Disruption of ER ion homeostasis maintained by an ER anion channel CLCC1 contributes to ALS-like pathologies.
Although anion channel activities have been demonstrated in sarcoplasmic reticulum/endoplasmic reticulum (SR/ER), their molecular identities and functions remain unclear. Here, we link rare variants of Chloride Channel CLIC Like 1 (CLCC1) to amyotrophic lateral sclerosis (ALS)-like pathologies. We demonstrate that CLCC1 is a pore-forming component of an ER anion channel and that ALS-associated mutations impair channel conductance. CLCC1 forms homomultimers and its channel activity is inhibited by luminal Ca2+ but facilitated by phosphatidylinositol 4,5-bisphosphate (PIP2). We identified conserved residues D25 and D181 in CLCC1 N-terminus responsible for Ca2+ binding and luminal Ca2+-mediated inhibition on channel open probability and K298 in CLCC1 intraluminal loop as the critical PIP2-sensing residue. CLCC1 maintains steady-state [Cl-]ER and [K+]ER and ER morphology and regulates ER Ca2+ homeostasis, including internal Ca2+ release and steady-state [Ca2+]ER. ALS-associated mutant forms of CLCC1 increase steady-state [Cl-]ER and impair ER Ca2+ homeostasis, and animals with the ALS-associated mutations are sensitized to stress challenge-induced protein misfolding. Phenotypic comparisons of multiple Clcc1 loss-of-function alleles, including ALS-associated mutations, reveal a CLCC1 dosage dependence in the severity of disease phenotypes in vivo. Similar to CLCC1 rare variations dominant in ALS, 10% of K298A heterozygous mice developed ALS-like symptoms, pointing to a mechanism of channelopathy dominant-negatively induced by a loss-of-function mutation. Conditional knockout of Clcc1 cell-autonomously causes motor neuron loss and ER stress, misfolded protein accumulation, and characteristic ALS pathologies in the spinal cord. Thus, our findings support that disruption of ER ion homeostasis maintained by CLCC1 contributes to ALS-like pathologies.


[Location of Dctn1 in the mouse testis and sperm and its role in spermiogenesis].
To investigate the role of dynactin 1 (Dctn1) in the process of mouse spermiogenesis. Western blot and indirect immunofluorescence were used to analyze the expression and location of Dctn1 in the mouse testis and spermatozoa. The highest efficiency of small interference RNA (siRNA) was verified by GC2-spd cell line in vitro and in vivo studies, respectively. Dctn1 siRNA mixed with the indicator (0.4% trypan blue) was injected into the seminiferous tubules of 3-week-old ICR mice through rete testis microinjection, and negative control siRNA injected into the control testes. The normal group included 3-week-old ICR mice that did not receive any treatment. Spermatozoa were collected from the cauda epididymis 3 weeks after siRNA injection for morphological analysis. Dctn1 was mainly localized in the tail of spermatozoa. After interference, the sperm tail abnormality in the Dctn1 siRNA group was (23.57 +/- 0.55)%, significantly higher than (12.35 +/- 2.29)% in the control (P < 0.01, n = 3), and it was (3.37 +/- 0.69)% in the normal group. Dctn1 plays an important role in mouse spermiogenesis, and mainly affects the formation of the tail of spermatozoa.


Therapy-resistant psoriasis treated with alefacept and subsequent narrow band ultraviolet B phototherapy with total clearing of psoriasis.
Combinations of therapeutic modalities have been utilized to meet the challenge of difficult to treat psoriasis. We report two cases of psoriasis, resistant to methotrexate and monotherapy with UVB, treated effectively with alefacept and concomitant narrow band UVB resulting in total disease clear.


Cardiac sympathectomy for refractory ventricular arrhythmias in cardiac sarcoidosis.
Ventricular arrhythmias (VAs) in cardiac sarcoidosis (CS) are frequently refractory to both antiarrhythmic drug (AAD) therapy and catheter ablation (CA). Cardiac sympathetic denervation (CSD) has been shown to reduce VA burden and implantable cardioverter-defibrillator (ICD) shocks in patients with nonischemic cardiomyopathy. We aimed to report our center's preliminary experience with CSD in patients with known or presumed CS and refractory VAs. Patients with CS and refractory VAs who underwent CSD at our institution were included. Patient characteristics, procedural outcomes, and number of arrhythmic events including ICD shocks pre- and post-CSD are reported. Five patients with CS (mean age 53 ± 11 years; 2 men [40%]; mean left ventricular ejection fraction 38% ± 11%) underwent CSD for VA refractory to AAD therapy and CA. Four of 5 patients underwent bilateral CSD; 1 patient underwent right-sided sympathectomy only because of poor intraoperative visualization on the left. Procedural complications included hemothorax in 1 patient and azygous vein injury in 1 patient. The median number of ICD shocks in the 6 months pre-CSD was 5. During a median follow-up of 26 months (range 5-29 months), the median number of ICD shocks post-CSD was 0; 1 patient had sustained VA that was below the threshold for device therapy, and 1 patient had symptomatic premature ventricular contractions; both underwent repeat CA. In addition, 1 patient required cardiac transplantation for progressive heart failure. CSD may be a feasible therapeutic adjunct for patients with CS and VA refractory to AAD therapy and CA.


Athletes Report Favorable Outcomes at 5-Year Minimum Follow-Up After Primary Hip Arthroscopy: A Systematic Review.
To evaluate minimum 5-year outcomes and survivorship after primary hip arthroscopy in athletes. A systematic review of current literature was performed with the following key words: "hip arthroscopy," "long-term," "outcomes," "ten-year," "survivorship," "10-year," "5-year," "five-year," "midterm," "athlete," "sport," and "femoroacetabular impingement" in PubMed, Cochrane Trials, and Scopus in April 2022 using the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines. Study and demographic variables such as title, author, publication date, study design, demographic, number of hips, follow-up time, study period, indications for hip arthroscopy, patient-reported outcome scores, and rates of secondary surgeries and conversion to total hip arthroplasty. Radiographic, intraoperative surgical treatment, and clinical benefit was documented as well. Ten studies that captured data on 691 hips were included in this study. Eight studies reported mean and standard deviation for the modified Harris Hip Score. Significant improvements were reported in all studies with postoperative scores ranging from 83.3 to 94.4 (standardized mean difference [SMD] 1.65; 95% confidence interval [CI] 1.39-1.91, P < .00001). Eight studies reported significant improvement in Hip Outcome Score - Sports Specific Scale with postoperative scores ranging from 59.2 to 94.6 (SMD 1.66; 95% CI 1.33-1.98, P < .00001). Seven studies reported significant improvement in Nonarthritic Hip Score with postoperative scores ranging from 79.6 to 95.3 (SMD 1.41; 95% CI 1.16-1.65, P < .00001). Seven studies also reported significant improvement in VAS with postoperative scores ranging from 0.7 to 3.1 (SMD -1.57; 95% CI -1.88, -1.26, P < .00001). Nine of 10 studies reported achieving at least 1 psychometric threshold at a rate of least 75%. Rates of secondary arthroscopy and conversion to total hip arthroplasty varied from 0% to 15.2% and 0% to 33.3%. Athletes who underwent primary hip arthroscopy demonstrated favorable outcomes and high rates of clinical benefit at 5-year follow-up. In addition, they demonstrated high rates of survivorship and achieving psychometric thresholds. Level IV, systematic review of Level III and IV studies.


Reduction structured rhinoplasty.
The reduction structured rhinoplasty has evolved based on an increased understanding of nasal function and structural anatomy. The discussion begins with the philosophic origins of this technique. Next, the preoperative visit is outlined including the history, physical examination, basic facial analysis, and surgical planning. The procedure is detailed and the postoperative care is described.


Use of the subclavian vessels for microvascular reconstruction in the vessel-depleted neck.
The previously treated neck is a serious problem if further free tissue transfer reconstruction is required. Vessel depletion is often a terrifying finding that creates the need for novel approaches to access vessels that lie outside the anatomical region of the head and neck. We present what is, to our knowledge, the first case of utilising the subclavian artery and vein in head and neck reconstruction. We describe the anatomical details and a step-by-step access approach, together with tips on how to perform safe and effective microvascular anastomoses. The quality of the subclavian vessels is excellent, and access to the area is straightforward provided it is carefully planned. The technique could be considered more often in the vessel-depleted neck.


