Quantitative Determination of Ethyl Glucuronide and Ethyl Sulfate in Postmortem and Antemortem Whole Blood Using Phospholipid Removal 96-Well Plate and UHPLC-MS-MS.
Postmortem ethanol formation is a well-known problem in forensic toxicology. Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are ethanol metabolites that can be used to distinguish antemortem alcohol intake from postmortem formation of ethanol and in addition can be a helpful tool in assessment of the hip-flask defense. To an aliquot of 100 µL whole blood, internal standard (IS) and water was added before protein precipitation treatment (PPT) with ice-cold acetonitrile (ACN). The supernatants were filtered through a 96-well phospholipid removal plate, evaporated to dryness and reconstituted in 150 µL water/ACN/formic acid (FA). Identification of compounds was performed using multiple reaction monitoring (MRM) in negative mode. Gradient elution was performed on a C18 column with methanol (MeOH) and 0.1% FA. The run time was 4.5 min, and 0.5 µL was injected on an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS-MS) instrument. Linearity was achieved (coefficient of determination (R2) ≥ 0.999) for EtG in the range of 0.089 to 22 mg/L (0.40-100 µM) and EtS 0.025 to 6.3 mg/L (0.20-50 µM). The limit of quantification (LOQ) was 0.067 mg/L (0.30 µM) for EtG and 0.019 mg/L (0.15 µM) for EtS. Between assay accuracy was -15% to 8% and precision reported as relative standard deviation (RSD) was ≤ 4.5%. Precision, estimated as the RSD of the concentration difference between results from two independent analyses of authentic whole blood samples, was ≤ 6.7%. Recovery was ≥ 61% for EtG and ≥ 77% for EtS and matrix effects (ME) were 99% to 103%. Method comparison was carried out with a previously used UHPLC-MS-MS method, and satisfactory agreement was achieved, and external proficiency testing control samples had z-score < ± 1. The method has been used in routine work for more than 4 years analyzing about 6,000 antemortem and postmortem whole blood samples and has proven to be robust and reliable.


Treatment of epidural cord compressions from Hodgkin's disease with chemotherapy. A report of two cases and a review of the literature.
Epidural cord compressions are a frequent and serious problem for patients with cancer. Standard treatment for these lesions includes local irradiation with or without surgery. Herein are reported two cases of epidural cord compression from Hodgkin's disease that responded dramatically to systemic chemotherapy. A review of the literature reveals reports of successful chemotherapy in the treatment of seven patients with epidural metastases secondary to lymphomas and 15 secondary to a variety of other tumors. The use of systemic chemotherapy is an increasingly important therapeutic option for treatment of epidural cord compressions in carefully selected patients.


Ethyl caffeate suppresses NF-kappaB activation and its downstream inflammatory mediators, iNOS, COX-2, and PGE2 in vitro or in mouse skin.
Ethyl caffeate, a natural phenolic compound, was isolated from Bidens pilosa, a medicinal plant popularly used for treating certain inflammatory syndromes. The purpose of this study was to investigate the structural activity, and the anti-inflammatory functions and mechanism(s) of ethyl caffeate. Ethyl caffeate was found to markedly suppress the lipopolysaccharide (LPS)-induced nitric oxide (NO) production (IC(50) = 5.5 microg ml(-1)), mRNA and protein expressions of inducible nitric oxide synthase (iNOS), and prostaglandin E(2) (PGE(2)) production in RAW 264.7 macrophages. Transient gene expression assays using human cox-2 promoter construct revealed that ethyl caffeate exerted an inhibitory effect on cox-2 transcriptional activity in 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated MCF-7 cells. Immunohistochemical studies of mouse skin demonstrated that TPA-induced COX-2 expression was significantly inhibited by ethyl caffeate with a superior effect to that of celecoxib, a nonsteroidal anti-inflammatory drug. The phosphorylation and degradation of inhibitor kappaB (IkappaB) and the translocation of nuclear transcription factor-kappaB (NF-kappaB) into the nucleus, as well as the activation of mitogen-activated protein kinases (MAPKs) induced by LPS in macrophages, were not affected by ethyl caffeate. Ethyl caffeate, however, could inhibit NF-kappaB activation by impairing the binding of NF-kappaB to its cis-acting element. These results suggest that ethyl caffeate suppresses iNOS and COX-2 expressions partly through the inhibition of the NF-kappaB.DNA complex formation. Structure-activity relationship analyses suggested that the catechol moiety and alpha,beta-unsaturated ester group in ethyl caffeate are important and essential structural features for preventing NF-kappaB.DNA complex formation. This study provides an insight into the probable mechanism(s) underlying the anti-inflammatory and therapeutic properties of ethyl caffeate.


Infections in prosthetic devices.
Infection in prosthetic devices is a rare but potentially serious complication of prosthesis implant surgery. Infections associated with a variety of permanently implanted devices are reviewed in the context of recent knowledge of the host-prosthesis interaction.


L-NAME inhibits tumor cell progression and pulmonary metastasis of r/m HM-SFME-1 cells by decreasing NO from tumor cells and TNF-alpha from macrophages.
Highly metastatic ras/myc-transformed serum-free mouse embryo (r/m HM-SFME-1) cells were injected subcutaneously to mice and the effects of Nomega-nitro-L-arginine methyl ester (L-NAME) on the tumor progression and pulmonary metastasis were investigated. In addition, production of nitric oxide (NO), matrix metalloproteinases (MMPs) and tumor necrosis factor-alpha (TNF-alpha) in the tumor cells and in a mouse macrophage-like cell line, J774.1 cells, was analyzed. The increase in footpad thickness was significantly smaller in the mice which were fed the L-NAME containing water (4.24+/-0.39 mg/day/mouse). The number of the tumor cells metastasized to the lungs was smaller in the L-NAME treated mice, although statistical significance was not found. Co-treatment of r/m HM-SFME-1 cells with interferon-gamma (IFN-gamma; 100 U/ml) and lipopolysaccharide (LPS; 0.5 microg/ml) significantly enhanced NO production, and the presence of L-NAME at 1 mM significantly decreased this response. In r/m HM-SFME-1 cells, MMP-2 was undetectable and MMP-9 was also very little in the basal level, and both MMPs were unaffected by the IFN-gamma and/or LPS treatments, not to mention by the L-NAME treatment. In J774.1 cells, any treatment including LPS appeared to enhance MMP-9 production, however, this upregulation was not inhibited by the additional presence of L-NAME. Production of TNF-alpha by J774.1 cells was markedly enhanced with LPS treatment, and this enhancement was significantly reduced in the presence of L-NAME. These results indicate that the inhibitory effects of L-NAME on the tumor cell progression and pulmonary metastasis could be due to suppression of NO from tumor cells and TNF-alpha from macrophages (Mol Cell Biochem, 2007).


Controversies in in rheumatology and autoimmunity: Is CORA meeting a good educational tool to increase the scientific knowledge?
Rheumatology and Autoimmunity are closely related fields and are among the most exciting areas in Medicine. Many conditions once regarded as mysterious and incurable are better understood and managed nowadays. Part of the great interest in these subjects derives from the fact that many controversial issues have arisen due to the rapid progression of knowledge, which means they are debatable. Over the years, the Controversies in Rheumatology and Autoimmunity (CORA) meetings promoted critical discussions not as an end but as a tool to increase the scientific knowledge of Rheumatologists and Clinical Immunologists. Beyond pursuing knowledge, being critical means questioning our inveterate beliefs and evaluating new conjectures and hypotheses. Thus, the approach to the debates in Medicine should be done with an open mind and free from all prejudices. Freedom of thought and speech are the fundamental values of our University, as exemplified by the motto "Universa Universis Patavine Libertas" which means "Padua freedom is universal for everyone". Patavine libertas initially referred to freedom from political and religious power but also freedom in research and teaching. For these reasons, Galileo Galilei moved to Padua in 1592, where he spent the most prosperous years of his life before being accused of heresy by the catholic church. We aim for the CORA congress to be an open-minded forum where active participation and exchange of ideas are promoted without prejudice. This special Issue of Autoimmunity Reviews is devoted to some controversies debated during the 7th CORA conference held in Turin, Italy, on March 16-18, 2023. Here we will discuss controversial entities, the use of old and new drugs, and insights into the classification, assessment of disease activity, and management of rheumatic diseases.


Hepatitis tuberculosis presenting as tumoral disease on ultrasonography.
Macronodular involvement of the liver is a rare manifestation of hepatitis tuberculosis. Two cases of this pseudotumoral form are reported on ultrasonography, demonstrating multiple hypoechoic nodules distributed throughout the liver. The authors state the difficulty in differentiating this form in its atypical presentation from lymphomatous or secondary malignancies. They stress the importance of the bacteriological and/or histological diagnosis which can be performed with fine-needle percutaneous biopsy under ultrasound guidance and which allows effective therapy.


Periodic vs constant high glucose in inducing pro-inflammatory cytokine expression in human coronary artery endothelial cells.
Fluctuating hyperglycemia exerts a more deleterious effect than constant hyperglycemia on cardiovascular outcome in diabetic patients. We investigated the inflammatory responses of human coronary artery endothelial cells (HCAECs) to constant and periodic high glucose in vitro. HCAECs were incubated for 72 h continuously either in normal glucose (5.5 mmol/L), constant high glucose (25 mmol/L glucose), periodic high glucose (5.5 and 25 mmol/L glucose alternating every 6 h) or mannitol. Concentrations of interleukin (IL)-6, tumor necrosis factor (TNF)-α and intercellular adhesion molecule (ICAM)-1 in the supernatants of cell culture were measured using ELISA kits. The mRNAs of IL-6, TNF-α and ICAM-1 were evaluated by real-time quantitative PCR. Periodic high glucose caused a more intense inflammatory response than normal glucose and constant high glucose in HCAECs, with a marked increase in IL-6, TNF-α and ICAM-1 in supernatants of cell culture (P < 0.05). The concentrations of the three pro-inflammatory cytokine mRNAs were higher in cells exposed to periodic high glucose than those exposed to constant high glucose and normal glucose (P < 0.05). In cultured HCAECs, periodic high glucose evoked a more intense inflammatory response than constant high glucose.


Evaluation of Platelet Indices in Lung Cancer Patients.
In this study, we aimed to determine platelet indices such as platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), Plateletcrit (PCT) platelet count (PLT) in lung cancer cases, and evaluate any relationships between these parameters and stage or histologic types. This retrospective study covered 44 lung cancer patients and 47 healthy subjects. Platelet indices including PLT, PCT, MPV, PDW were estimated and compared with normal subjects. The results were evaluated statistically. The PDW value was significantly higher in the cancer group compared to the control group; however, the values for PCT and MPV were lower. We suggest potential use of platelet indices in diagnosis of lung cancer.


