Diagnostic Capacity for Invasive Fungal Infections in the Greek Paediatric Haematology-Oncology Units: Report from the Infection Working Group of the Hellenic Society of Paediatric Haematology-Oncology.
An audit based on a specific questionnaire was attempted, in order to investigate the mycology laboratory diagnostic capacity for invasive fungal diseases (IFDs) in Greek Paediatric Haematology-Oncology departments/units. The study provided the relevant information for the years 2019 and 2020 and included data from all units, concerning culture-based methods and direct microscopy, phenotypic and molecular identification, sensitivity testing, serology and molecular diagnosis, as well as therapeutic drug monitoring. The target was mostly to reveal the level of laboratory coverage for hospitalised paediatric patients, independently of the possibility of performing the tests in the host hospital, or otherwise to refer the specimens elsewhere. In total, the current study demonstrated that the most important facilities and services regarding the IFD diagnostics for paediatric haematology-oncology patients in Greece are available and relatively easily accessible, with a reasonable turnaround time. Acting as an initial registry for further improvements, the audit can serve as a valuable approach to the actual situation and future perspectives. A national clinical mycology network under the auspices of the relevant scientific societies will probably facilitate collaboration between all the departments (clinical and laboratory) involved in invasive fungal infections and provide an easier approach to any necessary test for any hospitalised patient.


Clearance of viscoelastic mucus simulant with airflow in a rectangular channel, an experimental study.
Interaction of mucus simulant with airflow in a rectangular channel is investigated experimentally. Two different viscoelastic gel mucus simulants are prepared by cross linking Borax with Locust Bean Gum (LBG) solution; liquid-like (LM) with lower storage modulus and semi-solid (SM). The rheological difference between LM and SM represent the qualitative change from liquid-like healthy mucus to the one with higher storage modulus found in a person with lung disease. The study concentrates on the effect of viscoelastic layer thickness and rheology on the wave formation and clearance due to its interaction with airflow. The results indicate that the onset air velocity for wave initiation reduces by increasing layer thickness. This effect is more pronounced for SM. Slowly propagating waves initiate at a lower air velocity for LM compared to SM for thinner layer thickness and this behavior reverses for a thicker layer. Although SM clears at a critical air velocity, LM does not show clearance behavior, defined as separation of layer section from rest and movement in the downstream flow direction. This seems to suggest that thicker mucus with higher elastic modulus, similar to the mucus for a person with lung disease, may clear easier with a two-phase air-liquid flow, as in cough.


SPL8, a local regulator in a subset of gibberellin-mediated developmental processes in Arabidopsis.
Gibberellins (GAs) are important plant growth regulators, regulating many plant developmental processes, including seed germination, root and stem elongation, rosette expansion, floral induction and anther development. The diverse effects of GAs on plant development make it critical to maintain an appropriate endogenous GA level and a fine-tuned GA signalling. Some global regulators in GA signalling have been identified but little is known about genes specifically involved in local implementation of GA signalling. Here we report that the Arabidopsis thaliana SBP-box gene SQUAMOSA-PROMOTER-BINDING-PROTEIN-LIKE8 (SPL8) acts as a local regulator in a subset of GA-dependent developmental processes. Previous knowledge holds that SPL8 is involved in reproductive development as deduced from its loss-of-function phenotype (Unte et al. (2003) Plant Cell 15:1009-1019). We now determined that constitutive overexpression of SPL8 affects fertility due to non-dehiscent anthers, likely resulting from a constitutive GA response, suggesting a positive role of SPL8 in GA-mediated anther development. On the other hand, SPL8 gain- and loss-of-function mutants showed opposite responses to GA and its biosynthetic inhibitor paclobutrazol (PAC) with respect to seed germination and root elongation during the seedling stage. Genes involved in GA biosynthesis and signalling are transcriptionally affected by altered SPL8 expression. Our study uncovered a tissue-dependent regulatory role for SPL8 in the response to GA signalling in plant development.


Dispensability of zinc and the putative zinc-binding domain in bacterial glutamyl-tRNA synthetase.
The putative zinc-binding domain (pZBD) in Escherichia coli glutamyl-tRNA synthetase (GluRS) is known to correctly position the tRNA acceptor arm and modulate the amino acid-binding site. However, its functional role in other bacterial species is not clear since many bacterial GluRSs lack a zinc-binding motif in the pZBD. From experimental studies on pZBD-swapped E. coli GluRS, with Thermosynechoccus elongatus GluRS, Burkholderia thailandensis GluRS and E. coli glutamyl-queuosine-tRNA(Asp) synthetase (Glu-Q-RS), we show that E. coli GluRS, containing the zinc-free pZBD of B. thailandensis, is as functional as the zinc-bound wild-type E. coli GluRS, whereas the other constructs, all zinc-bound, show impaired function. A pZBD-tinkered version of E. coli GluRS that still retained Zn-binding capacity, also showed reduced activity. This suggests that zinc is not essential for the pZBD to be functional. From extensive structural and sequence analyses from whole genome database of bacterial GluRS, we further show that in addition to many bacterial GluRS lacking a zinc-binding motif, the pZBD is actually deleted in some bacteria, all containing either glutaminyl-tRNA synthetase (GlnRS) or a second copy of GluRS (GluRS2). Correlation between the absence of pZBD and the occurrence of glutamine amidotransferase CAB (GatCAB) in the genome suggests that the primordial role of the pZBD was to facilitate transamidation of misacylated Glu-tRNA(Gln) via interaction with GatCAB, whereas its role in tRNA(Glu) interaction may be a consequence of the presence of pZBD.


Rehospitalisation for neonatal jaundice: risk factors and outcomes.
This case-control study sought to determine whether rehospitalisation for jaundice in newborns is associated with the length of hospital stay after birth and to identify risk factors for and outcomes of rehospitalisation for jaundice. It was carried out among women who delivered a normal, term infant vaginally at any of 10 medical centres from 1992 to 1994. Cases were infants rehospitalised with jaundice within 14 days of birth. Controls were randomly selected from normal, term infants delivered vaginally but not rehospitalised within 90 days of birth. Maternal medical records for pregnancy, labour and delivery care, records for all the birth hospitalisations, and rehospitalisations for the cases were abstracted. The length of birth hospitalisation did not differ between case and control infants, whether length was measured as a categorical variable or as a continuous measure (median = 22.8 h for cases and 23.3 h for controls, P = 0.931). Rehospitalisation for jaundice was associated with race/ethnicity, primiparity, preterm birth, breast feeding and suspicion of jaundice during the birth hospitalisation. None of the rehospitalised infants died, were diagnosed with kernicterus or required resuscitation. Attention to risks associated with jaundice rehospitalisation might reduce this undesirable, but not commonly severe, outcome.


Reorganization of perylene bisimide J-aggregates: from delocalized collective to localized individual excitations.
Water-induced reorganization of individual one-dimensional J-aggregates of perylene bisimide (PBI) dyes was observed by fluorescence microscopy. Fluorescence spectra and decay kinetics of individual J-aggregates immobilized on glass surfaces were measured under a dry nitrogen atmosphere and under humid conditions. The fluorescence properties of PBI J-aggregates arisen from collective excitons under dry nitrogen atmosphere were changed to those of non-interacting dye monomers when water vapor was introduced into the environment (sample chamber). Time-dependent changes of the fluorescence spectra and lifetimes upon exposure to water vapor suggest an initial coordination of water molecules at defect sites leading to the formation of H-type dimer units that act as exciton quenchers, and a subsequent slower disintegration of the hydrogen-bonded J-aggregate into monomers that lack resonance coupling. Our present studies resulted in a direct demonstration of how drastically the optical properties of molecular ensembles and characteristics of their excited states can be changed by delicate reorganization of dye molecules at nanometre scales.


5-hydroxytryptamine stimulates phosphorylation of p44/p42 mitogen-activated protein kinase activation in bovine aortic endothelial cell cultures.
5-Hydroxytryptamine (5-HT) is sequestered and released by endothelial cells, acts as an endothelial cell mitogen, promotes the release of nitric oxide (NO), and has been associated with the p44/p42 mitogen-activated protein kinase (MAPK) cascade. NO also acts as a cell mitogen and promotes signals that culminate in the phosphorylation of MAPK. The aim of this study was to test whether endothelial 5-HT receptors stimulate dual (tyrosyl- and threonyl-) phosphorylation of MAPK through a mitogen-activated protein kinase kinase-1 (MEK-1) and eNOS-dependent pathway in bovine aortic endothelial cells (BAECs). As shown by Western blot analysis, 5-HT and the 5-HT1B-selective agonist 5-nonyloxytryptamine (5-NOT) stimulate time- and concentration-dependent (0.001-10 microM) phosphorylation of MAPK in these cells. The agonist-stimulated phosphorylation of MAPK was blocked by the 5-HT1b-receptor antagonist isamoltane (0.01-10 p3M) and the MEK-1 inhibitor PD 098059 ([2-(2'-amino-3'-methoxy-phenyl)-oxanaphthalen-4-one]; 0.01-10 microM¿. The eNOS inhibitor L-N(omega)-iminoethyl-L-ornithine (L-NIO; 0.01-10 microM) failed to block the 1 microM 5-NOT-stimulated responses. Our findings suggest that the 5-HT receptors (specifically 5-HT1B) mediate signals to MEK-1 and subsequently to MAPK through an eNOS-independent pathway in BAECs.


[Research on acting mechanism of rhubarb on aquaporin-4 in rats with blood-brain barrier injury after acute cerebral hemorrhage].
To investigate the mechanism of rhubarb in regulating aquaporin-4 in rats with blood-brain barrier damage after acute cerebral hemorrhage (CH). CH model was induced by stereospecific injection of auto-blood into caudate nucleus of rats, and the brain water content and neurological defect were detected to evaluate cerebral edema and neurological defect level. Also, the blood-brain barrier damage was observed by Evan's blue staining; the changes of blood-brain barrier tight junction and astrocyte end feet at different time points were observed with electron microscope; and the AQP-4 mRNA and protein expression were measured with RT-PCR and Western blot. Rhubarb showed effects in reducing cerebral edema. Evan's blue result indicated the blood-brain barrier was evidently damaged at the 12th hour after CH, with blood-brain barrier tight junction damaged and astrocyte end feet process swelled obviously, but these changes could be relieved by rhubarb. The AQP-4 mRNA and protein expression in rats increased significantly 24 hrs after modeling (P < 0.05) and reached the peak value at 72 hrs, and decreased gradually after then. Rhubarb also showed inhibiting transcription and translation of AQP-4 gene. Rhubarb could alleviate cerebral edema via reducing blood-brain barrier tight junction damage and astrocyte end feet process swelling, which might be realized by the inhibition on transcription and translation of AQP-4 gene.


