Growth, survival, proliferation and pathogenesis of Listeria monocytogenes under low oxygen or anaerobic conditions: a review.
Listeria monocytogenes is a Gram positive facultative anaerobe that causes listeriosis, a disease that mainly affects the immune-compromised, the elderly, infants and pregnant women. In the susceptible immune challenged population, listeriosis is very severe and has a fatality rate of up to 30%. Control of L. monocytogenes is difficult due to its: 1) widespread presence in the environment, 2) intrinsic physiological resistance, 3) ability to adapt to external stresses and 4) ability to grow at a wide range of temperatures. L. monocytogenes encounters anaerobic conditions in the external environment as well as during pathogenesis. Although L. monocytogenes is a facultative anaerobe, the differential effects of O(2) and oxidation-reduction potential on the multiplication of L. monocytogenes have not been established. In addition, most laboratory studies to determine the growth, survival and persistence of this pathogen in foods as well as in the environment have emphasized the response of this pathogen under aerobic conditions. Consequently, this has led to a limited understanding of the metabolic and physiological responses of L. monocytogenes in low oxygen environments. Therefore, the objective of our review was to highlight the progress that has been made in L. monocytogenes research with emphasis on the role of low oxygen and/or anaerobiosis in the growth, survival and proliferation of this pathogen in the environment as well as during pathogenesis.


Expression of a secretory protein gene during androgen-induced cell growth.
Guinea pig seminal vesicle epithelium is an androgen-dependent tissue composed of tall columnar cells which synthesize four major secretory proteins designated SVP-1-4. Castration promptly causes the seminal vesicle epithelium cells to undergo major morphological changes. By the fifth day after castration, cell number decreases to 12% of normal, and the surviving cells are greatly involuted structurally. An injection of testosterone was sufficient to stimulate individual cell growth to the normal tall columnar configuration by 48 h without increasing cell number. Cell growth following testosterone was preceded by a 4-fold increase between 0-12 h in total cellular RNA signifying a large increase in rRNAs. To aid in the analysis of gene expression during androgen repletion, we identified a cDNA plasmid clone representing the 3'-end of the mRNA of SVP-4. The clone pAT 76 was identified by hybrid-select translation and characterization of the product of in vitro translation. The steady-state levels of SVP-4 mRNA in blots of total cellular RNA increased between 0-24 h; this appeared to be due mainly to a general nonselective increase in all or most of the abundant mRNAs. We demonstrated by in vitro translation studies that the corresponding transcript was still present during the 5th and 36th days of castration without having undergone a selective loss in abundance and during hormone repletion without undergoing a selective increase. Testosterone exerted an early and more conspicuous effect on rRNA synthesis. Whether direct or indirect this may be a key event underlying the ultimate action of testosterone, namely maintenance of cell structure.


Gender aspects of survival after abdominoperineal resection for low rectal cancer: a retrospective study.
The difference in prognosis between genders after abdominoperineal resection (APR) for low rectal cancer (LRC) is unclear. This study aimed to compare survival outcomes between genders in patients with LRC who underwent curative APR. This retrospective cohort study used a database of consecutive colorectal resections. Patients who received curative APR with LRC were grouped according to their gender. Female patients were frequency-matched 1:1 on American Joint Committee on Cancer (AJCC) stage to male patients. Overall survival (OS), disease-free survival (DFS), and their independent risk factors were examined. A total of 140 patients with APR for LRC were included after matching: 70 (50.0%) males and 70 (50.0%) females. No significant differences were found between the groups in terms of age, operation methods, AJCC stage, and adjuvant therapy (all P > 0.05). Median follow-up was 39 (range: 3-128) months. Male gender was independently associated with worse OS (adjusted hazard ratio [HR] = 2.755, 95% CI: 1.507-5.038, P = 0.001) and worse DFS (adjusted HR = 2.440, 95% CI: 1.254-4.746, P = 0.009). Subgroup analysis revealed that female patients with stage III disease had better OS (P = 0.001) and DFS (P < 0.001) than male patients. Gender affects survival after a curative APR for LRC. Compared with females, male patients with LRC after curative APR had worse prognosis, especially for stage III disease.


The value of novel oximes for treatment of poisoning by organophosphorus compounds.
Poisoning by organophosphorus compounds (OP) still is a major therapeutic problem. Intentional OP pesticide poisoning results in up to 300.000 deaths each year and highly toxic OP nerve agents pose a permanent threat for the civilian population and military forces. The therapeutic value of clinically used oximes, pralidoxime, obidoxime and TMB-4, in human OP pesticide poisoning is under debate. Moreover, these oximes lack efficacy in poisoning by various nerve agents. An innumerable number of novel oximes have been synthesized in the past five decades to provide more effective oximes and compounds with improved blood-brain-barrier penetration. Novel compounds were tested with largely different experimental protocols in vitro and in animals in vivo. The lack of comparable experimental conditions and the absence of human in vivo studies hamper a well-founded evaluation of the available data. At present, it appears that only a small number of (bispyridinium) oximes show superior potency and efficacy against individual OP. However, until now, no oxime with sufficient broad-spectrum activity against structurally different OP pesticides and nerve agents is available. An interim solution may be the combination of two oximes with overlapping reactivation spectrum. In conclusion, the unsatisfying situation calls for studies with standardized and comparable experimental conditions in order to allow a sound assessment of available and novel oximes.


Veteran Perspectives on Phantom Limb Pain and Non-Drug Interventions.
Phantom limb pain (PLP) commonly occurs post-amputation and can negatively affect the daily functioning of persons with amputation. Best practices for medication and non-drug management remain unclear. To better understand the PLP experience and patients' familiarity with treatments, phone interviews were conducted at the Minneapolis Veterans Affairs Regional Amputation Center in Veterans with amputations. Fifty Veteran participants (average age 66, 96% male) with lower limb amputation were recruited for phone-based data collection of patient-reported outcomes (ie, demographics using the Trinity Amputation and Prosthesis Experience Scales-Revised (TAPES-R) and pain experience using the Phantom Phenomena Questionnaire) to characterize the population and a semi-structured interview. Notes taken during interviews were analyzed using the Krueger and Casey constant comparison analysis method. Participants had an average of 15 years since amputation, and 80% reported PLP as identified with the Phantom Phenomena Questionnaire. Investigators identified several core themes from the qualitative interviews including 1) high variability in the experience of PLP, 2) acceptance and resilience, and 3) PLP treatment perceptions. The majority of participants reported trying common non-drug treatments with none endorsed consistently as highly effective. More research is needed to inform identification and implementation of clinical best practices for non-drug interventions for PLP and understand the factors that influence engagement in non-drug interventions. The participants in this study were largely male, so these results may not be generalizable to females.


Pain and hyperpathia in pontine haemorrhage.
A 56-year-old man had acute onset of left sided weakness, spontaneous pain and hyperpathia in the left limbs. CT demonstrated a small pontine haemorrhage, in the floor of the fourth ventricle.


Comparative studies on the electric nature of amphibian gonadotropin.
The isoelectric focusing (IEF) profiles of pituitary gonadotropic (GTH) activity in six species of adult male amphibians were examined in addition to the newt and bullfrog. The GTH activity was measured by two radioreceptor assay (RRA) systems; one with Xenopus laevis testicular homogenate as the receptor preparation, and the other with Anolis carolinensis testicular homogenate, both employing 125I-labeled rat FSH as the radioligand, or by a radioimmunoassay (RIA) system for bullfrog basic GTH (LH). The IEF profiles were classified into four groups: (1) Cynops pyrrhogaster pyrrhogaster, Onychodactylus japonicus, (2) X. laevis, (3) Rana japonica, Bufo japonicus formosus, (4) Rana nigromaculata, Hyla arborea japonica, Rana catesbeiana. In Onychodactylus japonicus (Group 1), Anolis RRA-positive peaks and Xenopus RRA-positive peaks were found in almost the same fractions. They were in the alkaline to neutral pH region. This IEF profile was very similar to that of newt pituitary GTH in July when spermatogenesis was active. In X. laevis (Group 2), the IEF profile was similar to that of urodeles. However, the most alkaline peak was found at pH 9.02, which was lower than those of urodeles. These peaks showed some differences in the reactivity to the RRA systems. In R. japonica (Group 3), the major peak obtained by Xenopus RRA was found at pH 10.27, which corresponded to the only positive peak obtained by the RIA system for bullfrog basic GTH. Furthermore, this peak was also recognized by Anolis RRA as one of the eight peaks, which appeared in the alkaline to acidic region. In B. japonicus formosus (Group 3), similar results were observed. In H. arborea japonica, R. nigromaculata and R. catesbeiana (Group 4), Anolis RRA-positive and Xenopus RRA-positive peaks were quite independent, indicating distinct separation of LH and FSH, though pIs of FSH-like GTH in the frogs were not so acidic as those of mammalian and avian FSH.


Serotonin induces peripheral mechanical antihyperalgesic effects in mice.
The role of serotonin (5-HT) in nociception will vary according to the subtypes of receptors activated. When administered peripherally, it induces pain in humans and in rats by activation of 5-HT1, 5-HT2 and 5-HT3 receptors. In addition, endogenous 5-HT produced in situ, is involved in the nociceptive response induced by formalin in rat's paw inflammation, possibly via 5-HT3 receptors. Moreover, it has been shown that 5-HT released in the dorsal horn of the spinal cord by stimulation of the periaqueductal gray causes activation of inhibitory interneurons, resulting in inhibition of spinal neurons. In the present study we evaluated the effect of serotonin and its receptors at peripheral antinociception. The mice paw pressure test was used in animals that had increased sensitivity by an intraplantar injection of PGE2 (2 µg). We used selective antagonists of serotonin receptors (isamoltan 5-HT1B, BRL 15572 5-HT1D, ketanserin 5-HT2A, ondansetron 5-HT3 and SB-269970 5-HT7). Administration of serotonin into the right hind paw (62.5, 125, 250 and 500 ng and 1 µg) produced a dose-dependent peripheral mechanical antihyperalgesic effect of serotonin in mice. Selective antagonists for 5-HT1B, 5-HT2A, 5-HT3 receptors at doses of 0.1, 1 and 10 µg, reversed the antihyperalgesic effect induced by 250 ng serotonin. In contrast, selective antagonists for 5-HT1D and 5-HT7 receptors were unable to reverse the antihyperalgesic effect induced by serotonin. These results demonstrated for the first time, the peripheral mechanical antihyperalgesic effect of serotonin, and participation of 5-HT1B, 5-HT2A and 5-HT3 receptors in this event.