Magnetic Trapping of Bacteria at Low Magnetic Fields.
A suspension of non-magnetic entities in a ferrofluid is referred to as an inverse ferrofluid. Current research to trap non-magnetic entities in an inverse ferrofluid focuses on using large permanent magnets to generate high magnetic field gradients, which seriously limits Lab-on-a-Chip applications. On the other hand, in this work, trapping of non-magnetic entities, e.g., bacteria in a uniform external magnetic field was studied with a novel chip design. An inverse ferrofluid flows in a channel and a non-magnetic island is placed in the middle of this channel. The magnetic field was distorted by this island due to the magnetic susceptibility difference between this island and the surrounding ferrofluid, resulting in magnetic forces applied on the non-magnetic entities. Both the ferromagnetic particles and the non-magnetic entities, e.g., bacteria were attracted towards the island, and subsequently accumulate in different regions. The alignment of the ferrimagnetic particles and optical transparency of the ferrofluid was greatly enhanced by the bacteria at low applied magnetic fields. This work is applicable to lab-on-a-chip based detection and trapping of non-magnetic entities bacteria and cells.


Nonmembrane associated dolichol in rat liver.
The distribution of dolichol in rat liver was studied. Upon high-speed centrifugation, 9% of the total tissue dolichol was recovered in the supernatant. Dolichol was enclosed in vesicles and in lipidic particles which were isolated by gel filtration and density gradient centrifugation. The particles had a diameter of 20 nm and contained dolichol, ubiquinone, cholesterol, phospholipid and some protein. Similar particles were recovered upon incubation of isolated hepatocytes with liposomes containing dolichol. From the lysosomal lumen, lipid particles containing dolichol, ubiquinone, cholesterol and phospholipid, but no protein, were isolated. The diameter of the particles was 20-40 nm with a molecular weight of 130 kDa. Puromycin treatment inhibited protein synthesis, but did not affect dolichol transfer from the endoplasmic reticulum to lysosomes, suggesting that the transfer is not mediated by newly synthesized apoprotein. The results indicate that a sizeable portion of the total cellular dolichol is present in cytoplasm and in lysosomal lumen. Furthermore, dolichol probably participates in the translocation process.


Influence of compartmental localization on the function of yeast NADP+-specific isocitrate dehydrogenases.
Three differentially compartmentalized isozymes of isocitrate dehydrogenase (mitochondrial IDP1, cytosolic IDP2, and peroxisomal IDP3) in the yeast Saccharomyces cerevisiae catalyze the NADP(+)-dependent oxidative decarboxylation of isocitrate to form alpha-ketoglutarate. These enzymes are highly homologous but exhibit some significant differences in physical and kinetic properties. To examine the impact of these differences on physiological function, we exchanged promoters and altered organellar targeting information to obtain expression of IDP2 and IDP3 in mitochondria and of IDP1 and IDP3 in the cytosol. Physiological function was assessed as complementation by mislocalized isozymes of defined growth defects of isocitrate dehydrogenase mutant strains. These studies revealed that the IDP isozymes are functionally interchangeable for glutamate synthesis, although mitochondrial localization has a positive impact on this function during fermentative growth. However, IDP2, whether located in mitochondria or in the cytosol, provided the highest level of defense against endogenous or exogenous oxidative stress.


Identification of stem cell-related subtypes and risk scoring for gastric cancer based on stem genomic profiling.
Although numerous studies demonstrate the role of cancer stem cells in occurrence, recurrence, and distant metastases in gastric cancer (GC), little is known about the evolving genetic and epigenetic changes in the stem and progenitor cells. The purpose of this study was to identify the stem cell subtypes in GC and examine their clinical relevance. Two publicly available datasets were used to identify GC stem cell subtypes, and consensus clustering was performed by unsupervised machine learning methods. The cancer stem cell (CSC) typing-related risk scoring (RS) model was established through multivariate Cox regression analysis. Cross-platform dataset-based two stable GC stem cell subtypes, namely low stem cell enrichment (SCE_L) and high stem cell enrichment (SCE_H), were prudently identified. Gene set enrichment analysis revealed that the classical oncogenic pathways, immune-related pathways, and regulation of stem cell division were active in SCE_H; ferroptosis, NK cell activation, and post-mutation repair pathways were active in SCE_L. GC stem cell subtypes could accurately predict clinical outcomes in patients, tumor microenvironment cell-infiltration characteristics, somatic mutation landscape, and potential responses to immunotherapy, targeted therapy, and chemotherapy. Additionally, a CSC typing-related RS model was established; it was strongly independent and could accurately predict the patient's overall survival. This study demonstrated the complex oncogenic mechanisms underlying GC. The findings provide a basis and reference for the diagnosis and treatment of GC.


Systematic review of the CUP trials characteristics and perspectives for next-generation studies.
Research on therapeutic strategies for patients with unknown primary cancer (CUP) has been underwhelming. This paper summarized and evaluated the CUP therapeutic research over the previous five years. Based on this evaluation, recommendations for clinical trial designs are made to improve the impact of CUP research on patients. Published and ongoing research were evaluated. PubMed was searched from January 1, 2015, to November 1, 2021. The start date of 2015 was chosen to identify research published after ESMO issued new diagnostic and therapeutic guidelines. The US National Library of Medicine indexed ongoing clinical trials. Of the 244 CUP studies indexed in PubMed, 11.9% were prospective studies, and 4.9% were clinical trials. The review protocol deemed 65 publications eligible for full-text review. Eleven studies evaluating therapeutic regimens were retained. The two prospective studies and non-randomized trials showed promising outcomes for site-specific treatments. Randomized clinical trials were less promising; however, the trials had recruitment challenges resulting in biased accrual and the inability to keep pace with advancing diagnostics and therapeutics. Most of the 35 ongoing studies were phase II single-arm trials assessing immune checkpoint inhibitors (ICI) or site-specific therapies among CUP patients with suspected favorable prognoses. Our evaluation suggests two prospective clinical trial designs that addressed recent study design and recruitment challenges. A visionary approach uses a multi-arm, multistage randomized trial to address rapid advancements in diagnosis and therapy. A pragmatic approach utilizes a single-arm trial with historical controls to overcome comparison group and recruitment challenges.


An integrative analysis of Qingfei Paidu Decoction for its anti-HCoV-229E mechanism in cold and damp environment based on the pharmacokinetics, metabolomics and molecular docking technology.
The novel coronavirus pneumonia (COVID-19) has spread rapidly around the world. As a member against the epidemic, Qingfei Paidu Decoction (QFPDD) has been approved for the treatment of COVID-19 in China. However, its antiviral mechanism was still largely unclear. An integrated strategy was used to explore the antiviral mechanisms of QFPDD in cold and damp environment, including pharmacokinetic (PK), network pharmacology, metabolomics and protein verification. Firstly, the pharmacokinetic study of the prototype absorbed ingredients were analyzed by UHPLC-QqQ-MS. Secondly, the metabolomics analysis of the endogenous constituents was carried out. Based on the aforementioned results, an integrated network was constructed to identify the curative components, crucial endogenous differential metabolites and related pathways. Finally, the validation tests were implemented by molecular docking and western blotting (WB). According to the pharmacokinetic behaviors analysis of 31 components in vivo, the flavonoids presented more longer residence time and higher exposure compared with the other compounds. The efficacy and antiviral mechanism of QFPDD were verified by the poly-pharmacology, metabolomics, molecular docking and WB. For the occurrence of metabolic disorder, the change of amino acid transporters should not be neglected. Afterward, 8 curative compounds, 6 key genes and corresponding metabolic pathways were filtered by compound-reaction-enzyme-gene network. The molecular docking verified that the active ingredients bound to the relevant targets well. In the present study, an in vivo comprehensive pharmacokinetic behaviors of QFPDD was analyzed for the first time. The results illustrated that QFPDD could exhibit immune regulation, anti-infection, anti-inflammation and metabolic disorder to perform a corresponding therapeutic effect. Moreover, our findings highlighted the roles of amino acid transporters in the coronavirus infection situation.