A multi-group confirmatory factor analysis of the revised children's anxiety and depression scale (RCADS) in Spain, Chile and Sweden.
There is a need for a measure that can be used across countries and cultures to advance cross-cultural research about internalizing mental health symptoms in children and adolescents. The Revised Children's Anxiety and Depression Scale (RCADS) is a potential candidate, but no study has examined whether its scales are measured similarly in youth populations from different countries. In this study, we use confirmatory factor analysis (CFA) and multi-group CFA to examine the cross-cultural properties of a short and free to use 30-item version of RCADS that assesses social, generalized, panic, and separation anxiety alongside depression and obsessive-compulsive symptoms. We tested the factor structure of RCADS in children and adolescents from Chile, Spain, and Sweden, recruited using different research designs (i.e., school-based studies and an anonymous web survey), and whether the factor structure showed measurement invariance across the three countries. The proposed factor structure of RCADS showed good model/data fit in all three countries and was superior to a unidimensional model in which correlations among scale items were explained by a single broad internalizing factor. Each RCADS subscale showed adequate to excellent internal consistency in all three countries and multi-group CFA supported scalar invariance across the three countries. No clinical sample was included. This study provides an important first step in supporting the use of RCADS in cross-cultural research on depression, anxiety and obsessive-compulsive symptoms in children and adolescents, but more work on validity aspects of the scale across cultures is needed.


Adsorption and photodegradation of reactive red 120 with nickel-iron-layered double hydroxide/biochar composites.
Layered double hydroxide (LDH) materials were widely applied for adsorption and photodegradation of pollutants for wastewater treatment. New efficient LDH materials with adsorption and photodegradation abilities will be promising candidates for pollutants removal. Hence, a series of NiFe-LDH/biochar (NiFe/BC) were fabricated by the coprecipitation method for synergistic adsorption and photodegradation anionic dyes of reactive red 120 (RR120). The removal experiment showed that the addition of an appropriate amount of biochar into NiFe-LDH enhanced the adsorption capacity and its photocatalytic ability. The optimized NiFe/BC2 composite can remove 88.5 % of RR120 under visible light by adsorption and photocatalysis, which was much better than NiFe-LDH (63.3 %) and biochar (2.6 %). The photodegradation kinetic constant of the NiFe/BC2 composite was 3.1 and 104.8 times that of NiFe-LDH and BC. In addition, active species capture experiments and electron spin resonance (ESR) tests revealed the removal mechanisms of NiFe/BC composites for RR120 removal. This work affords a feasible strategy for preparing LDH-based photocatalyst with excellent adsorption and photocatalytic performance for wastewater treatment.


Rethinking the combination treatment of fulvestrant and anastrozole for metastatic breast cancer: an integrated reanalysis of aromatase-estrogen receptor axis.
Aberrant expression or hyperactivation of aromatase (CYP19A1)-estrogen receptor (ESR) axis is well identified as one of the major causes of breast cancer. Lots of drugs have been developed for targeting CYP19A1 or ESR respectively, such as anastrozole and fulvestrant. Recently, Mehta et al. reported in NEJM that the combined treatment of anastrozole and fulvestrant increased long-term survival of patients with metastatic breast cancer, especially for those without receiving endocrine therapy. However, the integrated prognostic analyses of CYP19A1 and ESR1/ESR2 indicated some contradictory outcomes to the recent clinical trial. Moreover, immunological investigation further revealed that targeting the whole CYP19A1-ESR axis might cause the inactivation of anti-tumor immune response, which largely attenuated its application prospects in breast cancer. Considered the pathophysiologic functions of CYP19A1 and ESR1/ESR2-mediated signaling pathway in breast cancer seem as more complicated than what we have already known, more precise evaluation will be needed in urgent.


The cerebellar projection from the raphe nuclei in the cat as studied with the method of retrograde transport of horseradish peroxidase.
Following injections of horseradish peroxidase (HRP) in the cerebellar cortex and nuclei of the cat, the distribution of labeled cells in the raphe nuclei was mapped. The findings confirm those made previously in studies of retrograde cell degeneration following cerebellar ablations (Brodal et al., 1960a), and in addition reveal new details in the projection of the raphe nuclei onto the cerebellar cortex and nuclei. All the raphe nuclei except nucleus linearis intermedius and nucleus linearis rostralis project onto the cerebellar cortex. The nuclei raphe obscurus and pontis contribute the greatest number of afferents to the cerebellum. With the exception of lobule VI which probably is the recipient of a weak projection, all parts of the cerebellar cortex receive afferents from the raphe nuclei. The heaviest projection is to the vermis of lobules VIIA and X, and to crus II. The afferents to the cerebellar nuclei are few in number (Tables 2-6). The observations indicate that each raphe neuron probably projects to more than one terminal site in the cerebellum. The findings are discussed with reference to other efferent and afferent studies of the raphe nuclei. All these studies indicate that the raphe nuclei have widespread efferent and afferent connections, making them capable to participate in a variety of regulatory functions.


Agenesis of Anterior Falx Cerebri in Patient with Planned Interhemispheric Approach to Third Ventricle Mass.
Complete or partial agenesis of the falx cerebri may occur in pediatric patients with developmental anomalies. However, isolated agenesis of the falx in a developmentally normal adult is exceptionally rare. We describe the first reported case of a patient with a third ventricular mass associated with partial agenesis of the anterior falx cerebri, a circumstance that influenced surgical access to a third ventricular epidermoid cyst. A 60-year-old developmentally normal woman presented with progressively worsening aphasia and altered mental status. Brain magnetic resonance imaging showed obstructive hydrocephalus from a third ventricular mass. An anterior interhemispheric transcallosal approach was planned to remove the tumor. However, upon dural opening there was no evidence of a falx cerebri, an anomaly visible but not reported on the prior imaging studies. An interhemispheric fissure was present, but the medial frontal lobes were densely adherent, with multiple traversing veins within the superficial arachnoid of the fissure. Therefore, a left frontal transcortical approach was performed for microsurgical resection of the tumor. Histopathologic analysis identified the lesion to be an epidermoid cyst. Partial agenesis of the falx cerebri is exceedingly rare in a developmentally normal adult, particularly in the presence of an anatomically normal superior sagittal sinus. If present, however, it is important to note this association preoperatively because partial agenesis of the falx cerebri precludes an interhemispheric transcallosal approach to the lateral and third ventricles.


A renaissance for Freud's Papers on Technique.
The analyst depicted in Freud's Papers on Technique (1911-1915) struggles to split the patient's stubborn desire by reframing it as memory. In contrast, after the structural theory was adopted, analysts could picture themselves as liberating rather than struggling with patients, since their design was to show that the patient's fears are unrealistic. But North American analysts have come to doubt their ability to rule on what is and isn't realistic. They thus find themselves again struggling with stubborn desires, not just easing fears. Their solution has been to emphasize--and thus detach from--enactments. These analysts would seem to be even more in need of the old ideals of procedure than were "classical" analysts, who thought they could use "reality" as a standard.


Genotype and phenotype study of 34 Spanish patients diagnosed with oculopharyngeal muscular dystrophy.
Oculopharyngeal muscular dystrophy is an autosomal dominant adult-onset disease with several clinical features. The genetic cause is an expanded (GCN)n mutation coding for polyalanine. Severity and the age of onset are variable and may depend on the size of the unstable triplet. Our objectives were to correlate the genotypic and phenotypic features in 34 affected patients, and to complete the molecular analysis for a control Spanish population in order to confirm the (GCN)n polymorphism frequency observed in other populations. We found a correlation between impaired CPK levels and sex. No statistical differences were found when comparing the length in triplet expansion and other variables. The (GCN)n polymorphism's frequency observed in other countries could not be proven in ours. Moreover, no correlation was observed amongst the size of the mutation, the age of onset, and the phenotype. This fact suggests that other conditions apart from the already known genotype could influence the age of onset and the severity of the symptoms.


Chromosome Painting Using Chromosome-Specific BAC Clones.
Chromosome painting (CP) refers to visualization of large chromosome regions, chromosome arms or entire chromosomes via fluorescence in situ hybridization (FISH) of chromosome-specific DNA sequences. For CP in crucifers (Brassicaceae), typically contigs of chromosome-specific bacterial artificial chromosomes (BAC) from Arabidopsis thaliana are applied as painting probes on chromosomes of A. thaliana or other species (comparative chromosome painting, CCP). CP/CCP enables to identify and trace particular chromosome regions and/or chromosomes throughout all mitotic and meiotic stages as well as corresponding interphase chromosome territories. However, extended pachytene chromosomes provide the highest resolution of CP/CCP. Fine-scale chromosome structure, structural chromosome rearrangements (such as inversions, translocations, centromere repositioning), and chromosome breakpoints can be investigated by CP/CCP. BAC DNA probes can be accompanied by other types of DNA probes, such as repetitive DNA, genomic DNA, or synthetic oligonucleotide probes. Here, we describe a robust step-by-step protocol of CP and CCP which proved to be efficient across the family Brassicaceae, but which is also applicable to other angiosperm families.


Anaplastic astrocytoma following radiation for a glomus jugular tumor.
Evaluation of radiation therapy for a given neoplasm includes consideration of possible treatment complications as well as potential benefit. A 43-year-old male with a glomus jugular tumor or the right middle ear had received 4480 rad to the right middle and inner ear and temporal bone. Eight years later, he developed an anaplastic astrocytoma of the right cerebellar hemisphere. At this time a third neoplasm, a left carotid body tumor, was demonstrated angiographically. Although radiation can be implicated in the genesis of the glial neoplasm, the presence of two neural crest derived tumors suggests that a lowered threshold for neoplastic transformation in neuroectodermal cells may have been an additional factor. Long-term follow-up of large numbers of patients with glomus jugulare tumors will be necessary to determine if multiple paragangliomas predispose to radiation-associated gliomas.


A new radio-opaque injection technique for tissue preservation.
This study has investigated the action of two commonly available preservatives, chlorbutol and chlorocresol, on fresh cadaver tissue. If the arterial system of the subject is perfused first with either preservative then the progression of tissue necrosis can be delayed for up to 1 month. The preservative can be combined with the lead oxide-gelatin mixture described by Rees and Taylor without compromising the results of radiographic studies. Both chlorocresol and chlorbutol have the advantage over formalin of retaining the normal colour and texture of the tissues. A regimen is suggested which has all of the advantages of the mixture originally described by Salmon, but is much simpler and cheaper to produce. It has proved successful when used in whole cadavers but the visceral contents must be removed at an early stage.