Integrating transcriptomics and genomics to identify fibroblast growth factor/receptor candidate genes for non-syndromic orofacial cleft in Chinese.
The objective of this study was to explore the relationship between fibroblast growth factor/receptor (FGF/FGFR) and non-syndromic orofacial cleft (NSOC) in individuals of Han Chinese. Initially, we performed RNA-Seq between non-syndromic cleft lip only (NSCLO) or non-syndromic cleft palate only (NSCPO) and control groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were carried out to evaluate the functions of differentially expressed genes (DEGs) of FGF/FGFR. Then, we selected the most significant DEG FGFR2 and performed an association analysis in Chinese. Linkage disequilibrium (LD) and haplotype analyses were performed with HaploView and PLINK. Additional bioinformatics functional prediction for the notable single nucleotide polymorphisms was performed with HaploReg V4.1 and 3DSNP. Finally, we identified 32 mRNAs related to FGF/FGFR via RNA-Seq and chose FGFR2 in the subsequent association analysis. Results indicated that the single nucleotide polymorphism (SNP) rs2288336 in FGFR2 contributed significantly to both non-syndromic cleft lip with or without cleft palate (NSCL/P) and NSCLO, with p values of 5.00E-05 (OR = 0.79, 95% CI: 0.70-0.88) and 1.38E-04 (OR = 0.76, 95% CI: 0.65-0.87), respectively. In addition, rs3793893 in FGFR2 was found to be associated with NSCLO, with a p value of 1.02E-04 (OR = 0.67, 95% CI: 0.55-0.82). Our research demonstrated that FGFR2 is significantly more involved in NSOC than other FGF/FGFRs in Chinese and further identified rs2288336 and rs3793893 in FGFR2 associated with NSOC subtypes, which provide further evidence for the genetic etiology of NSOC in Han Chinese.


Complete chloroplast genomes of Chinese wild-growing Vitis species: molecular structures and comparative and adaptive radiation analysis.
As a basalmost family of Vitaceae, Chinese wild Vitis species offer key insights into the demographic history of grapes. In this study, we obtained 10 complete chloroplast (cp) genomes from Chinese wild-growing Vitis species based on our whole genome re-sequencing data. These chloroplast genomes ranged from 160,838 to 232,020 bp in size and exhibited typical quadripartite structures. Comparative analyses revealed that inverted repeat (IR) regions are especially abundant and contribute to cp genome arrangements. Phylogenetic analysis of the whole Vitis cp genomes supported three clearly partitioned main origins, in keeping with their geographic distributions, among which East Asian species from China were found to be sister species with Eurasian Vitis species but exhibited significant divergence from the North American group. Two well-supported subgroups were observed within the Chinese wild-growing Vitis species. Among these species, Vitis piasezkii and Vitis betulifolia were closely related species, exhibiting a support rate of 100%. The molecular clock-based divergence time suggested that the earliest split subspecies was Vitis pseudoreticulata, which further indicated that the origin and initial gene pool are located in southern China (the habitat of V. pseudoreticulata is located in the region). Coincidentally, the divergence time was during the Pleistocene period (2.6-0.1 Ma). Due to glacial/interglacial temperature fluctuations, cold-adapted subspecies, e.g., Vitis amurensis, could re-colonize new habitats. Our results may help to elucidate the adaptive radiation of Chinese wild Vitis species in different environments.


Single isotopic probe for gastro-esophageal reflux diagnosis in children.
Gastro-esophageal reflux (G.E.R.) in children has been implicated in various recurring respiratory diseases. Several techniques including oesophageal pH testing and scintigraphy have been devised to detect and quantify G.E.R. Limitations have been found for each test: short duration with gamma-camera and restricted acceptability of the pH probe by children. A single isotopic probe was designed for a non-invasive screening test of G.E.R. in infants. This device was checked by comparison with oesophageal scintigraphy using a gamma camera. Only 1 discrepancy was detected in 19 reflux episodes. This method, using a relatively inexpensive detector and data acquisition module, seems to be well accepted by children, and may be associated with a pH probe for GER diagnosis.


Strategies to improve genomic predictions for 35 duck carcass traits in an F2 population.
Carcass traits are crucial for broiler ducks, but carcass traits can only be measured postmortem. Genomic selection (GS) is an effective approach in animal breeding to improve selection and reduce costs. However, the performance of genomic prediction in duck carcass traits remains largely unknown. In this study, we estimated the genetic parameters, performed GS using different models and marker densities, and compared the estimation performance between GS and conventional BLUP on 35 carcass traits in an F2 population of ducks. Most of the cut weight traits and intestine length traits were estimated to be high and moderate heritabilities, respectively, while the heritabilities of percentage slaughter traits were dynamic. The reliability of genome prediction using GBLUP increased by an average of 0.06 compared to the conventional BLUP method. The Permutation studies revealed that 50K markers had achieved ideal prediction reliability, while 3K markers still achieved 90.7% predictive capability would further reduce the cost for duck carcass traits. The genomic relationship matrix normalized by our true variance method instead of the widely used [Formula: see text] could achieve an increase in prediction reliability in most traits. We detected most of the bayesian models had a better performance, especially for BayesN. Compared to GBLUP, BayesN can further improve the predictive reliability with an average of 0.06 for duck carcass traits. This study demonstrates genomic selection for duck carcass traits is promising. The genomic prediction can be further improved by modifying the genomic relationship matrix using our proposed true variance method and several Bayesian models. Permutation study provides a theoretical basis for the fact that low-density arrays can be used to reduce genotype costs in duck genome selection.


Haploinsufficiency of A20 caused by a novel pathogenic missense variant of TNFAIP3 and successfully treated with anti-TNF and immunosuppressive therapies.
Loss-of-function of protein A20, encoded by TNFAIP3, leads to an early-onset haploinsufficiency of A20 (HA20). This study reports one Chinese child with HA20 and explores the genetic etiology of TNFAIP3 variant. The patient exhibited transient recurrent episodes of fever, intermittent signs of arthritis, gastrointestinal symptoms and multiple colonic ulcers. Laboratory tests revealed elevated inflammatory indicators and mild to moderate anemia. Genetic analysis identified a heterozygous de novo variant in his TNFAIP3 gene (c.740C>T, p. P247L), which had never been reported before. The novel missense variation was validated to be pathogenic through causing insufficient expression of A20, over-activation of NF-κB signaling pathway and elevated levels of proinflammatory cytokines in response to stimulation by lipopolysaccharide. A combination of oral corticosteroids, TNF-α inhibitors and thalidomide freed him from symptoms and abnormal inflammatory indicators. Furthermore, continual improvement of the patient's condition was observed during a follow-up period of five months. We demonstrate a case with a de novo missense variant resulting in a loss-of-function of TNFAIP3, which expands the clinical spectrum of HA20. Cytokine antagonists and immunosuppressants may be effective drugs.


Renin-like (angiotensinogenase) activity in human eccrine sweat.
The presence of renin or renin-like activity (RLA) was demonstrated in human eccrine sweat incubated with purified sheep angiotensinogen, using rat bioassay and angiotensin I radioimmunoassay. Following cholinergic stimulation, sweat RLA was found to range between 0 (unmeasurable) and 266 ng/ml.h, i.e. RLA-values of sweat can be about 10 times higher than those of plasma. Therefore, renin synthesis in sweat glands could be assumed. RLA following activation of beta-adrenergic receptors by the administration of isoprenaline (Aludrin) did not exceed the mean values obtained by cholinergic activation. After beta-adrenergic receptor blockade by propranolol (Dociton), RLA became unmeasurably low. Higher RLA-values were found after local injection of dibutyryl-c-AMP (90--210 ng/ml.h). The results indicate a beta-adrenergic regulation of RLA-release in human sweat glands. Human sweat glands appear to be useful for studying extrarenal renin release.


Unmasking of severe mitral regurgitation after pericardiectomy.
Mitral valve prolapse in the young is the most common cause of mitral regurgitation (MR). Constrictive pericarditis (CP) is a chronic and end-stage manifestation of the inflammatory process of the pericardium. Apart from equalization of diastolic pressure of the chambers as well as increased ventricular interdependence, the thickened pericardial sheath in end-stage pericarditis fails to accommodate the intrathoracic venous return due to restricted cardiac chamber filling. Intracardiac valvular pathologies might co-exist in CP. This report presents a case in which effusive CP was associated with MR and highlights the pivotal role of intraoperative transesophageal echocardiography (TEE) in the decision-making of such co-existent lesions.


Eosinophilic meningitis due to Angiostrongylus cantonensis in a Belgian traveller.
Eosinophilic meningitis is a rare clinical entity. The most frequent cause in travellers to the tropics is infection with the rat lungworm Angiostrongylus cantonensis. In this report, we describe a case of eosinophilic meningitis due to infection with this nematode in a traveller who presented with slight headache, diarrhoea, general malaise and thoracic radicular pain after a trip through Latin America and the Fiji Islands. She responded less than optimally to repeated steroid and albendazole treatments, but finally recovered completely.


Quantitative studies of ciliary and sphenopalatine ganglia in familial dysautonomia.
In 3 patients with familial dysautonomia (FD) sphenopalatine ganglia were less than one fifth normal in volume and their total neuronal content was reduced to a mean of 1,510 (control mean 56,500). Parasympathetic denervation accounts for absence of overflow tears in FD. Ciliary ganglia were not reduced in volume and in 4 patients the mean neuron total was only slightly diminished to 2,900 (control 3,670, P less than or equal to 0.05). This does not appear sufficient to account for pupillary supersensitivity to methacholine in FD on the basis of denervation. Temporal differences in the embryogenesis of sphenopalatine and ciliary ganglia may underlie the observed differences in neuronal populations and suggest that the developmental anomaly in FD does not occur early in gestation.


Process Evaluation of Two Interventions to Improve Health Environments in Family Child Care Homes.
Early care and education (ECE) environments influence children's lifelong health behaviors, growth, and development. Although the number of interventions to improve health in ECE environments is increasing, few have been designed for and tested in family child care homes (FCCHs). This study reports the process evaluation of two interventions to improve FCCH health environments, both part of Happy Healthy Homes, a matched-attention randomized-controlled intervention trial conducted in Oklahoma FCCHs. Participating child care providers received one of two multicomponent interventions: (a) an intervention focused on enhancing the quality of the nutrition environment, self-efficacy, and practices or (b) an intervention focused on improving providers' environmental health literacy, self-efficacy, and practices. Guided by "Implementation" constructs of the RE-AIM framework (i.e., reach, effectiveness, adoption, implementation, maintenance), intervention report forms and participant tracking were used to assess intervention dose delivered and interventionist-perceived challenges and successes to implementation. Interviews were conducted to obtain participant feedback after the intervention. Dose delivered was high for both interventions overall and across individual sessions, and provider feedback was positive. Implementation challenges and strengths identified for both interventions may be useful for further enhancing intervention appropriateness and success, particularly for interventions with FCCHs. Process evaluation findings indicate that the two Happy Healthy Home interventions can be conducted with high delivery and are well attended and considered to be valuable to FCCH providers.