Morphometric study of the korean adult pituitary glands and the diaphragma sellae.
To investigate the morphometric characteristics of the pituitary gland and diaphragma sellae in Korean adults. Using the 33 formaline fixed adult cadavers (23 male, 10 female), the measurements were taken at the diaphragma sellae and pituitary gland. The authors investigated the relationship between dura and structures surrounding pituitary gland, morphometric aspects of pituitary gland and stalk, and morphometric aspect of central opening of diaphragma sellae. The boundary between the lateral surface of pituitary gland and the medial wall of cavernous sinus was formed by the thin dural layer and pituitary capsule. The pituitary capsule adherent tightly to the pituitary gland was observed to continue from the diaphragma sellae. Mean width, length, and height of the pituitary gland were 14.3 +/- 2.1, 7.9 +/- 1.3, and 6.0 +/- 0.9 mm in anterior lobes, and 8.7 +/- 1.7, 2.9 +/- 1.1, and 5.8 +/- 1.0 mm in posterior lobes, respectively. Although all dimensions of anterior lobe in female were slightly larger than those in male, statistical significance was noted in only longitudinal dimension. The ratio of posterior lobe to the whole length of pituitary gland was about 27%. The mean thickness of pituitary stalk was 2 mm. The diaphragmal opening was 5 mm or more in 26 (78.8%) of 33 specimen. The opening was round in 60.6% of the specimen, and elliptical oriented in an anterior-posterior or transverse direction in 39.4%. These results provide the safe anatomical knowledge during the transsphenoidal surgery and may be helpful to access the possibility of the development of empty sella syndrome.


Hyperendemicity of onchocerciasis in ovia northeast local government area, edo state, Nigeria.
Onchocerciasis is a chronic parasitic infection caused by the filarial nematode, Onchocerca volvulus. The objective of this study was to determine the prevalence, endemicity, and symptomatic effects of the disease in Ovia Northeast Local Government Area. The prevalence of onchocerciasis was investigated in Ovia Northeast Local Government Area of Edo State, Nigeria, between March 2008 and June 2009 using the standard skin-snip method. A total of 2020 subjects, who had visited various primary health centres located in each community, were enlisted using randomised sampling, and the data were analysed using the Chi-squared (χ(2)) test and logistic regression. A Of the 2020 individuals examined, 1674 (83%) harboured microfilaria in their skin tissues. On the basis of the standardised scale for microfilaria prevalence-less than 10% is considered sporadic, 10%-29% is considered hypoendemic, 30%-59% is considered mesoendemic, and 60% and above is considered hyperendemic-the prevalence (83%) reported in this study led to the disease being classified as hyperendemic. Females were more frequently infected than were males, and this was statistically significant (P < 0.001). Prevalence was also found to increase with age, and this correlation was significant (P < 0.001). The prevalence of the clinical features of the disease in the local government area was 87.5% for leopard skin, 84.16% for itching, and 75.42% for nodules. A prevalence of 83% was observed and considered hyperendemic. Female gender and age (50 years or more) were significant risk factors that affected the prevalence of onchocerciasis. The findings demonstrated the hyperendemicity of infection and the need for urgent attention with ivermectin treatment and other control measures.


Drug-induced alterations in tumour perfusion yield increases in tumour cell radiosensitivity.
The perfusion of human tumour xenografts was manipulated by administration of diltiazem and pentoxifylline, and the extent that observed changes in tumour perfusion altered tumour radiosensitivity was determined. 2 tumour systems having intrinsically different types of hypoxia were studied. The responses of SiHa tumours, which have essentially no transient hypoxia, were compared to the responses of WiDr tumours, which contain chronically and transiently hypoxic cells. We found that relatively modest increases in net tumour perfusion increased tumour cell radiosensitivity in WiDr tumours to a greater extent than in SiHa tumours. Moreover, redistribution of blood flow within WiDr tumours was observed on a micro-regional level that was largely independent of changes in net tumour perfusion. Through fluorescence-activated cell sorting coupled with an in vivo-in vitro cloning assay, increases in the radiosensitivity of WiDr tumour cells at intermediate levels of oxygenation were observed, consistent with the expectation that a redistribution of tumour blood flow had increased oxygen delivery to transiently hypoxic tumour cells. Our data therefore suggest that drug-induced changes in tumour micro-perfusion can alter the radiosensitivity of transiently hypoxic tumour cells, and that increasing the radiosensitivity of tumour cells at intermediate levels of oxygenation is therapeutically relevant.


Nystatin and valinomycin induce tubuloglomerular feedback.
The macula densa expresses a luminal Na(+)-K(+)-2Cl(-) cotransporter and a basolateral Cl(-) conductance. Although it is known that cotransport of Na(+), K(+), and Cl(-) is the first step in tubuloglomerular feedback (TGF), subsequent steps are unclear. We hypothesized that Na(+)-K(+)-2Cl(-) entry via the luminal Na(+)-K(+)-2Cl(-) cotransporter elevates intracellular Cl(-), increases electrogenic Cl(-) efflux across the basolateral membrane, and depolarizes the macula densa, initiating TGF. We perfused afferent arterioles with macula densa attached. The macula densa was perfused with solutions containing either 5 mM Na(+) and 3 mM Cl(-) (low NaCl) or 80 mM Na(+) and 77 mM Cl(-) (high NaCl). When the macula densa perfusate was changed from low to high NaCl, afferent arteriole diameter decreased from 15.8 +/- 0.8 to 13.1 +/- 0.7 mm (P < 0.05). Adding 10 microM furosemide to the macula densa lumen blocked TGF. When nystatin, a group I cation ionophore, was added to the macula densa lumen together with furosemide in the presence of low NaCl, it induced TGF (from 18.0 +/- 1.5 to 15.6 +/- 1.6 mm; P = 0.003). When valinomycin, a K(+)-selective ionophore, was added to the macula densa lumen together with furosemide in the presence of low NaCl containing 5 mM K(+), it did not induce TGF. Subsequent addition of 50 mM KCl to the macula densa perfusate induced TGF (from 21.7 +/- 0.8 to 17.5 +/- 1.3 mm; P = 0.0047; n = 6). Adding 50 mM KCl without valinomycin did not induce TGF. When 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB; 1 microM), a Cl(-) channel blocker, was added to the bath, it blocked TGF induced by high NaCl, but did not block TGF induced by valinomycin plus 50 mM KCl. NPPB did not alter afferent arteriole constriction induced by norepinephrine. We concluded that increased NaCl in the lumen of the macula densa leads to influx of Cl(-) via the Na(+)-K(+)-2Cl(-) cotransporter. The accelerated transport increases intracellular Cl(-). The subsequent exit of Cl(-) across the basolateral membrane via Cl( -) channels in turn leads to depolarization of the macula densa and thereby induces TGF.


Middle ear cholesteatoma treated with a mastoidectomy: a systematic review of the measures used.
The inconsistent measures used to report outcomes after mastoidectomy to treat cholesteatoma make it impossible for clinicians to compare results and apply them appropriately to patient care. We sought to identify and assess the type and relative frequency of the reported measures. PubMed, EMBASE, the Web of Science. We searched 3 independent databases for articles that reported outcomes of middle ear cholesteatoma treated with mastoidectomy. Articles were assessed for eligibility using Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) protocol and data were extracted by 2 independent investigators. Observational studies and randomized controlled trials from the past 30 years were included. Articles with less than 50 subjects and nonsurgical studies were excluded. The measures used by each study were recorded, and the relative frequency of each measure was calculated. Forty-three of 380 articles met criteria. Time to follow up was inconsistent. Most articles reported on preoperative qualitative parameters (77%); however, few reported on the severity of disease (19%) or formally staged the disease (12%). Not all studies reported on the presence of recurrent or residual disease (88%) or the postoperative audiogram results (70%). Only 12% presented results as a Kaplan-Meier disease-free curve. Most studies reported on complications (70%). Reported measures were inconsistent among the studies, which makes comparisons between studies unreliable. Standardization will optimize future reporting and will allow for the establishment of best practices.


Effects of 1,25-dihydroxycholecalciferol and phytase on zinc utilization in broiler chicks.
Studies were conducted with corn-soybean meal diets to evaluate the effects of phytate phosphorus utilization on zinc absorption and retention in broiler chicks. In the first two experiments, zinc-65 was used to determine zinc absorption. Experiment 1 was a 2 x 2 factorial with 0 or 5 micrograms/kg dihydroxycholecalciferol and 0 or 40 ppm supplemental zinc. In Experiment 2, 5 micrograms/kg 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] or 750 units/kg phytase or both were added to a diet containing 35 ppm zinc. The diets in Experiment 3 were similar to Experiment 2 except that 600 units/kg phytase was fed. Experiment 4 was similar to Experiment 3 except that dietary phosphorus was decreased by .15%. There were no treatment effects on body weight in Experiments 1 and 2. Zinc absorption was higher in zinc-deficient birds in Experiment 1, but there were no other effects on zinc-65 absorption or retention. Body weight was increased by 1,25-(OH)2D3 in Experiments 3 and 4 and by phytase in Experiment 4. Phytate phosphorus retention was increased by phytase and 1,25-(OH)2D3 and was increased additively when both sources were fed. Dietary 1,25-(OH)2D3 increased zinc retention at times during Experiments 3 and 4, but this response was inconsistent. Phytase did not affect zinc retention. Phytase plus 1,25-(OH)2D3 increased zinc retention synergistically in Experiment 3. Bone zinc was increased by 1,25-(OH)2D3 and phytase, and there was an additive effect in Experiment 3. Plasma zinc and alkaline phosphatase were not affected. The results suggest that supplemental zinc may be decreased in a corn-soybean meal diet when phytate phosphorus utilization is enhanced.