Natural history of duodenal lesions in Japanese patients with familial adenomatosis coli (Gardner's syndrome).
The natural history of duodenal lesions was studied in 20 patients with familial adenomatosis coli/Gardner's syndrome. These patients were followed for an average of 7.1 yr (range 1 yr, 7 mo to 12 yr, 2 mo) and repeatedly examined by hypotonic duodenography, duodenofiberscopy, and biopsy. Tubular adenomas of the duodenum (less than 8 mm in diameter) were present in 18 (90%). During the follow-up period, there was a slight increase or decrease in the number of duodenal adenomas in 4 patients but no distinct change in 13. In the remaining patient (aged 49 yr), a polypoid lesion (17 mm in diameter) of the duodenal bulb, which had been overlooked at the initial examination, became an advanced cancer (50 x 30 mm in size) during 22 mo. Tubular adenomatous tissue of the duodenal papilla was detected in 11 patients (55%). Follow-up study of the lesions revealed no change in endoscopic and histologic findings in all but 1 patient, in whom there was a slight enlargement of the duodenal papilla, as seen at endoscopy. These findings suggest to us that in patients with this disease, duodenal adenomas do not require prophylactic surgery. However, careful repeated follow-up examinations with endoscopic biopsy should be performed, with endoscopic removal of larger polyps when possible.


Denitrification performance and microbial diversity in a packed-bed bioreactor using biodegradable polymer as carbon source and biofilm support.
A novel kind of biodegradable polymer, i.e., starch/polycaprolactone (SPCL) was prepared and used as carbon source and biofilm support for biological denitrification in a packed-bed bioreactor. The denitrification performances and microbial diversity of biofilm under different operating conditions were investigated. The results showed that the average denitrification rate was 0.64 ± 0.06 kg N/(m(3)d), and NH3-N formation (below 1mg/L) was observed during denitrification. The nitrate removal efficiency at 15°C was only 55.06% of that at 25°C. An initial excess release of DOC could be caused by rapid biodegradation of starch in the surfaces of SPCL granules, then it decreased to 10.08 mg/L. The vast majority of species on SPCL biofilm sample (99.71%) belonged to six major phyla: Proteobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Spirochaetes and Actinobacteria. Proteobacteria were the most abundant phylum (85.50%) and mainly consisted of β-proteobacteria (82.39%). Diaphorobacter and Acidovorax constituted 52.75% of the identified genera which were denitrifying bacteria.


Achondroplasia in Latin America: practical recommendations for the multidisciplinary care of pediatric patients.
Achondroplasia is the most common bone dysplasia associated with disproportionate short stature, and other comorbidities, such as foramen magnum stenosis, thoracolumbar kyphosis, lumbar hyperlordosis, genu varum and spinal compression. Additionally, patients affected with this condition have higher frequency of sleep disorders, ear infections, hearing loss and slowed development milestones. Considering these clinical features, we aimed to summarize the regional experts' recommendations for the multidisciplinary management of patients with achondroplasia in Latin America, a vast geographic territory with multicultural characteristics and with socio-economical differences of developing countries. Latin American experts (from Argentina, Brazil, Chile and Colombia) particiáted of an Advisory Board meeting (October 2019), and had a structured discussion how patients with achondroplasia are followed in their healthcare centers and punctuated gaps and opportunities for regional improvement in the management of achondroplasia. Practical recommendations have been established for genetic counselling, prenatal diagnosis and planning of delivery in patients with achondroplasia. An outline of strategies was added as follow-up guidelines to specialists according to patient developmental phases, amongst them neurologic, orthopedic, otorhinolaryngologic, nutritional and anthropometric aspects, and related to development milestones. Additionally, the role of physical therapy, physical activity, phonoaudiology and other care related to the quality of life of patients and their families were discussed. Preoperative recommendations to patients with achondroplasia were also included. This study summarized the main expert recommendations for the health care professionals management of achondroplasia in Latin America, reinforcing that achondroplasia-associated comorbidities are not limited to orthopedic concerns.


Making Medical Decisions for Incapacitated Patients Without Proxies: Part II.
In the United States, there is no consensus about who should make decisions in acute but non-emergent situations for incapacitated patients who lack surrogates. For more than a decade, our academic medical center has utilized community volunteers from the hospital ethics committee to engage in shared decision-making with the medical providers for these patients. In order to add a different point of view and minimize conflict of interest, the volunteers are non-clinicians who are not employed by the hospital. Using case examples and interviews with the community members, this paper describes how the protocol has translated into practice over the years since its inception. Members reported comfort with the role as well as satisfaction with the thoroughness of their discussions with the medical team. They acknowledged feelings of moral uncertainty, but expressed confidence in the process. Questions raised by the experience are discussed. Overall, the protocol has provided oversight, transparency, and protection from conflict of interest to the decision-making process for this vulnerable patient population.


Smoking patterns and cancer of the oral cavity and pharynx: a case-control study in Uruguay.
A case-control study of cancer of the oral cavity and pharynx was conducted in Uruguay, between 1992 and 1996. 425 patients microscopically diagnosed with squamous cell carcinoma of the oral cavity and pharynx were frequently matched on age, residence, and urban/rural status with 427 hospitalised controls. The study was restricted to males. Smokers of black tobacco cigarettes were associated with an increased risk of 12.1 (95% confidence interval (CI) 7.6-19.4), when compared with non-smokers after fitting a model which included the matching variables, birthplace, education, and total alcohol consumption. Lifelong smokers of hand-rolled cigarettes displayed an odds ratio (OR) of 8.7 (95% CI 5.6-13.4), compared with non-smokers. When smokers were excluded from the calculations, the OR for smokers of black tobacco cigarettes was 3.0 (95% CI 2.0-4.6), compared with smokers of blond tobacco cigarettes, after controlling for the same variables mentioned above, plus pack-years, years since stopping, and filter use. Hand-rolling appears to be less important than smoking black tobacco in this study (OR 1.6, 95% CI 0.9-2.5). Thus, smoking black tobacco cigarettes appears to be an important habit in oral and pharyngeal carcinogenesis.


Winning confidence.
Nursing Standard Nurse Awards 2008 winner Karen Logan improved intermittent self-catheterisation services, provided evidence-based guidance for nurses and ensured a trust-wide approach.


TBRG-1 a Ras-like protein in Trichoderma virens involved in conidiation, development, secondary metabolism, mycoparasitism, and biocontrol unveils a new family of Ras-GTPases.
Ras-GTPases are nucleotide hydrolases involved in key cellular processes. In fungi, Ras-GTPases regulate conidiation, development, virulence, and interactions with other fungi or plants. Trichoderma spp. are filamentous saprophytic fungi, widely distributed along all latitudes, characterized by their rapid growth and metabolic diversity. Many species of this genus interact with other fungi, animals or plants. Furthermore, these fungi are used as biocontrol agents due to their ability to antagonize phytopathogenic fungi and oomycetes, through competence, antibiosis, and parasitism. However, the genetic and molecular regulation of these processes is scarcely described in these fungi. In this work, we investigated the role of the gene tbrg-1 product (GenBank accession number XP_013956100; JGI ID: Tv_70852) of T. virens during its interaction with other fungi and plants. Sequence analyses predicted that TBRG-1 bears the characteristic domains of Ras-GTPases; however, its size (1011 aa) is 3- to 4-times bigger compared with classical GTPases. Interestingly, phylogenetic analyses grouped the TBRG-1 protein with hypothetical proteins of similar sizes, sharing conserved regions; whereas other known Ras-GTPases were perfectly grouped with their respective families. These facts led us to classify TBRG-1 into a new family of Ras-GTPases, the Big Ras-GTPases (BRG). Therefore, the gene was named tbrg-1 (TrichodermaBigRas-GTPase-1). Quantification of conidia and scanning electron microscopy showed that the mutants-lacking tbrg-1 produced less conidia, as well as a delayed conidiophore development compared to the wild-type (wt). Moreover, a deregulation of conidiation-related genes (con-10, con-13, and stuA) was observed in tbrg-1-lacking strains, which indicates that TBRG-1 is necessary for proper conidiophore and conidia development. Furthermore, the lack of tbrg-1 affected positively the antagonistic capability of T. virens against the phytopathogens Rhizoctonia solani, Sclerotium rolfsii, and Fusarium oxysporum, which was consistent with the expression patterns of mycoparasitism-related genes, sp1 and cht1, that code for a protease and for a chitinase, respectively. Furthermore, the antibiosis effect of mycelium-free culture filtrates of Δtbrg-1 against R. solani was considerably enhanced. The expression of secondary metabolism-related genes, particularly gliP, showed an upregulation in Δtbrg-1, which paralleled an increase in gliotoxin production as compared to the wt. These results indicate that TBRG-1 plays a negative role in secondary metabolism and antagonism. Unexpectedly, the biocontrol activity of Δtbrg-1 was ineffective to protect the tomato seeds and seedlings against R. solani. On the contrary, Δtbrg-1 behaved like a plant pathogen, indicating that TBRG-1 is probably implicated in the recognition process for establishing a beneficial relationship with plants.