Involvement of Elevated Intracellular and Extracellular ATP in the Regulation of Insulin Secretion: Therapeutic Targets in Non-Insulin-Dependent Diabetes Mellitus.
Non-insulin-dependent diabetes mellitus (NIDDM) is a heterogeneous disease resulting primarily from a variety of pancreatic beta-cell disorders and insulin resistance. Whereas insulin resistance, which constitutes a defect in insulin action, increases the risk of developing NIDDM and, as such, is a predictor of the onset of this disease, it is mostly the beta-cell dysfunction in regulating insulin secretion which yields the chronic hyperglycemia with all its associated clinical complications. The individual steps in the secretory pathway of insulin which is induced primarily by blood plasma glucose have now been identified. The transport of the sugar into the beta-cell is followed by its phosphorylation as the rate-determining step. The glycolytic metabolism of glucose-6-phosphate leads to the generation of ATP resulting in increases in beta-cell ATP pools (steady-state-levels) as well as ATP/ADP ratios, which, in turn, produce the closure of ATP-sensitive K(+) channels, thus depolarizing the beta-cell membrane and opening of Ca(2+) channels. The resulting influx of extracellular Ca(2+) and the increase in recruitment of Ca(2+) from intracellular stores in response to extracellular signals yield an increase in total [Ca(2+)](i) which activates the granular insulin secretory machinery. The intracellular beta-cell ATP pools have a key role in transducing the signals of the stimulus-secretion coupling pathway and toxins such as alloxan and streptozotocin which produce experimental diabetes in animals act by damaging mitochondrial oxidative phosphorylation, leading to permanent decreases in cellular ATP pools which, due to the sensitivity of beta-cell function to these pools, manifest itself as a form of diabetes. In addition to the major effects of blood plasma glucose in the regulation of insulin secretion, a variety of hormonal and neural factors producing endocrine and paracrine effects modulate and fine-tune beta-cell insulin secretion. The enteroinsular axis provides a linkage between the gastrointestinal tract and pancreatic beta-cells stimulus-secretion pathway. Although a powerful effect of ATP on insulin secretion was demonstrated more than 30 years ago, only recently has it been shown that beta-cells possess P(2)-purinoceptors. Extracellular ATP and its synthetic agonists are insulin secretagogues by virtue of their activation of membrane purinergic receptors which is coupled to increases in extracellular Ca(2+) influx and mobilization of Ca(2+) from internal stores resulting in insulin release from beta-cell granules. The physiological significance of extracellular ATP regulation of insulin secretion as well as the physiological source of these ATP pools have not yet been established. It has been recently demonstrated that the administration of adenine nucleotides in vivo can yield significant increases in tissue, blood (red blood cell), and blood plasma ATP pools. Increasing pancreatic beta-cell intracellular and blood plasma (extracellular) pools of ATP is a new therapeutic modality in non-insulin-dependent diabetes mellitus.


In their own words: Safety and quality perspectives from families of hospitalized children with medical complexity.
Children with medical complexity (CMC) experience adverse events due to multiorgan impairment, frequent hospitalizations, subspecialty care, and dependence on multiple medications/equipment. Their families are well-versed in care and can help identify safety/quality gaps to inform improvements. Although previous studies have shown families identify important safety/quality gaps in hospitals, studies of inpatient safety/quality experience of CMC and their families are limited. To address this gap and identify otherwise unrecognized, family-prioritized areas for improving safety/quality of CMC, we conducted a secondary qualitative analysis of safety reporting surveys among families of CMC. Explore safety reports from families of hospitalized CMC to identify areas to improve safety/quality. We analyzed free-text responses from predischarge safety reporting surveys administered to families of CMC at a quaternary children's hospital from April 2018 to November 2020. Using a qualitative descriptive approach, we categorized responses into standard clinical categories. Three team members inductively generated an initial codebook to apply iteratively to responses. Reviewers coded responses collaboratively, resolved discrepancies through consensus, and generated themes. Outcomes: family-reported areas of safety/quality improvement. pre-discharge family surveys. Two hundred and eight/two hundred and thirty-seven (88%) families completed surveys; 83 families offered 138 free-text safety responses about medications, feeds, cares, and other categories. Themes included unmet expectations of hospital care/environment, lack of consistency, provider-patient communication lapses, families' expertise about care, and the value of transparency. To improve care of CMC and their families, hospitals can manage expectations about hospital limitations, improve consistency of care/communication, acknowledge family expertise, and recognize that family-observed quality concerns can have safety implications. Soliciting family input can help hospitals improve care in meaningful, otherwise unrecognized ways.


A concise synthesis and in vitro cytotoxicity of new labdane diterpenes.
A new series of labdane-related diterpenes have been synthesized from (-)-sclareol and assayed in vitro cytotoxicity against mouse and human cancer cells. A key intermediate, homodrimane and furanolabdane derivatives show good in vitro cytotoxicity comparable to those of mitomycin C and adriamycin.


[Congenital dislocation of the hip: current concepts as a preliminary for diagnosis and treatment].
Congenital hip dislocation has been regularly reviewed, in the wake of important progress made over the last thirty years, and more precisely in the field of clinical and ultrasonographic screening, as well as in early treatment. The search of a consensus on the best screening methods (who? how?) should start from one initial point: a thorough knowledge of anatomical, clinical, pathological and therapeutic definitions and concepts underlying this far-reaching question. This is the purpose of the present review of current ideas (from a historical perspective). The two authors, members of French Society for Paediatric Orthopaedics (SOFOP), have extensively researched this subject and contributed to the implementation of national "good practice".


Transarticular invasion of bone tumours across the sacroiliac joint.
The purpose of this study was to evaluate the pattern of tumour spread across the SI articulation, correlating with cadaveric anatomic observations, in order to better understand the local spread of tumour and to assist in the assessment of local staging. Twenty-four consecutive patients (14 male, 10 female; age range 22-89 years, mean 52 years) with primary bone tumours of the iliac bone or sacrum abutting the SI joint, in whom surgical resection of the SI joint was performed, were studied following institutional ethics approval. In all patients, preoperative magnetic resonance (MR) imaging studies of the pelvis and SI joint were reviewed for imaging evidence of transarticular extension across the SI joint. Gross pathologic and histologic assessment of possible transarticular SI joint tumour extension was performed in all patients. Nine cadaveric pelvic specimens without pelvic neoplastic disease (4 male, 5 female; age range 20-84 years, mean 59 years, median 58 years) were anatomically dissected and the articular anatomy of the SI joint examined macroscopically. Twelve of the twenty-four patients demonstrated imaging and histological evidence of transarticular SI joint invasion. Eight tumours infiltrated only the interosseous ligamentous aspect of the SI joint. In the remaining four cases, extensive tumour infiltrated both the cartilaginous and ligamentous aspects of the joint. No case showed tumour involvement isolated to the cartilaginous aspect of the joint. Among the cadaveric specimens studied, degenerative changes were found involving the majority of cases (6/9), with cartilage thinning and fibrillation and antero-superior marginal osteophytes seen involving the cartilaginous portion of the SI joint articulation. Four of the nine specimens demonstrated central ossification bridging the iliac and sacral aspects of the ligamentous (interosseous) SI joint. Tumour invasion across the SI articulation favours its interosseous ligamentous portion. Factors influencing transarticular tumour extension and its preferential course across the interosseous component of the articulation have been discussed. It is particularly important to assess the interosseous ligamentous portion of the SI articulation when locally staging primary pelvic bone tumours subjacent to the SI articulation, as this will have a significant impact on planning surgical management.


Ant parasite queens revert to mating singly.
Multiple mating (polyandry) is widespread among animal groups, particularly insects. But the factors that maintain it and underlie its evolution are hard to verify because benefits and costs are not easily quantified and they tend to be similar in related species. Here we compare the mating strategies of the leaf-cutting ant Acromyrmex echinatior and its recently derived social parasite Acromyrmex insinuator, which is also its closest relative. We find that although the host queens mate with up to a dozen different males, the social parasite mates only singly. This rapid and surprising reversion to single mating in a socially parasitic ant indicates that the costs of polyandry are probably specific to a free-living lifestyle.


In vitro isolation and characterization of biolarvicidal compounds from micropropagated plants of Spilanthes acmella.
Spilanthes acmella (Family: Asteraceae) commonly known as "toothache plant" is known to possess strong insecticidal and larvicidal properties. Experiments have been conducted to isolate and characterise the biolarvicidal compounds from the flower head extract of micropropagated S. acmella plants employing various tools like FT-IR, TLC, CC, NMR. FT-IR spectroscopy of the crude hexane extract sample revealed the presence of amide (secondary metabolite) as functional group in S. acmella flower heads. The crude extract was separated into 85 fractions (100 ml each) through silica gel column chromatography using hexane-ethyl acetate mobile phase. All fractions were tested for their larvicidal activity against late III/early IV instar Anopheles stephensi larvae and fraction showing maximum bioefficacy against aforesaid larvae was further resolved into three separate bands on Preparative TLC plate, the respective R (f) values being (a) 0.18, (b) 0.23 and (c) 0.27. Based on Proton NMR spectrum of the eluted compounds and their comparison with published results, three different compounds were identified: N-isobutyl-2,6,8-decatrienamide (compound 1), undeca-2E,7Z,9E-trienoic acid isobutylamide (compound 2) from band a and (2E)-N-(2-methylbutyl)-2-undecene-8,10-diynamide (compound 3) from band b. The amount of the compounds obtained were 338 mg (compounds 1 and 2) and 188.4 mg (compound 3), respectively. This is the first report of biolarvicidal compounds isolation and characterisation from micropropagated S. acmella plants.