Streptococcus thermophilus is able to produce a beta-galactosidase active during its transit in the digestive tract of germ-free mice.
This work presents data on the application of a bacterial luciferase used to monitor gene expression of Streptococcus thermophilus in the digestive tract. The main result is that the bacterium was able to produce an active beta-galactosidase in the digestive tract, although it did not multiply during its transit. This production was enhanced when lactose (the inducer) was added to the diet.


Short-term fasting and prolonged semistarvation have opposite effects on 2-AG levels in mouse brain.
2-Arachidonoyl glycerol (2-AG) levels in whole mouse brain and two of its regions-hippocampus and hypothalamus-were determined after diet restriction (between 60 and 40%) lasting 12 days. The diet restriction lowered the level of 2-AG, which in the hypothalamus depended on the severity of the diet restriction, while the level in the hippocampus was not dependent on the diet regimen. As these observations differ from previously published data showing elevation of 2-AG levels in rat brain after 24 h of severe food restriction, we measured 2-AG levels in whole mouse brain after a comparable period of full starvation (fasting). We confirmed the elevation of 2-AG levels. It seems possible that these time-dependent variations of 2-AG levels may be of importance as a general coping strategy by animals during periods of starvation.


Quantification of low levels of amorphous content in crystalline celecoxib using dynamic vapor sorption (DVS).
A minor amount of amorphous phase, especially present on the surface of crystalline pharmaceutical actives, can have a significant impact on their processing and performance. Despite the presence of sophisticated analytical tools, detection and quantification of low levels of amorphous content pose significant challenges owing to issues of sensitivity, suitability, limit of detection and limit of quantitation. Current study encompasses the quantification of amorphous content in the crystalline form of celecoxib (CLB) using a dynamic vapor sorption (DVS) based method. Water, used as the solvent probe, achieved equilibration within a very short period of time (i.e. 6h) due to hydrophobic nature of CLB, thus allowing development of a rapid quantification method. The study included optimization of instrument and sample related parameters for the development of an analytical method. The calibration curve for amorphous CLB in crystalline CLB was prepared in the concentration range of 0-10% w/w. The analytical method was validated for linearity, range, accuracy and precision. The method for quantification was found to be linear with R(2) value of 0.999, rapid and sensitive for quantification of low levels of amorphous CLB content. It was able to detect the presence of amorphous phase in a predominantly crystalline phase at concentrations as low as 0.3% w/w. The limit of quantitation was found to be 0.9% w/w. Moreover, the influence of mechanical processing on the amorphous content in crystalline CLB was also investigated.


Modifications of the axon initial segment during the hibernation of the Syrian hamster.
Mammalian hibernation is a natural process in which the brain undergoes profound adaptive changes that appear to protect the brain from extreme hypoxia and hypothermia. In addition to a virtual cessation of neural and metabolic activity, these changes include a decrease in adult neurogenesis; the retraction of neuronal dendritic trees; changes in dendritic spines and synaptic connections; fragmentation of the Golgi apparatus; and the phosphorylation of the microtubule-associated protein tau. Furthermore, alterations of microglial cells also occur in torpor. Importantly, all of these changes are rapidly and fully reversed when the animals arouse from torpor state, with no apparent brain damage occurring. Thus, hibernating animals are excellent natural models to study different aspects of brain plasticity. The axon initial segment (AIS) is critical for the initiation of action potentials in neurons and is an efficient site for the regulation of neural activity. This specialized structure-characterized by the expression of different types of ion channels and adhesion, scaffolding and cytoskeleton proteins-is subjected to morpho-functional plastic changes upon variations in neural activity or in pathological conditions. Here, we used immunocytochemistry and 3D confocal microscopy reconstruction techniques to measure the possible morphological differences in the AIS of neocortical (layers II-III and V) and hippocampal (CA1) neurons during the hibernation of the Syrian hamster. Our results indicate that the general integrity of the AIS is resistant to the ischemia/hypoxia conditions that are characteristic of the torpor phase of hibernation. In addition, the length of the AIS significantly increased in all the regions studied-by about 16-20% in torpor animals compared to controls, suggesting the existence of compensatory mechanisms in response to a decrease in neuronal activity during the torpor phase of hibernation. Furthermore, in double-labeling experiment, we found that the AIS in layer V of torpid animals was longer in neurons expressing phospho-tau than in those not labeled for phospho-tau. This suggests that AIS plastic changes were more marked in phospho-tau accumulating neurons. Overall, the results further emphasize that mammalian hibernation is a good physiological model to study AIS plasticity mechanisms in non-pathological conditions.


Environmental fluctuations and host skin bacteria shift survival advantage between frogs and their fungal pathogen.
Fluctuating environments can modulate host-pathogen interactions by providing a temporary advantage to one of the interacting organisms. However, we know very little about how environmental conditions facilitate beneficial interactions between hosts and their microbial communities, resulting in individual persistence with a particular pathogen. Here, we experimentally infected Eleutherodactylus coqui frogs with the fungal pathogen Batrachochytrium dendrobatidis (Bd) under environmental conditions known to confer the survival advantage to the host during the warm-wet season, or alternatively to the pathogen during the cool-dry season. We used 16S rRNA amplicon sequencing to quantify changes in bacterial richness and phylogenetic diversity, and identified operational taxonomic units (OTUs) that became overrepresented or suppressed as a consequence of Bd infection. During the warm-wet season, frogs limited Bd infections, recruited putatively beneficial bacteria and returned to pre-infection levels of richness and phylogenetic diversity. In contrast, during the cool-dry season, Bd infections kept increasing through time, and bacterial diversity remained constant. Our findings confirm that infection outcome not only depends on abiotic factors, but also on biotic interactions between hosts and their associated bacterial communities.


Comparison of cyclizine and ondansetron for the prevention of postoperative nausea and vomiting in laparoscopic day-case gynaecological surgery.
Seventy-four patients undergoing laparoscopic gynaecological surgery were randomly allocated to two groups receiving cyclizine 50 mg or ondansetron 4 mg at induction of anaesthesia. Anaesthetic and postoperative analgesia regimens were standardised. Approximately half of the patients in each group experienced some degree of postoperative nausea and vomiting (cyclizine, 56%; ondansetron, 54%). There was no difference between groups in respect of pre- and postdischarge incidence. Mean (SD) time to eye opening was significantly prolonged in the cyclizine group [10 (4) min vs. 8 (2) min; p < 0.001], but this had no influence on discharge times. Cyclizine and ondansetron appear equally effective in preventing postoperative nausea and vomiting but the 10-fold price differential favours cyclizine.


Skeletal rearrangement of O-propargylic formaldoximes by a gold-catalyzed cyclization/intermolecular methylene transfer sequence.
Skeletal rearrangement of O-propargylic formaldoximes, in the presence of gold catalysts, afforded 4-methylene-2-isoxazolines in good to excellent yields by an intermolecular methylene transfer. In addition, the cascade reaction with maleimide in the presence of a gold catalyst afforded isoxazole derivatives by cyclization/methylene transfer and a subsequent ene reaction, whereas that using a copper catalyst gave oxazepines through a 2,3-rearrangement.


Oral Dirofilariasis-a Rare Case Report from South India.
Human dirofilariasis is a mosquito-borne/zoonotic nematode parasitic infestation caused by the genus Dirofilaria. From the forty isolated species of dirofilariasis, six are known to cause human infections. Intraoral occurrence is rare. We are reporting a case of oral dirofilariasis from Kerala, South India, with the view of acquainting Keralite dental professionals and Oral and Maxillofacial surgeons with the possibility of the parasitic infection in case of recurrent swellings and when the patient hails from a tropical climate and region where animal husbandry is the mainstay of livelihood.


Body composition and dietary intake in patients with head and neck cancer during radiotherapy: a longitudinal study.
To investigate the body composition and dietary intake in the patients with head and neck cancer (HNC) during radiotherapy (RT), and explore the relationship between them. This was a prospective, longitudinal observational study. Adult patients with HNC undergoing RT between March 2017 and August 2018 were recruited. Patients' body compositions were evaluated by bioelectrical impedance analysis, and dietary intake was recorded by 24-hour dietary recall at three time points, including baseline (T1), mid-treatment (T2) and post-treatment (T3). Patients were divided into low, middle and high energy intake groups based on the average daily energy intake (DEI). Changes in body weight (BW), fat mass (FM), fat-free mass (FFM) and skeletal muscle mass (SMM) among these three groups were compared. From T1 to T3, the median loss of patients' BW, FM, FFM and SMM was 4.60, 1.90, 2.60 and 1.50 kg, respectively. The loss of BW was more dramatic from T2 to T3 than that from T1 to T2. BW loss was mainly contributed by SMM loss from T1 to T2 and by FM loss from T2 to T3. Meanwhile, patients' dietary intake reduced during treatment. High DEI group had a significantly attenuated loss of patients' BW, FFM, SMM and FM compared with the low DEI group. Patients' BW, FM, FFM and SMM all significantly reduced, especially from T2 to T3, with decreased DEI during RT, which stresses the importance of nutrition intervention during the whole course of RT.


[Production of antibodies to influenza virus A by human lymphoid cells in vitro].
The data are presented on studying the production of antibodies to influenza A (H3N2) virus by lymphoid cells of the amygdaline tonsils and mediastinal lymph nodes in vitro during stimulation of these cells with influenza virus or lipopolysaccharide of typhoid bacteria.


Follicle-stimulating hormone receptor polymorphisms in a population of infertile women.
There are two known polymorphisms of clinical relevance in the follicle-stimulating hormone (FSH) receptor exon 10, alanine or threonine at position 307, and asparagine or serine at position 680, giving rise to two discrete allelic variants: Thr307/Asn680 and Ala307/Ser680. At position 680, three FSH receptor variants are possible: Asn/Asn, Asn/Ser, and Ser/Ser. We hypothesized an association between FSH receptor polymorphisms and ovarian reserve. FSH receptor polymorphisms at position 680 were studied in a population of 68 infertile women. We used serum FSH level at cycle day 3 as a screening for ovarian reserve. DNA was extracted from peripheral leukocytes in whole blood by using PCR and DNA sequencing in order to determine the type of FSH receptor. The distribution of FSH receptor variants was Asn/Asn (AA) 35%, Asn/Ser (AS) 24%, and Ser/Ser (SS) 41%. In women with normal ovarian reserve, FSH levels at cycle day 3 were 5.6 +/- 1.9 (AA group), 6.7 +/- 1.3 (AS group), and 5.7 +/- 1.7 (SS group) (non-significant). Corresponding FSH levels at cycle day 10 were 6.9 +/- 1.9, 6.3 +/- 1.7, and 8.3 +/- 2.8 (P < 0.01, AA and AS vs. SS group). In the SS group, FSH levels at cycle day 10 were significantly higher than they were at cycle day 3 (P < 0.05). The results show that Ser/Ser-680 predominates in the studied infertile population. Furthermore, women with normal ovarian reserve and the Ser/Ser FSH receptor variant had significantly higher FSH levels, compared to women with Asn/Asn and Asn/Ser variants. FSH receptor genotyping may, thus, be interesting as an adjunct indicator of ovarian reserve for infertile women undergoing assisted reproduction, and may be helpful in the determination of the starting dosage of FSH in in vitro fertilization.