A phase I/II study of nevirapine for pre-exposure prophylaxis of HIV-1 transmission in uninfected subjects at high risk.
To evaluate the safety, tolerability, and trough levels of three pre-exposure prophylaxis regimens of nevirapine among HIV-1-uninfected subjects at high risk for HIV-1 infection. A phase I/II trial (HIVHOP 101) in which 33 such uninfected subjects received a 200 mg tablet of nevirapine once weekly (cohort A, n = 12), twice weekly (cohort B, n = 12), or every other day (cohort C, n = 9) for 12 weeks. Clinical signs/symptoms, laboratory parameters, and nevirapine trough levels were assessed at entry and at 1, 2, 4, 6, 9, and 12 weeks, with a follow-up sample at 16 weeks. No subject experienced clinical symptoms attributed to nevirapine, including rash. There were no significant changes in liver enzyme levels from baseline to week 12 in the three cohorts, except for glutamyl transpeptidase in cohort B. Median nevirapine trough levels at weeks 1 and 12 were 119 ng/ml (range, < 25-205) and 135 ng/ml (range, < 25-1065), respectively, for cohort A, 569 ng/ml (range, 135-2641) and 431 ng/ml (range, 42-2454) for cohort B, and 1942 ng/ml (range, 1214-2482) and 943 ng/ml (range, 262-5281) for cohort C. No subject became HIV-1 antibody positive by week 16. A single dose of nevirapine taken once weekly, twice weekly, or every other day for 12 weeks was safely tolerated by the subjects in this small study, and resulted in nevirapine levels well above the IC (inhibitory concentration of 50%: 10 ng/ml) over the 12-week period in nearly all evaluable subjects. (50)


Delayed subaponeurotic fluid collection on an infant's head: Underreported case and review of the literature.
Delayed subaponeurotic fluid collection (DSFC) is a relatively uncommon problem, probably under reported soft swelling in the scalp which usually develops in infancy that occurs weeks to months after birth. Although the exact etiology remains unclear, several theories have been postulated such as (i) cerebrospinal fluids (CSF) leak from microfractures of the skull and (ii) disrupted lymphatic drainage, gradually liquefying subaponeurotic bleeding. Here, we reported typical clinical findings of DSFC and analysis of the fluid aspirate from our patient. To the best of our knowledge, this is the first case reported from Asia, particularly from Indonesia. A healthy 2-month-old girl infant presented with 2 weeks history of occipital painless fluctuant scalp mass with no swelling. She was born at term from a nulliparous mother; by emergency cesarean delivery following failed induction of labor. There was no history of scalp injury at birth nor recent head trauma; ultrasonography showed translucent fluid in subaponeurotic or subgaleal space. The fluid collection was noted to be fluctuant, free-flowing across suture lines, without discoloration or bruising; when placed supine, the fluid collected at her midocciput. A diagnostic tap confirmed the presence of serosanguinous CSF led to a diagnosis of DSFC. Herein, we reported the first DSFC case from Indonesia. With no previous experience of the condition, a definitive diagnosis was possible through a very carefully physical examination and history taking, along with a good communication among the neurosurgeon, pediatrician, and the clinical pathologist.


A comparison of tests for antifibrillatory action.
THE OBJECT OF THE EXPERIMENTS WAS TWOFOLD: first, to choose the most satisfactory test for antifibrillatory action; secondly, to place several drugs with reputed antifibrillatory activity in an order of potency as a preliminary to investigating their mode of action. Measurements were made on isolated rabbit atria at 34 degrees C of (1) the maximum driving frequency the atria would follow, (2) conduction velocity, (3) contractions, and of the threshold for the production of (4) extrasystoles, (5) flutter and (6) fibrillation. Log dose-response curves were plotted for quinidine, papaverine, procaine, dibenamine and procaine amide. The maximum frequency test and fibrillation threshold test gave similar results with all the drugs, and the results gave the order quinidine 1.0, procaine 0.53, Dibenamine 0.47, papaverine 0.43 and procainamide 0.26. Thresholds for extrasystoles and flutter were much more variable. The regressions relating changes in conduction velocity and contraction to log dose were different from those for maximum driving frequency and fibrillation threshold for procaine, papaverine and dibenamine, but the regressions for quinidine and procaine amide were nearly parallel in all tests. Serpajmaline contained a substance with antifibrillatory activity as great as that of quinidine and with no greater depressant action on contractions.


Irritability, Externalizing, and Internalizing Psychopathology in Adolescence: Cross-Sectional and Longitudinal Associations and Moderation by Sex.
Irritability is a common feature of many psychiatric disorders, including both externalizing and internalizing disorders. There is little research, however, examining associations between irritability and these symptom domains, particularly during the important developmental period of adolescence, characterized by sex differences in the prevalence of disorders. We examined the cross-sectional associations between irritability, measured with the Affective Reactivity Index, and symptoms of externalizing and internalizing domains of psychopathology, measured with the Youth Self Report, in a volunteer community sample (N = 183) of 9- to 13-year-old (M = 11.39, SD = 1.07) boys and girls (37% White/Caucasian, 8% Asian, 11% Hispanic, 8% African American, 2% Native American, 2% Pacific Islander, 28% Other, and 3% not reported). A subset of the sample (n = 112) provided data at a 2-year follow-up, used to extend these associations. There were no sex differences in levels of irritability; however, the associations between irritability and symptom domains were moderated by sex. Specifically, in girls, irritability was associated equally with externalizing and internalizing symptoms. In contrast, in boys, irritability was associated more strongly with externalizing symptoms than with internalizing symptoms. Thus, across both sexes, irritability was moderately associated with externalizing symptoms, but the association between irritability and internalizing symptoms was stronger in girls than in boys. At follow-up, sex moderated the association between baseline irritability and later externalizing and internalizing symptoms. These findings indicate that irritability is associated with both externalizing and internalizing symptoms in early adolescence and that irritability is associated with internalizing symptoms more strongly in girls than in boys.


A unified strategy for the asymmetric total syntheses of diversonol and lachnone C.
A unified synthetic strategy for the asymmetric syntheses of the natural products diversonol and lachnone C was developed by using the domino vinylogous aldol-oxa-Michael reaction as the enantioselective key step. Further transformations include dihydroxylation, lactol-opening by a Wittig-reaction, and lactonization. The obtained chromone lactones, a class of mycotoxins, can further be converted to tetrahydroxanthones by a Dieckmann condensation. This general method allows for the first time the enantioselective access to these classes of natural products and should be applicable to other members of the tetrahydroxanthone and chromone lactone families.


[Palliative treatment of esophageal carcinoma].
The choice of the most beneficial method of the treatment of patients with esophageal cancer still remains a matter of debate. According majority of epidemiological reports only 10% of esophageal cancer patients has a chance for 5-year survival. Unfortunately, the curative surgical treatment is possible in relatively small number of patients, nevertheless esophagectomy is regarded as a principle method of treatment. Usually it referrers to the group of 50% of patients with locally advanced cancer. Thus, the management of the majority of esophageal cancer patients is palliative. Its main objective is to improve esophageal passage compromised by tumor narrowing esophageal lumen. Maintenance of esophageal passage facilitates food intake and usually postpones the stage of terminal cachexia and thus results in quality of life improvement. As it has been evident from up-to-date research currently used methods of palliative treatment do not significantly improve survival. In this paper we demonstrate a review of currently available methods of palliative treatment of patients with esophageal cancer and brief discussion on our own experience from last four years.


Immunomodulation of transgene responses following naked DNA transfer of human factor VIII into hemophilia A mice.
A robust humoral immune response against human factor VIII (hFVIII) following naked DNA transfer into immunocompetent hemophilia A mice completely inhibits circulating FVIII activity despite initial high-level hFVIII gene expression. To prevent this undesirable response, we compared transient immunomodulation strategies. Eight groups of mice (n = 4-9 per group) were treated with naked DNA transfer of pBS-HCRHPI-hFVIIIA simultaneously with immunosuppressive reagents that included cyclosporine A (CSA), rapamycin (RAP), mycophenylate mofetil (MMF), a combination of CSA and MMF, a combination of RAP and MMF, a monoclonal antibody against murine CD40 ligand (MR1), recombinant murine Ctla4Ig, and a combination of MR1 and Ctla4Ig. All animals except those receiving only CSA exhibited delayed or absent immune responses against hFVIII. The most effective immunosuppressive regimen, the combination of Ctla4Ig and MR1, prevented inhibitor formation in 8 of 9 animals; the ninth had transient low-titer antibodies. All 9 mice of this group produced persistent, therapeutic levels of hFVIII for more than 6 months. When challenged with the T-dependent antigen bacteriophage Phix174, tolerized mice exhibited normal primary and secondary antibody responses, suggesting that transient immunomodulation to disrupt B/T-cell interaction at the time of plasmid injection effectively promoted long-term immune tolerance specific for hFVIII.


Season of birth and schizophrenia in Northeast Brazil: relationship to rainfall.
Although the association of schizophrenia and winter birth has been replicated many times in the Northern hemisphere, studies in the Southern hemisphere have been less consistent in their findings. A study from NE Brazil indicated the period between May and July, 3 months after the peak in rainfall, as a risk period for schizophrenia birth. We report findings from a random selection of charts (N = 1789) from the only public inpatient psychiatric facility in Ceará, Brazil (2 degrees to 5 degrees S). We compared the seasonality of birth in the schizophrenia group (N = 406) to multiple control groups: (1) psychosis not otherwise specified (N = 868), (2) other psychiatric diagnoses (N = 515), and (3) the average monthly birth rate in the general population. Patients with schizophrenia had a significantly greater risk of being born in the risk period compared with any of the control groups. There is a significant association between rainfall in a month and schizophrenia births 3 (p = 0.03) and 4 months (p = 0.01) later. This study corroborates findings of a significant seasonality in schizophrenia births in northeast Brazil.


Transient Axial Length Change during the Accommodation Response in Young Adults.
To measure the degree of transient axial elongation during the accommodation response in emmetropic and myopic young adults. To evaluate the effect of refractive error and accommodative demand on transient axial elongation of the eye. Axial length of the right eye was measured in 30 emmetropes and 30 myopes, by using the IOLMaster (Carl Zeiss Meditec, Inc., Dublin, CA), while accommodative stimuli of 0, 2, 4 and 6 D were presented with a Badal optometer. Axial length increased in both emmetropic and myopic subjects during short periods of accommodative stimulation. Greater transient increases in axial length were observed in myopic than in emmetropic subjects. The mean axial elongation with a 6-D stimulus to accommodation was 0.037 mm in emmetropes and 0.058 mm in myopes (P = 0.02). The degree of transient axial elongation correlated well with the stimulus to accommodation in emmetropes and myopes. Anterior chamber depth decreased, on average, by 0.19 mm in emmetropes and 0.18 mm in myopes when observing a 6-D stimulus to accommodation. During relatively short periods of accommodative stimulation, axial length increases in both emmetropic and myopic young adults. At higher levels of accommodative stimulation, a significantly greater transient increase in axial length is observed in myopic subjects than in their emmetropic counterparts.