Development of an organic acid compound disinfectant to control food-borne pathogens and its application in chicken slaughterhouses.
During poultry slaughter, cross-contamination of chicken carcasses with microorganisms (including drug-resistant bacteria) can occur because of incomplete disinfection during the pre-cooling process, and surface contact with contaminated tools and equipment. The use of disinfectants is the most common way to reduce the risk of cross-contamination and bacterial spread, as they can effectively reduce the number of bacteria. We developed a disinfectant consisting of organic acids and sodium dodecyl sulfate (SDS) and tested its bactericidal effects at different concentrations against Salmonella and Campylobacter. The main effective components in the disinfectant were citric acid, lactic acid, and SDS, and together they exerted a synergistic bactericidal effect. The bactericidal efficacy of the disinfectant increased with increasing concentrations of the 3 active ingredients. To reach a 100% reduction rate during a 15-s treatment in vitro, for Salmonella, the lowest concentrations of citric acid, lactic acid, and SDS were 0.06, 0.08, and 0.02%, respectively; and for Campylobacter, the lowest concentrations were 0.02, 0.025, and 0.0125%, respectively. The disinfectant remained effective in presence of interfering substances (e.g., 15% fetal bovine serum). Further experiments showed that the disinfectant inactivated sensitive bacteria as well as 23 drug-resistant strains of Salmonella and Campylobacter. Treatment with the disinfectant for 15 s decreased the concentrations of all tested strains by more than 4.7 log colony forming units per mL, and the reduction rate was as high as 100%. In on-site disinfection tests in chicken slaughterhouses, the disinfectant significantly reduced the number of pathogenic bacteria on carcasses during the pre-cooling process, and on tools (such as knives and gloves) during the segmentation process. Thus, this disinfectant has potential uses in preventing cross-contamination of food-borne pathogens (including resistant bacteria) in slaughterhouses.


Maintenance of luminal NADPH in the endoplasmic reticulum promotes the survival of human neutrophil granulocytes.
The present study demonstrates the expression of hexose-6-phosphate dehydrogenase and 11 beta-hydroxysteroid dehydrogenase type 1 in human neutrophils, and the presence and activity of these enzymes in the microsomal fraction of the cells. Their concerted action together with the previously described glucose-6-phosphate transporter is responsible for cortisone-cortisol interconversion detected in human neutrophils. Furthermore, the results suggest that luminal NADPH generation by the cortisol dehydrogenase activity of 11 beta-hydroxysteroid dehydrogenase type 1 prevents neutrophil apoptosis provoked by the inhibition of the glucose-6-phosphate transporter. In conclusion, the maintenance of the luminal NADPH pool is an important antiapoptotic factor in neutrophil granulocytes.


Cooling efficiency of vests with different cooling concepts over 8-hour trials.
As frequency and severity of heat waves are increasing, personal cooling systems are being considered as a tool to mitigate heat strain in workers in various occupational settings. This study assessed cooling capacities (C; W·h·m-2) of various commercially available vests using different cooling concepts. Measurements were conducted over 8 h in a climatic chamber (Ta: 35 °C, RH: 35 %) using a thermal manikin (Ts: 35 °C). Cooling power (P) and duration of efficient cooling (tc) determined the C value of each vest. Among the cooling concepts the active cooling vests were the most efficient, extracting 331 W·h·m-2, followed by the vests with phase change material (PCM) inserts, hybrid and evaporative vests, extracting a maximum of 164 W·h·m-2, 146 W·h·m-2 and 113 W·h·m-2, respectively. While some vests with PCM inserts provided intense but shorter cooling, evaporative vests provided mild but longer cooling throughout. Practitioner summary: The study assessed the cooling capacity of commercially available vests, using a thermal manikin. The vests present an affordable solution in various occupational settings where air-conditioning is not an option. A range of cooling capacities among different cooling concepts and vests of the same category were noted. Abbreviations: ACVs: air-cooled vests; LCVs: liquid-cooled vests; ECVs: evaporative cooling vests; HCVs: hybrid cooling vests; PCVs: phase-change cooling vests; PCM: phase change material; C: cooling capacity; Rt: thermal resistance; Re: evaporative resistance; Re (%): relative evaporative resistance; P: cooling power; Pmax: maximal cooling power; Pavg: average cooling power; tc: cooling duration; AUC: area under the curve; Ta: ambient temperature; RH: relative humidity; va: chamber air flow; Ts: manikin surface temperature.


Chlorin e6 and CRISPR-Cas9 dual-loading system with deep penetration for a synergistic tumoral photodynamic-immunotherapy.
Photodynamic therapy (PDT) is a relatively safe and clinically promising treatment to combat primary tumors, especially epidermal carcinoma, while has negligible effects on distant metastasis. Therefore, this work reports a multifunctional nanosystem (HPR@CCP) exerting a combined photodynamic and immunotherapy to amplify the therapeutic effect on primary tumors and distant metastasis. Specifically, this nanosystem was obtained by electrostatic adsorption of a negatively charged hyaluronic acid "shell" with a positively charged "core" consisting of the CRISPR-Cas9 system targeting the Ptpn2 gene (Cas9-Ptpn2) and a modified mitochondria-targeting chlorin e6 (TPP-PEI-Ce6). Cell experiments demonstrated that the HPR@CCP nanoparticles possessed very high transfection efficiency on B16F10 cells, and TPP-PEI-Ce6 in the nanoparticles resulted in a significant PDT efficacy due to the efficient singlet oxygen generation in mitochondria under laser-irradiation. The accumulation of the nanoparticles in the tumor by active and passive tumor-targeting in vivo led to the disruption of the Ptpn2 gene by the Cas9-Ptpn2 plasmids in the nanocarriers, thus sensitizing tumors to immunotherapy by the increase of the IFN-γ and TNF-α signaling and the promotion of the proliferation of CD8+ T cells. In addition, Hyaluronidase was administered in advance to destroy the hyaluronic acid in the condensed extracellular matrix and to remove the hyaluronic acid "shell" from the nanosystem, subsequently leading to an enhanced penetration of oxygen and therapeutic agents. Fortunately, the primary and distant tumors in the experimental animals were remarkably inhibited after the combination of PDT-immunotherapy, thus, this easy-to-built nanomedicine could be used as a potential combination therapy against tumors.


Screening and management of dyslipidemia in oncologic patients undergoing cardiotoxic therapies: results from an Italian survey.
Baseline cardiovascular risk factors correction is recommended in all cancer patients undergoing potentially cardiotoxic therapies. Despite available guidelines, real-world data on dyslipidemia prevalence and management in the oncologic population are still sparse. This survey was an Italian, investigator-initiated survey initially designed and drafted by the Cardio-Oncology section of the Associazione Nazionale Medici Cardiologi Ospedalieri (ANMCO), comprising 10 individual multi-choice questions and spread after validation through the ANMCO mailing list. The survey was sent to cardiologists working in cardio-oncology units and/or managing patients with cancer. Our survey included 139 Italian cardiologists. The majority of them routinely ask for the baseline lipidic profile of their patients, regardless of previous clinical history and planned treatment. According to our participants, the estimated prevalence of dyslipidemia in this population is between 20% and 60%. Although this high prevalence, our results highlight that there is poor harmony in terms of scores for CV risk prediction used in clinical practice to guide drug prescription and baseline therapy optimization. On the same line, coronary artery calcium score is poorly used in this setting. At the same time, more than 30% of interrogated physicians do not prescribe adequate statin doses, even though necessary, and have uncertainties on the use of other anti-dyslipidemic drugs in this population. Our results highlight the necessity of strong evidences on dyslipidemia screening and management in the cancer population, as well as the need of knowledge diffusion from scientific societies to clinicians treating these patients.


Cryo-injury procedure-induced cardiac regeneration shows unique gene expression profiles in the newt Pleurodeles waltl.
Cardiac regeneration in the adult mouse is not substantial. Some vertebrates, such as newts and zebrafish, regenerate the heart throughout their lives. To understand how regenerative abilities differ among animal species, comparative research has been conducted in animals like mouse, zebrafish, and newt. For those purposes, cryo-injury is suitable as an experimental model for the pathological condition of human myocardial infarction. In fact, cryo-injury procedures are common in mouse and zebrafish. In the present study, we induced cryo-damage on the ventricle in Iberian ribbed newts using a liquid nitrogen-chilled probe. We observed that the injured area recovered within 8 weeks, with remodeling of scar tissue and proliferation of cardiomyocytes. We investigated the subsequent recovery of cryo-injured and amputated tissues by comparative analysis of the gene expression profiles following these two procedures. Notably, we established a cryo-injury procedure for the newt and confirmed that regeneration of the cryo-damaged myocardial tissue is achieved by changes in gene expression that are milder than those observed in the amputation model. Our results suggest that the cryo-injury method is suitable for comparing the process of cardiac regeneration in the newt with that in other animal models.


Organophosphorous pesticide exposures and sperm quality.
Many Americans are exposed to low levels of organophosphorous (OP) pesticides. It is unclear whether these exposures impact sperm production. We investigated whether there was an association between urinary OP insecticide metabolites and sperm concentration and motility in newly married men from a rural area of eastern People's Republic of China. Ninety-four cases and 95 controls were included based on their median residual value of sperm concentration and motility after adjusting for relevant covariates. Their urine was analyzed for six dialkylphosphate (DAP) compounds. After adjustment for demographic and exposure variables, the odds of being a case were greater (odds ratio=1.30, 95% confidence interval 1.02-1.65) in men with higher urinary concentrations of dimethylphosphate (DMP) compared to men with lower levels. No significant differences between cases and controls were found among the other DAP concentrations. DMP exposure and sperm concentration and motility should be explored further in environmental exposure studies.


Shortening time to reperfusion after transfer from a primary to a comprehensive stroke center.
This study assessed whether a quality improvement (QI) process to streamline transfer from a primary stroke center (PSC) to a comprehensive stroke center (CSC) could reduce the delay of reperfusion by mechanical thrombectomy (MT). From 2015 to 2017, a QI process was implemented with specific interventions to reduce door-in-to-door-out (DIDO) time in a high volume PSC, and speed up interhospital transfer and inhospital processes at the CSC. Clinical characteristics and time metrics were compared in the QI (2015-2017; n = 157) and pre-QI cohorts (2012-2014; n = 121). During the QI process, the median symptom onset to reperfusion time was reduced by 50 minutes (367 vs 417 minutes in the pre-QI cohort, p < 0.04), with a substantial 40-minute DIDO reduction (78 vs 118 minutes, p < 0.01), related to the faster administration of IV thrombolysis (median door-to-needle time: 49 vs 82 minutes, p = 0.0001). The door-to-door time was shortened (170 vs 205 minutes, p = 0.002), but not the transfer time (92 vs 87 minutes, p = 0.5). The QI process had no effect on the prehospital phase (77 vs 76 minutes, p = 0.83) and on the time from MRI imaging at the PSC to reperfusion (252 vs 288 minutes, p = 0.12). The rate of modified Rankin Scale score 0-2 at 90 days was comparable in the pre-QI and QI cohorts. A QI process can reduce the reperfusion therapy delay in a distant CSC; however, we could not demonstrate that it can also improve the outcome of patients who undergo MT.


The use of patient-reported outcomes after routine arthroplasty: beyond the whys and ifs.
The limitations and benefits of patient-reported outcome measures, in defining the merits of arthroplasty surgery, are discussed.