A haplotypic variant at the IRGM locus and rs11747270 are related to the susceptibility for chronic periodontitis.
Immunity-regulated GTPase M (IRGM) plays a critical role in the defense against intracellular bacteria by regulating autophagy formation. This direct genetic association study aimed to determine whether variants at the IRGM genetic locus are associated with chronic periodontitis. Using PCR and melting curve analysis 390 periodontitis patients and 770 healthy controls have been genotyped regarding six polymorphisms in the IRGM gene (rs13361189, rs10065172, rs4958847, rs1000113, rs11747270, rs931058). Frequency distribution of alleles and genotypes for the six polymorphisms were not significantly different between the periodontitis and the control group. Also following stratification according to gender and smoking no significant linkage was found for any of the IRGM variants with periodontitis. Analysis of a subsample of patients revealed a significant association for rs11747270 with severe periodontitis (p = 0.003). Pairwise linkage analysis revealed one block composed of rs13361189, rs10065172, rs4958847, rs1000113 and 11747270 with strong or even complete linkage disequilibrium (r 2 > 0.9). Four haplotypes showed a frequency of > 1%, among which the haplotype C-T-A-T-G was significantly associated with chronic periodontitis (p = 0.0051; OR 4.66, 95% CI 1.41-15.42). One rare haplotype of the IRGM locus is significantly associated with chronic periodontitis in a German cohort.


Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica.
The enteric protozoan parasite Entamoeba histolytica was shown to possess cysteine synthase (CS) activity. The cDNA and genomic clones that encode two isoforms of the E. histolytica CS were isolated and characterized from a clonal strain of E. histolytica by genetic complementation of the cysteine-auxotrophic Escherichia coli NK3 with an E. histolytica cDNA library. The two types of the E. histolytica CS genes differed from each other by three nucleotides, two of which resulted in amino acid substitution. Deduced amino acid sequences of the E. histolytica CS, with a calculated molecular mass of 36721 Da and an isoelectric point of 6.39, exhibited 38-48% identity with CS of bacterial and plant origins. The absence of the amino-terminal transit peptide in the deduced protein sequences and the presence of the CS protein mainly in the supernatant fraction of the amoebic lysate after cellular fractionation suggested that the identified E. histolytica CS genes encoded cytosolic isoforms. Substrate specificity of the recombinant E. histolytica CS was similar to that of plant CS. Phylogenetic analysis indicates that the amoebic CS, first described in Protozoa, does not belong to any families of the CS superfamily, and represents a new family.


Isoprene in the Exhaled Breath is a Novel Biomarker for Advanced Fibrosis in Patients with Chronic Liver Disease: A Pilot Study.
Analysis of volatile organic compounds (VOCs) in the exhaled breath can identify markers for alcoholic and nonalcoholic fatty liver disease. The aim of this pilot study was to investigate the utility of breath VOCs measured by mass spectrometry to diagnose advanced fibrosis in patients with chronic liver disease (CLD). Patients undergoing liver biopsy were recruited. Fibrosis was determined by an experienced pathologist (F0-4) and advanced fibrosis was defined as F3-4. Exhaled breath and plasma samples were collected on the same day of the biopsy. Selective ion flow tube mass spectrometry (SIFT-MS) was used to analyze breath samples. Bonferroni correction was applied to decrease the false discovery rate. In all, 61 patients were included with a mean age of 50.7±9.9 years and 57% were male. Twenty patients (33%) had advanced fibrosis (F3-4), 44% had chronic hepatitis C, 30% had nonalcoholic fatty liver disease, and 26% had other CLD. SIFT-MS analysis of exhaled breath revealed that patients with advanced fibrosis had significantly lower values of six compounds compared with those without advanced fibrosis, P value <0.002 for all. Isoprene was found to have the highest accuracy for the prediction of advanced fibrosis with an area under the receiver operating characteristics curve of 0.855 (95% confidence interval: 0.762, 0.948). The median breath isoprene level in patients with F3-4 was 13.5[8.7, 24.7] p.p.b. compared with 40.4[26.2, 54.1] for those with F0-2, P value <0.001. Isoprene is an endogenous VOC that is a byproduct of cholesterol biosynthesis. Isoprene is a potential biomarker for advanced fibrosis that deserves further validation.


Reproductive Microbiomes: A New Thread in the Microbial Network.
Almost every part of our body has a coevolved microbial community. The expressed microbial genes comprise the various microbiomes that play important roles in normal physiology and development. The various microbiomes are separate, yet often connected, with the species composition of one affecting others. The female reproductive system microbiomes (eg, vaginal, placental, and mammary/milk) remain less well explored than the gut microbiome although they comprise a large proportion of the female microbial network. This review examines the evidence for interconnectivity between the female reproductive microbiomes, other maternal microbiomes, and developing infant microbiomes and the potential roles of each in health and disease. Disruptions in maternal microbiomes may be linked to pregnancy complications and maternal, fetal, and neonatal health. The diversity of the vaginal microbiome's makeup, which appears to vary across ethnicity, has led researchers to reconsider the idea of a "healthy" or "normal" vaginal microbial community. Less is known about the possible placental microbiome, although an association between the placenta's bacterial makeup and preterm labor and other pregnancy complications is being investigated. The mammary/milk microbiome appears to be influenced by maternal characteristics and may play a role in inoculating the infant but may also be affected by the infant's oral microbiome. Probiotic therapies such as "vaginal seeding" offer potential health benefits but require more rigorous testing. Exploring the reproductive microbiomes in detail and pairing this information with an individual's detailed medical history will provide a more complete picture of the status and importance of the microbial network to health.


Pharmaceutical innovation: possibilities and limits of personalised medicine.
In the framework of the German EU Council Presidency, the Federal Ministry of Health and the Federal Institute for Drugs and Medical Devices, an expert conference on Pharmaceutical Innovation - Possibilities and Limits of Personalised Medicine was held in Bonn, on June 11-12, 2007. It was financially supported by the European Commission. The conference aimed to intensify a sustainable networking among the participants and develop robust concepts for the future design of pharmaceutical research and production within the EU. To this end, various stakeholders from the fields of pharmaceutical research, production and licensing were brought together. This publication summarises the main results of the conference. It may reflect the personal views of the individual contributors and is not necessarily the perspective of the Federal Ministry of Health or of the Federal Institute for Drugs and Medical Devices.


Visual outcomes and predictors in optic pathway glioma: a single centre study.
Optic pathway gliomas (OPGs) may cause progressive visual loss despite chemotherapy. Newer, less toxic treatments might be given earlier, depending on visual prognosis. We aimed to investigate the prognostic value of visual evoked potentials (VEP) and optical coherence tomography (OCT). A retrospective study of OPG patients (treated 2003-2017) was conducted. Primary outcome was PEDIG category visual acuity in better and worse eyes (good < = 0.2, moderate 0.3-0.6 and poor > = 0.7 logMAR). Binary logistic regression analysis was used to identify predictors of these outcomes. 60 patients (32 Neurofibromatosis type 1 [NF1] and 28 sporadic) had median presentation age 49 months (range 17-183) (NF1) and 27 months (range 4-92) (sporadic). Median follow up was 82 months (range 12-189 months). At follow up 24/32 (75%) of NF1 children and 14/28 (50%) of sporadic children had good better eye visual acuity and 11/32 (34%) of NF1 children and 15/28 (54%) of sporadics had poor worse eye acuity. Mean peripapillary retinal nerve fibre layer (RNFL) thickness predicted good better eye final acuity (OR 0.799, 95%CI 0.646-0.987, p = 0.038). Presenting with visual symptoms (OR 0.22 95% CI 0.001-0.508, p = 0.017) and poorer VEP scores (OR 2.35 95% CI 1.1-5.03, p = 0.027) predicted poor worse eye final acuity. 16 children had homonymous hemianopias at follow up, predicted by poor presenting binocular VEP score (OR 1.449 95%CI 1.052-1.995, p = 0.02). We found that both RNFL thickness on OCT and VEP were useful in predicting future visual acuity and vision and potentially in planning treatment. We had a high prevalence of homonymous hemianopia.


Corrigendum: Machine learning for the detection of social anxiety disorder using effective connectivity and graph theory measures.
[This corrects the article DOI: 10.3389/fpsyt.2023.1155812.].


Enhancement of ε-poly-L-lysine (ε-PL) production by a novel producer Bacillus cereus using metabolic precursors and glucose feeding.
Epsilon poly-L-lysine (ε-PL) is a homo-biopolymer with approximately 25-30 L-lysine residues. It is a promising natural biopolymer widely used in food and pharmaceutical industry. The present work reports enhanced production of ε-PL with a novel producer Bacillus cereus using amino acids and TCA cycle intermediates in the fermentation medium. Among the various amino acids and TCA cycle intermediates tested 2 mM L-aspartic acid and 5 mM citric acid gave ε-PL yield of 145.5 and 230 mg/L, respectively. A combination of citric acid after 24 h and L-aspartic acid after 36 h improved ε-PL yield from 85 mg/L (control) to 335 mg/L. Glucose feeding strategy along with metabolic precursors was employed which further enhanced ε-PL yield to 565 mg/L. Thus, more than sixfold increase in ε-PL yield was achieved suggesting the potential of Bacillus cereus as a novel ε-PL producer.