Tight control of mycotoxin biosynthesis gene expression in Aspergillus flavus by temperature as revealed by RNA-Seq.
To better understand the effect of temperature on mycotoxin biosynthesis, RNA-Seq technology was used to profile the Aspergillus flavus transcriptome under different temperature conditions. This approach allowed us to quantify transcript abundance for over 80% of fungal genes including 1153 genes that were differentially expressed at 30 and 37 °C. Eleven of the 55 secondary metabolite clusters were upregulated at the lower temperature, including aflatoxin biosynthesis genes, which were among the most highly upexpressed genes. On average, transcript abundance for the 30 aflatoxin biosynthesis genes was 3300 times greater at 30 °C as compared with 37 °C. The results are consistent with the view that high temperature negatively affects aflatoxin production by turning down transcription of the two key transcriptional regulators, aflR and aflS. Subtle changes in the expression levels of aflS to aflR appear to control transcription activation of the aflatoxin cluster.


[Significance of iliac crest biopsy in the diagnosis of immunovasculitis in neurologic disease pictures].
In the present study we evaluated and analysed 81 patients with mainly neurological symptoms in whom iliac crest biopsy was performed because of a suspected immunovasculitis. In 20 of these patients, an immunopathy could be verified via the iliac crest biopsy. The relevance, reliability and accuracy of iliac crest biopsy is compared with the corresponding properties of other conventional methods of biopsy and neurological examination. The clinical syndromes of patients with immunovasculitis are analysed and compared with those of a group without detectable immunovasculitis in the iliac crest biopsy sample. For immunovasculitides with neurological symptomatology, no typical complex of symptoms was seen compared with the control group which would support a suspected diagnosis of immunovasculitis. Four selected case histories are described in keyword style. It follows from the result of this study that iliac crest biopsy is a low-risk examination method which the patient can be expected to tolerate, which is far more accurate in respect of the specific question of an immunodisease such as immunovasculitis with a strongly predominant neurological syndrome, compared with all other conventional neurological examination methods including other biopsy methods.


Premenstrual Syndrome and Childbirth Fear Prior to Pregnancy in Young Women: An Association and Cross-Sectional Study.
The aim of the study was to determine the association between premenstrual syndrome and the childbirth fear prior to pregnancy. This was an association and cross-sectional study conducted on 327 university students. Data were collected using "Participant Information Form," "Premenstrual Syndrome Scale," and "Childbirth Fear-Prior to Pregnancy Scale." It was found that the childbirth fear had increased in students with premenstrual syndrome. The Women Childbirth Fear-Prior to Pregnancy Scale score was statistically significantly higher among students who preferred caesarean section than those who preferred vaginal delivery. There was a weak, positive, and statistically significant correlation between the students' depressive sensation, anxiety, fatigue, nervousness, depressive thoughts, pain, appetite changes, sleep pattern changes, and bloating subscales of Premenstrual Syndrome Scale and Women Childbirth Fear-Prior to Pregnancy Scale. The score of the Women Childbirth Fear-Prior to Pregnancy Scale increases with an increase in the score of the Premenstrual Syndrome subscale. It should be evaluated whether or not women experiencing premenstrual syndrome have the childbirth fear prior to pregnancy.


Flexible Video Fetoscopy: Feasibility and Outcomes of a Novel Modality for Laser Therapy in Twin-to-Twin Transfusion Syndrome Presenting with Inaccessible Anterior Placenta.
Identification of intertwin anastomosis may be challenging during fetoscopy in cases with complete anterior placenta. The aim of this study was to describe the technique, feasibility, and outcomes of flexible video fetoscopy for laser coagulation in monochorionic (MC) twin pregnancies with twin-to-twin transfusion syndrome (TTTS) presenting with inaccessible anterior placenta. From April 2021 to March 2022, a prospective cohort of consecutive MC twin pregnancies complicated with TTTS presenting with anterior placenta after 20 weeks was recruited. Cases with inaccessible anterior placenta during standard technique were converted into flexible video fetoscopy for completion of laser coagulation of placental anastomoses using a 270° flexible video endoscope through the same uterine port. Descriptive analysis includes feasibility, remaining anastomoses requiring laser photocoagulation, and perinatal outcomes. A total of 45 pregnancies with TTTS were treated with fetoscopic laser therapy during the 1-year study period. Twenty-one pregnancies presented with anterior placenta after 20 weeks, in which an inaccessible vascular equator was observed in 33.3% (7/21). Flexible video fetoscopy was successfully performed in all 7 cases at a median gestational age of 22+2 (20+0-27+1) weeks+days. Visualization of the entire placental surface, coagulation of selected vessels, and exploration of the entire vascular equator were achieved in all cases. Six cases (85.7%) required additional laser coagulation due to either vascular patency despite initial coagulation with conventional fetoscopy (1/6, 16.7%) and/or remaining noncoagulated anastomoses (5/6, 83.3%). Perinatal survival of at least one twin and both twins was achieved in 85.7% and 57.1%, respectively. Flexible video fetoscopy for completion of laser coagulation of placental anastomoses is feasible and represents a good option for TTTS cases presenting after 20 weeks with inaccessible anterior placenta.


Assessment of relative importance of tablet computer features in supporting direct electronic documentation of encounters by eye care professionals.
Extensive utilization of mobile devices at the point of care will depend on device acceptance by the providers. We conducted focus groups involving nine eye care professionals to evaluate and elucidate the most important features of a tablet Personal Computer (PC) for data entry at the point of care. Ease of use, and quality and size of display were considered to be the most critical features of such a mobile device by the majority of the participants. Keyboard and weight of device were deemed to be the least important features of a tablet PC.


Reye's syndrome associated with respiratory syncytial virus infection.
An upper respiratory tract infection in a 22-month-old boy was followed by rapid loss of consciousness, hypoglycaemia, uraemia, and death. Necropsy examination showed fatty change of liver and kidneys, severe cerebral oedema, bronchiolitis, and endocardial fibroelastosis affecting the left ventricle. Immunofluorescence staining showed infection with respiratory syncytial virus (RSV). The clinical and pathological findings were those of Reye's syndrome, not previously reported accompanying RSV infection.


Sternal healing after coronary artery bypass grafting using bilateral internal thoracic arteries: assessment by computed tomography scan.
This study aimed to investigate sternal healing over time and the incidence of poor sternal healing in patients undergoing coronary artery bypass graft (CABG) surgery using bilateral internal thoracic arteries. This study enrolled 197 patients who underwent isolated CABG using skeletonized bilateral internal thoracic arteries (sBITA) from 2006 through 2009. Postoperative computed tomography (CT) angiography was performed on all patients at monthly intervals for three to six months after surgery. In 108 patients, an additional CT study was performed 24 to 48 months after surgery. The axial CT images were used to score sternal fusion at the manubrium, the upper sternum, and the lower sternum. These scores were added to evaluate overall healing: a score of 0 to 1 reflected poor healing, a score of 2 to 4 was defined as fair healing, and a score of 5 to 6 indicated complete healing. Medical records were also retrospectively reviewed to identify perioperative variables associated with poor early sternal healing. Three to six months after surgery, the average total score of sternal healing was 2.07±1.52 and 68 patients (34.5%) showed poor healing. Poor healing was most frequently found in the manubrium, which was scored as zero in 72.6% of patients. In multivariate analysis, the factors associated with poor early healing were shorter post-surgery time, older age, diabetes mellitus, and postoperative renal dysfunction. In later CT images, the average sternal healing score improved to 5.88±0.38 and complete healing was observed in 98.2% of patients. Complete sternal healing takes more than three months after a median sternotomy for CABG using sBITA. Healing is most delayed in the manubrium.


The conserved and divergent roles of Prdm3 and Prdm16 in zebrafish and mouse craniofacial development.
The formation of the craniofacial skeleton is a highly dynamic process that requires proper orchestration of various cellular processes in cranial neural crest cell (cNCC) development, including cell migration, proliferation, differentiation, polarity and cell death. Alterations that occur during cNCC development result in congenital birth defects and craniofacial abnormalities such as cleft lip with or without cleft palate. While the gene regulatory networks facilitating neural crest development have been extensively studied, the epigenetic mechanisms by which these pathways are activated or repressed in a temporal and spatially regulated manner remain largely unknown. Chromatin modifiers can precisely modify gene expression through a variety of mechanisms including histone modifications such as methylation. Here, we investigated the role of two members of the PRDM (Positive regulatory domain) histone methyltransferase family, Prdm3 and Prdm16 in craniofacial development using genetic models in zebrafish and mice. Loss of prdm3 or prdm16 in zebrafish causes craniofacial defects including hypoplasia of the craniofacial cartilage elements, undefined posterior ceratobranchials, and decreased mineralization of the parasphenoid. In mice, while conditional loss of Prdm3 in the early embryo proper causes mid-gestation lethality, loss of Prdm16 caused craniofacial defects including anterior mandibular hypoplasia, clefting in the secondary palate and severe middle ear defects. In zebrafish, prdm3 and prdm16 compensate for each other as well as a third Prdm family member, prdm1a. Combinatorial loss of prdm1a, prdm3, and prdm16 alleles results in severe hypoplasia of the anterior cartilage elements, abnormal formation of the jaw joint, complete loss of the posterior ceratobranchials, and clefting of the ethmoid plate. We further determined that loss of prdm3 and prdm16 reduces methylation of histone 3 lysine 9 (repression) and histone 3 lysine 4 (activation) in zebrafish. In mice, loss of Prdm16 significantly decreased histone 3 lysine 9 methylation in the palatal shelves but surprisingly did not change histone 3 lysine 4 methylation. Taken together, Prdm3 and Prdm16 play an important role in craniofacial development by maintaining temporal and spatial regulation of gene regulatory networks necessary for proper cNCC development and these functions are both conserved and divergent across vertebrates.


Adhesion of synthetic organic polymer on soft tissue. I. A fast setting polyurethane adhesive.
Conventional polyurethane prepolymers have been shown to adhere to living biological tissues. However, their setting is not sufficiently expedient to permit convenient applications in vivo. A prepolymer prepared from the highly reactive 6-chloro-2,4,5-trifluoro-1,3-phenylene diisocyanate, castor oil, and a trace of pyridine has afforded an adhesive which sets in about 2 min in vivo. The fast setting has resulted in poor adhesion on biological tissue. The bonding has been improved by the inclusion of tolylene diisocyanate in the composition without affecting the fast curing rate of the prepolymer. The dispersion of the adhesive and its cohesion after solidification have been adjusted by other minor additives. Preliminary evaluation on animals indicates that this adhesive is most useful as a hemostatic coating in hepatic lacerations.