A novel silicon complex is as effective as sodium metasilicate in enhancing the collagen-induced inflammatory response of silicon-deprived rats.
An experiment was conducted with rats to determine whether silicon deprivation affects the inflammatory response to the injection of type II collagen, and to compare the effectiveness of the organic complex arginine silicate inositol (ASI) with inorganic silicon (NaSiO(3)) in mitigating any observed change in response. Dark Agouti rats were fed a ground corn-casein-safflower-based diet containing about 2.8 mg Si/kg. The experimental variables were supplemental 0 and 35 mg Si/kg as either ASI or NaSiO3. After five weeks on their respective treatments, each rat was injected with type II collagen and euthanized four weeks later. Urine was collected before injection during week five and week nine before euthanasia. The silicon-supplemented rats generally exhibited a more marked inflammatory response than the silicon-deprived rats. The circulating number of lymphocytes was higher (p<0.003) and number of neutrophils was lower (p<0.008) in silicon-deprived than silicon-supplemented rats. ASI and NaSiO3 were about equally effective in enhancing these changes. Post-injection of tibial release of prostaglandin E(2) (PGE(2)) (p<0.04), urinary excretion of magnesium (p<0.03) and deoxypyridinoline (p<0.009), and plasma osteopontin (p<0.009), magnesium (p<0.0007) and copper (p<0.004) were higher in silicon-supplemented than silicon-deprived rats. The increases in plasma magnesium (Si x sex, p<0.04) and copper (Si x sex, p<0.02) were more marked in male than female rats. One but not the other silicon supplement when compared to silicon deprivation significantly affected the tibial release of PGE(2), and plasma copper and iron concentrations. However, with the exception of the pre-injection urinary excretion of helical peptide, no other of the variables determined was significantly different between rats fed ASI and those fed NaSiO3. The findings suggest that, in rodents, physiological amounts of silicon promote the immune response, sex may influence the response to dietary silicon, and that both organic silicon complexes and inorganic silicon are similarly effective in preventing changes in inflammation induced by silicon deprivation.


The indusium griseum in the mouse: architecture, Timm's histochemistry and some afferent connections.
The Indusium griseum (IG) is an enigmatic cortical field classically felt to be a part of the hippocampus (HC). In the mouse, IG lies just dorsal to the corpus callosum at the base of the anterior half of cingulate cortex. In coronal sections the field is small but constitutes a fairly long rostro-caudal strip. The connections of the IG are poorly understood. The Timm's staining pattern of the IG is reminiscent of a mini-dentate gyrus (DG) comprising a layer of granule cells with two bands of staining in the molecular layer. In the DG there are three bands which correspond to inputs from the lateral and medial entorhinal area (LEA and MEA) and the ipsi- and contralateral association systems. Using anterograde transport of HRP we have found that the LEA and MEA also terminate in the molecular layer of the IG. This suggests that the IG is a displaced portion of the DG. The olfactory system is known to have a strong indirect influence on the HC via primary and secondary bulbar projections to the LEA. Wheat germ agglutinin-HRP injections confined to the main olfactory bulb (MOB) show a direct projection from the MOB to IG. Both the olfactory bulb itself and retrobulbar structures such as the piriform cortex (PC) convey olfactory information to the LEA; the LEA supplies a major input to the DG. Our results suggest that there is a more direct pathway whereby olfactory information may influence a cortical region, IG, whose histochemistry and direct afferents from the entorhinal cortex suggest that it is part of or closely related to the DG. Thus, IG may represent a phylogenetically old olfacto-recipient outpost of the hippocampus.


Clinical characteristics of acute respiratory syndrome with SARS-CoV-2 infection in children in South China.
A retrospective study was conducted to summarize the clinical information of childhood infections during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemic. Children with SARS-CoV-2 infection in 11 hospitals from three provinces of South China were included in the study. Clinical information was collected and compared with children and adults infected by SARS-CoV-2 in Wuhan. In total, 52 children were enrolled, including 28 boys. The median age was 9 years (interquartile range [IQR], 4-12); 44.2% cases were of clustered occurrences, 40.4% patients had fever, 48.1% had cough, and 46.2% had a high lymphocyte count. No abnormalities were found in the liver and kidney function. Also, 82.7% of patients received antiviral therapy, but such therapy did not shorten the time to virus negativity or hospital stay (P = .082). The time to virus negativity was 12.0 days (IQR, 8.0-16.8) and hospital stay was 14.5 days (IQR, 10.3-17.9). Compared with reports in Wuhan, there were more acute upper respiratory tract infection (AURTI) and fewer pneumonia cases (P = .000). Compared with the non-ICU adult COVID-19 in Wuhan, these children's diseases were relatively mild, with fewer complications. Children with SARS-CoV-2 infection had a mild fever, lymphocyte elevation was more common than reduction, and antiviral treatment had no obvious effect. The overall clinical manifestations were mild, and the prognosis was good.


Two of four alternatively spliced isoforms of RUNX2 control osteocalcin gene expression in human osteoblast cells.
Runx2 is a Runt domain transcription factor that transcriptionally regulates osteoblast differentiation and bone formation. In this study, we show that human chondro- and osteosarcoma cell lines, human mesenchymal stem cells (hMSC) and a human primary chondrocytes (HC), osteoblst cells (HOb) express an intact isoform (RUNX2wt) and 3 alternatively spliced isoforms (RUNX2Delta5, Delta7, and Delta5Delta7) that are generated by skipping exon 5 and/or exon 7. Two of the truncated forms of RUNX2 (RUNX2Delta5 and RUNX2Delta5Delta7) did not localize in the nucleus and had lost their DNA binding activity. In cotransfection experiments with an osteocalcin (OC) promoter construct, we confirmed that only RUNX2wt and RUNX2Delta7 could upregulate the OC promoter activity in the osteosarcoma cell line. In addition, the coactivator CBP/p300 enhanced the transcriptional activity of the OC promoter when coexpressed with RUNX2wt or RUNX2Delta7, but not when coexpressed with RUNX2Delta5 or RUNX2Delta5Delta7. In contrast, the corepressor HDAC3 only repressed the activation from the OC promoter when coexpressed with RUNX2wt. These results support the hypothesis that RUNX2 both up- and downregulates its target gene promoters, as exemplified by the OC gene, using various isoforms and context-dependent formation of transcriptional complexes.


Green synthesis of zero valent iron using tannins to activate persulfate for sulfamethoxazole degradation.
Majority zero-valent iron (ZVI) materials are prepared by reducing agents in liquid phase, resulting in the high environmental pollution and poor particle size distribution uniformity. Therefore, this study employed a green synthesis method to prepare ZVI. Tannins (TA) with phenolic hydroxyl groups that are characterized by strong reducing capacity were employed to synthesize ZVI (TA@ZVI). The dispersity and stability of ZVI was improved by TA, which inhibited the agglomeration of ZVI. Meanwhile, the specific surface area of TA@ZVI was higher than chemical prepared ZVI, increasing the reactive sites. The organic matter components enriched on TA could promote the adsorption of pollutants and complex with Fe(II/III) to enhance the reactivity of TA@ZVI. Also, the polyphenol structure in TA was oxidized to quinone, which facilitated electron transport. In order further test the performance of TA@ZVI, SMX was chosen as a target pollutant to study the oxidative degradation performance of TA@ZVI. SO4•- degraded about 16.4%-25.5% SMX and •OH degraded about 49.8%-63.9% SMX in the pH range of 4-6 while •OH played a dominant role in the neutral and alkaline conditions. Moreover, the presence of TA reduced Fe(III) to Fe(II) and promoted the release of Fe(II), providing a continuous source of •OH for the oxidative degradation of SMX. Besides, the conversion of Fe(II/III) was accelerated due to TA, which delayed the formation of passivation layer. Thus, TA enhanced the antioxidant capacity of ZVI. Generally, this study provided an environmental-friendly technology to synthesize and improve the reactivity of ZVI.


Descriptions of Finlaya Theobald, a genus in tribe Aedini (Diptera: Culicidae), and its type species Fl. kochi (Dönitz).
Genus Finlaya Theobald is characterized based on the morphology of females, female genitalia, males, male genitalia, pupae, and 4th-stage larvae. Distinctive characters of the genus are discussed and included species are listed. Finlaya kochi, the type species, is described in detail and a syntype female is selected as neotype. An extensive list of previous literature pertaining to the genus is provided.