Maternal levels of acute phase proteins in early pregnancy and risk of autism spectrum disorders in offspring.
Previous research supports a contribution of early-life immune disturbances in the etiology of autism spectrum disorders (ASD). Biomarker studies of the maternal innate (non-adaptive) immune status related to ASD risk have focused on one of the acute phase proteins (APP), C-reactive protein (CRP), with conflicting results. We evaluated levels of eight different APP in first-trimester maternal serum samples, from 318 mothers to ASD cases and 429 mothers to ASD-unaffected controls, nested within the register-based Stockholm Youth Cohort. While no overall associations between high levels of APP and ASD were observed, associations varied across diagnostic sub-groups based on co-occurring conditions. Maternal levels of CRP in the lowest compared to the middle tertile were associated with increased risk of ASD without ID or ADHD in offspring (OR = 1.92, 95% CI 1.08-3.42). Further, levels of maternal ferritin in the lowest (OR = 1.78, 95% CI 1.18-2.69) and highest (OR = 1.64, 95% CI 1.11-2.43) tertiles were associated with increased risk of any ASD diagnosis in offspring, with stronger associations still between the lowest (OR = 3.81, 95% CI 1.91-7.58) and highest (OR = 3.36, 95% CI 1.73-6.53) tertiles of ferritin and risk of ASD with ID. The biological interpretation of lower CRP levels among mothers to ASD cases is not clear but might be related to the function of the maternal innate immune system. The finding of aberrant levels of ferritin conferring risk of ASD-phenotypes indicates a plausibly important role of iron during neurodevelopment.


Hemodynamic changes in the portal vein with age: evaluation using four-dimensional flow MRI.
Aging process is associated with gradual change of liver function and structure. The goal of this study was to evaluate age-related hemodynamic changes in the portal vein (PV) using four-dimensional (4D) flow MRI in healthy adults. A total of 120 healthy subjects were enrolled and categorized into groups A (n = 25, 30-39 years), B (n = 31, 40-49 years), C (n = 34, 50-59 years), and D (n = 30, 60-69 years). All subjects underwent 4D flow data acquisition using a 3-T MRI system to measure the hemodynamic parameters in the main PV. The clinical characteristics and 4D flow parameters were compared among the groups using analysis of variance and analysis of covariance after controlling for significant covariates, accordingly. The outcome metric applying the age-related quadratic model to estimate the age at which 4D flow parameters are the highest (the peak age) as well as the rates of age-related 4D flow changes was estimated. The average area, average through-plane velocity, peak velocity magnitude, average net flow, peak flow, and net forward volume in group D were significantly lower than those in groups A, B and C (P < 0.05). Group C showed significantly lower values of the average through-plane velocity and peak velocity magnitude than those of group B (P < 0.05). The peak age computed was approximately 43-44 years of age for all 4D flow parameters. The rates of age-related 4D flow changes for all 4D flow parameters were negatively correlated with age (P < 0.05). The volume and velocity of the blood flow through the PV peaked at approximately 43-44 years of age and decreased significantly after 60 years of age.


The micromechanics of fluid-solid interactions during growth in porous soft biological tissue.
In this paper, we address some modelling issues related to biological growth. Our treatment is based on a formulation for growth that was proposed within the context of mixture theory (J Mech Phys Solids 52:1595-1625, 2004). We aim to make this treatment more appropriate for the physics of porous soft tissues, paying particular attention to the nature of fluid transport, and mechanics of fluid and solid phases. The interactions between transport and mechanics have significant implications for growth and swelling. We also reformulate the governing differential equations for reaction-transport of solutes to represent the incompressibility constraint on the fluid phase of the tissue. This revision enables a straightforward implementation of numerical stabilisation for the advection-dominated limit of these equations. A finite element implementation with operator splitting is used to solve the coupled, non-linear partial differential equations that arise from the theory. We carry out a numerical and analytic study of the convergence of the operator splitting scheme subject to strain- and stress-homogenisation of the mechanics of fluid-solid interactions. A few computations are presented to demonstrate aspects of the physical mechanisms, and the numerical performance of the formulation.


Gene duplications are extensive and contribute significantly to the toxic proteome of nematocysts isolated from Acropora digitifera (Cnidaria: Anthozoa: Scleractinia).
Gene duplication followed by adaptive selection is a well-accepted process leading to toxin diversification in venoms. However, emergent genomic, transcriptomic and proteomic evidence now challenges this role to be at best equivocal to other processess . Cnidaria are arguably the most ancient phylum of the extant metazoa that are venomous and such provide a definitive ancestral anchor to examine the evolution of this trait. Here we compare predicted toxins from the translated genome of the coral Acropora digitifera to putative toxins revealed by proteomic analysis of soluble proteins discharged from nematocysts, to determine the extent to which gene duplications contribute to venom innovation in this reef-building coral species. A new bioinformatics tool called HHCompare was developed to detect potential gene duplications in the genomic data, which is made freely available ( https://github.com/rgacesa/HHCompare ). A total of 55 potential toxin encoding genes could be predicted from the A. digitifera genome, of which 36 (65 %) had likely arisen by gene duplication as evinced using the HHCompare tool and verified using two standard phylogeny methods. Surprisingly, only 22 % (12/55) of the potential toxin repertoire could be detected following rigorous proteomic analysis, for which only half (6/12) of the toxin proteome could be accounted for as peptides encoded by the gene duplicates. Biological activities of these toxins are dominatedby putative phospholipases and toxic peptidases. Gene expansions in A. digitifera venom are the most extensive yet described in any venomous animal, and gene duplication plays a significant role leading to toxin diversification in this coral species. Since such low numbers of toxins were detected in the proteome, it is unlikely that the venom is evolving rapidly by prey-driven positive natural selection. Rather we contend that the venom has a defensive role deterring predation or harm from interspecific competition and overgrowth by fouling organisms. Factors influencing translation of toxin encoding genes perhaps warrants more profound experimental consideration.


Human metabolome variation along the upper intestinal tract.
Most processing of the human diet occurs in the small intestine. Metabolites in the small intestine originate from host secretions, plus the ingested exposome1 and microbial transformations. Here we probe the spatiotemporal variation of upper intestinal luminal contents during routine daily digestion in 15 healthy male and female participants. For this, we use a non-invasive, ingestible sampling device to collect and analyse 274 intestinal samples and 60 corresponding stool homogenates by combining five mass spectrometry assays2,3 and 16S rRNA sequencing. We identify 1,909 metabolites, including sulfonolipids and fatty acid esters of hydroxy fatty acids (FAHFA) lipids. We observe that stool and intestinal metabolomes differ dramatically. Food metabolites display trends in dietary biomarkers, unexpected increases in dicarboxylic acids along the intestinal tract and a positive association between luminal keto acids and fruit intake. Diet-derived and microbially linked metabolites account for the largest inter-individual differences. Notably, two individuals who had taken antibiotics within 6 months before sampling show large variation in levels of bioactive FAHFAs and sulfonolipids and other microbially related metabolites. From inter-individual variation, we identify Blautia species as a candidate to be involved in FAHFA metabolism. In conclusion, non-invasive, in vivo sampling of the human small intestine and ascending colon under physiological conditions reveals links between diet, host and microbial metabolism.


Multi-variate analysis of burns patients in the Singapore General Hospital Burns Centre (2003-2005).
The Burns Centre at the Singapore General Hospital (SGH) serves as a tertiary referral centre for burns management for Singapore's 4 million residents as well as the Southeast Asia region. Our study is a multivariate analysis of all burns patients admitted between 2003 and 2005. A total of 482 patients were admitted during this period with an average annual admission of 161. This represents a low incidence of 0.04 per 1000 admissions for the Singapore population. 13.3% of the study population were children, which is lower than previous studies. The mean age at admission was 35 years old and the male:female ratio was 1.9:1. We found a significant difference in age between the local and foreign patients, with the latter being younger. Our study demonstrated a 7.3% increase in cases of occupational burns. The bulk of our patients (57.3%) were directly admitted from SGH's Accident and Emergency Department. The patient characteristics of the various referral sources were found to be very different. GP referrals had significantly lower TBSA while overseas patients had significantly higher TBSA and longer length of stay. The mean and median time to admission was 3.05 days (+/-6.26) and 0 (0-60) day, respectively and the mean and median time to surgery was 7.33 days (+/-8.18) and 5 (0-22) days, respectively. The most common cause of burns was due to scalding. The mean extent of burn (TBSA) was 13.5% (+/-18.0), with significant correlation with the social background. Length of stay was dependent on the need for surgery. The overall mortality rate in this study population was 4.5%, with inhalation injury the main aetiological factor. In addition, the mean duration of the first surgery that patients undergo was significantly longer than that of the second one. This information will be useful for estimating operation times in the future. Finally, Acinetobacter baumannii was the most common bacteria in wound cultures. There is a need for periodic reviews of wound cultures in burns patients in order to modify the preventive and therapeutic strategies against these bacteria.


Integrating a Machine Learning System Into Clinical Workflows: Qualitative Study.
Machine learning models have the potential to improve diagnostic accuracy and management of acute conditions. Despite growing efforts to evaluate and validate such models, little is known about how to best translate and implement these products as part of routine clinical care. This study aims to explore the factors influencing the integration of a machine learning sepsis early warning system (Sepsis Watch) into clinical workflows. We conducted semistructured interviews with 15 frontline emergency department physicians and rapid response team nurses who participated in the Sepsis Watch quality improvement initiative. Interviews were audio recorded and transcribed. We used a modified grounded theory approach to identify key themes and analyze qualitative data. A total of 3 dominant themes emerged: perceived utility and trust, implementation of Sepsis Watch processes, and workforce considerations. Participants described their unfamiliarity with machine learning models. As a result, clinician trust was influenced by the perceived accuracy and utility of the model from personal program experience. Implementation of Sepsis Watch was facilitated by the easy-to-use tablet application and communication strategies that were developed by nurses to share model outputs with physicians. Barriers included the flow of information among clinicians and gaps in knowledge about the model itself and broader workflow processes. This study generated insights into how frontline clinicians perceived machine learning models and the barriers to integrating them into clinical workflows. These findings can inform future efforts to implement machine learning interventions in real-world settings and maximize the adoption of these interventions.


Specific detection of anti-idiotypic immune responses in cancer patients treated with murine monoclonal antibody.
Procedures are described for the specific detection in cancer patients' sera of human antibodies raised during the course of monoclonal antibody (MAb) immunotherapy against common and idiotypic determinants of monoclonal anti-colon carcinoma antibody 17-1A. The possible interference of serum factors in these assays other than human anti-murine immunoglobulin antibodies has been virtually excluded. Minimal detectable concentrations of antibody in the various assays ranged between 0.1 and 2.8 micrograms antibody per ml serum. Thirty-five of 41 patients who raised an immune response against the administered MAb produced anti-idiotypic antibodies in concentrations of up to 42 micrograms per ml serum. Anti-idiotypic antibodies persisted in the blood for up to 475 days following the administration of a single dose of MAb. A significant proportion of the anti-idiotypic antibodies were directed against the antigen-combining site of the MAb. The importance of these results in light of possible immunoregulatory functions of the anti-idiotypic antibodies is discussed.


