteroatom doping SACs of Fe-N/P-C for the catalysis of cathodic luminol ECL. The doping of P could lower the reaction energy barrier of the OH* reduction, and promote catalytic efficiency toward ORR. The formation of reactive oxygen species (ROS) during ORR triggered cathodic luminol ECL. Greatly enhanced ECL emission catalyzed by SACs proved that Fe-N/P-C exhibited higher catalytic activity to ORR compared with Fe-N-C. Since the system was highly dependent on oxygen, an ultra-sensitive detection of a typical antioxidant, ascorbic acid, was achieved with detection limit of 0.03 nM. This study provides possibility to greatly enhance the performance of ECL platform through rational tailoring of SACs via heteroatom doping.


New Developments in the Management of Recurrent Pericarditis.
Recurrent pericarditis is a common and troublesome complication that affects 15%-30% of patients with a previous episode of pericarditis. However, the pathogenesis of these recurrences is not well understood, and most cases remain idiopathic. Recent advances in medical therapy, including the use of colchicine and anti-interleukin-1 agents like anakinra and rilonacept, have suggested an autoinflammatory rather than an autoimmune mechanism for recurrences with an inflammatory phenotype. As a result, a more personalized approach to treatment is now recommended. Patients with an inflammatory phenotype (fever and elevated C-reactive protein level) should receive colchicine and anti-interleukin-1 agents as first-line therapy, whereas those without systemic inflammation should receive low to moderate doses of corticosteroids (eg, prednisone 0.2-0.5 mg/kg/d as an initial dose) and consider azathioprine and intravenous human immunoglobulins in the case of corticosteroid failure. Tapering of corticosteroids should be slow after achieving clinical remission. In this article, we review the new developments in the management of recurrent pericarditis.


Flap-in-flap method for enhancing the advancement of a V-Y flap.
The V-Y principal is used in a number of flap reconstruction techniques. V-Y flaps suffer from design problems of the advancing edge, which is usually concave, trying to match another concave edge on the opposite side of the defect. The flap-in-flap technique uses a second V-Y flap at the advancing edge of the main flap to overcome this mismatch, and also improves the amount of advancement possible. The technique has been used in three patients, demonstrating reliability and a further advantage, which is the ability of the second flap to rotate at an angle to the main flap to aid in reconstructing a complex defect such as the alar base of the nose. Further experience with this technique should widen its applications.


Electroretinography in seasonal affective disorder.
Retinal mechanisms have been hypothesized in the pathophysiology of seasonal affective disorder (SAD). Electroretinography (ERG) is a noninvasive electrophysiologic test that provides an objective measure of photoreceptor and retinal function. We conducted dark-adapted ERG examinations with a bright white light stimulus in a group of depressed, drug-free patients with seasonal affective disorder (6 men, 18 women) diagnosed by DSM-III-R criteria, and a group of sex- and age-matched control subjects (6 men, 16 women) during the winter. A significant difference was found between SAD patients and controls, but female SAD patients had lower ERG b-wave amplitudes than female controls, while male SAD patients had higher amplitudes than the matched controls. The ERG b-wave implicit times (times from onset of stimulus to peak of b-wave) were significantly longer in the left eyes of the male control subjects. These data may indicate subtle retinal changes in patients with SAD, but the results must be considered preliminary because of the small number of subjects studied and the large intersubject variability in the ERG procedure.