Intraoperative recording of electroneurographic signals from cuff electrodes on extradural sacral roots in spinal cord injured patients.
A safe and reliable method for monitoring intravesical pressure on a long-term basis is needed for conditional electrical stimulation to be feasible as a treatment option for neurogenic detrusor overactivity in patients with a spinal cord injury. Therefore, we investigated the possibility of recording afferent nerve activity related to mechanical activity of the bladder and other pelvic organs from the extradural sacral nerve root in human. Nerve cuff electrodes were temporary placed on the extradural S3 sacral root in 6 spinal cord injured patients who underwent implantation of an extradural FineTech-Brindley Bladder System (Finetech Medical Lt., Welwyn Garden City, United Kingdom). The dorsal penile/clitoral nerve was electrically stimulated to evoke compound action potentials. Electroneurographic signals were recorded together with bladder and rectal pressure during mechanical stimulation of the dermatome rapid bladder filling and rectal distention, and during bladder contraction evoked by electrical stimulation of the contralateral sacral root. Compound action potentials and electroneurographic responses during stimulation of the dermatome and rectum were present in all 6 patients and during bladder filling in 5 of 6. However, recorded responses from the bladder and rectum were small and mainly phasic in nature. Nerve responses following bladder contractions were present in 4 of 5 stimulated patients. Afferent nerve activity from the dermatome, bladder and rectum can be recorded using cuff electrodes placed on the extradural S3 sacral root in humans but improvements in recording quality and sophisticated signal processing methods are needed for chronic application.


Enhancement of transepidermal skin clearing agent delivery using a 980 nm diode laser.
Patient compliant optical skin clearing requires non-invasive topical delivery of clearing agents such as glycerol. This requires reducing the skin barrier function by disrupting stratum corneum integrity, which was achieved using a 980 nm diode laser with artificial absorption substrates on the skin surface. Reduction of light scattering has the potential to improve many current and novel diagnostic and therapeutic applications of lasers in medicine. In vivo hamster and rat skin was used to test optical skin clearing. Absorption substrates were applied to the skin after shaving. These included black ink, dark children's rub-on tattoo, and carbon paper. 980 nm cw laser light was used to ablate these substrates and to heat the skin surface to enhance the diffusion of topically applied glycerol for optical skin clearing. Increased light penetration was determined from amplitude optical coherence tomography data. Results indicate an improvement of the ability to measure an OCT signal at a wavelength of 1,290 nm up to 42% deeper into in vivo rodent skin using a 980 nm laser with a fluence of less than 0.96 J/mm(2) to alter the stratum corneum. The use of an inexpensive diode laser can significantly enhance the delivery of topically applied glycerol for optical skin clearing. The laser use involves application of an absorption substrate onto the skin surface. Using carbon paper left no unwanted residue behind and is considered optimal for this purpose.


Comparison between ESD and RFA in patients with total or near-total circumferential early esophageal squamous cell neoplasia.
Patients with early esophageal squamous cell neoplasias (ESCNs) that are totally or nearly totally circumferential face challenges in their clinical work. Endoscopic submucosal dissection (ESD) frequently leads to esophageal strictures. Endoscopic radiofrequency ablation (RFA), which stands out for its simplicity of use and low rate of stenosis, is a rapidly evolving therapeutic strategy for early ESCNs. We contrast ESD with RFA in order to find which method is best for the treatment of a wide range of esophageal diseases. Patients who had flat-type, early, large ESCNs (extending more than 3/4 of the esophageal circumference) treated endoscopically were enrolled retrospectively. The primary outcome measurements were adverse events and local control of the neoplastic lesion. A total of 105 patients received treatment; 60 had ESD and 45 received RFA. Despite the patients receiving RFA typically having larger tumors (14.27 vs. 5.70 cm, P < 0.05), the local control of the neoplastic lesion and procedure-related complications were comparable between the ESD and RFA groups. A considerably higher risk of esophageal stenosis was observed in patients with extensive lesions in the ESD group compared to the RFA group (60% vs. 31%; P < 0.05), and the rate of refractory stricture is also higher than that of the RFA. Both RFA and ESD are effective in treating large, flat, early ESCNs; however, ESD is more likely to cause side effects, such as esophageal stricture, particularly in lesions that are larger than 3/4 of the diameter. Before RFA, a more precise and thorough pretreatment examination should be performed. A more accurate pretreatment evaluation will be an important development direction for early esophageal cancer in future. After surgery, a strict routine review is crucial.


The influence of nicotine in healing of small bowel anastomoses in rats: angiogenesis and miofibroblasts.
to know the effect of nicotine on angiogenesis and myofibroblast formation in anastomoses of the small bowel of rats. we randomly divided 60 Wistar rats into the groups Nicotine (N) and control (C), according to the proposed treatment. Each group was subdivided into three subgroups according to the time interval used for the evaluation (7, 14 or 28 days). The N group with 30 animals received nicotine subcutaneously at a dose of 2mg/kg body weight, diluted in 0.3ml of 0.9% saline, twice daily for 28 days prior to the operation, and for more 7, 14 or 28 days, depending on the subgroup. The C group (also 30 animals) received only saline on the same conditions and time intervals. After 28 days we carried out an end-to-end anastomosis 10cm distal to the duodenojejunal flexure in each rat. After 7, 14 or 28 days after surgery, we euthanized ten animals of each group, sent specimens of the anastomosis areas, 1cm proximal to 1cm distal, to counting of blood vessels and myofibroblasts through immunohistochemical staining by the application of monoclonal anti-factor VIII antibodies and anti-smooth muscle alpha-actin. the administration of nicotine led to the decrease in the number of blood vessels measured on the 28th postoperative day and the number of myofibroblasts measured on the seventh day following completion of the anastomoses. administration of nicotine was deleterious on angiogenesis and myofibroblast formation in rats' small intestine anastomoses. conhecer o efeito da nicotina sobre a angiogênese e formação de miofibroblastos em anastomoses do intestino delgado de ratos. sessenta ratos Wistar foram divididos de maneira aleatória em grupos Nicotina(N) e Controle (C), conforme o tratamento proposto. Cada grupo foi subdividido em três subgrupos, de acordo com o intervalo de tempo utilizado para a avaliação (7, 14 ou 28 dias). O grupo N, com 30 animais, recebeu nicotina por via subcutânea, na dose de 2mg/Kg de peso, diluída em 0,3ml de solução salina a 0,9%, em duas aplicações diárias, durante 28 dias prévios à operação e por mais 7, 14 ou 28 dias, conforme o subgrupo. O grupo C (igualmente com 30 animais) recebeu somente a solução salina nas mesmas condições e intervalos de tempo. Após 28 dias efetuou-se, em cada rato, anastomose término-terminal a 10cm da flexura duodenojejunal. Após 7, 14 ou 28 dias da cirurgia, os dez animais de cada subgrupo foram eutanasiados, sendo que as áreas anastomosadas, 1cm proximal a 1cm distal, foram encaminhadas para contagem de vasos sanguíneos e miofibroblastos, através de coloração imuno-histoquímica por aplicação dos anticorpos monoclonais antifator VIII e anti-alfa-actina muscular lisa. a administração de nicotina levou à diminuição do número de vasos sanguíneos aferidos no 28o dia pós-operatório e do número de miofibroblastos aferidos no sétimo dia após a realização das anastomoses. a administração de nicotina foi deletéria sobre a angiogênese e formação de miofibroblastos em anastomoses do intestino delgado de ratos.


Patients with gastric volvulus recurrence have high incidence of wandering spleen requiring laparoscopic gastropexy and splenopexy.
Though gastric volvulus in neonates and infants resolves by conservative therapy and aging, some cases require surgical intervention. This study aimed to review the cases of gastric volvulus requiring surgical intervention and evaluate their characteristics. We retrospectively reviewed gastric volvulus cases requiring surgical intervention. Surgical indication was persistent acute gastric volvulus and repeated hospitalization for gastric volvulus. We evaluated the characteristics of those cases requiring surgical intervention and the surgical results of laparoscopic gastropexy. The median age of patients included was 4 years (range: 1-6 years). All eight cases of gastric volvulus requiring sugery had congenital spleen diseases. Six of the eight cases suffered from a wandering spleen, while two cases presented with situs inversus with asplenia. Both splenopexy (preperitoneal distension balloon [PDB] or blunt separaion methods) and gastropexy were performed in cases with wandering spleen. No postoperative complications were reported in any of the eight cases, except the recurrence of gastric volvulus due to suture shedding in one case. Laparoscopic gastropexy for gastric volvulus and splenopexy for cases concomitant with wandering spleen were found to be effective surgical approaches. Both PDB and blunt separation methods for making extraperitoneal pockets for the spleen were employed successfully.


A Novel Redox Modulator Induces a GPX4-Mediated Cell Death That Is Dependent on Iron and Reactive Oxygen Species.
Redox modulators have been developed as an attractive approach to treat cancer. Herein, we report the synthesis, identification, and biological evaluation of a quinazolinedione reactive oxygen species (ROS) inducer, QD394, with significant cytotoxicity in pancreatic cancer cells. QD394 shows a transcriptomic profile remarkably similar to napabucasin, a cancer stemness inhibitor. Both small molecules inhibit STAT3 phosphorylation, increase cellular ROS, and decrease the GSH/GSSG ratio. Moreover, QD394 causes an iron- and ROS-dependent, GPX4 mediated cell death, suggesting ferroptosis as a major mechanism. Importantly, QD394 decreases the expression of LRPPRC and PNPT1, two proteins involved in mitochondrial RNA catabolic processes and both negatively correlated with the overall survival of pancreatic cancer patients. Pharmacokinetics-guided lead optimization resulted in the derivative QD394-Me, which showed improved plasma stability and reduced toxicity in mice compared to QD394. Overall, QD394 and QD394-Me represent novel ROS-inducing drug-like compounds warranting further development for the treatment of pancreatic cancer.