1,4-Benzoquinone as a Highly Efficient Dopant for Enhanced Ionization and Detection of Nitramine Explosives on a Single-Quadrupole Mass Spectrometer Fitted with a Helium-Plasma Ionization (HePI) Source.
Previous investigations have evaluated the efficacy of anions such as NO3-, Cl-, Br-, CH3COO-, and CF3COO- as additives to generate or enhance mass spectrometric signals from explosives under plasma ionization conditions. The results of this study demonstrate that for detecting nitramine-class explosives, such as 1,3,5-trinitroperhydro-1,3,5-triazine (RDX) and 1,3,5,7-tetranitro-1,3,5,7-tetrazacyclooctane (HMX), 1,4-benzoquinone (BQ) is a highly effective and efficient dopant. When used in conjunction with ambient-pressure negative-ion helium-plasma ionization (HePI), 1,4-benzoquinone readily captures an electron, forming an abundant molecular anion (m/z 108), which upon exposure to vapors of RDX and HMX generates adduct ions of m/z 330 and 404, respectively. The signal level recorded for RDX upon adduction to the radical anion of 1,4-benzoquinone under our experimental conditions was significantly higher than that realized by chloride adduction using dichloromethane (DCM) as the dopant.


Isolation of proteolytic rumen bacteria by use of selective medium containing leaf fraction 1 protein (ribulosebisphosphate carboxylase).
The principle proteolytic bacteria isolated from bovine rumen contents by virtue of the ability to obtain nitrogen from the proteolysis of leaf fraction 1 protein (ribulosebisphosphate carboxylase, EC 22.214.171.124) were identified as Streptococcus bovis and Butyrivibrio spp. Substitution of fresh fodder, rich in soluble protein, for a hay-concentrates diet resulted in enhanced ruminal proteolytic activity and a significant increase in the number of bacteria able to use fraction 1 protein as the sole nitrogen source. Isolated proteolytic bacteria degraded fraction 1 protein and casein readily. Bovine serum albumin was attacked by Butyrivibrio spp. but was resistant to proteolysis by the streptococci.


Patient knowledge about biosimilars and satisfaction with the education provided by rheumatologists or nurse specialists in a biosimilar multiswitch scenario - The perception study.
To study the knowledge of patients with chronic inflammatory rheumatic diseases (CIRD) about biosimilars (bsDMARDs), assess patients' satisfaction after being educated about switching of bsDMARDs by rheumatologists compared to nurse specialists, and to explore the impact of multiple switches on patient satisfaction. Adult patients with CIRD who underwent a non-medical switch from the adalimumab bsDMARDs GP2017 to the adalimumab bsDMARDs MSB 11022 were 1:1 randomized with randomly selected block sizes into two groups in which information about multiple switching of bsDMARDs was provided by either a nurse specialist or a rheumatologist. Validated outcome tools and standardized parameters for disease activity and function were assessed at baseline and 12 weeks after the switch. The primary endpoint was to evaluate whether satisfaction with care differs when education about switching is provided by rheumatologists or nurse specialists. Secondary endpoints were patients' knowledge about bsDMARDs and the efficacy and safety of switching in routine care. Patients' satisfaction with care was assessed by the Leeds Satisfaction Questionnaire. A structured questionnaire was used to assess the patient's knowledge. A total of 102 patients was randomized, with 40 educated by rheumatologists (39.2%) and 62 by nurse specialists (60.8%). Patients had moderate to low disease activity and limited impairment of physical function without progression on follow-up, implying that switching did not affected disease activity. Almost half of the patients (n = 50, 49%) had undergone one and 52 multiple switches (51%), respectively. Less than one-third of patients were able to correctly answer questions on manufacturing, effectiveness, clinical trial evidence, and cost of bsDMARDs. Patients were generally satisfied with the education - irrespective of whether the information had been provided by nurses or rheumatologists. No relevant differences in the outcomes assessed were observed. Efficacy and safety results were consistent with previously published data. Patient satisfaction after education about bsDMARDs and multiple switching by nurses and rheumatologists was equally good. Multiple switches had no negative impact on patient satisfaction, and outcomes after switching of bsDMARDs did not significantly worsen. Patients' knowledge about bsDMARDS was limited.


Expression of the chondrogenic phenotype by mineralizing cultures of embryonic chick calvarial bone cells.
Cells released by sequential enzymatic digestion of 18-day chick calvariae were cultured over a 4-5 week period in Alpha modified Eagles medium. In some cultures the medium was supplemented with ascorbate and/or Na-beta-glycerophosphate. Microscopic examination of these cultures showed both polygonal and spindle-shaped cells. The biochemical nature of these cells was investigated by incubating the cultures with radiolabelled proline and subsequently analysing the medium and cell layer proteins by SDS/PAGE and fluorography. Osteoblast and chondrocyte-containing cultures were clearly distinguished in this way as the former cells secreted type I collagen while the latter secreted types II and X collagens as the major medium macromolecules. Type X collagen synthesis occurred after 14 days, but only in cultures supplemented with both ascorbate and Na-beta-glycerophosphate, and was maintained for the duration of the culture period. Unsupplemented cultures and those containing either ascorbate alone or Na-beta-glycerophosphate alone failed to synthesize type X collagen after 28 days. Isolated cells pulsed with radiolabelled proline at confluence and organ cultures of embryonic chick calvaria synthesized types I and V collagens only. These data demonstrate that the expression of phenotype by heterogeneous populations of bone cells could be modulated by a combination of culture conditions including the length of time in culture and conditions favourable for the formation of a mineralized matrix.


Effects of traditional Chinese medicine-based exercises on cognitive function in older people with mild cognitive impairment: A systematic review and meta-analysis.
This study evaluated the effects of traditional Chinese medicine based exercise (TCE) on cognitive outcomes for patients with mild cognitive impairment (MCI), and tried to identify the most effective TCE modality. A systematic review and meta-analysis of randomized controlled trials were conducted. Ten studies met the inclusion criteria. TCE interventions were classified into three types: (a) Tai Chi, (b) Baduanjin, and (c) Qigong. The pooled analysis showed that, overall, TCE had beneficial effects on global cognition and memory. Subgroup analysis further demonstrated that Baduanjin had a larger effect size on global cognition than the other TCE modalities. By contrast, Tai Chi had a larger effect size on memory than the other modalities. This study implied that TCE is a promising exercise option for improving cognition in MCI. However, further studies with a more rigorous study design are warranted to support or falsify the findings of the present review.


Quantitative measurements of binary amino acids mixtures in yellow foxtail millet by terahertz time domain spectroscopy.
Terahertz time domain spectroscopy (THz-TDS) combined with chemometrics has been utilized for the qualitative and quantitative analysis of binary mixtures of l-glutamic acid and l-glutamine which have similar chemical structures and properties. The binary mixtures of amino acids were prepared with yellow foxtail millet matrix, substituted for polyethylene (PE) as previously reported. After proper pretreatment of absorption spectra, quantitative analysis was achieved by partial least squares (PLS) and interval partial least squares (iPLS) regressions. The performance of models was evaluated based on the root mean square error of prediction (RMSEP) and correlation coefficient (R(2)) of cross-validations with bootstrapped Latin partitions as criterion. The iPLS yielded better results with low RMSEP (0.39±0.02%, 0.39±0.02%), and higher R(2) values (0.9904, 0.9906) for glutamine and glutamic acid comparing to the conventional PLS models. Multivariate curve resolution alternating least squares (MCR-ALS) was successfully applied for resolution of pure THz spectra and concentration profiles of two amino acids components from mixtures.


BMP-2 modulates the proliferation and differentiation of normal and cancerous gastric cells.
Bone morphogenetic protein 2 (BMP-2), a member of the transforming growth factor beta super-family, has been shown to act as an antiproliferative agent for a variety of cell lines by activating signaling cascades that cause cell cycle arrest. However, the biological effect and mechanism of action of BMP-2 on gastric cells remain unknown. In the present study, we showed that recombinant human BMP-2 dose-dependently inhibited the growth of OUMS37 rat gastric cells and MKN74 human gastric cancer cells. The antiproliferation seems to be due to cell cycle arrest in the G1-phase, which was revealed by flow cytometric assays. BMP-2 increased the level of p21/WAF1/CIP1, suggesting that BMP-2-mediated inhibition of cell proliferation may be induced through p21/WAF1/CIP1. In addition, BMP-2 increased the expression of pepsinogen II, a differentiation marker of the stomach, in MKN74 cells. These results indicate that BMP-2 plays important roles in modulating the proliferation and differentiation of gastric epithelial cells.


Subspace-based Identification Algorithm for characterizing causal networks in resting brain.
The resting brain has been extensively investigated for low frequency synchrony between brain regions, namely Functional Connectivity (FC). However the other main stream of brain connectivity analysis that seeks causal interactions between brain regions, Effective Connectivity (EC), has been little explored. Inherent complexity of brain activities in resting-state, as observed in BOLD (Blood Oxygenation-Level Dependant) fluctuations, calls for exploratory methods for characterizing these causal networks. On the other hand, the inevitable effects that hemodynamic system imposes on causal inferences in fMRI data, lead us toward the methods in which causal inferences can take place in latent neuronal level, rather than observed BOLD time-series. To simultaneously satisfy these two concerns, in this paper, we introduce a novel state-space system identification approach for studying causal interactions among brain regions in the absence of explicit cognitive task. This algorithm is a geometrically inspired method for identification of stochastic systems, purely based on output observations. Using extensive simulations, three aspects of our proposed method are investigated: ability in discriminating existent interactions from non-existent ones, the effect of observation noise, and downsampling on algorithm performance. Our simulations demonstrate that Subspace-based Identification Algorithm (SIA) is sufficiently robust against above-mentioned factors, and can reliably uncover the underlying causal interactions of resting-state fMRI. Furthermore, in contrast to previously established state-space approaches in Effective Connectivity studies, this method is able to characterize causal networks with large number of brain regions. In addition, we utilized the proposed algorithm for identification of causal relationships underlying anti-correlation of default-mode and Dorsal Attention Networks during the rest, using fMRI. We observed that Default-Mode Network places in a higher order in hierarchical structure of brain functional networks compared to Dorsal Attention Networks.