Acute hemodynamic effects of coronary artery ligation in conscious rats.
Because of the growing interest in the use of coronary artery ligation (CAL) in rats as a model for studies on heart failure, we have investigated the acute hemodynamic changes following CAL in conscious rats. Animals were equipped for measurement of cardiac output (CO), arterial pressure (MAP), and central venous pressure (CVP). These parameters were measured before CAL, immediately after, and 24 h after. Furthermore, peak CO, obtained by rapid infusion of 12 ml Ringer's solution (in 1 min) was measured 2 days before and 1 day after CAL. CAL resulted in immediate reduction of CO, because of reduced stroke volume (SV). CO as well as SV were inversely correlated with infarct size as determined 24 h after CAL. Heart rate (HR) and MAP did not change. Twenty-four hours later, CO was still reduced. MAP was now reduced, possibly as a result from resetting of nervous reflex mechanisms. Before CAL, peak CO and SV were similar in CAL and sham animals. At 24 h after CAL, these parameters were greatly reduced in CAL rats. Peak values were strongly correlated to infarct size. Results indicate that CAL in rats leads to hemodynamic changes similar to the ones observed following myocardial infarction in man. Cardiac function is related to infarct size and is altered both at rest and during maximal stimulation.


Identification of azurocidin as a potential periodontitis biomarker by a proteomic analysis of gingival crevicular fluid.
The inflammatory disease periodontitis results in tooth loss and can even lead to diseases of the whole body if not treated. Gingival crevicular fluid (GCF) reflects the condition of the gingiva and contains proteins transuded from serum or cells at inflamed sites. In this study, we aimed to discover potential protein biomarkers for periodontitis in GCF proteome using LC-MS/MS. We identified 305 proteins from GCF of healthy individuals and periodontitis patients collected using a sterile gel loading tip by ESI-MS/MS coupled to nano-LC. Among these proteins, about 45 proteins were differentially expressed in the GCF proteome of moderate periodontitis patients when compared to the healthy individuals. We first identified azurocidin in the GCF, but not the saliva, as an upregulated protein in the periodontitis patients and verified its increased expression during periodontitis by ELISA using the GCF of the classified periodontitis patients compared to the healthy individuals. In addition, we found that azurocidin inhibited the differentiation of bone marrow-derived macrophages to osteoclasts. Our results show that GCF collection using a gel loading tip and subsequent LC-MS/MS analysis following 1D-PAGE proteomic separation are effective for the analysis of the GCF proteome. Our current results also suggest that azurocidin could be a potential biomarker candidate for the early detection of inflammatory periodontal destruction by gingivitis and some chronic periodontitis. Our data also suggest that azurocidin may have an inhibitory role in osteoclast differentiation and, thus, a protective role in alveolar bone loss during the early stages of periodontitis.


Epidemiology of dyslipidemia in Tunisia, HSHS 3 study (Hammam Sousse Sahloul Heart Study).
Dyslipidemia is a major cause of morbidity and mortality worldwide because it increases the risk of cardiovascular diseases. To determine the prevalence of dyslipidemia and its components in the general population of Hammam Sousse (Tunisia) and to identify its risk factors. This was an analysis of the HSHS database (Hammam Sousse Sahloul Heart Study), a «community-based» cross-sectional study on cardiovascular risk factors including dyslipidemia, with a random sample in two-stages, proportional-probability clusters. All subjects above 20 years underwent a lifestyle interview, clinical examination with anthropometric measurements, and blood sampling. Dyslipidemia was defined by: total cholesterol ≥5.2 mmol/l, hyper LDL (Low Density Lipoprotein) cholesterolemia ≥4.1 mmol/l, hypo HDL (High Density Lipoprotein) cholesterolemia <1.03 mmol/l for men and <1.29 mmol/l for women and hyper-triglyceridaemia: ≥2.26 mmol/l. Multivariate logistic regression analysis was conducted to determine factors independently associated with dyslipidemia. The study population was composed of 481 males (M) and 960 females (F), with a sex ratio of 0.5. Mean cholesterol was higher in women (5 mmol/l±1.01) than in men (4.8 mmol/l±0.92). Only 24.9% of men and 29.1% of women had normal HDL cholesterol levels. Women had higher levels of hypercholesterolemia and LDL cholesterolemia than men (p<10-6). Multivariate analysis showed that dyslipidemia was independently and statistically significantly associated with age ≥40 years (p<10-3), physical inactivity (p<10-3) and obesity (p=0.025). As a result of the epidemiological situation of dyslipidemia, the promotion of an active lifestyle seems essential. In addition, nutritional education improves the lipid profile by promoting weight loss and balancing lipid consumption.