Uveal melanoma in the Saudi Arabian population: Two decades of management at the King Khaled Eye Specialist Hospital.
To present the experience of King Khaled Eye Specialist Hospital (KKESH) with uveal melanoma over the last two decades in a fashion similar to the result of the Collaborative Ocular Melanoma Study (COMS). Retrospective, non-comparative, interventional, case series. All patients were diagnosed with uveal melanoma at the King Khaled Eye Specialist Hospital (KKESH), Riyadh, Saudi Arabia from June 1983 to July 2005 and met the inclusion criteria of the COMS. A medical record review of clinical history, imaging studies, surgical procedures and treatment outcome was performed. Forty patients (24 males and 16 females) with uveal melanoma (average age 50 years; range 24-77 years) were included in the study; 28 (70%) were of Saudi Arabian descent and the remaining 12 (30%) patients were from neighboring Arab countries. Decreased vision was the main presenting complaint of 29 (72.5%) patients; the duration of this symptom was 3 months or more in 27 (67.5%) patients. The apical height of the tumor was 10 mm or more in nine (22.5%) of the affected eyes and the largest basal dimension was more than 16 mm in nine (22.5%) of the affected eyes. The posterior border of the tumor was 1-2 mm from the optic disc in three (7.5%) affected eyes. Primary enucleation was performed for 33 (82.5%) eyes, episcleral radiation plaque therapy for six (15%) of the eyes and endo resection of the uveal melanoma in one (2.5%) eye. Adjunct external beam radiation therapy was performed in two (5%) orbits for extrascleral extension. The histopathological diagnosis was available for 34 (84%) eyes in which surgery had been performed (33 patients underwent primary enucleation and one patient underwent endo resection of the uveal melanoma); 24 (70.6%) eyes had spindle cell and the remaining 10 (29.4%) had epithelioid or mixed cell types. Evidence of extraocular tumor extension was found in three eyes. The average follow-up was 33.7 months with a median of 19 months (range 0.5 months to 10 years). Two (5%) patients developed metastasis after 2 years and 5 years from the initial treatment of large and medium-sized uveal melanomas, respectively. Individuals of Saudi Arabian ancestry appear to have a low incidence of uveal melanoma. Further studies are required to estimate the true incidence of uveal melanoma in the larger Arab population. Early detection is essential for improving the patient outcomes. Regular communication between the tertiary care eye centers and the local ophthalmic care providers is required to enhance the understanding about uveal melanoma behavior in Arab population.


Notch signaling contributes to the expression of cardiac markers in human circulating progenitor cells.
It has been demonstrated that adult human circulating endothelial progenitor cells (EPCs) can differentiate to a cardiomyogenic phenotype. Notch signaling promotes epithelial-to-mesenchymal transformation and plays a role in heart and vessel development. Here, we investigated the role of Notch activation for cardiac differentiation of EPCs in a coculture system with neonatal cardiomyocyte. After coculture, Notch activation was transiently detected in EPCs, as determined by immunohistochemical detection of NICD (the intracellular cleavage fragment of Notch-1) and expression of human Notch target genes. Inhibition of gamma-secretase blocked Notch cleavage and NICD translocation. Furthermore, the expression of the cardiac marker protein alpha-sarcomeric actinin and troponin T was significantly suppressed by gamma-secretase inhibition or addition of soluble recombinant Jagged-1, indicating that Notch activation facilitates cardiac marker gene expression. Because noncanonical Wnts have previously been shown to promote cardiac differentiation, we additionally determined the influence of Notch activation on the expression of Wnt5a and Wnt11. Wnt5a and Wnt11 expression in the human cells was induced by the coculture and was blocked by gamma-secretase inhibition. Likewise, stimulation of Notch signaling by immobilized Jagged-1 promoted Wnt5a expression in EPCs. These data suggest that Notch is activated upon coculture of EPCs with neonatal rat cardiac myocytes. Gamma-secretase-dependent Notch activation is required for cardiac gene expression in human cells and induces the expression of noncanonical Wnt proteins, which may act in a paracrine manner to further amplify cardiac differentiation.


Abruptio placentae and perinatal death: a prospective study.
Abruptio placentae caused 3.96 perinatal deaths per 1,000 births in a large prospective study. Intrapartum but not prepartum maternal hypertension was observed in the fatal cases. Decidual necrosis at the placental margin and large placental infarcts were the most characteristic placental abnormalities. The decidual necrosis was correlated with maternal cigarette smoking and low pregnancy weight gains in the abruption placentae cases. The fetuses and neonates who died had a pattern of growth retardation characteristic of antenatal undernutrition, indicating that poor maternal nutrition during pregnancy may have contributed to the genesis of the abruptio placentae.


Whole-genome allelotyping using laser microdissected tissue.
Laser-based microdissection technologies have been recently developed and applied to procure homogenous populations of tumor cells from paraffin-embedded and frozen tissue sections. When combined with whole-genomic amplification techniques, sufficient amounts of DNA can be generated from a small number of tumor cells procured by laser-based microdissection. Amplified DNA can then be used to perform high-throughput genome-wide allelotyping using fluorescent-labeled microsatellite markers spanning the whole genome. Loss of heterozygosity can be assessed by Genescan and Genotyper software (ABI Prism).


Immunohistochemistry for Cancer Stem Cells Detection: Principles and Methods.
Cancer stem cells (CSCs) are rare immortal cells within a tumor that can self-renew and drive tumorigenesis. CSCs play a pivotal role in the tumor development, progression and relapse, as well as in the resistance of anticancer therapy. Different tools could help in the analysis of CSCs, especially Immunohistochemistry (IHC) represents a useful technique able to identify several specific CSC markers. The main aims of this chapter are the description of the explain immunohistochemical methods used in the characterization of CSCs. Furthermore, focus on the most common troubleshooting in CSCs IHC is provided, especially the pitfalls of the CSCs markers IHC on tissue microarrays.


No evidence to support use of glass ionomer as a fissure sealant in primary molars.
Randomised controlled trial in primary care setting. 508 children aged 18-30 months from high caries areas of South Wales with caries-free first primary molars were recruited to the trial after informed consent. All children (n=508) received a standard package of dental health education. Children in the test group (n=241) had their first primary molars sealed with glass ionomer. All the children were re-examined once at varying intervals between 12 and 30 months. Decayed, missing and filled teeth (dmft) and presence of glass ionomer sealant. Analysis of the caries data revealed no significant difference between test and control groups for any of the parameters examined. Examination of the confidence intervals showed no indication that there might be a difference even if the sample size had been increased. There is no evidence that the intervention as used in this population had any effect on caries incidence and it cannot be recommended as a clinical procedure.


The system approach to the identification of aflatoxigenic fungi in foodstuffs and feedstuffs.
Aspergillus flavus, A parasiticus, A nomius, A tamarii andA pseudotamarii are important microorganisms capable of producing aflatoxins and further mycotoxins. Aflatoxigenic Aspergillus species are morphologically similar species belonging to the Aspergillus section Flavi. The aflatoxigenic fungal strains were isolated from foods (cereals, pulses, oilseeds, dried fruit, spices), soil, air and water. Mycological analyses are based on valid standards and recommendations of the International Commission for Food Mycology (ICFM). The identification of isolated aflatoxigenic fungi in foodstuffs and feedstuffs can be proved by using classical mycological cultivation methods, diagnostic nutrient media, chemotaxonomy and molecular biological methods (PCR). The system approach to the identification of aflatoxigenic fungi combines these four methods. Thirty strains of the aflatoxigenic fungi were tested.


Partial purification of a ribosomal ribonucleic acid methylase from rat liver nuclei and methylation of undermethylated nuclear ribonucleic acid from regenerating liver of ethionine-treated rat.
S-Adenosylmethionine-dependent ribosomal RNA (rRNA) methylase has been purified approx. 90-fold from rat liver nuclei. The partially purified methylase catalyzes the methylation of base and ribose in hypomethylated nuclear rRNA prepared from the regenerating rat liver after treatment with ethionine and adenine. The enzyme has an apparent molecular weight of about 3 x 10(4) and a sedimentation coefficient of 3.0 S. The enzyme is optimally active at pH 9.5 and sensitive to p-chloromercuribenzoate. Thiol-protecting reagents, such as dithiothreitol, are necessary for its activity, and the enzyme requires no divalent cations for its full activity. This enzyme did not efficiently transfer the methyl group to nuclear rRNA from normal rat liver, compared with hypomethylated nuclear rRNA. Methyl groups were mainly incorporated into pre-rRNA larger than 28 S, and the extent of 2'-O-methylation of ribose by this enzyme was greater than that of base methylation in the hypomethylated rRNA. No other nucleic acids, including transfer RNA (tRNA) and microsomal RNA from normal as well as ethionine-treated rat livers, tRNA from Escherichia coli, yeast RNA, and DNA from rat liver and calf thymus, were significantly methylated by this methylase. These results suggest that partially purified rRNA methylase from rat liver nuclei incorporates methyl groups into hypomethylated pre-rRNA from S-adenosylmethionine.


The many faces of semaphorins: from development to pathology.
The semaphorin family is a large group of proteins controlling cell migration and axonal growth cone guidance. These proteins are bi-functional signals capable of growth promotion or growth inhibition. Initially described in the nervous system, the majority of studies related to semaphorins and semaphorin signalling are nowadays performed in model systems outside the nervous system. Here, we provide an exhaustive review of the many faces of semaphorins both during developmental, regulatory and pathological processes. Indeed, because of their crucial fundamental roles, the semaphorins and their receptors represent important targets for the development of drugs directed at a variety of diseases.


Comparison of the treadmill exercise score and single-photon emission computed tomographic thallium imaging in risk assessment.
This study compared the prognostic value of exercise single-photon emission computed tomographic (SPECT) thallium imaging with that of treadmill exercise score in medically treated patients with coronary artery disease (CAD). The treadmill exercise score was derived from exercise duration, degree of ST segment depression, and the treadmill anginal index. There were 121 patients with no CAD and 316 patients with angiographically defined CAD (> or = 50% diameter stenosis of one or more vessels). During a mean follow-up of 29 months, there were 35 cardiac deaths or nonfatal myocardial infarctions. Multivariate Cox survival analysis showed the extent of thallium imaging abnormality and CAD to be independent predictors of prognosis. On the other hand, the treadmill exercise score was not a significant predictor even on univariate analysis. The results of thallium uptake were the strongest independent predictors of prognosis and in addition provided incremental prognostic power to coronary angiography (chi 2 = 29 for SPECT, 27 for coronary angiography, and 37 for both). Thus exercise SPECT thallium imaging is significantly better than the treadmill exercise score in risk assessment. The size of the perfusion abnormality is an important predicator of prognosis.


Synthesis of Some Benzimidazole Derivatives Bearing 1,3,4-Oxadiazole Moiety as Anticancer Agents.
In an effort to establish new benzimidazole related structural leads with improved anticancer activity, several new benzimidazole derivatives (5a-i) with 1,3,4-oxadiazole scaffold incorporated were synthesized and studied for their anticancer activity. The anticancer screening against MDA-MB-231 breast cancer cell lines showed that compound (5c) exhibited moderate cytotoxicity.