The role of gas chromatography/mass spectrometry. Instrumental techniques in forensic urine drug testing.
Appropriate GC/MS analytic techniques can provide conclusive confirmatory evidence for the presence of drugs and metabolites in forensic urine drug testing. Although forensic laboratories are required to utilize GC/MS, they have flexibility and freedom in selecting instrumentation, modes of operation, and actual analytic methods. It is incumbent on the laboratory to optimize analytic procedures so that reliability and accuracy are maximized. The laboratory needs to be aware of potential pitfalls in both the handling and the analyses of specimens. Specificity can be enhanced by utilizing mass spectrometric operating modes such as CI and full scan, which serve to complement SIM analyses. They are useful in solving difficult identification problems and in enhancing sensitivity as well as specificity. Newer types of GC/MS systems utilizing ion trapping or tandem mass spectrometry offer the promise of enhanced analytic performance and productivity gains. Knowledge of the advantages and potential pitfalls of different GC/MS systems as well as ionization and detection modes is helpful in optimizing the accuracy of compound identification.


Trends and Risk Factors for Vaginal Cuff Dehiscence after Laparoscopic Hysterectomy.
The primary objective was to assess the effect of the route of closure of the vaginal cuff on the incidence of vaginal cuff dehiscence (VCD) in laparoscopic hysterectomy (LH). The secondary objective was to assess patient- and surgical-risk factors associated with VCD, rate of perioperative complications by route of closure, and impact of surgeon volume on complications. Retrospective chart review with case-control component. Tertiary care center (main hospital and regional hospitals). A total of 1278 women underwent LH or robot-assisted hysterectomy in 2016, and met the inclusion criteria. Independently, 26 cases of VCD were identified from 2009 through 2016. A retrospective comparison of patients with vaginal cuff closure and laparoscopic cuff closure (LCC) undergoing LH or robot-assisted hysterectomy in 2016. Patients with VCD from 2009 through 2016 (n = 26) were matched by route of cuff closure to the next 7 patients who underwent hysterectomies (n = 182), who became controls. In 2016, there were 9 cases of VCD (0.70%). There was no significant difference in VCD between LCC (8/989; 0.81%) and vaginal cuff closure (1/289; 0.35%; p = .41). Seven VCD cases were performed by high-volume surgeons (>30 hysterectomies per year) who were more likely to perform LCC and use barbed suture. There were no significant differences in the rates of perioperative complications or surgeon volume between routes of cuff closure. The case-control patients differed in smoking status (p = .010) and history of prior laparotomy (p = .017). Logistic regression showed that increasing age (odds ratio 0.95; 95% confidence interval, 0.91-0.99) and increasing body mass index (odds ratio 0.98; 95% confidence interval, 0.83-0.97) were protective for VCD. VCD is a rare but serious complication of LH. Despite previous studies, we did not find a significant difference in VCD or intra- and perioperative complications by route of cuff closure or surgeon volume. Given the lack of evidence favoring one route of cuff closure, we recommend that, to optimize patient outcomes, surgeons employ the closure technique that they are best accustomed to.


Imaging features associated with H3 K27-altered and H3 G34-mutant gliomas: a narrative systematic review.
Advances in molecular diagnostics accomplished the discovery of two malignant glioma entities harboring alterations in the H3 histone: diffuse midline glioma, H3 K27-altered and diffuse hemispheric glioma, H3 G34-mutant. Radiogenomics research, which aims to correlate tumor imaging features with genotypes, has not comprehensively examined histone-altered gliomas (HAG). The aim of this research was to synthesize the current published data on imaging features associated with HAG. A systematic search was performed in March 2022 using PubMed and the Cochrane Library, identifying studies on the imaging features associated with H3 K27-altered and/or H3 G34-mutant gliomas. Forty-seven studies fulfilled the inclusion criteria, the majority on H3 K27-altered gliomas. Just under half (21/47) were case reports or short series, the remainder being diagnostic accuracy studies. Despite heterogeneous methodology, some themes emerged. In particular, enhancement of H3 K27M-altered gliomas is variable and can be less than expected given their highly malignant behavior. Low apparent diffusion coefficient values have been suggested as a biomarker of H3 K27-alteration, but high values do not exclude this genotype. Promising correlations between high relative cerebral blood volume values and H3 K27-alteration require further validation. Limited data on H3 G34-mutant gliomas suggest some morphologic overlap with 1p/19q-codeleted oligodendrogliomas. The existing data are limited, especially for H3 G34-mutant gliomas and artificial intelligence techniques. Current evidence indicates that imaging-based predictions of HAG are insufficient to replace histological assessment. In particular, H3 K27-altered gliomas should be considered when occurring in typical midline locations irrespective of enhancement characteristics.


Frequency of night shift and menstrual cycle characteristics in Japanese nurses working under two or three rotating shifts.
In Japan, the prevalence of irregular menstrual cycles and its association with the frequency of night shifts have scarcely assessed. The present study aimed to evaluate the relationship between irregular menstrual cycles and the frequency of night shifts in Japanese female nurses. We conducted a cross-sectional web-based self-administered questionnaire survey in 2019. An irregular menstrual cycle was defined as a cycle length of ≤21 days or ≥39 days at least a few times over the past year or amenorrhea for at least 3 months. We used Poison regression analysis with a robust error variance to calculate the prevalence ratios adjusted for age, body mass index, hospital size, and the department in which they worked. A total of 1249 women were included, and 679 (54.4%) and 195 (15.6%) of them worked under two and three rotating shifts. The prevalence of irregular menstrual cycles was 24.8%, 37.4%, and 35.9% in the no night, two rotating, and three rotating shifts groups, respectively. While the frequency of night shifts had a dose-responsive relationship with irregular menstrual cycles in the two rotating shifts group, it was not observed in the three rotating shifts group. However, the risk of work getting affected by dysmenorrhea or premenstrual symptoms increased in the three rotating shifts group. Over 30% of Japanese female nurses working under night shifts had irregular menstrual cycles. The high frequency of night shifts increased the risk of irregular menstrual cycles and secondary amenorrhea in the two rotating shifts group.


ISR II study: a long-term evaluation of sirolimus-eluting stent in the treatment of patients with in-stent restenotic native coronary artery lesions.
The aim of this pilot study was to determine the safety and long-term efficacy of treating intrastent restenosis (ISR) with the slow-release sirolimus-eluting stent Bx Velocity (Cypher stent) without intravascular ultrasound (IVUS) guidance. Of patients who received a bare metal stent implantation and presented an ISR, 30-80% of the patients will develop a second restenosis within the stent, at the stent edges or both. To date, intravascular brachytherapy using beta- and gamma-radiation has been the only effective treatment for ISR. Twenty-three patients with ISR and evidence of ischemia were treated with Cypher stent. Clinical information was collected 1, 8, 12, and 24 months after stent implantation. During the first 8 months of the study, in-stent lumen diameter remained essentially unchanged from postprocedure in 80% of the case. The target lesion repeat revascularization (TLR) was 17%, of which 50% were oculostenotic reflexes. Only one patient presented a restenosis greater than 70%. During the 2-year study period, the TLR rate was 17%; the major adverse coronary event rate was 26%, and the non-Q-wave myocardial infarction (MI) rate was 9%. There were no reports of death, coronary artery bypass grafting, or Q-wave MI during the study. This study demonstrates the feasibility of using sirolimus-eluting stents without IVUS guidance for the treatment of ISR, providing long-term stability of immediate results.


Human gaze stability in the horizontal, vertical and torsional direction during voluntary head movements, evaluated with a three-dimensional scleral induction coil technique.
The stability of gaze in three dimensions (horizontal, vertical and torsion) was measured with a new type of scleral search coil in eight emmetropic observers. Subjects held the head still or oscillated it at 0.16-0.67 Hz (amplitude about 10 deg) in the horizontal, vertical or torsional plane while fixating a point target at optical infinity. Veridical gaze and head coordinates were calculated with full correction for non-linear goniometric relations and for cross-coupling artifacts due to misalignments of the coil on the eye. The amount of gaze instability in the horizontal and vertical direction was virtually identical. With the head still, in either of these directions the mean standard deviation of gaze position (inclusive saccades) was about 7 min arc; mean non-saccadic retinal image speeds were 20-30 min arc/sec. During head oscillation these values increased to about 16 min arc and 1 deg/sec; a mean of about 2.5% of the head motion remained uncorrected by the compensatory eye movements. These findings agree well with our earlier results for the horizontal plane; the effect of the corrections was relatively small because the adventitious cross-coupling of horizontal and vertical to torsional head movements proved to be usually smaller than 10%. However, the corrections were important when head torsion was deliberately produced. Gaze stability in the torsional plane was considerably inferior to that in the horizontal and vertical plane. With the head held still, the mean SD of torsional gaze position was about 17 min arc; mean torsional non-saccadic retinal image speed was about 46 min arc/sec. Gain of the torsional compensatory eye movements was frequency dependent and rose from about 0.26 in static conditions (0 Hz) to about 0.42 at 0.16 Hz and 0.64 at 0.67 Hz. Accordingly, position instability and speed of the retinal image in torsion were about an order of magnitude larger than in the horizontal and vertical direction.


Diagnosis and treatment of malignant tumors using integrated traditional and western medicine: progress, challenges and reflections.
Malignant tumors are one of the leading causes of death in the world. Considerable progresses have been made on the treatment of tumors in recent decades, especially in the prevention, early diagnosis and the model changing of therapeutics. But we are still facing tough challenges, including the increasing treatment burden and limited improvement of efficacy. In China, Chinese medicine (CM) provides a powerful arsenal to fight against tumors. CM can be well applied to the onset and progression of tumors in China, bearing the characteristics of multi-target, multi-phase and multi-effect. But there are also many problems demanding urgent attention in the use of CM. Some most debated problems in this field were summarized. We should upgrade our concepts in using CM, find its position scientifically, and establish evidence of its effect by high quality clinical research.


Hydrothermal conversion of biomass to fuels, chemicals and materials: A review holistically connecting product properties and marketable applications.
Biomass is a renewable and carbon-neutral resource with good features for producing biofuels, biochemicals, and biomaterials. Among the different technologies developed to date to convert biomass into such commodities, hydrothermal conversion (HC) is a very appealing and sustainable option, affording marketable gaseous (primarily containing H2, CO, CH4, and CO2), liquid (biofuels, aqueous phase carbohydrates, and inorganics), and solid products (energy-dense biofuels (up to 30 MJ/kg) with excellent functionality and strength). Given these prospects, this publication first-time puts together essential information on the HC of lignocellulosic and algal biomasses covering all the steps involved. Particularly, this work reports and comments on the most important properties (e.g., physiochemical and fuel properties) of all these products from a holistic and practical perspective. It also gathers vital information addressing selecting and using different downstream/upgrading processes to convert HC reaction products into marketable biofuels (HHV up to 46 MJ/kg), biochemicals (yield >90 %), and biomaterials (great functionality and surface area up to 3600 m2/g). As a result of this practical vision, this work not only comments on and summarizes the most important properties of these products but also analyzes and discusses present and future applications, establishing an invaluable link between product properties and market needs to push HC technologies transition from the laboratory to the industry. Such a practical and pioneering approach paves the way for the future development, commercialization and industrialization of HC technologies to develop holistic and zero-waste biorefinery processes.