Systemic NK cell ablation attenuates intra-abdominal adipose tissue macrophage infiltration in murine obesity.
Natural killer (NK) cells are understudied in the context of metabolic disease and obesity. The goal of this study was to define the effect of NK cell ablation on systemic inflammation and glucose homeostasis in murine obesity. A transgenic murine model was used to study the effect of NK cell ablation on systemic inflammation and glucose homeostasis in the context of diet-induced obesity using flow cytometry, QRTPCR, and glucose tolerance and insulin sensitivity testing. NK cell ablation achieved a three to fourfold decrease in NK cells but had no effect on T-cell levels in adipose tissues and spleen. NK cell ablation was associated with decreased total macrophage infiltration in intra-abdominal adipose tissue, but macrophage infiltration in subcutaneous adipose tissue and spleen was unaffected. NK cell ablation was associated with modest improvement in insulin sensitivity but had no effect on tissue transcript levels of inflammatory cytokines. NK cells play a role in promoting intra-abdominal adipose tissue macrophage infiltration and systemic insulin resistance in obesity.


The occurrence of rare rpoB mutations in rifampicin-resistant clinical Mycobacterium tuberculosis isolates from Kuwait.
Mutations associated with rifampicin (RIF) resistance in two regions of the rpoB gene were studied by line probe (INNO-LiPA Rif. TB) assay (LiPA) and/or DNA sequencing in 32 RIF-resistant and 21 RIF-susceptible Mycobacterium tuberculosis strains isolated from 53 tuberculosis patients in Kuwait. The LiPA identified all susceptible strains as RIF sensitive, and the DNA sequences of the 81bp rifampicin resistance-determining region (RRDR) and the N-terminal region of the rpoB gene from five isolates were identical to the wild-type sequences from M. tuberculosis H(37)Rv. The LiPA identified 24 of 32 (75%) phenotypically documented RIF-resistant M. tuberculosis isolates as RIF resistant, with specific detection of mutation in 19 isolates, whilst 8 strains were identified as RIF susceptible. DNA sequencing of the RRDR confirmed the results for the 19 RIF-resistant isolates and identified precise mutations in five isolates for which specific base changes could not be determined by LiPA, as well as identifying a mutation (insertion 514TTC) in three isolates and no mutation in five of the eight isolates that were detected as RIF susceptible by LiPA. Two of the latter five isolates contained a V146F mutation in the N-terminal region of the rpoB gene and were recovered from patients of Middle Eastern origin. These analyses identified 11 different mutations in two regions of the rpoB gene. The majority of the isolates carrying identical or no mutations in the rpoB gene exhibited unique DNA banding patterns in fingerprinting studies. The occurrence of rare mutations in the rpoB gene, namely insertion 514TTC in 3 (9%) and V146F in 2 (6%) of 32 RIF-resistant M. tuberculosis isolates from Kuwait, is the highest reported so far.


Cytomorphologic findings of intraosseous notochordal rest on touch preparation: a case report.
Intraosseous notochordal rest is a rare intravertebral lesion of notochord origin which is presumably benign. It is usually an incidental finding in microscopic examination of vertebrectomy due to unrelated lesions or autopsy cases. Chordoma is a malignant neoplasm originating from notochord with a different clinicoradiographic presentation, prognosis, and treatment. However, the histology of intraosseous notochordal rest and chordoma is almost identical. Herein, we report cytomorphologic findings of a case of intraossous notochordal rest on touch preparation.