Changes in the prevalence of metabolic syndrome in Korean adults after the COVID-19 outbreak.
This study sought to reveal changes in the prevalence of metabolic syndrome (MetS) after the start of the coronavirus disease 2019 (COVID-19) pandemic and to identify the groups showing the greatest changes. We compared the prevalence of MetS between 2017-2019 (i.e., the 3-year period before the COVID-19 pandemic) and soon after the initial outbreak of COVID-19 in 2020 among 24,564 adults ≥ 19 years of age using data from the Korea National Health and Nutrition Examination Survey. The prevalence of MetS increased steadily between 2017 and 2020 (29.4 to 35.3%, p for trend < 0.001), with annual percent changes of 2.0%p, 2.2%p, and 1.7%p, respectively. Compared to 3 years before the COVID-19 pandemic, the prevalence of MetS significantly increased during the COVID-19 pandemic in males (6.2%p; 95% confidence interval [CI], 3.5 to 8.9) compared to females (1.5%p; 95% CI, -1.2 to 4.1). The age groups with the greatest increases in MetS prevalence after the COVID-19 pandemic were those in their 40s (4.6%p; 95% CI, 0.9 to 8.4) and 50s (5.8%p; 95% CI, 2.2 to 9.4). By educational background, the increase in MetS prevalence was greatest among those with a college degree or higher (5.1%p). The prevalence of MetS in high-income (5.3%p) and low-income (4.6%p) groups significantly increased. The increasing trend in the overall prevalence of MetS continued during the COVID-19 pandemic. In particular, the prevalence of MetS among adult males in their 40s and 50s increased significantly after the COVID-19 outbreak in Korea.


Long-term chemical fertilization-driving changes in soil autotrophic microbial community depresses soil CO2 fixation in a Mollisol.
Soil is the largest C pool in the terrestrial ecosystem. Numerous studies have been devoted to the decomposition of soil organic C as influenced by agricultural management. However, little is known about the effect of fertilization on the microbial CO2 fixation potential. Here, we examined the atmospheric CO2 fixation rates and structure of autotrophic cbbL-containing bacterial communities and accA-containing archaeal communities in response to 38 years of chemical and/or organic fertilizer application in a Mollisol. The autotrophic microbial abundance and community composition were analyzed by quantitative polymerase chain reaction and high throughput sequencing, respectively. Our results showed that chemical fertilization additions significantly decreased CO2 fixation rates by 57%, but organic manure use resulted in no notable differences compared to no fertilizer regimes (0.38 mg CO2 kg-1 soil d-1) through stable isotope methods. The declining soil pH and increasing Olsen-phosphorus in soils with chemical fertilization dramatically reduced the cbbL gene diversity and accA gene abundances and altered both the autotrophic bacterial and archaeal community compositions. The changes in CO2-fixation rate were more greatly attributed to the shifts in autotrophic bacterial community composition than to the diversity and abundance. The C fixation potentials were positively correlated with the relative abundances of Acidiphilium and Methylibium but were negatively related to those of Azospirillum and Nitrosospira. Both composition and abundance of the autotrophic archaeal community contributed together to the CO2 fixation activities. Our finding suggests that long-term chemical fertilization has a strong impact on the soil microbial CO2 fixation activity and autotrophic microorganisms in upland soils and highlight the important roles of the CO2 fixing process in soil organic carbon sequestration.


Blood cancer survivorship in NCI-Designated Cancer Centers: a study of services, gaps, and access barriers.
This study explored survivorship services provided at National Cancer Institute (NCI)-Designated Cancer Centers for patients with blood cancer to identify gaps in services, unmet needs, and barriers to access. Qualitative interviews with national experts and blood cancer survivors aided the development of an online survey distributed to a survivorship clinic director or staff at 63 NCI-Designated Cancer Centers. Staff at 71% of centers participated (n = 45). Survivorship needs identified as most important-follow-up for health issues due to treatment side effects, healthy behavior programming, and individual mental health services-were reported as offered at 91%, 84%, and 84% of centers, respectively. However, respondents indicated that satellite centers offered these services 20% of the time or less. Forty-five percent reported that they believed the majority of survivorship needs of patients with blood cancer who have completed treatment were not adequately met by their center or other organizations. Most frequently indicated barriers to accessing services were distance from the patient's home and lack of primary care physician adherence to best practices or knowledge of late/long-term effects. To enhance survivorship services for patients with blood cancer, NCI-Designated Cancer Centers and non-profits should focus on increasing access to services for survivors who do not reside near the main center and improving coordination between oncology, mental health, and primary care. Understanding survivorship service delivery is an important step toward developing solutions that help survivors seek and access support at their site of care and from other non-profit organizations.


Case report: an attendant-administered contingency management programme for the treatment of a toileting phobia.
A fifteen-year-old mentally retarded child, institutionalised for eleven years, displayed an intense fear of urinating in a toilet. As a result, he wet his pants at a frequent rate. In an attempt to eliminate this fear a variety of response-contingent consequences were programed (token reinforcement, social reinforcement, time-out). The effects of introducing structured contingencies resulted in a steady reduction in the frequency of pants wettings. During a follow-up phase of evaluation, when programmed contingencies were discontinued, the problem behaviour had been completely eliminated. At that time the child was initiating his own bathroom visits and toileting himself in an appropriate manner.


Synergistic effect of nano-sized mackinawite with cyano-cobalamin in cement slurries for reductive dechlorination of tetrachloroethylene.
Experiments were conducted to investigate the reductive dechlorination of tetrachloroethylene (PCE) by nano-Mackinawite (nFeS) with cyano-cobalamin (Cbl(III)) in cement slurries. Almost complete degradation of PCE by nFeS-Cbl(III) was observed in cement slurries in 5 h and its degradation kinetics (k(obs-PCE)=0.57 h(-1)) was 6-times faster than that of nFeS-Cbl(III) without the cement slurries. PCE was finally transformed to non-chlorinated organic compounds such as ethylene, acetylene, and C3-C4 hydrocarbons by nFeS-Cbl(III) in cement slurries. X-ray photoelectron spectroscopy and PCE degradation by cement components (SiO2, Al2O3, and CaO) revealed that both the reduced Co species in Cbl(III) and the presence of Ca in cement played an important role for the enhanced reductive dechlorination of PCE. The increase in the concentration of Cbl(III) (0.005-0.1 mM), cement ratio (0.05-0.2), and suspension pH (11.5-13.5) accelerated the PCE degradation kinetics by providing more favorable environments for the production of reactive Ca species and reduction of Co species. We also observed that the degradation efficiency of PCE by nFeS-Cbl(III)-cement lasted even at high concentration of PCE. The experimental results obtained from this study could provide fundamental knowledge of redox interactions among nFeS, Cbl(III), and cement, which could significantly enhance reductive dechlorination of chlorinated organics in contaminated natural and engineered environments.


Coordinate-based versus structural approaches to brain image analysis.
A basic issue in neurosciences is to look for possible relationships between brain architecture and cognitive models. The lack of architectural information in magnetic resonance images, however, has led the neuroimaging community to develop brain mapping strategies based on various coordinate systems without accurate architectural content. Therefore, the relationships between architectural and functional brain organizations are difficult to study when analyzing neuroimaging experiments. This paper advocates that the design of new brain image analysis methods inspired by the structural strategies often used in computer vision may provide better ways to address these relationships. The key point underlying this new framework is the conversion of the raw images into structural representations before analysis. These representations are made up of data-driven elementary features like activated clusters, cortical folds or fiber bundles. Two classes of methods are introduced. Inference of structural models via matching across a set of individuals is described first. This inference problem is illustrated by the group analysis of functional statistical parametric maps (SPMs). Then, the matching of new individual data with a priori known structural models is described, using the recognition of the cortical sulci as a prototypical example.


8-hydroxy-2'-deoxyguanosine in colorectal adenocarcinoma--is it a result of oxidative stress?
8-hydroxy-2'-deoxyguanosine (8-OHdG) is one of the most abundant oxidatively modified lesions in DNA and is a marker of the oxidative stress. 8-OHdG is a mutagenic lesion and it can mispair with adenine, causing G:C→T: A transversion. Our task was to determine the 8-OHdG level in patients with colorectal adenocarcinoma directly in tumor tissues and corresponding normal mucosa. Samples of tumor tissues and corresponding normal mucosa of 47 patients undergoing surgery for colorectal cancer were analyzed. DNA was isolated from both tumor and normal tissues. Then, DNA was hydrolyzed to nucleotides using nuclease P1 and alkaline phosphatase. The 8-OHdG and 2'-dG (2'-deoxyguanosine) were determined in hydrolysates by high-performance liquid chromatography (HPLC) with electrochemical (EC) and UV detector. The levels of 8-OHdG in colorectal adenocarcinoma tissues were higher than in corresponding normal mucosa. No significant differences were shown in 8-OHdG levels in the cancerous and cancer-free tissues between age and sex and stages A/B and C/D of Duke's classification. 8-OHdG reflects the local oxidative stress in colon adenocarcinoma tissue together with ageing processes, but not the intensity of tumorigenesis itself. Because of many factors that could influence the oxidative modification of DNA bases, its role as a diagnostic and/or prognostic factor in colon adenocarcinoma seems to be limited.


A new methodology for automated diagnosis of mild cognitive impairment (MCI) using magnetoencephalography (MEG).
Mild cognitive impairment (MCI) is a cognitive disorder characterized by memory impairment, greater than expected by age. A new methodology is presented to identify MCI patients during a working memory task using MEG signals. The methodology consists of four steps: In step 1, the complete ensemble empirical mode decomposition (CEEMD) is used to decompose the MEG signal into a set of adaptive sub-bands according to its contained frequency information. In step 2, a nonlinear dynamics measure based on permutation entropy (PE) analysis is employed to analyze the sub-bands and detect features to be used for MCI detection. In step 3, an analysis of variation (ANOVA) is used for feature selection. In step 4, the enhanced probabilistic neural network (EPNN) classifier is applied to the selected features to distinguish between MCI and healthy patients. The usefulness and effectiveness of the proposed methodology are validated using the sensed MEG data obtained experimentally from 18 MCI and 19 control patients.


Intracellular analysis of the development of responses of caudate neurons to stimulation of cortex, thalamus and substantia nigra in the kitten.
Intracellular recordings were made from caudate neurons in anesthetized kittens of 2-72 days of age. In adult cats, results of intracellular recordings indicate that caudate neurons respond most frequently to stimulation of their major afferents from cortex, thalamus and substantia nigra with a sequence of excitation followed by inhibition (EPSP-IPSP sequence). The results of the present study show that the prominent IPSP of this sequence is not well developed in young kittens and does not reach adult values in terms of frequency of occurrence until beyond 40 days of age. Amplitude and duration of the IPSP evoked by cortical stimulation also did not reach adult values until beyond 40 days of age. In contrast, EPSPs can be evoked in the youngest kittens by stimulation of afferents to the caudate. These findings suggest that the caudate nucleus may alter its role during development. In early postnatal periods it functions as a simple relay system transmitting incoming information to its outputs in a relatively unaltered fashion. Later in development it becomes a system capable of complex modulation and filtering of neural information.