Observations of the pineal region of non-eutherian mammals.
A survey has been made of the pineal region of the brain of 11 species of marsupials belonging to 5 families and a species from both families of monotremes. The results show that the pineal body of non-eutherian mammals, although well-defined in all species, has a very varied morphology. Three types of pineal recess occur: (i) a pineal recess in sensu stricto, (ii) an intercommissural pineal recess, and (iii) an infrapineal recess. The existence of nerve fibres which pass through the pineal body and form a spatial link between the habenular and posterior commissures, has been demonstrated in marsupials and monotremes. It is also likely that these animals as well as eutherian mammals possess a nervus conarii. Nerve cells are not a constant feature of the non-eutherian pineal body. The subcommissural organ (SCO) is present in all species. It does not exhibit the same degree of morphological variation as the pineal body. Horizontal sections available for 4 species within 3 families of marsupials show it to be composed of a median portion jointed to bilateral protuberances. Large nerve cells occur within the SCO in all marsupial species; they are absent from the monotreme SCO. Tentatively, the relationship of these neurons to the SCO is considered to be merely one of association. The importance of an extended comparative study of this region in non-eutherian mammals in order to add insight into its phylogeny and function is emphasized.


Psychrobacter fjordensis sp. nov., a psychrotolerant bacterium isolated from an Arctic fjord in Svalbard.
A Gram-negative, non-motile, non-spore-forming, psychrotolerant and halotolerant bacterium designated BSw21516B(T), was obtained from seawater in Kongsfjorden, a glacial fjord in the Arctic Svalbard and subjected to taxonomic analysis using a polyphasic approach. This bacterium was observed to optimally grow at 25-29 °C; between at 4 and 34 °C, but not at >35 °C; and in the presence of 0-8 % (w/v) NaCl at an optimum concentration of 2-5 % (w/v) NaCl. Strain BSw21516B(T) was found to contain Ubiquinone-8 (Q-8) as a predominant respiratory lipoquinone and C18:1 ω9c and summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH) as predominant cellular fatty acids. Phylogenetic analysis of 16S rRNA and gyrB gene sequences showed that this isolate belongs to the genus Psychrobacter and is closely related to Psychrobacter fozii LMG 21280(T), which was isolated from a sediment sample in Antarctica. DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 58.6 %) between strain BSw21516B(T) and its closest relatives. Based on these results a new species Psychrobacter fjordensis sp. nov. is proposed (type strain BSw21516B(T) = KCTC 42279(T) = CCTCC AB 2014020(T)).


Access to Derivatizable Octahydrofluorenyl Ligand: Half-Sandwich Rare-Earth Metal Complexes for Highly Syndiospecific (Co)Polymerization of Styrene.
A simple and facile synthetic pathway for accessing new derivatizable bulky-demanding octahydrofluorenyl (OHF) ligands has been developed, and a series of half-sandwich rare-earth metal (Sc, Y, Lu) complexes bearing the OHF ancillary ligands have been synthesized. In conjunction with a borate, the OHF-ligated Sc complexes exhibited high catalytic activity for styrene (co)polymerization to afford polymers with highly syndiotactic polystyrene sequence (>99% rrrr).


RNAlater facilitates microdissection of sensory cell-enriched samples from the mouse cochlea for transcriptional analyses.
Molecular analyses of cochlear pathology rely on the acquisition of high-quality cochlear samples. For small rodents, isolating sensory cell-enriched samples with well-preserved RNA integrity for transcriptional analyses poses a significant challenge. Here, we report a microdissection technique for isolating sensory cell-enriched samples from the cochlea. We found that treating the tissue with RNAlater, a RNA preservation medium, alters the physical properties of the tissue and facilitates the dissection. Unlike previous samples that have been isolated from the sensory epithelium, our samples contain defined cell populations that have a consistent ratio of sensory cells to supporting cells. Importantly, the RNA components were well preserved. With this microdissection method, we collected three types of samples: sensory cell-enriched, outer hair cell-enriched, and inner hair cell-enriched. To demonstrate the feasibility of the method, we screened multiple reference genes in the sensory cell-enriched samples and identified stable genes in noise-traumatized cochleae. The method described here balances the need for both quality and purity of sensory cells and also circumvents many limitations of the currently available techniques for collecting cochlear tissues. With our approach, the collected samples can be used in diverse downstream analyses, including qRT-PCR, microarray, and RNA sequencing.


Substance P in the amygdaloid complex, bed nucleus and stria terminalis of the rat brain.
Radioimmunoassay and immunohistochemical techniques have demonstrated the presence of substance P in the medial and central nuclei of the amygdala and the bed nucleus of the stria terminalis. Hemisections and micro-knife cuts transecting the anterior, posterior, medial, lateral and ventral connections to the amygdala did not modify the content of substance P in the amygdala. In addition knife cuts totally isolating the medial amygdaloid nucleus from lateral and anterior-posterior connections did not reduce the substance P content of the medial nucleus, but produced a 70% reduction in the substance P content of the central nucleus. These results suggest that substance P containing neurones in the medial and central amygdaloid nuclei do not receive substance P projections originating outside the amygdala. However, there appears to be a short substance P projection from the medial nucleus to the central nucleus.


Motor-sparing peripatellar plexus block provides noninferior block duration and complete block area of the peripatellar region compared with femoral nerve block: a randomized, controlled, noninferiority study.
Developing adequate regional anaesthesia for knee surgeries without affecting lower limb mobilization is crucial to perioperative analgesia. However, reports in this regard are limited. We proposed a technique for ultrasound-guided peripatellar plexus (PP) block. Compared with the femoral nerve (FN) block, we hypothesized that this technique would provide a noninferior block duration and a complete cutaneous sensory block in the peripatellar region without affecting lower limb mobilization. An investigation was conducted to verify our hypothesis in cadavers and volunteers. The study was designed in two parts. First, eight cadaveric lower limbs were dissected to verify the feasibility of PP block after methylene blue injection under ultrasound. Second, using a noninferiority study design, 50 healthy volunteers were randomized to receive either a PP block (PP group) or an FN block (FN group). The primary outcome was the duration of peripatellar cutaneous sensory block, with the prespecified noninferiority margin of -3.08 h; the secondary outcome was the area of peripatellar cutaneous sensory block; in addition, the number of complete anaesthesias of the incision line for total knee arthroplasty and the Bromage score 30 min after block were recorded. The PP was successfully dyed, whereas the FN and saphenous nerve were unstained in all cadaveric limbs. The mean difference of the block duration between the two groups was - 1.24 (95% CI, -2.81 - 0.33) h, and the lower boundary of the two-sided 95% CI was higher than the prespecified noninferiority margin (Pnoninferiority = 0.023), confirming the noninferiority of our technique over FN block. The cutaneous sensory loss covered the entire peripatellar region in the PP group. PP block achieved complete anaesthesia of the incision line used for total knee arthroplasty and a Bromage score of 0 in 25 volunteers, which differed significantly from that of volunteers who underwent FN block. Ultrasound-guided PP block is a feasible technique. Compared with FN block, PP block provides noninferior block duration and complete blocking of the peripatellar region without affecting lower limb mobilization. This study was registered in the Chinese Clinical Trial Register (registration no. ChiCTR2000041547, registration date 28/12/2020).


Interception of 1,2-cyclohexadiene with TEMPO radical.
Transient strained cyclic intermediates, such as strained cyclic allenes, are useful building blocks for the synthesis of structurally and stereochemically complex scaffolds. Trappings of strained cyclic allenes are thought to occur primarily through either two or one electron processes. Regarding the latter, diradical intermediates have been invoked in (2 + 2) cycloadditions and (3 + 2) nitrone cycloadditions. The present study questions if a monoradical pathway could exist for strained cyclic allene reactivity, as examined in the reaction of 1,2-cyclohexadiene and TEMPO radical. Our findings suggest the viability of this monoradical pathway.


Early use of high efficacy therapies in pediatric forms of relapsing-remitting multiple sclerosis: A real-life observational study.
Pediatric forms of multiple sclerosis are more active than those in adults. Yet, the effectiveness of different therapeutic approaches is not well studied in this population. Our objective was to compare the effectiveness of the early use of high efficacy therapies (HETs) with the effectiveness of moderate efficacy therapies (METs) in children with MS. This observational study included patients diagnosed with pediatric MS, at 4 hospital centers in France, during a 10-year period. METs included: interferon β-1a, glatiramer acetate, dimethyl fumarate, teriflunomide; HETs included: fingolimod, natalizumab, ocrelizumab, alemtuzumab. The primary endpoint was the occurrence of a new relapse, the secondary endpoint was EDSS worsening. Sixty-four patients were included in the analysis (80% women; mean age 15.5 years, 81% treated with MET) with a median follow-up of 22.5 months. At baseline, 52 patients were on MET (interferon β-1a, glatiramer acetate, dimethyl fumarate, teriflunomide) and 12 patients were on HET (natalizumab, ocrelizumab). The cumulative probability of being relapse-free at 6.5 years was 23.3% on MET, vs 90.9% on HET (p = 0.013). The cumulative probability of no EDSS worsening did not differ between the 2 groups. Patients starting with METs had much higher clinical disease activity than those starting early with HETs. Rapid initiation of more aggressive treatment may allow better disease control; however, the data on EDSS worsening are not conclusive.


A serological survey of toxocariasis in patients and healthy donors in Barcelona (Spain).
The ELISA test, using excretory-secretory antigen from larvae II of Toxocara canis, was applied on 1018 sera (793 from adults and 225 from pediatrics) distributed in: A) patients with an hypereosinophilia where the ethiological agent was undetermined (99); B) patients with ocular complaints compatible with an ocular toxocariasis (116); C) patients with hidatidosis (97); D) patient with other non-toxocaral helminthiasis (34); E) patients with other clinical features (468) and F) healthy donors (204). Over 3,6% of sera showed elevated levels of antibodies reacting with T. canis antigen. The prevalence of seropositivity was statistically higher in patients with eosinophilia (14,1%) (a less than 0,001) and ocular complaints (6%) (0,025 greater than alpha greater than 0,01) than in the control group (1%). In the overall seropositivity from pediatrics did not differ from that of the adults.