Stereocontrolled syntheses of the nemorensic acids using 6-diazoheptane-2,5-dione in carbonyl ylide cycloadditions.
Levulinic acid-derived 6-diazoheptane-2,5-dione (9) serves as a common precursor in a formal synthesis of frontalin 19, and in syntheses of cis-nemorensic acid 1, 4-hydroxy-cis-nemorensic acid 2, 3-hydroxy-cis-nemorensic acid 3, and nemorensic acid 4. The key step in these syntheses is the Rh(2)(OAc)(4)-catalyzed tandem carbonyl ylide formation-intermolecular 1,3-dipolar cycloadditions of diazodione 9 with formaldehyde, alkynes or allene, which occur with high regioselectivity. Subsequent oxidative cleavage of the ring originally derived from the cyclic carbonyl ylide intermediate provides a straightforward access to polysubstituted tetrahydrofurans, and in particular an efficient entry to the nemorensic acids. Enantioselective cycloadditions with diazodione 9, using chiral rhodium catalysts, gave cycloadducts in up to 51% ee.


HAMPLE: deciphering TF-DNA binding mechanism in different cellular environments by characterizing higher-order nucleotide dependency.
Transcription factor (TF) binds to conservative DNA binding sites in different cellular environments and development stages by physical interaction with interdependent nucleotides. However, systematic computational characterization of the relationship between higher-order nucleotide dependency and TF-DNA binding mechanism in diverse cell types remains challenging. Here, we propose a novel multi-task learning framework HAMPLE to simultaneously predict TF binding sites (TFBS) in distinct cell types by characterizing higher-order nucleotide dependencies. Specifically, HAMPLE first represents a DNA sequence through three higher-order nucleotide dependencies, including k-mer encoding, DNA shape and histone modification. Then, HAMPLE uses the customized gate control and the channel attention convolutional architecture to further capture cell-type-specific and cell-type-shared DNA binding motifs and epigenomic languages. Finally, HAMPLE exploits the joint loss function to optimize the TFBS prediction for different cell types in an end-to-end manner. Extensive experimental results on seven datasets demonstrate that HAMPLE significantly outperforms the state-of-the-art approaches in terms of auROC. In addition, feature importance analysis illustrates that k-mer encoding, DNA shape, and histone modification have predictive power for TF-DNA binding in different cellular environments and are complementary to each other. Furthermore, ablation study, and interpretable analysis validate the effectiveness of the customized gate control and the channel attention convolutional architecture in characterizing higher-order nucleotide dependencies. The source code is available at https://github.com/ZhangLab312/Hample.


Theoretical study of catalytic dinitrogen reduction under mild conditions.
Density functional theory results on key steps of Schrock's catalytic cycle are presented. The quantum chemical modeling of the dinitrogen-reducing reaction steps is conducted without simplifying the bulky HIPT [HIPT = 3,5-(2,4,6-iPr(3)C(6)H(2))(2)C(6)H(3)] substituents at the triamidoamine ligand.


Dynamic features of water molecules in superconcentrated aqueous electrolytes.
An existence of ions dissolved in water has significant effects on bulk properties of water. Superconcentrated conditions have been recently proposed to provide a new concept of lithium ion batteries in order to overcome limitations for practical applications. In those conditions, water would undergo significant changes in structure and dynamics compared to its bulk properties. However, little is known about water in superconcentrated aqueous electrolytes. Here we study the properties of water in aqueous electrolytes with various salt concentrations via molecular dynamics simulations. We find that new dynamic features of water arise in the limit of an extremely high salt concentration. In particular, we observe a decoupled temporal character of water molecules exhibiting a subdiffusive translation and a diffusive rotation in the superconcentrated condition. Furthermore, we find that the rotational dynamics for each principal axis of a water molecule differently responds to the salt concentration, resulting in an occurrence of anisotropy in the rotation as the salt concentration increases. The superconcentrated environments also invoke new features in the hydrogen-bonding characteristics of water such as an emergence of two time scales in the hydrogen bond dynamics of water with respect to the salt concentration.


Formation of nitrotyrosine by methylene blue photosensitized oxidation of tyrosine in the presence of nitrite.
Methylene blue photosensitized oxidation of tyrosine in the presence of nitrite produces 3-nitrotyrosine, with maximum yield at pH 6. The formation of 3-nitrotyrosine requires oxygen and increases using deuterium oxide as solvent, suggesting the involvement of singlet oxygen in the reaction. The detection of dityrosine as an additional reaction product suggests that the first step in the interaction of tyrosine with singlet oxygen generates tyrosyl radicals which can dimerize to form dityrosine or react with a nitrite-derived species to produce 3-nitrotyrosine. Although the chemical identity of the nitrating species has not been established, the possible generation of nitrogen dioxide (*NO(2)) by indirect oxidation of nitrite by intermediately produced tyrosyl radical, via electron transfer, is proposed. One important implication of the results of this study is that the oxidation of tyrosine by singlet oxygen in the presence of nitrite may represent an alternative or additional pathway of 3-nitrotyrosine formation of potential importance in oxidative injures such as during inflammatory processes.


Induction of Mitochondrial Fragmentation and Mitophagy after Neonatal Hypoxia-Ischemia.
Hypoxia-ischemia (HI) leads to immature brain injury mediated by mitochondrial stress. If damaged mitochondria cannot be repaired, mitochondrial permeabilization ensues, leading to cell death. Non-optimal turnover of mitochondria is critical as it affects short and long term structural and functional recovery and brain development. Therefore, disposal of deficient mitochondria via mitophagy and their replacement through biogenesis is needed. We utilized mt-Keima reporter mice to quantify mitochondrial morphology (fission, fusion) and mitophagy and their mechanisms in primary neurons after Oxygen Glucose Deprivation (OGD) and in brain sections after neonatal HI. Molecular mechanisms of PARK2-dependent and -independent pathways of mitophagy were investigated in vivo by PCR and Western blotting. Mitochondrial morphology and mitophagy were investigated using live cell microscopy. In primary neurons, we found a primary fission wave immediately after OGD with a significant increase in mitophagy followed by a secondary phase of fission at 24 h following recovery. Following HI, mitophagy was upregulated immediately after HI followed by a second wave at 7 days. Western blotting suggests that both PINK1/Parkin-dependent and -independent mechanisms, including NIX and FUNDC1, were upregulated immediately after HI, whereas a PINK1/Parkin mechanism predominated 7 days after HI. We hypothesize that excessive mitophagy in the early phase is a pathologic response which may contribute to secondary energy depletion, whereas secondary mitophagy may be involved in post-HI regeneration and repair.


Application of kriging models for a drug combination experiment on lung cancer.
Combinatorial drugs have been widely applied in disease treatment, especially chemotherapy for cancer, due to its improved efficacy and reduced toxicity compared with individual drugs. The study of combinatorial drugs requires efficient experimental designs and proper follow-up statistical modeling techniques. Linear and nonlinear models are often used in the response surface modeling for such experiments. We propose the use of kriging models to better depict the response surfaces of combinatorial drugs. We illustrate our method via a drug combination experiment on lung cancer and further show how proper experimental designs can reduce the necessary run size. We demonstrate that only 27 runs are needed to predict all 512 runs in the original experiment and achieve better precision than existing analyses.


[Method of quantitative estimation of computer tomograms of inner organs and its application to intravital topographic-anatomical investigations].
This article describes the method of quantitative estimation of computer tomograms, which served as the basis for description of morphometric and skeletotopic parameters of the pancreas and thyroid gland that were obtained as a result of study of 320 persons aged from 20 to 70 without pathology of these glands.


Anoxemia and brain disease.
The author advances the concept that anoxemia, either in its general or restricted form, or both, is probably responsible for a considerable portion of "degenerative diseases," whose etiologic delineation has not yet been traced. It is necessary, he believes, to enlarge greatly the comprehension of the disordered circulatory states to include oxygen want and thereby account for a number of conditions hitherto considered to be of unknown cause. More than this, he finds in oxygen want an explanation of the mechanism of a number of individual lesions or details of lesions otherwise not well understood. The author believes it is very likely that an understanding of cerebral anoxia in its ultimate ramifications will open still wider doors to the understanding of certain clinical syndromes the cause of which remains obscure.


Biomarkers of monocrotophos in a freshwater fish Channa punctatus (Bloch).
Activity of a few biomarkers have been investigated on freshwater fish Channa punctatus treated with monocrotophos for acute exposure to 18.56 ppm at 96 hr and subacute exposure viz. 0.46 ppm, 0.96 ppm and 1.86 ppm for 30 days. Biomarkers such as total protein, lipid peroxidation and acetylcholinesterase have been measured in different tissues of fish viz. gills, liver, brain and muscles. The protein levels were found to be depleted in all the tissues after pesticide exposure to lethal and sublethal concentration over the control, where as the lipid levels showed an increase under the stress of pesticide monocrotophos. The increased lipid level may be due to inhibition of lipase activity and other biomarkers of lipid metabolism. A significant inhibition of brain acetylcholinesterase (AChE) indicating its effects on nervous system have also been observed. These parameters can be used as biomarkers to predict the early toxicity of monocrotophos added to aquatic ecosystem.


Electrochemical oxidation of 2,4,5-trichlorophenoxyacetic acid by metal-oxide-coated Ti electrodes.
Electrochemical oxidation of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) over metal-oxide-coated Ti anodes, i.e., Ti/SnO2-Sb/Ce-PbO2, Ti/SnO2-Sb and Ti/RuO2, was examined. The degradation efficiency of over 90% was attained at 20 min at different initial concentrations (0.5-20 mg L(-1)) and initial pH values (3.1-11.2). The degradation efficiencies of 2,4,5-T on Ti/SnO2-Sb/Ce-PbO2, Ti/SnO2-Sb and Ti/RuO2 anodes were higher than 99.9%, 97.2% and 91.5% at 30 min, respectively, and the respective total organic carbon removal ratios were 65.7%, 54.6% and 37.2%. The electrochemical degradation of 2,4,5-T in aqueous solution followed pseudo-first-order kinetics. The compounds, i.e., 2,5-dichlorohydroquinone and 2,5-dihydroxy-p-benzoquinone, have been identified as the main aromatic intermediates by liquid chromatography-mass spectrometry. The results showed that the energy efficiencies of 2,4,5-T (20 mg L(-1)) degradation with Ti/SnO2-Sb/Ce-PbO2 anode at the optimal current densities from 2 to 16 mA cm(-2) ranged from 8.21 to 18.73 kWh m(-3).