Protective effects of hesperetin on lipopolysaccharide-induced acute lung injury by targeting MD2.
Inflammation plays an important role in acute lung injury (ALI). Hesperetin (HES), a natural flavanone and an aglycone of hesperidin, has established potent anti-inflammatory activity. The aim of this study was to evaluate the potential protective effect of HES on lipopolysaccharide (LPS)-induced ALI in mice and to illuminate its possible directly target. Results indicated that HES pretreatment significantly attenuated LPS-induced pulmonary pathological injury, total protein concentration, markedly decreased the number of neutrophils and the levels of inflammatory cytokines, TNF-α and IL-6, in ALI model in vivo and in vitro. Meanwhile, pretreatment with HES dramatically reduced myeloperoxidase (MPO) activity in LPS-induced ALI mice. Additionally, using molecular docking and co-immunoprecipitation assay, HES showed a directly bind with myeloid differentiation 2 (MD2), in which HES could inhibit MAPK activation, regulate IκB degradation, block the interaction MD2 and its co-receptor Toll-like receptor 4 (TLR4). Taken together, HES showed a significantly protective effect against LPS-induced ALI, which might be associated with MD2 protein. These results attested HES worthy of further progress into an adjunctive potential drug for the treatment for ALI.


Tooth-pulp-evoked rostral spinal trigeminal neuronal excitation is attenuated by the activation of 5-HT3 receptors via GABAergic interneurons in the rat.
The effect of iontophoretic application of the 5-HT3 receptor agonist, phenylbiguanide (PBG), on the excitation of the trigeminal spinal nucleus oralis (TSNO) neurons to tooth-pulp (TP) stimulation was examined. The PBG application inhibited the TP-evoked TSNO neuronal excitation, and this inhibition was completely blocked by co-application of a GABAA receptor antagonist, bicuculline. The results suggest that the activation of 5-HT3 receptors elicits GABA release in the TSNO.


Beneficial effect of oral administration of zinc sulfate on 5-fluorouracil-induced gastrointestinal mucositis in rats.
This experimental study explored the potential of oral zinc sulfate to protect the gut mucosa from 5-fluorouracil (5-FU)-induced degenerative lesions in Wistar rats. Female Wistar rats were used and divided into 2 interventional groups (Z with 6 animals and F with 5 animals) and one control group (M with 5 rats). After 2 hours of fasting, group Z received via oral gavage 1.5 ml of solution, corresponding to 15 mg zinc sulfate for 9 consecutive days. Groups F and M received only the vehicles. On day 3, 400 mg/kg of 5-FU was administered intraperitoneally to groups Z and F. Tissue samples were collected from the duodenum, jejunum, colon and liver. Histological assessment for each gastrointestinal tract segment was determined semi-quantitatively by rating 11 histological features from normal (0) to severe (3). The independent groups were analyzed using the Kruskal-Wallis test and the Mann-Whitney U-test, with a Bonferroni correction for alpha (p ≤ 0.016). In group F the jejunum was the most affected area with a mean histological score of 27 (25-32). In the Z group, significantly lower histological scores were obtained compared with group F (duodenum Z vs. F: U = 0, p = 0.004; jejunum Z vs. F: U = 0, p = 0.006 and colon: Z vs. F: U = 0, p = 0.005). Graded liver necro-inflammatory lesions were significantly lower in group Z compared with group F (U = 0, p = 0.004), suggesting fewer bacterial intestinal translocation processes. Zinc sulfate has a beneficial role, decreasing the severity of gut mucosal injuries induced by 5-FU in Wistar rats.


On the molecular-weight-dependence of the anticoagulant activity of heparin.
The inactivation of thrombin and factor Xa by antithrombin was determined in the presence of heparin fractions of different molecular weights and with high affinity for antithrombin. The ability to potentiate the inactivation of either coagulation factor increased with increasing length of the polysaccharide chain.


Selenoprotein W serves as an antioxidant in chicken myoblasts.
Selenoprotein W (SelW) was thought to play an antioxidant role in mammals. Because chicken SelW has no cysteine (Cys) at the residue 37 (Cys37) that is required for the presumed antioxidant function in mammals, this study was conducted to determine whether chicken SelW possessed the same function. Small interfering RNAs (siRNAs) technology was applied to suppress the SelW expression in chicken embryonic myoblasts. Thereafter, these myoblasts were treated with different concentrations of H2O2 and assayed for cell viability, apoptosis rate, reactive oxygen species (ROS) status, and expression levels of apoptosis-related genes and proteins (Bax, Bcl-2, and caspase-3). Silencing of the myoblast SelW gene decreased their cell viability, and increased their apoptosis rate and susceptibility to H2O2. While the knockout down of SelW up-regulated Bax and caspase-3 and down-regulated Bcl-2, the induced oxidative injuries were alleviated by treatment with a ROS scavenger, N-acetyl-l-cysteine (NAC). Chicken SelW protected embryonic myoblasts against cell apoptosis mediated by endogenous and exogenous H2O2. Chicken SelW possesses antioxidant function similar to the mammalian homologues despite the lack of Cys37 in the peptide.


Quantitation of protein by alkaline extraction of naphthol blue black-stained polypeptides in sodium dodecyl sulfate-polyacrylamide slab-gels.
A sodium dodecyl sulfate-polyacrylamide slab-gel system was used to study the alkaline extraction of Naphthol Blue Black B-stained polypeptides. A simple, rapid, and reproducible method for quantitative assessment of 1- to 5-micrograms quantities of individual polypeptides in the molecular weight range of 14,000 to 120,000 is described. The working range of the assay may be extended to the 20-micrograms level at the expense of rapidity.


Use of patient portals to support recruitment into clinical trials and health research studies: results from studies using MyChart at one academic institution.
Electronic health records (EHRs) are often used for recruitment into research studies, as they efficiently facilitate targeted outreach. While studies increasingly are reaching out to potential participants through the EHR patient portal, there is little available information about which approaches are most effective. We surveyed all investigators at one academic medical center who had used the Epic MyChart patient portal for recruitment. We found that messages were typically adapted for a large group, but not tailored further for individual subgroups. The vast majority of studies sent a message only once. Recruitment costs were modest, averaging $431/study. The results show some promise for recruiting through the patient portal but also identified ways in which messages could be optimized.


In utero treatment of myelomeningocele with allogenic umbilical cord-derived mesenchymal stromal cells in an ovine model.
The purpose of our study was to investigate the effects of ovine umbilical cord-derived mesenchymal stromal cells (UC-MSCs) seeded in a fibrin patch as an adjuvant therapy for fetal myelomeningocele repair in the ovine model. MMC defects were surgically created at 75 days of gestation and repaired 15 days later with UC-MSCs patch or an acellular patch. At birth, motor function, tail movements, and voiding abilities were recorded. Histological and immunohistochemical analysis included study of MMC defect's healing, spinal cord, UC-MSCs survival, and screening for tumors. Six lambs were born alive in each group. There was no difference between the two groups on the median sheep locomotor rating score but all lambs in the control group had a score between lower than 3 compared to 50% in UC-MSCs group. There were more lambs with tail movements and voiding ability in UC-MSCs group (83% vs 0% and 50% vs 0%, respectively). gray matter area and large neurons density were higher in UC-MSCs group (2.5 vs 0.8 mm2 and 19.3 vs 1.6 neurons/mm2 of gray matter, respectively). Fibrosis thickness at the myelomeningocele scar level was reduced in UC-MSCs group (1269 µm vs 2624 µm). No tumors were observed. Fetal repair of myelomeningocele using allogenic UC-MSCs patch provides a moderate improvement in neurological functions, gray matter and neuronal preservation and prevented from fibrosis development at the myelomeningocele scar level.


Reshaping of global gene expression networks and sex-biased gene expression by integration of a young gene.
New genes originate frequently across diverse taxa. Given that genetic networks are typically comprised of robust, co-evolved interactions, the emergence of new genes raises an intriguing question: how do new genes interact with pre-existing genes? Here, we show that a recently originated gene rapidly evolved new gene networks and impacted sex-biased gene expression in Drosophila. This 4-6 million-year-old factor, named Zeus for its role in male fecundity, originated through retroposition of a highly conserved housekeeping gene, Caf40. Zeus acquired male reproductive organ expression patterns and phenotypes. Comparative expression profiling of mutants and closely related species revealed that Zeus has recruited a new set of downstream genes, and shaped the evolution of gene expression in germline. Comparative ChIP-chip revealed that the genomic binding profile of Zeus diverged rapidly from Caf40. These data demonstrate, for the first time, how a new gene quickly evolved novel networks governing essential biological processes at the genomic level.