The effectiveness of smart home technologies to support the health outcomes of community-dwelling older adults living with dementia: A scoping review.
To map the current state-of-knowledge about the effectiveness of smart home technologies to support the health outcomes of community-dwelling older adults living with dementia. A scoping review following the methodological frameworks described by Arksey and O'Malley (2005) and Levac et al. (2010). Electronic databases and online sources were searched in April 2020 using database specific medical subject headings and keywords about 'smart homes' and 'dementia'. Empirical peer-reviewed articles were included if they were written in English; used a quantitative, qualitative, or mixed method design; and presented the effects of a smart home technology on the health outcomes of community-dwelling adults living with dementia. Methodological and reporting quality of studies was assessed using the Mixed Methods Appraisal Tool - Version 2018. Five studies described evaluations of five smart home technology systems with a total of n = 617 community-dwelling people living with dementia. Collectively, studies showed potential effectiveness of the technologies on a range of health outcomes (physical activity, activities of daily living, sleep, anxiety, depression, agitation, irritability, risk of falls, cognitive functioning, night-time injury, unattended home exits). However, the overall methodological and reporting quality of studies was low and profiled a research field lacking in rigorous evaluation. Based on current evidence, the success of smart home technologies to support people with dementia to live at home remains unclear. Recommendations are provided to inform future research into smart home technologies for community-based dementia care.


Movement-related cerebral potentials in schizophrenics.
Measures concerning the P2 component of movement-related cerebral potentials (MPs) (the time from the trigger produced by key-tapping to the onset of P2: T-N latency, the width of P2: N-P interval, and the amplitude of P2: N-P amplitude) and various clinical factors of 36 schizophrenics were studied and statistically compared with those of 35 normal subjects. Concerning the P2 component, 22/36 of the schizophrenics showed abnormal findings. These three measures, especially the N-P interval, are significantly related to various clinical symptoms at the stage when the MPs were recorded but are not related to the previous worst stage. The chronicity of schizophrenic illness influenced each measure of the T-N latency and N-P interval, both of which were significantly longer than those of the normal subjects. The pressure strength, the velocity of key-tapping, the duration of muscular contraction, (namely the manner of key-tapping) and the daily dosage of neuroleptics did not affect the P2 component. These results suggest that 1) the P2 component of MPs is related to certain brain functions as a signal of information processing concerning action and not to the peripheral feed-back mechanism and 2) abnormal waveforms of the MPs found in pathological psychiatric conditions of schizophrenia may be a reflection of disturbances of the central mechanism concerning action, attention and volition.


[Examination of the antibacterial action of metronidazole against vibrios and Campylobacter (author's transl)].
The in vitro antibacterial activity of metronidazole was tested against 70 strains of aerobic vibrios (V. cholerae biotype cholerae, V. cholerae biotype eltor, NAG-vibrios, V. parahaemolyticus, v. alginolyticus) and 30 strains of microaerophilic Campylobacter (C. fetus subsp. fetus, C. fetus subsp. intestinalis and C. fetus subsp. jejuni). All strains of aerobic vibrios proved to be resistant (MIC 100 micrograms/ml) in contrast to campylobacter strains which were sensitive (MIC 1-4 micrograms/ml) to the drug. The findings confirm that metronidazole can be considered to be a selective inhibitor of anaerobic microorganisms, but its action is not restricted to obligate anaerobes.


Bio-syncretic robots composed of biological and electromechanical systems.
Potential approaches to developing bio-syncretic robots are proposed in different dimensions, including monomial functions and closed loop systems of living cell-based sensing, intelligence, and actuation.