Theoretical foundations of analytical psychology: recent developments and controversies.
This article gives an introductory overview of the papers in this volume originally given at the Joint Conference of the IAAP and the University of Basel, Basel, October 18-20, 2018. The aim of the conference was to bring core concepts of analytical psychology together with theorizing and research from academic sciences, at the very place where Jung started his academic career, the University of Basel. The conference focussed on three fields: the relationship of consciousness and the unconscious and the theory of complexes; the theory of archetypes; and the status of analytical psychotherapy in contemporary psychotherapy research. The aim of the conference was to further the development of theory in analytical psychology in relation to results and insights in contiguous areas of knowledge. In the first area, contributors pointed to the solid evidence especially from the neurosciences for the psychodynamic conceptualizations of the unconscious, and also for the concept of complexes. In contrast to this, the concept of archetypes is controversial, with a majority of contributors questioning Jung's biological conceptualizations of archetypes, and speaking instead for reformulations from the perspective of cultural theory, dynamic systems theory and other approaches. In the field of psychotherapy research, contributors pointed to the profound need for conducting more empirical studies on the outcome of Jungian psychotherapy, but also for a thorough reconsideration of standard research designs in the field.


Robotic surgery: an update.
Minimally invasive surgery techniques have revolutionized surgery. Robotic surgery may be the next revolution in surgical technology. Robotics coupled with minimally invasive surgery and microscopic surgery provides the potential to do more complex and more precise tasks. Robotic surgery offers tremor filtration, motion scaling, indexed movements, additional degrees of freedom, and improved ergonomics. We explore robotic history, the present surgical technology, the current clinical cases and research, and the future of robotics. We will look specifically at the birth and progress of our own problem.


Immobilization of Clostridium perfringens type D in calcium alginate beads: toxin production mimics free cell culture.
Cell-immobilization is used to maintain microbial culture to produce metabolites in repeated-batch or continuous fermentations, thereby reducing the time and resources spent on delivering mass production of microbe. The technique also enables shortening of the detoxification phase and the amount of formaldehyde required due to low incidence of viable bacteria in the extract. A solution of sodium alginate containing Clostridium perfringens cells was dropped into stirring CaCl solution via a sterile syringe needle. Optimizations resulted in reasonably uniform beads containing C. perfringens. Beads were externally stabilized by poly L-lysine, followed by immersion in a solution of Na-alginate to coat them with a new layer of alginate forming an alginate-PLL-alginate cortex. This study proved successful in immobilizing C. perfringens cells inside uniform alginate microspheres. Cell loading and cell propagation inside the beads were measured. The cell loaded beads were cultivable in liquid media producing 550 minimum lethal doses per milliliter (MLD/ml) in a 72 h. The research paved the way for further investigations to optimize and establish an efficient bacterial encapsulation method. Thus, it seems possible to produce toxins from beads engulfing C. perfringens on larger scales via repeated-batch or continuous fermentation processes.


[Aortic valve replacement. Apropos of a series of 117 patients].
With a series of 117 aortic valve replacements, the authors have examined the results in relation to the method of protecting the myocardium while the aorta is clamped off. There does not appear to be much difference between coronary perfusion and the technique of profound local hypothermia by the perfusion of a chilled solution into the pericardium. Because of this, the authors remain in favour of the latter technique, which provides effective protection of the myocardium during periods of aortic occlusion sufficient for monovalvular replacements. An occlusion time of up to 90 minutes has been achieved without any major problems. Nevertheless, perfusion of the two coronary arteries is sometimes indicated, notably when profound local hypothermia cannot be employed at revision surgery, feeling of the pericardial cavity being neither possible nor desirable.


Chemosensory protein regulates the behavioural response of Frankliniella intonsa and Frankliniella occidentalis to tomato zonate spot virus-Infected pepper (Capsicum annuum).
Many herbivorous insects rely on plant volatiles to locate their host plants. Vector-borne viral infections induce changes in plant volatiles, which render infected plants more attractive to insect vectors. However, the detailed mechanisms underlying the olfactory responses of insect vectors induced by the volatiles produced by virus-infected plants are poorly understood. Here, we show that volatiles emitted by pepper (Capsicum annuum) plants infected with tomato zonate spot virus (TZSV), particularly the volatile cis-3-hexenal, which is recognized by chemosensory protein 1 of the thrips Frankliniella intonsa (FintCSP1), are more attractive to F. intonsa than the volatiles emitted by non-infected pepper plants. FintCSP1 is highly abundant in the antenna of F. intonsa. Silencing of FintCSP1 significantly decreased electroantennogram responses of F. intonsa antennae to cis-3-hexenal and impaired thrips' responses to TZSV-infected pepper plants and cis-3-hexenal, as assessed using a Y-tube olfactometer. Three-dimensional model predictions indicated that FintCSP1 consists of seven α-helixes and two disulfide bridges. Molecular docking analysis suggested that cis-3-hexenal is positioned deep inside the binding pocket of FintCSP1 and binds to residues of the protein. We combined site-directed mutagenesis and fluorescence binding assays and identified three hydrophilic residues, Lys26, Thr28, and Glu67, of FintCSP1 as being critical for cis-3-hexenal binding. Furthermore, CSP of F. occidentalis (FoccCSP) is also a key olfactory protein involved in modulating the behaviour of F. occidentalis to TZSV-infected pepper. This study revealed the specific binding characteristics of CSPs to cis-3-hexenal and confirmed the general hypothesis that virus infections induce changes in host volatiles, which can be recognized by the olfactory proteins of the insect vector to enhance vector attraction and this may facilitate viral spread and transmission.


Comparison of mersilene mesh-methyl metacrylate sandwich and polytetrafluoroethylene grafts for chest wall reconstruction.
We report the outcomes of patients who underwent reconstruction with Mersilene mesh-methyl methacrylate (MM-MM) sandwich and polytetrafluoroethylene (PTFE) grafts after a large chest wall resection. Between June 1990 and September 2001, 59 consecutive patients (37 men, 22 women; mean age, 48.1 +/- 11.8 years; range 22-74 years) underwent large chest wall resection (greater than 5 cm diameter) and reconstruction with prosthetic material in our department. Twenty-one patients (33%) underwent reconstruction with a PTFE graft (group 2) between 1990 and 1994, and 38 patients (67%) underwent reconstruction with an MM-MM sandwich graft (group 1) between 1994 and 2001. Operative morbidity ratios were 5.2% (2/38) in group 1 and 24% (5/21) in group 2 (p = .036). The paradoxical respiration ratio was significantly higher (p = .018) in group 2 (5/21: 24%) than it was in group 1 (1/38: 2.6%). The operative mortality ratio was 4.5% (1/21) in group 2 and 0% in group 1. Mean hospital stay was 10.6 days (range 5-21 days) in group 1 and 13.3 days (range 7-36 days) in group 2 (p = .015). The MM-MM graft is inexpensive and easy to apply, provides better cosmetic options, and offers minimal morbidity. We therefore recommend that the MM-MM sandwich graft be used rather than the PTFE graft for large defects of the anterolateral chest wall and sternum where successful prevention of paradoxical respiration is required.


Left Frontal EEG Power Responds to Stock Price Changes in a Simulated Asset Bubble Market.
Financial bubbles are a result of aggregate irrational behavior and cannot be explained by standard economic pricing theory. Research in neuroeconomics can improve our understanding of their causes. We conducted an experiment in which 28 healthy subjects traded in a simulated market bubble, while scalp EEG was recorded using a low-cost, BCI-friendly desktop device with 14 electrodes. Independent component (IC) analysis was performed to decompose brain signals and the obtained scalp topography was used to cluster the ICs. We computed single-trial time-frequency power relative to the onset of stock price display and estimated the correlation between EEG power and stock price across trials using a general linear model. We found that delta band (1-4 Hz) EEG power within the left frontal region negatively correlated with the trial-by-trial stock prices including the financial bubble. We interpreted the result as stimulus-preceding negativity (SPN) occurring as a dis-inhibition of the resting state network. We conclude that the combination between the desktop-BCI-friendly EEG, the simulated financial bubble and advanced signal processing and statistical approaches could successfully identify the neural correlate of the financial bubble. We add to the neuroeconomics literature a complementary EEG neurometric as a bubble predictor, which can further be explored in future decision-making experiments.


Metagenomics and Single-Cell Omics Data Analysis for Human Microbiome Research.
Microbes are ubiquitous on our planet, and it is well known that the total number of microbial cells on earth is huge. These organisms usually live in communities, and each of these communities has a different taxonomical structure. As such, microbial communities would serve as the largest reservoir of genes and genetic functions for a vast number of applications in "bio"-related disciplines, especially in biomedicine. Human microbiome is the area in which the relationships between ourselves as hosts and our microbiomes have been examined.In this chapter, we have first reviewed the researches in microbes on community, population and single-cell levels in general. Then we have focused on the effects of recent metagenomics and single-cell advances on human microbiome research, as well as their effects on translational biomedical research. We have also foreseen that with the advancement of big-data analysis techniques, deeper understanding of human microbiome, as well as its broader applications, could be realized.


Hepatitis E virus co-infection in HIV-infected patients in Foggia and Naples in southern Italy.
Hepatitis E virus (HEV) infection represents an emerging infection in developed countries and is thought to be a zoonotic infection. It has recently been described as a new causative agent of acute and chronic hepatitis in immunosuppressed subjects, including HIV-infected patients. The aim of this study was to assess the sero-virological prevalence of HEV in HIV patients and in the general population as control group. A prospective and observational cohort study was carried out in two hospitals in southern Italy. The seroprevalence of HEV was determined in a cohort of 959 subjects, 509 (53%) of whom were HIV-positive patients and 450 were from the general population. Serum samples were tested for anti-HEV antibodies; repeatedly positive results were confirmed by a Western blot assay. In positive patients HEV RNA and genotypes were also determined. A total of 46 (4.8%) of the 959 serum samples examined were reactive to anti-HEV Ig and confirmed by Western blotting. The prevalence of HEV antibodies (IgG and/or IgM) was 2.7% in the control group and 6.7% in HIV-infected patients. Anti-HEV IgM was found in 6/46 (13.0%) of the anti-HEV Ig-positive serum samples, in 5/34 HIV patients and in 1/12 of the general population. No HIV-infected patient presented chronic hepatitis with HEV infection alone. This study indicates a higher circulation of HEV in HIV-infected patients, whereas a low prevalence of HEV antibodies in the general Italian population was shown. Chronic hepatitis with HEV alone was absent, while it was present in subjects with HIV-HEV, co-infected with hepatitis B virus (HBV) and/or hepatitis C virus (HCV).