Experiments with Seasonal Forecasts of ocean conditions for the Northern region of the California Current upwelling system.
Resource managers at the state, federal, and tribal levels make decisions on a weekly to quarterly basis, and fishers operate on a similar timeframe. To determine the potential of a support tool for these efforts, a seasonal forecast system is experimented with here. JISAO's Seasonal Coastal Ocean Prediction of the Ecosystem (J-SCOPE) features dynamical downscaling of regional ocean conditions in Washington and Oregon waters using a combination of a high-resolution regional model with biogeochemistry and forecasts from NOAA's Climate Forecast System (CFS). Model performance and predictability were examined for sea surface temperature (SST), bottom temperature, bottom oxygen, pH, and aragonite saturation state through model hindcasts, reforecast, and forecast comparisons with observations. Results indicate J-SCOPE forecasts have measurable skill on seasonal timescales. Experiments suggest that seasonal forecasting of ocean conditions important for fisheries is possible with the right combination of components. Those components include regional predictability on seasonal timescales of the physical environment from a large-scale model, a high-resolution regional model with biogeochemistry that simulates seasonal conditions in hindcasts, a relationship with local stakeholders, and a real-time observational network. Multiple efforts and approaches in different regions would advance knowledge to provide additional tools to fishers and other stakeholders.


Voices from the Margins: Reflections of an Oncology Chaplain.
Michael Eselun, BCC, who serves as the oncology chaplain for the Simms-Mann/UCLA Center for Integrative Oncology, recently gave a lecture as part of the Insights Into Cancer lecture series at UCLA called 'Cancer: "It Could Be Worse" - A Spiritual Ponzi Scheme?'. Michael has written a personal reflection to provide a fuller context for his chaplaincy, as he views it, and to provide some background to the lecture. A link to a video of the lecture is included.


Epidemiology of lower gastrointestinal bleeding.
Lower GI bleeding is a very broad topic, which can encompass situations from a small amount of red blood on tissue paper associated with formed brown stool, to life-threatening severe haemorrhage. Much of the literature on this topic focuses on acute bleeding necessitating hospitalisation and urgent intervention. The literature that is available focuses primarily on medical intervention and support, which will be covered in another review in this series. Causes for lower GI bleeding include diverticular disease, vascular ectasia, ischemic, inflammatory or infectious colitis, colonic neoplasia (including post polypectomy bleeding), anorectal causes (including haemorrhoids, fissures and rectal varices), and small bowel lesions (Crohn's, vascular ectasia, Meckel's diverticula, and small bowel tumours). Different clinical series identified these lesions in varying frequencies. Factors associated with the development of acute lower GI bleeding include advanced age and use of non-steroidal anti-inflammatory medication. Colonoscopy is the single most frequent intervention in evaluating all the patients with lower GI bleeding. Determining the precise impact of colonoscopy on the outcome of lower GI bleeding is difficult due to the retrospective nature of many studies, and the frequent inability to definitively establish the exact bleeding site.


An Algorithm for Correction of Mild Tuberous Breast Deformity in Augmentation Mammaplasty: Camouflage or Correct?
Tuberous breast deformity (TBD) is a common abnormality, particularly in patients presenting for breast augmentation. Failure to correct each regional abnormality, including the inframammary fold, lower pole deficiency, nipple-areola complex widening or herniation, or any degree of ptosis, will result in exaggeration of the deformity and a poor aesthetic outcome. To describe an algorithm, including novel techniques, to address each region of mild TBD in patients undergoing breast augmentation. This is a retrospective review and description of the senior author's (K.T.) techniques for correction of early-stage TBD from 2016 to 2021. One hundred forty-two patients underwent a stepwise approach to correct milder TBD features when undergoing breast augmentation. The authors propose a regional algorithm for management of TBD, to allow mostly single-stage correction, except in cases with marked ptosis, severe asymmetry, or marked macroareola.


Immunohistochemical localization of gamma-aminobutyric acid in the rat pituitary gland and related hypothalamic regions.
The distribution of gamma-aminobutyric acid (GABA) containing neurons in the rat pituitary gland and related hypothalamic areas was immunohistochemically investigated using antibodies raised against GABA conjugated to bovine serum albumin by glutaraldehyde. A dense network of GABA-like immunoreactive fine varicose nerve fibers was observed within the posterior and intermediate lobes of the pituitary gland, surrounding endocrine cells and capillaries, but not in the anterior lobe. In the pituitary stalk, the dense varicose fibers ran along the anterior wall of the posterior lobe into the posterior and intermediate lobes. A small number of GABA-like immunoreactive cell bodies were evident in the intermediate lobe. GABA-like immunoreactive fibers occurred at low to high density in most parts of the hypothalamus. GABA-like immunoreactive neurons were observed in some regions related to the pituitary gland (such as periventricular nucleus, paraventricular nucleus, arcuate nucleus and accessory magnocellular nucleus). These results provide morphological evidence for the presence of GABAergic neurons in the rat hypothalamo-pituitary system.


The Slovenian food composition database.
The preliminary Slovenian food composition database was created in 2003, through the application of the Data management and Alimenta nutritional software. In the subsequent projects, data on the composition of meat and meat products of Slovenian origin were gathered from analyses, and low-quality data of the preliminary database were discarded. The first volume of the Slovenian food composition database was published in 2006, in both electronic and paper versions. When Slovenia joined the EuroFIR NoE, the LanguaL indexing system was adopted. The Optijed nutritional software was developed, and later upgraded to the OPEN platform. This platform serves as an electronic database that currently comprises 620 foods, and as the Slovenian node in the EuroFIR virtual information platform. With the assimilation of the data on the compositions of foods of plant origin obtained within the latest project, the Slovenian database provides a good source for food compositional values of consistent and compatible quality.


Combination chemotherapy for advanced breast cancer: two regimens containing adriamycin.
Forty-eight women with advanced metastatic carcinoma of the breast were treated with one of two combination chemotherapy regimens: 1) adriamycin and cyclophosphamide or 2) adriamycin, cyclophosphamide, methotrexate and 5-fluorouracil. The response rate in the two-drug treatment group was 50% and in the four-drug treatment group, 55%. The median duration of response was ten months in both treatment groups. Dramatic responses were seen in patients with visceral metastases. Patients who responded to chemotherapy had a significantly longer survival than nonresponders (p less than 0.01). The long interval between adriamycin doses (six weeks) in the four drug regimen did not adversely effect the response rate--an important finding in view of the dose-related cardiac toxicity of this agent.


Merkel cell polyomavirus, a highly prevalent virus with tumorigenic potential.
Merkel cell polyomavirus (MCPyV) is the only human polyomavirus known to be involved in tumorigenesis. Like other human polyomaviruses, MCPyV is highly prevalent in the healthy population, yet the MCPyV-associated Merkel cell carcinoma (MCC) is a very rare disease. Although in vitro and in vivo models have provided significant details regarding molecular functions of viral oncoproteins during cellular transformation, many open questions about the natural life cycle of the virus, its mechanisms of persistence and the precise role of MCPyV during MCC pathogenesis remain. This review will carve out the specifics of MCPyV biology and discuss unresolved issues to help the reader gain a better understanding of what may differentiate MCPyV from other polyomaviruses.


Sequential changes of the T lymphocytes following ER rosette formation in guinea pigs. I. Expression of Fc and complement receptors.
Changes in proportions of the Fc and complement receptor (FcR, CR) positive T lymphocytes from guinea pigs following their interaction with rabbit erythrocytes (ER) were studied using EA and EAC rosette forming assays. Significant increases in the percentages of EA and EAC rosette forming cell (RFC) were observed when thymocytes or lymph node cells were assayed after ER rosette formation. Furthermore, T-enriched fraction by the ER monolayer adherence technique also showed similar or somewhat higher increases in the proportions of both EA and EAC RFC than those of unfractionated cells after contact with ER. The double rosette assay by ER with EA or EAC showed that 50-80% of the Fc and/or complement receptor positive lymphocytes bound rabbit erythrocytes simultaneously. These findings strongly suggest that at least a subset of the guinea pig T cells is altered to express Fc and/or complement receptors on their surfaces following the interaction with ER.