Histamine release-neutralization assay for sera of patients with atopic dermatitis and/or cholinergic urticaria is useful to screen type I hypersensitivity against sweat antigens.
We previously reported that about 80 % of patients with atopic dermatitis and 60 % with cholinergic urticaria revealed type I allergy against sweat, by means of skin test against autologous sweat and/or histamine-release test for peripheral blood basophils with semi-purified sweat antigen. In this study, we developed an assay for sera to neutralize histamine-releasing activity of semi-purified sweat antigen. The semi-purified sweat antigen was pre-incubated with serially diluted sera for 30 min at 37 °C and was subjected to histamine-release activity. Histamine release-neutralization (HRN) activities were calculated by measuring the amount of histamine release from basophils in the presence or absence of semi-purified sweat antigen. Of 62 subjects, 39 showed positive histamine release (≥5 %) from their basophils in response to semi-purified sweat antigen, and sera of 34 out of 39 subjects (87.2 %) were also positive in HRN activity (≥10 %). The specificity of the HRN assay was 0.522. Moreover, HRN activities in sera were largely correlated with degrees of histamine release from peripheral blood basophils of the same donors in response to sweat antigen. To identify the substance that neutralizes histamine-release activity, we removed IgE and IgG from the sera of HRN (+) subjects by column chromatography. The HRN activities in 30 out of 42 sera were largely reduced by the removal of IgG. On the other hand, sera of four subjects lost HRN activity by the removal of IgE, suggesting that the majority of HRN (+) subjects have serum IgG against the sweat antigen as well as IgE bound to peripheral basophils. Thus, the HRN assay maybe useful for the screening of type I allergy against sweat antigen.


Redesigning immunization supply chains: Results from three country analyses.
Immunization supply chains (iSC) are essential for ensuring access to vaccines that prevent diseases. Guinea, Madagascar, and Niger initiated iSC system design efforts to conduct analysis of alternative supply chain scenarios to identify areas for improvement. Key stakeholders from Ministries of Health and immunization programs identified bottlenecks in the current iSC and prioritized five general design scenarios to model in each country. Scenarios included aspects of integration, changing supply chain levels and delivery frequency, ignoring administrative boundaries, and direct delivery. Primary and secondary data were collected and cleaned. Analysis was completed using Supply Chain Guru (Madagascar and Niger) and AnyLogistix (Guinea) modeling software to build a virtual representation of the iSC physical components and operating policies. Modeling results were compared using both quantitative and qualitative criteria (total operating costs, cost per dose, cold chain capacity, risk of mishandling, logistics burden on healthcare workers, feasibility to implement, and equity). Using a Stop Light Analysis for user-friendly understanding of positive, negative or minimal impact on scenarios, cost savings were realized in most scenarios in Madagascar (except using autonomous aerial vehicles); proposed scenarios in Guinea increased costs or had minimal impact; in Niger, eliminating regional tiers reduced costs. Facility level cold chain was sufficient in all countries. Effect on qualitative indicators largely depended on the scenario and country context. Similarities in scenarios selected were seen across the three countries. Results suggest that efficiencies can be found through changes to the iSC design, but the benefits of each scenario must be considered in the country context. Results of the analysis do not provide "the right answer" but rather options and guidance which then must be grounded in the country context and used as evidence for decision making to ensure reliable availability of vaccines.


Large area arrays of discrete single-molecule junctions derived from host-guest complexes.
The desire to continually reduce the lower limits of semiconductor integrated circuit (IC) fabrication methods continues to inspire interest in unimolecular electronics as a platform technology for the realization of future (opto)electronic devices. However, despite successes in developing methods for the construction and measurement of single-molecule and large-area molecular junctions, exercising control over the precise junction geometry remains a significant challenge. Here, host-guest complexes of the wire-like viologen derivative 1,1'-bis(4-(methylthio)-phenyl)-[4,4'-bipyridine]-1,1'-diium chloride ([1][Cl]2) and cucurbit[7]uril (CB[7]) have been self-assembled in a regular pattern over a gold substrate. Subsequently, ligandless gold nanoparticles (AuNPs) synthesized in situ are deposited over the host-guest array. The agreement between the conductance of individual mono-molecular junctions, appropriately chosen as a function of the AuNP diameter, within this array determined by conductive probe atomic force microscope (c-AFM) and true single-molecule measurements for a closely similar host-guest complex within a scanning tunneling microscope break-junction (STM-BJ) indicates the formation of molecular junctions derived from these host-guest complexes without deleterious intermolecular coupling effects.