Targeting Pathogenic Biofilms: Newly Developed Superhydrophobic Coating Favors a Host-Compatible Microbial Profile on the Titanium Surface.
Polymicrobial infections are one of the most common reasons for inflammation of surrounding tissues and failure of implanted biomaterials. Because microorganism adhesion is the first step for biofilm formation, physical-chemical modifications of biomaterials have been proposed to reduce the initial microbial attachment. Thus, the use of superhydrophobic coatings has emerged because of their anti-biofilm properties. However, these coatings on the titanium (Ti) surface have been developed mainly by dual-step surface modification techniques and have not been tested using polymicrobial biofilms. Therefore, we developed a one-step superhydrophobic coating on the Ti surface by using a low-pressure plasma technology to create a biocompatible coating that reduces polymicrobial biofilm adhesion and formation. The superhydrophobic coating on Ti was created by the glow discharge plasma using Ar, O2, and hexamethyldisiloxane gases, and after full physical, chemical, and biological characterizations, we evaluated its properties regarding oral biofilm inhibition. The newly developed coating presented an increased surface roughness and, consequently, superhydrophobicity (contact angle over 150°) and enhanced corrosion resistance (p < 0.05) of the Ti surface. Furthermore, proteomic analysis showed a unique pattern of protein adsorption on the superhydrophobic coating without drastically changing the biologic processes mediated by proteins. Additionally, superhydrophobic treatment did not present a cytotoxic effect on fibroblasts or reduction of proliferation; however, it significantly reduced (≈8-fold change) polymicrobial adhesion (bacterial and fungal) and biofilm formation in vitro. Interestingly, superhydrophobic coating shifted the microbiological profile of biofilms formed in situ in the oral cavity, reducing by up to ≈7 fold pathogens associated with the peri-implant disease. Thus, this new superhydrophobic coating developed by a one-step glow discharge plasma technique is a promising biocompatible strategy to drastically reduce microbial adhesion and biofilm formation on Ti-based biomedical implants.


Integrated analysis of the gut microbiome and metabolome in a mouse model of inflammation-induced colorectal tumors.
Colorectal cancer (CRC) is a common malignancy worldwide, and the gut microbiota and metabolites play an important role in its initiation and progression. In this study, we constructed a mouse model of inflammation-induced colorectal tumors, with fixed doses of azoxymethane/dextran sulfate sodium (AOM/DSS). We found that colorectal tumors only formed in some mice treated with certain concentrations of AOM/DSS (tumor group), whereas other mice did not develop tumors (non-tumor group). 16S rDNA amplicon sequencing and liquid chromatography-mass spectrometry (LC-MS)/MS analyses were performed to investigate the microbes and metabolites in the fecal samples. As a result, 1189 operational taxonomic units (OTUs) were obtained from the fecal samples, and the non-tumor group had a relatively higher OTU richness and diversity. Moreover, 53 different microbes were identified at the phylum and genus levels, including Proteobacteria, Cyanobacteria, and Prevotella. Furthermore, four bacterial taxa were obviously enriched in the non-tumor group, according to linear discriminant analysis scores (log10) > 4. The untargeted metabolomics analysis revealed significant differences between the fecal samples and metabolic phenotypes. Further, the heatmaps and volcano plots revealed 53 and 19 dysregulated metabolites between the groups, in positive and negative ion modes, respectively. Styrene degradation and amino sugar-nucleotide sugar metabolism pathways were significantly different in positive and negative ion modes, respectively. Moreover, a correlation analysis between the metabolome and microbiome was further conducted, which revealed the key microbiota and metabolites. In conclusion, we successfully established a tumor model using a certain dose of AOM/DSS and identified the differential intestinal microbiota and characteristic metabolites that might modulate tumorigenesis, thereby providing new concepts for the prevention and treatment of CRC.


[Psychoanalytic therapy for a child with somnambulism: a case report].
Somnambulism is defined as a state of dissociated consciousness triggered by impaired arousal, which results in partial wakefulness and partial sleep. No effective therapy or medication has been available for treating children with somnambulism. Herein we present a case in a 4.5-year-old girl, who presented with somnambulism associated with separation anxiety disorder every night in a week. The girl received formal assessment and appropriate interventions, and the symptoms disappeared within a week. The treatment was carried out in 5 stages: diagnosis of the disease, establishment of trust, information collection, supervision, and individualized psychotherapy. Although dreams in childhood can be difficult to interpret, close observation of the behaviors in sleepwalking, as a special form of dream, in addition to the more precise description by the guardians, still provides useful clues to understand those dreams. For children with somnambulism, early intervention with psychotherapy can significantly decrease the false revival of the unconscious desires, and thus may serve as a treatment option other than medications.


Solvent effects on supramolecular gel-phase materials: two-component dendritic gel.
The self-assembly of diaminododecane with dendritic l-lysine-based peptides to form gel-phase materials was investigated in a range of different solvents. The degree of structuring was modulated by the solvent employed, an effect which induced subtle changes in the mesoscale aggregate morphology and macroscopic behavior of the self-assembled state. In this paper a range of different solvent parameters are investigated, and it is clearly shown that macroscopic gelation can be related to a solvent polar solubility parameter for this system. The results also show a dependence on Kamlet-Taft hydrogen bonding parameters, and this clearly demonstrates the role of the solvent environment in terms of dendron--dendron intermolecular hydrogen bonding and its impact on the supramolecular chiral organization of the assembled superstructure.


Hiding in plain sight-wildlife as a neglected reservoir and pathway for the spread of antimicrobial resistance: a narrative review.
Antimicrobial resistance represents a global health problem, with infections due to pathogenic antimicrobial resistant bacteria (ARB) predicted to be the most frequent cause of human mortality by 2050. The phenomenon of antimicrobial resistance has spread to and across all ecological niches, and particularly in livestock used for food production with antimicrobials consumed in high volumes. Similarly, hospitals and other healthcare facilities are recognized as significant 'hotspots' of ARB and antimicrobial resistance genes (ARGs); however, over the past decade, new and previously overlooked ecological niches are emerging as hidden reservoirs of ARB/ARGs. Increasingly extensive and intensive industrial activities, degradation of natural environments, burgeoning food requirements, urbanization, and global climatic change have all dramatically affected the evolution and proliferation of ARB/ARGs, which now stand at extremely concerning ecological levels. While antimicrobial resistant bacteria and genes as they originate and emanate from livestock and human hosts have been extensively studied over the past 30 years, numerous ecological niches have received considerably less attention. In the current descriptive review, the authors have sought to highlight the importance of wildlife as sources/reservoirs, pathways and receptors of ARB/ARGs in the environment, thus paving the way for future primary research in these areas.


Electrosynthesis of Well-Defined Metal-Organic Framework Films and the Carbon Nanotube Network Derived from Them toward Electrocatalytic Applications.
An efficient and controllable method to synthesize continuous metal-organic framework (MOF) films is highly desired. Herein, we demonstrate a facile and universal electrochemical method to synthesize homogeneous and uniform zeolitic imidazolate framework (ZIF, a typical class of MOF family) films on various macroconductive substrates (e.g., conductive glass, Ni foam and carbon cloth) as well as nanostructured substrates (one-dimensional nanorod array, two-dimensional nanowall array, and three-dimensional nanoframework). Particularly, the MOF film can be easily transformed into hierarchical ordered carbon nanotube networks, which display multifunctional electrocatalytic performances, such as excellent activity and good long-term stability in overall water splitting and oxygen reduction. It is worth mentioning that this is the first reported work for electrosynthesis of ZIF films on various conductive substrates, illustrating the great potential of the fast and economical method for constructing functional and integrated films or electrodes toward energy-related applications.


A literature review of autograft and allograft anterior cruciate ligament reconstruction.
Knee anterior cruciate ligament reconstructive surgery has significantly evolved and now includes the option of using an allograft. This has resulted in numerous studies evaluating the advantages and disadvantages of allografts. The purpose of this literature review is to evaluate this research and present important findings to allow the selection of the most appropriate graft source when considering allograft versus autograft reconstruction.


In vitro analog of classical conditioning of feeding behavior in aplysia.
The feeding behavior of Aplysia californica can be classically conditioned using tactile stimulation of the lips as a conditioned stimulus (CS) and food as an unconditioned stimulus (US). Moreover, several neural correlates of classical conditioning have been identified. The present study extended previous work by developing an in vitro analog of classical conditioning and by investigating pairing-specific changes in neuronal and synaptic properties. The preparation consisted of the isolated cerebral and buccal ganglia. Electrical stimulation of a lip nerve (AT4) and a branch of the esophageal nerve (En2) served as the CS and US, respectively. Three protocols were used: paired, unpaired, and US alone. Only the paired protocol produced a significant increase in CS-evoked fictive feeding. At the cellular level, classical conditioning enhanced the magnitude of the CS-evoked synaptic input to pattern-initiating neuron B31/32. In addition, paired training enhanced both the magnitude of the CS-evoked synaptic input and the CS-evoked spike activity in command-like neuron CBI-2. The in vitro analog of classical conditioning reproduced all of the cellular changes that previously were identified following behavioral conditioning and has led to the identification of several new learning-related neural changes. In addition, the pairing-specific enhancement of the CS response in CBI-2 indicates that some aspects of associative plasticity may occur at the level of the cerebral sensory neurons.


Primatologist: A modular segmentation pipeline for macaque brain morphometry.
Because they bridge the genetic gap between rodents and humans, non-human primates (NHPs) play a major role in therapy development and evaluation for neurological disorders. However, translational research success from NHPs to patients requires an accurate phenotyping of the models. In patients, magnetic resonance imaging (MRI) combined with automated segmentation methods has offered the unique opportunity to assess in vivo brain morphological changes. Meanwhile, specific challenges caused by brain size and high field contrasts make existing algorithms hard to use routinely in NHPs. To tackle this issue, we propose a complete pipeline, Primatologist, for multi-region segmentation. Tissue segmentation is based on a modular statistical model that includes random field regularization, bias correction and denoising and is optimized by expectation-maximization. To deal with the broad variety of structures with different relaxing times at 7 T, images are segmented into 17 anatomical classes, including subcortical regions. Pre-processing steps insure a good initialization of the parameters and thus the robustness of the pipeline. It is validated on 10 T2-weighted MRIs of healthy macaque brains. Classification scores are compared with those of a non-linear atlas registration, and the impact of each module on classification scores is thoroughly evaluated.


Interface controlled solid-state lithium storage performance in free-standing bismuth nanosheets.
Bismuth (Bi) has recently been discovered as a potential lithium-ion anode material for batteries with high Li capacity and suitable equilibrium potential, and without dendrite formation. However, the reversible electrochemical stability remains insufficient for applications. Herein, it is demonstrated that two-dimensional free-standing Bi nanosheets (Bi-NSs) have superior anode performance using either liquid or solid electrolytes. The Bi-NSs with a uniform thickness of ∼40 nm prepared by aqueous methods exhibit a record high capacity of ∼287 mA h g-1 at a current density of 250 mA g-1 with the LiBH4 solid electrolyte even after 100 cycles. Fast and stable solid-state lithium plating and stripping occur without side reactions. The 2D layered nanostructure has more active sites and a shorter diffusion length, and forms stable interfaces with the electrolyte. The present work reveals a facile synthesis route of novel 2D materials and paves an efficient pathway for high-capacity and safe bismuth-based anodes for lithium batteries.