Borderline Boolean states improve the biosensing applications of DNA circuits.
Molecular circuits have been used in a wide range of diagnosis applications, from the detection of chemical molecules in solution to the complex processing of cell surface receptors. One of the most important challenges of these systems is the lack of distinguishability between different circuit states when each circuit state represents a specific disease. In this work, we designed a molecular amplification circuit with borderline Boolean states that each state can be distinguished with different color intensity. For this purpose, two DNA complexes and four DNA hairpin structures were designed to detect miR-218 and miR-215 biomarkers. One of the designed DNA complexes has two G-quadruplex structures and the other has only one G-quadruplex structure. In the absence of the inputs, all three G-quadruplex structures are active and produce a high-intensity signal, while in the other three states, including the presence of miR-218, the presence of miR-215, and the presence of both inputs, respectively, one, two, and zero G-quadruplex structures are active. Therefore, the designed system can identify two different biomarkers simultaneously with different signal ratios, which can easily distinguish the different states of the circuit. This strategy is very promising to identify diseases in which any combination of biomarkers leads to a particular disease.


Aurora-B/AIM-1 kinase activity is involved in Ras-mediated cell transformation.
Aurora-B, previously known as AIM-1, is a conserved eukaryotic mitotic protein kinase. In mammals, this kinase plays an essential role in chromosomal segregation processes, including chromosome condensation, alignment, control of spindle checkpoints, chromosome segregation, and cytokinesis. Aurora-B is overexpressed in various cancer cells, suggesting that the kinase activity perturbs chromosomal segregation processes. Its forced overexpression induces chromosomal number instability and progressive tumorigenicity in rodent cells in vitro and in vivo. Nevertheless, based on focus formation in BALB/c 3T3 A31-1-1 cells, Aurora-B is not oncogenic. Here, we show that Aurora-B kinase activity augments Ras-mediated cell transformation. RNA interference with short hairpin RNA inhibits transformation by Ras and its upstream oncogene Src, but not by the downstream oncogene Raf. In addition, the inner centromere protein, which is a passenger protein associated with Aurora-B, has a similar ability to potentiate the activity of oncogenic Ras. These data indicate that elevated Aurora-B activity promotes transformation by oncogenic Ras by enhancing oncogenic signaling and by converting chromosome number-stable cells to aneuploid cells.


Erratum: DCF RENTZ, YOU NING SU amp; NORIHIRO UESHIMA (2021) Studies in Australian Tettigoniidae: New short-winged Agraeciini from Australia (Orthoptera: Tettigoniidae; Conocephalinae; Agraeciini). Zootaxa 5059: 001072.
n/a.


Management and patient survival in hepatocellular carcinoma: does the physician's level of experience matter?
The prognostic determinants of hepatocellular carcinoma (HCC) depend on tumor stage, liver function reserve, and treatments offered. The clinical impact of the physician's experience on HCC management and the survival outcome is unknown. A total of 103 patients were managed by one high-volume physician and 249 patients by seven low-volume physicians. The experience of high-volume physician in HCC management was five times more than that of low-volume physicians. Patient survival was the single end point for this study. Compared to the low-volume physician group, more of the patients allocated to the high-volume physician had early stage HCC on the date of diagnosis (66/103, 64.1%; vs 119/249, 47.8%; P = 0.008), and they received curative therapies including radiofrequency ablation or liver resection (66/103, 64.1% vs 54/249, 21.7%, P < 0.001), and had significantly better survival outcome (median survival of 34 months, 95% confidence interval [CI], 17.6-50.4; vs 6 months, 95% CI, 4.3-7.7; P < 0.001) with a multivariable-adjusted hazard ratio (HR) for survival of 1.94 (95%, CI, 1.31-2.87, P < 0.001). A multivariate analysis of the pretreatment prognostic factors for these two groups identified alpha-fetoprotein (AFP) level (HR, 1.42; 95% CI, 1.01-1.99; P = 0.042), ascites (HR, 1.68; 95% CI, 1.15-2.46; P = 0.007), maximum tumor diameter (HR, 1.78; 95% CI, 1.16-2.74; P = 0.009), and portal vein thrombosis (PVT) (HR, 2.17; 95% CI, 1.49-3.17; P < 0.001) as independent factors for the low-volume physician group. However, only maximum tumor diameter (HR, 4.54; 95% CI, 1.77-11.67; P < 0.001) and PVT (HR, 5.73; 95% CI, 2.30-14.22; P = 0.002) were independent factors for the high-volume physician group. The survival of HCC patients was dependent on the level of experience of the physicians who oversaw these patients.


Gastrointestinal bleeding of undetermined origin: What diagnostic strategy to propose?
Gastrointestinal bleeding of undetermined origin (GBUO) is defined as gastrointestinal bleeding without an identified cause or location despite an endoscopic assessment including an esogastroduodenal endoscopy (EOGD) and a total colonoscopy. A distinction is made between exteriorized GBUO and non-exteriorized occult GBUO. The causes in the majority of cases (vascular, inflammatory and tumoral) are located in the small intestine. The diagnostic strategy aiming to locate the origin of the GBUO is a real challenge. Innovation in endoscopic and imaging techniques has enabled minimally invasive exploration of the small intestine. In Europe, there is a strong consensus to recommend a video-capsule endoscopy (VCE) as the first-intention study. If there is reason to suspect intestinal obstruction, VCE is contraindicated and a CT-enteroscopy is then performed as first intention. Enteroscopy is performed as a second-line treatment, either for therapeutic purposes after a positive VCE or CT-enteroclysis, or for diagnostic purposes after a negative VCE. Finally, intraoperative enteroscopy (IOE) coupled with surgical exploration should be reserved either for therapeutic purposes in the event of impossibility or failure of preoperative enteroscopy, or for diagnostic purposes in the event of recurrent GBUO after failure of all other studies and explorations of the small intestine.


Switching of subunit composition of muscle spectrin during myogenesis in vitro.
Spectrin comprises a family of polypeptides thought to be involved in mediating linkage of actin filaments to the plasma membrane in a wide variety of cell types (for reviews see refs 1-3). Spectrin is present as a tetramer composed of two non-identical subunits. Most cells express a common subunit with a molecular weight (Mr) of 240,000 (240 K; termed alpha-spectrin) in association with a polymorphic cell type-specific subunit: Mr 260 K in the intestinal terminal web (termed TW260), Mr 235 K in nervous tissue., liver, lymphocytes and fibroblasts (termed gamma-spectrin; also referred to as fodrin), and Mr 225/220 K in erythrocytes and adult cardiac and skeletal muscle (termed beta'- and beta-spectrin, respectively). We show here that primary chicken myoblasts express predominantly alpha gamma-spectrin, but on terminal differentiation in vitro the cells gradually switch to alpha beta-spectrin as a result of the onset of beta- and beta'-spectrin synthesis and by the subsequent differential stabilization of beta- and gamma-spectrin. This switching correlates with known changes in the biophysical properties and function of the developing muscle sarcolemma and cytoskeleton.


Proton Transfer Accounting for Anomalous Collision-Induced Dissociation of Proton-Bound Hoogsteen Base Pair of Cytosine and Guanine.
To understand the anomalous collision-induced dissociation (CID) behavior of the proton-bound Hoogsteen base pair of cytosine (C) and guanine (G), C:H+∙∙∙G, we investigated CID of a homologue series of proton-bound heterodimers of C, 1-methylcytosine, and 5-methylcytosine with G as a common base partner. The CID experiments were performed in an energy-resolved way (ER-CID) under both multiple and near-single collision conditions. The relative stabilities of the protonated complexes examined by ER-CID suggested that the proton-bound complexes produced by electrospray ionization in this study are proton-bound Hoogsteen base pairs. On the other hand, in contrast to the other base pairs, CID of C:H+∙∙∙G exhibited more abundant productions of C:H+, the fragment protonated on the moiety with a smaller proton affinity, than that of G:H+. This appeared to contradict general prediction based on the kinetic method. However, further theoretical exploration of potential energy surfaces found that there can be facile proton transfers in the proton-bound Hoogsteen base pairs during the CID process, which makes the process accessible to an additional product state of O-protonated C for C:H+ fragments. The presence of an additional dissociation channel, which in other words corresponds to twofold degeneracy in the transition state leading to C:H+ fragments, effectively doubles the apparent reaction rate for production of C:H+. In this way, the process gives rise to the anomaly, the observed pronounced formation of C:H+ in the CID of the proton-bound Hoogsteen base pair, C:H+∙∙∙G. Graphical Abstract .