Detection of Microcrystals for CryoEM.
Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The use of negative stain TEM allows visualization along the process and thus enables optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nanocrystals bearing perfect lattices are seeded and can be used for MicroED as well as growing larger crystals for X-ray and free electron laser (FEL) data collection.


Nile Red Incubation Time Before Reading Fluorescence Greatly Influences the Yeast Neutral Lipids Quantification.
High-throughput screening methodologies to estimate lipid content in oleaginous yeasts use Nile red fluorescence in a given solvent and optimized excitation/emission wavelengths. However, Nile red fluorescence stabilization has been poorly analyzed, and high variability occurs when relative fluorescence is measured immediately or a few minutes after dye addition. The aim of this work was to analyze the fluorescence of Nile red at different incubation times using a variety of solvents and oleaginous/non-oleaginous yeast strains. We showed that fluorescence stabilization occurs between 20 and 30 min, depending on the strain and solvent. Therefore, we suggest that fluorescence measurements should be followed until stabilization, where Relative Fluorescence Units should be considered after stabilization for lipid content estimation.


[Active pulmonary tuberculosis in the community. Current presentation].
To discover how patients in a provincial general hospital, diagnosed as having pulmonary tuberculosis had first clinically and radiologically presented. This was a retrospective observational study. SITE. The San Jorge de Huesca General Hospital. The period of study was between January, 1986, and December, 1989. Those studied were the 59 patients diagnosed as having active pulmonary tuberculosis during the study period. The average age of the patients was 45.6 +/- 19.25 SD. There was greater occurrence in males. The most common form in which the disease first appeared was thoracic symptomatology (59%), followed by constitutional syndrome (45.7%). 8 patients were addicted to drugs taken parenterally (ADVP), of whom 4 were HIV-positive (6.8%). The most common radiological pattern was ulcero-caseous. We underline the importance of diagnostical awareness at the Primary Care level in order to rapidly begin the correct treatment; and thus fight the adverse epidemiological situation caused by tuberculosis today.


Super-resolution biomolecular crystallography with low-resolution data.
X-ray diffraction plays a pivotal role in the understanding of biological systems by revealing atomic structures of proteins, nucleic acids and their complexes, with much recent interest in very large assemblies like the ribosome. As crystals of such large assemblies often diffract weakly (resolution worse than 4 A), we need methods that work at such low resolution. In macromolecular assemblies, some of the components may be known at high resolution, whereas others are unknown: current refinement methods fail as they require a high-resolution starting structure for the entire complex. Determining the structure of such complexes, which are often of key biological importance, should be possible in principle as the number of independent diffraction intensities at a resolution better than 5 A generally exceeds the number of degrees of freedom. Here we introduce a method that adds specific information from known homologous structures but allows global and local deformations of these homology models. Our approach uses the observation that local protein structure tends to be conserved as sequence and function evolve. Cross-validation with R(free) (the free R-factor) determines the optimum deformation and influence of the homology model. For test cases at 3.5-5 A resolution with known structures at high resolution, our method gives significant improvements over conventional refinement in the model as monitored by coordinate accuracy, the definition of secondary structure and the quality of electron density maps. For re-refinements of a representative set of 19 low-resolution crystal structures from the Protein Data Bank, we find similar improvements. Thus, a structure derived from low-resolution diffraction data can have quality similar to a high-resolution structure. Our method is applicable to the study of weakly diffracting crystals using X-ray micro-diffraction as well as data from new X-ray light sources. Use of homology information is not restricted to X-ray crystallography and cryo-electron microscopy: as optical imaging advances to subnanometre resolution, it can use similar tools.