Understanding who is and isn't involved and engaged in health research: capturing and analysing demographic data to diversify patient and public involvement and engagement.
Patient and public involvement and engagement (PPIE) can improve the relevance, quality, ethics and impact of research thus contributing to high quality research. Currently in the UK, people who get involved in research tend to be aged 61 years or above, White and female. Calls for greater diversity and inclusion in PPIE have become more urgent especially since the COVID-19 pandemic, so that research can better address health inequalities and be relevant for all sectors of society. Yet, there are currently no routine systems or requirements to collect or analyse the demographics of people who get involved in health research in the UK. The aim of this study was to develop to capture and analyse the characteristics of who does and doesn't take part in patient and public involvement and engagement (PPIE) activities. As part of its strategic focus on diversity and inclusion, Vocal developed a questionnaire to assess the demographics of people taking part in its PPIE activities. Vocal is a non-profit organisation which supports PPIE in health research across the region of Greater Manchester in England. The questionnaire was implemented across Vocal activities between December 2018 and March 2022. In that time. Vocal was working with approximately 935 public contributors. 329 responses were received: a return rate of 29.3%. Analysis of findings and comparison against local population demographic data, and available national data related to public contributors to health research, was performed. Results show that it is feasible to assess the demographics of people who take part in PPIE activities, through a questionnaire system. Further, our emerging data indicate that Vocal are involving people from a wider range of ages and with a greater diversity of ethnic backgrounds in health research, as compared to available national data. Specifically, Vocal involves more people of Asian, African and Caribbean heritage, and includes a wider range of ages in its PPIE activities. More women than men are involved in Vocal's work. Our 'learn by doing' approach to assessing who does and doesn't take part in Vocal's PPIE activities has informed our practice and continues influence our strategic priorities for PPIE. Our system and learning reported here may be applicable and transferable to other similar settings in which PPIE is carried out. We attribute the greater diversity of our public contributors to our strategic priority and activities to promote more inclusive research since 2018.


Cell wall biomechanics: a tractable challenge in manipulating plant cell walls 'fit for purpose'!
The complexity and recalcitrance of plant cell walls has contributed to the success of plants colonising land. Conversely, these attributes have also impeded progress in understanding the roles of walls in controlling and directing developmental processes during plant growth and also in unlocking their potential for biotechnological innovation. Recent technological advances have enabled the probing of how primary wall structures and molecular interactions of polysaccharides define their biomechanical (and hence functional) properties. The outputs have led to a new paradigm that places greater emphasis on understanding how the wall, as a biomechanical construct and cell surface sensor, modulates both plant growth and material properties. Armed with this knowledge, we are gaining the capacity to design walls 'fit for (biotechnological) purpose'!


Detection of intra-tumor self antigen recognition during melanoma tumor progression in mice using advanced multimode confocal/two photon microscope.
Determining how tumor immunity is regulated requires understanding the extent to which the anti-tumor immune response "functions" in vivo without therapeutic intervention. To better understand this question, we developed advanced multimodal reflectance confocal/two photon fluorescence intra-vital imaging techniques to use in combination with traditional ex vivo analysis of tumor specific T cells. By transferring small numbers of melanoma-specific CD8+ T cells (Pmel-1), in an attempt to mimic physiologic conditions, we found that B16 tumor growth alone was sufficient to induce naive Pmel-1 T cell proliferation and acquisition of effector phenotype. Tumor -primed Pmel-1 T cells, are capable of killing target cells in the periphery and secrete IFNγ, but are unable to mediate tumor regression. Within the tumor, Pmel-1 T cells have highly confined mobility, displaying long term interactions with tumor cells. In contrast, adoptively transferred non tumor-specific OT-I T cells show neither confined mobility, nor long term interaction with B16 tumor cells, suggesting that intra-tumor recognition of cognate self antigen by Pmel-1 T cells occurs during tumor growth. Together, these data indicate that lack of anti-tumor efficacy is not solely due to ignorance of self antigen in the tumor microenvironment but rather to active immunosuppressive influences preventing a protective immune response.


An Efficient Genome Editing Strategy To Generate Putative Null Mutants in Caenorhabditis elegans Using CRISPR/Cas9.
Null mutants are essential for analyzing gene function. Here, we describe a simple and efficient method to generate Caenorhabditis elegans null mutants using CRISPR/Cas9 and short single stranded DNA oligo repair templates to insert a universal 43-nucleotide-long knock-in cassette (STOP-IN) into the early exons of target genes. This STOP-IN cassette has stop codons in all three reading frames and leads to frameshifts, which will generate putative null mutations regardless of the reading frame of the insertion position in exons. The STOP-IN cassette also contains an exogenous Cas9 target site that allows further genome editing and provides a unique sequence that simplifies the identification of successful insertion events via PCR. As a proof of concept, we inserted the STOP-IN cassette at a Cas9 target site in aex-2 to generate new putative null alleles by injecting preassembled Cas9 ribonucleoprotein and a short synthetic single stranded DNA repair template containing the STOP-IN cassette and two ∼35-nucleotide-long homology arms identical to the sequences flanking the Cas9 cut site. We showed that these new aex-2 alleles phenocopied an existing loss-of-function allele of aex-2 We further showed that the new aex-2 null alleles could be reverted back to the wild-type sequence by targeting the exogenous Cas9 cut site included in the STOP-IN cassette and providing a single stranded wild-type DNA repair oligo. We applied our STOP-IN method to generate new putative null mutants for 20 additional genes, including three pharyngeal muscle-specific genes (clik-1, clik-2, and clik-3), and reported a high insertion rate (46%) based on the animals we screened. We showed that null mutations of clik-2 cause recessive lethality with a severe pumping defect and clik-3 null mutants have a mild pumping defect, while clik-1 is dispensable for pumping. We expect that the knock-in method using the STOP-IN cassette will facilitate the generation of new null mutants to understand gene function in C. elegans and other genetic model organisms.


Plasmid profiles of Mycobacterium avium complex isolated from swine.
Twenty strains of Mycobacterium avium complex (MAC) isolated from swine and five strains from humans were examined for drug susceptibility and plasmid content. Four strains of swine origin and two strains of human origin harbored plasmid DNAs differing in molecular weights. No relationship between plasmid contents and drug resistance was observed. Southern DNA-DNA hybridization showed that small plasmids from swine MAC strains were homologous to those from human origin at the nucleotide level.


Modified Nucleosides, Nucleotides and Nucleic Acids via Click Azide-Alkyne Cycloaddition for Pharmacological Applications.
The click azide = alkyne 1,3-dipolar cycloaddition (click chemistry) has become the approach of choice for bioconjugations in medicinal chemistry, providing facile reaction conditions amenable to both small and biological molecules. Many nucleoside analogs are known for their marked impact in cancer therapy and for the treatment of virus diseases and new targeted oligonucleotides have been developed for different purposes. The click chemistry allowing the tolerated union between units with a wide diversity of functional groups represents a robust means of designing new hybrid compounds with an extraordinary diversity of applications. This review provides an overview of the most recent works related to the use of click chemistry methodology in the field of nucleosides, nucleotides and nucleic acids for pharmacological applications.


Randomized Controlled Trial Comparing the Efficacy of Sustained-Release Formula of Mosapride-Plus-Esomeprazole Combination Therapy to Esomeprazole Monotherapy in Patients with Gastroesophageal Reflux Disease.
We aimed to evaluate whether adding a sustained-release (SR) formula of mosapride to proton-pump inhibitors (PPIs) would be more effective in controlling symptoms than PPI alone in patients with gastroesophageal reflux disease (GERD). Sixty patients with heartburn and/or regurgitation were randomly assigned to two groups: mosapride SR 15 mg combined with esomeprazole 20 mg once daily (ME group) and esomeprazole 20 mg once daily alone (E group). The primary endpoint was the complete-resolution rate of GERD symptoms after eight-week medication, and the secondary endpoints were the complete-resolution rate of GERD symptoms after four-week medication, symptom-improvement rates ≥ 50% after four- and eight-week medication, and change in reflux-disease-questionnaire (RDQ) and GERD-health-related quality-of-life (GERD-HRQL) scores from baseline at four- and eight-week medication. No significant differences in complete-symptom-resolution rates at eight weeks and four weeks or in the changes in RDQ and GERD-HRQL scores from baseline at four- and eight-week medication were observed between the ME and E groups. The symptom-improvement rate of ≥50% after four and eight weeks was comparable between both groups. Adding mosapride SR to esomeprazole in patients with GERD provides no additional benefits in controlling GERD symptoms.


Transconjunctival lateral cantholysis for closure of full-thickness eyelid defects.
To report a novel technique for closure of full-thickness eyelid defects too large for primary closure that avoids an external lateral canthotomy. Retrospective review of a consecutive case series of all patients undergoing transconjunctival lateral cantholysis. Thirteen eyelids of 12 patients underwent transconjuctival lateral cantholysis for closure of a full-thickness eyelid defect. Each defect occurred after excision of an eyelid cutaneous malignancy. Each defect would not close primarily without undue wound tension. Average horizontal defect size measured 13.7 mm (range, 11-20 mm). Average follow-up interval was 2.3 months (range, 0.5-5 months). All defects healed well with no evidence of dehiscence, notching or other complication related to closure. No patient requested or required further surgery for any reason. Transconjunctival lateral cantholysis allows for closure of larger full-thickness eyelid defects and avoids morbidity from lateral canthotomy/cantholysis.


Local tunneling spectroscopy across a metamagnetic critical point in the bilayer ruthenate Sr3Ru2O7.
The local spectroscopic signatures of metamagnetic criticality in Sr(3)Ru(2)O(7) were explored using scanning tunneling microscopy (STM). Singular features in the tunneling spectrum were found close to the Fermi level, as would be expected in a Stoner picture of itinerant electron metamagnetism. These features showed a pronounced magnetic field dependence across the metamagnetic critical point, which cannot be understood in terms of a naive Stoner theory. In addition, a pseudogap structure was observed over several tens of meV, accompanied by a c(2 x 2) superstructure in STM images. This result represents a new electronic ordering at the surface in the absence of any measurable surface reconstruction.