Nonlinear propagation and transformation of arbitrary laser beams by means of the generalized ABCD formalism.
The evolution of non-Gaussian and nonspherical high-power laser beams in cubic nonlinear media is described by means of their mean or gross parameters: width, mean curvature radius, and quality factor. The influence of the beam over its own propagation is contained in a new mean parameter that measures the ability of a beam to build its own waveguide. Beam quality and threshold power for self-focusing are connected. The ABCD and invariance laws for modified complex beam parameter and quality factor allow one to transform in one step the mean beam parameters through a sequence of nonlinear propagations, lenses, mirrors, and nonlinear quadratic graded index.


Correction to: Roles and action mechanisms of herbs added to the emulsion on its lipid oxidation.
[This corrects the article DOI: 10.1007/s10068-020-00800-z.].


Kinetic and dynamic interaction study of zolpidem with ketoconazole, itraconazole, and fluconazole.
Azole antifungal agents may impair hepatic clearance of drugs metabolized by cytochrome P450-3A isoforms. The imidazopyridine hypnotic agent zolpidem is metabolized in humans in part by P450-3A, as well as by a number of other cytochromes. Potential interactions of zolpidem with 3 commonly prescribed azole derivatives were evaluated in a controlled clinical study. In a randomized, double-blind, 5-way, crossover, clinical pharmacokinetic-pharmacodynamic study, 12 volunteers received (A) zolpidem placebo plus azole placebo, (B) 5 mg zolpidem plus azole placebo (C) zolpidem plus ketoconazole, (D) zolpidem plus itraconazole, and (E) zolpidem plus fluconazole. Mean apparent oral clearance of zolpidem when given with placebo was 422 mL/min, and elimination half-life was 1.9 hours. Clearance was significantly reduced to 250 mL/min when zolpidem was given with ketoconazole, and half-life was prolonged to 2.4 hours. Coadministration of zolpidem with itraconazole or fluconazole also reduced clearance (320 and 338 mL/min), but differences compared to the zolpidem plus placebo treatment did not reach significance. Zolpidem-induced benzodiazepine agonist effects (increased electrocardiographic beta activity, digit-symbol substitution test impairment, and delayed recall) during the first 4 hours after dosage were enhanced by ketoconazole but not by itraconazole or fluconazole. Coadministration of zolpidem with ketoconazole impairs zolpidem clearance and enhances its benzodiazepine-like agonist pharmacodynamic effects. Itraconazole and fluconazole had a small influence on zolpidem kinetics and dynamics. The findings are consistent with in vitro studies of differentially impaired zolpidem metabolism by azole derivatives.


Increased susceptibility of annexin-A1 null mice to nociceptive pain is indicative of a spinal antinociceptive action of annexin-A1.
Annexin-A1 (ANXA1), a glucocorticoid-regulated protein, mediates several of the anti-inflammatory actions of the glucocorticoids. Previous studies demonstrated that ANXA1 is involved in pain modulation. The current study, using ANXA1 knockout mice (ANXA1-/-), is aimed at addressing the site and mechanism of the modulatory action of ANXA1 as well as possible involvement of ANXA1 in mediating the analgesic action of glucocorticoids. The acetic acid-induced writhing response was performed in ANXA1-/- and wild-type (ANXA1+/+) mice with spinal and brain levels of prostaglandin E2 (PGE2) examined in both genotypes. The effect of the ANXA1 peptomimetic Ac2-26 as well as methylprednisolone on the writhing response and on spinal cord PGE2 of ANXA1+/+ and ANXA1-/- was compared. The expression of proteins involved in PGE2 synthesis, cytosolic phospholipase A2 (cPLA2) and cyclooxygenases (COXs), in the spinal cord of ANXA1+/+ and ANXA1-/- was also compared. ANXA1-/- mice exhibited a significantly greater writhing response and increased spinal cord levels of PGE2 compared with ANXA1+/+ mice. Ac2-26 produced analgesia and reduced spinal PGE2 levels in ANXA1+/+ and ANXA1-/- mice, whereas methylprednisolone reduced the writhing response and spinal PGE2 levels in ANXA1+/+, but not in ANXA1-/- mice. The expression of cPLA2, COX-1, COX-2 and COX-3 in spinal cord tissues was upregulated in ANXA1-/-compared with ANXA1+/+. We conclude that ANXA1 protein modulates nociceptive processing at the spinal level, by reducing synthesis of PGE2 by modulating cPLA2 and/or COX activity. The analgesic activity of methylprednisolone is mediated by spinal ANXA1.


A Systematic Review of the Physical, Physiological, Nutritional and Anthropometric Profiles of Soccer Referees.
The importance of soccer referees is widely accepted by international soccer organizations and governing bodies, although there is little research summarizing and appraising the literature on soccer referees. The aim of this study was to systematically review the information related to physical demands, nutrition and physiological profiling of soccer referees. Conforming with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, searches of three electronic databases (Web of Sciences, PubMed and Scopus) were conducted on 24 April 2022. The following search terms were used: (Soccer OR football AND refer*) AND (physical OR physiolo* OR load* OR "body composition" OR "fat mass" OR "fat free mass" OR "body size" OR "nutrition*" OR "nutritional assessment" OR "nutritional intake" OR "macronutrient*" OR "micronutrient*"). The inclusion criteria of the manuscripts written in English were that articles with male and/or female soccer referees and included relevant data concerning performance, physical testing, nutrition, body composition, body size and/or physiology of soccer referees. The tools developed by the National Institute of Health were used to assess risk of bias according to the study design. In total, 110 manuscripts were included in the present review. Match activities of soccer referees and assistant referees are not comparable. Variation in performance is influenced by competitive level and competitive schedules. Additionally, match performance is often used to validate field protocols. The associations between match indicators and field protocols were not constant across the included studies, particularly in short-maximal tests. An age decline in physical performance was not conclusive. Body size did not discriminate between referees and assistant referees, as well as referees of different competitive levels. Few studies focused on nutritional guidelines among referees, particularly exercise energy expenditure. Energy intake was comparable across studies, but referees did not follow the daily dietary recommendations. Aerobic output was frequently assessed, and it was not comparable to the values for soccer players. Although there are decreases in the running profiles of older referees, they maintain the same distances from the ball, and thus, age per se should not be used to define participation at the international level. The assessment of physical fitness warrants future consideration given the levels of fatigue that are apparent at the end of matches. In order to attain ideal levels of body composition, future studies need to provide guidelines for daily energy expenditure and nutritional intake. The protocol was published in INPLASY (International Platform of Registered Systematic Review and Meta-Analysis Protocols) with the registration number 202280052 and https://doi.org/10.37766/inplasy2022.8.005 .


PML nuclear bodies and neuronal intranuclear inclusion in polyglutamine diseases.
In polyglutamine diseases, accumulation in the nucleus of mutant proteins induces the formation of neuronal intranuclear inclusions (NIIs). The nucleus is compartmentalized into structural and functional domains, which are involved in NII formation. Promyelocytic leukemia protein (PML), a major component of nuclear bodies, and mSin3A, a component of the transcription co-repressor complex, were used to investigate how the intranuclear domains/sites relate to NII formation in SCA2, SCA3, SCA7, SCA17 and DRPLA brains. We demonstrate that the size of PML-positive intranuclear structures was larger in pathological brains than in control ones and that these structures contained mutant proteins. PML colocalized only with small NIIs, which maintained the ring-like structure of normal nuclear bodies. Enlarged ring-like PML-positive structures, devoid of mutant proteins, were also found and might represent structures where mutant polyglutamine proteins have been successfully processed. These data suggest that NIIs originate from nuclear bodies, where mutant proteins accumulate for degradation.


A description of spinal fatigue strength.
Understanding fatigue failure of the spine is important to establish dynamic loading limits for occupational health and safety. In this study experimental data were combined with published data to develop a description of the predictive parameters for spinal fatigue failure. 41 lumbar functional spinal units (FSUs) from cadaveric spines (age 49.0 ± 11.9 yr) where cyclically loaded. Three different levels of sinusoidal axial compression (0-3 kN, 0-2kN or 1-3kN) were applied for 300,000 cycles. Further, published data consisted of 70 thoracic and lumbar FSUs loaded in axial compression for 5000 cycles. Cyclic forces ranged from lower peaks (Fmin) of 0.7-1kN to upper peaks (Fmax) of 1.2-7.1 kN. Based on Wöhler analysis, a fatigue model was developed accounting for three parameters: I) specimen-specific scaling based on the endplate area, II) specimen-specific strength dependency on age or bone mineral density, III) load-specific correction factors based on Fmax and Fmin. The most predictive model was achieved for a combination of Fmax, endplate area and bone mineral density; this model explained 61% of variation (p<0.001). A model including Fmax, endplate area and age explained only 28% of variation (p<0.001). Inclusion of a load-specific correction factor did not significantly improve model prediction of fatigue failure. This analysis presents the basis for the prediction of specimen-specific fatigue failure of the lumbar spine, provided the endplate area and bone mineral density can be derived.


Tunable microphase-regulated silk fibroin/poly (lactic acid) biocomposite materials generated from ionic liquids.
One of the most effective and promising strategies to develop novel biomaterials with unique, tunable structure and physicochemical properties is by creating composite materials that combine synthetic polymers with natural proteins using ionic liquids. In this study, biodegradable poly(d,l-lactic acid) (PDLLA) was blended with silk fibroin (SF) to create biocompatible films using an ionic liquid-based binary solvent system (1-butyl-3-methylimidazolium chloride/N,N-dimethylformamide), which can maintain the molecular weights of the proteins/polymers and encourage intermolecular interactions between the molecules. The effects of varying the ratio of PLA to SF were studied using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), water contact angle testing, and cytotoxicity analysis as well as enzymatic degradation. Results showed that the composite films were homogeneously blended on the macroscopic scale and exhibited typical fully miscible polymer blend characteristics. By increasing the SF content in the composites, the amounts of β-sheets in the films were significantly increased, allowing for SF to act as a physical crosslinker to maintain the stability of the protein-polymer network. Additionally, SF significantly improved the hydrophilicity and biocompatibility of the material and promoted the self-assembly of micelle structures in the biocomposites. Different topologies in the films also provided beneficial surface morphology for cell adhesion, growth, and proliferation. Overall, this study demonstrated an effective fabrication method for a fine-tuned polymer blends combining synthetic polymer and protein for a wide variety of biomedical and green material applications.