Analysis of a Borrelia burgdorferi phosphodiesterase demonstrates a role for cyclic-di-guanosine monophosphate in motility and virulence.
The genome of Borrelia burgdorferi encodes a set of genes putatively involved in cyclic-dimeric guanosine monophosphate (cyclic-di-GMP) metabolism. Although BB0419 was shown to be a diguanylate cyclase, the extent to which bb0419 or any of the putative cyclic-di-GMP metabolizing genes impact B. burgdorferi motility and pathogenesis has not yet been reported. Here we identify and characterize a phosphodiesterase (BB0363). BB0363 specifically hydrolyzed cyclic-di-GMP with a K(m) of 0.054 microM, confirming it is a functional cyclic-di-GMP phosphodiesterase. A targeted mutation in bb0363 was constructed using a newly developed promoterless antibiotic cassette that does not affect downstream gene expression. The mutant cells exhibited an altered swimming pattern, indicating a function for cyclic-di-GMP in regulating B. burgdorferi motility. Furthermore, the bb0363 mutant cells were not infectious in mice, demonstrating an important role for cyclic-di-GMP in B. burgdorferi infection. The mutant cells were able to survive within Ixodes scapularis ticks after a blood meal from naïve mice; however, ticks infected with the mutant cells were not able to infect naïve mice. Both motility and infection phenotypes were restored upon genetic complementation. These results reveal an important connection between cyclic-di-GMP, B. burgdorferi motility and Lyme disease pathogenesis. A mechanism by which cyclic-di-GMP influences motility and infection is proposed.


Morphological, biological, and genomic characterization of a newly isolated lytic phage Sfk20 infecting Shigella flexneri, Shigella sonnei, and Shigella dysenteriae1.
Shigellosis, caused by Shigella bacterial spp., is one of the leading causes of diarrheal morbidity and mortality. An increasing prevalence of multidrug-resistant Shigella species has revived the importance of bacteriophages as an alternative therapy to antibiotics. In this study, a novel bacteriophage, Sfk20, has been isolated from water bodies of a diarrheal outbreak area in Kolkata (India) with lytic activity against many Shigella spp. Phage Sfk20 showed a latent period of 20 min and a large burst size of 123 pfu per infected cell in a one-step growth analysis. Phage-host interaction and lytic activity confirmed by phage attachment, intracellular phage development, and bacterial cell burst using ultrathin sectioning and TEM analysis. The genomic analysis revealed that the double-stranded DNA genome of Sfk20 contains 164,878 bp with 35.62% G + C content and 241 ORFs. Results suggested phage Sfk20 to include as a member of the T4 myoviridae bacteriophage group. Phage Sfk20 has shown anti-biofilm potential against Shigella species. The results of this study imply that Sfk20 has good possibilities to be used as a biocontrol agent.


Interim estimates in null models of COVID-19 vaccine effectiveness.
Recently released interim numbers from advanced vaccine candidate clinical trials suggest that a COVID-19 vaccine effectiveness (VE) of >90% is achievable. However, SARS-CoV-2 transmission dynamics are highly heterogeneous and exhibit localized bursts of transmission, which may lead to sharp localized peaks in the number of new cases, often followed by longer periods of low incidence. Here we show that, for interim estimates of VE, these characteristic bursts in SARS-CoV-2 infection may introduce a strong positive bias in VE. Specifically, we generate null models of vaccine effectiveness, i.e., random models with bursts that over longer periods converge to zero VE but that for interim periods frequently produce apparent VE near 100%. As an example, by following the relevant clinical trial protocol, we can reproduce recently reported interim outcomes from an ongoing phase 3 clinical trial of an RNA-based vaccine candidate. Thus, to avoid potential random biases in VE, it is suggested that interim estimates on COVID-19 VE should control for the intrinsic inhomogeneity in both SARS-CoV-2 infection dynamics and reported cases.


Large molecular fluorescence enhancement by a nanoaperture with plasmonic corrugations.
We investigate the influence of circular corrugations surrounding a central nanoaperture to further enhance the fluorescence count rate per emitter and control its emission directionality. Adding a single corrugation already allows to significantly increase the fluorescence signal as compared to a bare nanoaperture. A complete fluorescence characterization quantifies the excitation and emission gains contributing to the fluorescence enhancement process as the number of corrugations is increased.