A practical synthesis of 3-[(1R)-1-t-butyldimethylsilyloxyethyl]-4-[(2R)- 4-halo-3-oxo-2-butyl]azetidinone, a versatile intermediate for carbapenem antibiotics.
A key intermediate for 2-(functionalized-methyl)-1-beta-methylcarbapenems, 3-[(1R)-1-t-butyldimethylsilyloxyethyl]-4-[(2R)- 4-halo-3-oxo-2-butyl]azetidinone, was prepared efficiently from a commercially available carboxylic acid in 3 steps.


Breast Sentinel Lymph Node Frozen Section Practice: An Enterprise Audit as a Guide for Moving Forward.
In recent years, there has been a shift to less aggressive surgical management of the axilla in breast cancer. Consequently, sentinel lymph node evaluation by frozen section (FS) has declined. Additionally, there has been an impetus to decrease efforts in identifying small sentinel lymph node metastases. To critically evaluate our enterprise performance in evaluating axillary sentinel lymph nodes submitted for FS prior to considering changes in processing. A retrospective review (August 1, 2017-July 31, 2019) was conducted to identify sentinel and nonsentinel lymph nodes from 1 academic institution and 2 community sites. Cases were evaluated for grossing technique and discordance between FS and permanent section (PS) due to sampling and/or interpretive error. Clinicopathologic features were assessed. Lymph nodes from 426 patients with 432 neoplasms were sent for FS. Serial sectioning at 2-mm intervals was adhered to in 338 of 432 (78.2%). Serial sectioning was significantly lower at the community sites (14 of 60; 23.3%) versus at the academic institution (324 of 372; 87.1%; P < .001). Discordant cases were all false negatives (21 of 432; 4.8%). A total of 7 of 21 false negatives (33.3%) had macrometastatic (>2 mm) disease; of these, 3 were post-neoadjuvant chemotherapy, 3 were neither serially sectioned nor posttherapy, and 1 was a small (0.3-cm) focus. A total of 15 of 16 false negatives due to sampling error were detected on the first permanent section level. Standard serial sectioning of sentinel lymph nodes at 2-mm intervals resulted in infrequent false negatives due to macrometastatic disease. A single additional permanent section level is reasonable, given adherence to serial sectioning.


Metabolic responses of perfused rat livers to alpha- and beta-adrenergic agonists, glucagon and cyclic AMP.
1. The mechanism of action of glucagon and epinephrine was studied in perfused rat livers. Hormone-induced transitions from one metabolic steady state to another were followed in a non-recirculating perfusion system. Glucose and lactate production rates, oxygen uptake and K+ redistribution were measured. 2. Glucagon (3 nM), cyclic AMP (0.2 mM) and epinephrine (0.5 muM) had similar effects on K+ concentrations in the perfusate. Glycogenolysis responded more rapidly and O2 uptake was enhanced to a larger extent with epinephrine than with the other agents. alpha- and beta-receptor responses were differentiated by the use of phenylephrine (0.5 muM), isoproterenol (0.5 muM) and adrenergic blocking agents (phentolamine and beta-blocker Ro 3-4787 at 0.1 mM). 3. alpha-receptors mediated an activation of glucose production that was very rapid and was paralleled by a transient decrease of K+ concentrations in the effluent from the liver, lactate production rose gradually. Respiration was also enhanced, but fell again as lactate production increased. 4. beta-receptor stimulation was followed by an increase of glucose production that was less drastic and was paralleled by a K+ release, lactate production and respiration were only slightly enhanced. beta stimulation and glucagon both resulted in an inhibition of the alpha-adrenergic effect on lactate release and simultaneously increased O2 uptake. 5. We concluded that in perfused rat livers alpha- as well as beta-adrenergic receptor stimulation resulted in an activation of glycogenolysis, possibly by two different mechanisms.


Penile reflexes and copulatory behavior in male rats following lesions in the bed nucleus of the stria terminalis.
Male rats with large lesions in the bed nucleus of the stria terminalis (BNST) had more intromissions preceding ejaculation (if they ejaculated at all), longer intervals between intromissions, and longer postejaculatory refractory periods than control animals. In tests for penile reflexes, BNST males were similar to control males in every respect. The copulatory deficits in BNST males are probably not mediated by a change in penile reflex potential.


Experimental Study of the Validity of Entangled Two-Photon Absorption Measurements in Organic Compounds.
Entangled two-photon absorption (ETPA) has recently become a topic of lively debate, mainly due to the apparent inconsistencies in the experimentally reported ETPA cross sections of organic molecules obtained by a number of groups. In this work, we provide a thorough experimental study of ETPA in the organic molecules Rhodamine B (RhB) and zinc tetraphenylporphirin (ZnTPP). Our contribution is 3-fold: first, we reproduce previous results from other groups; second, we on the one hand determine the effects of different temporal correlations─introduced as a controllable temporal delay between the signal and idler photons to be absorbed─on the strength of the ETPA signal, and on the other hand, we introduce two concurrent and equivalent detection systems with and without the sample in place as a useful experimental check; third, we introduce, and apply to our data, a novel method to quantify the ETPA rate based on taking into account the full photon-pair behavior rather than focusing on singles or coincidence counts independently. Through this experimental setup we find that, surprisingly, the purported ETPA signal is not suppressed for a temporal delay much greater than the characteristic photon-pair temporal correlation time. While our results reproduce the previous findings from other authors, our full analysis indicates that the signal observed is not actually due to ETPA but simply to linear losses. Interestingly, for higher RhB concentrations, we find a two-photon signal that, contrary to expectations, likewise does not correspond to ETPA.


Synthesis of a Disulfonated Derivative of Cucurbit[7]uril and Investigations of its Ability to Solubilize Insoluble Drugs.
Cucurbit[7]uril (CB[7]) is currently being investigated as a solubilizing agent for insoluble drugs. We recently found that acyclic CB[n]-type receptors that bear sulfonate solubilizing groups are well suited for this application. Herein, we report cucurbit[7]uril derivative (1) that bears two sulfonate groups on its convex face that we hypothesized would be a superior solubilizing excipient for insoluble drugs. Before using 1 for drug solubilization experiments we showed that 1 does not self-associate and that it retained its ability to bind to diammonium compounds as common guests for CB[7] sized cavities. X-ray crystallography shows that 1 maintains the key structural features of CB[7] with only minor ellipsoidal deformations at the equator and carbonyl portals of 1. Unfortunately, the aqueous solubility of 1 (20 mM) is slightly lower than CB[7] (20-30 mM) which limits its potential as a solubilizing excipient for insoluble drugs. We created phase solubility diagrams for the solubilization of three drugs (camptothecin, albendazole, cinnarizine) with two different containers (1 and CB[7]). CB[7] and 1 exhibit comparable solubilization abilities (e.g. Ka and maximum solubility) toward camptothecin and albendazole but 1 is an inferior solubilizing agent for cinnarizine because of the low solubility exhibited by the 1•cinnarizine complex.


In vivo enzyme activity of purified human brain renin.
Cerebrospinal fluid (CSF) of rats contains high angiotensinogen concentrations. When 3500-fold purified renin from human brain was injected into the brain ventricles of rats, angiotensin I concentrations increased from undetectable levels to 147.9 +/- 18.8 fMol per ml CSF. In parallel, mean arterial blood pressure increased from 93 +/- 2.4 mm Hg to 107 +/- 3.7 mm Hg. The increase in blood pressure could be abolished by intraventricular administration of saralasin, a blocker of angiotensin II receptors. Intraventricular injection of cathepsin D had no effect on arterial blood pressure and the agiotensin I concentration in CSF remained below detection limits of the radioimmunoassay. We conclude that brain renin acts on endogenous brain angiotensinogen under physioloical in vivo conditions to form angiotensin I. The latter is converted to angiotensin II and leads to biological effects, i.e. increase of blood pressure.


Prevalence and Risk Factors for Lateral Femoral Cutaneous Nerve Palsy in the Beach Chair Position.
To report on the prevalence of lateral femoral cutaneous nerve (LFCN) palsy in patients who had undergone shoulder surgery in the beach chair position and to identify patient and surgical risk factors for its development. We retrospectively reviewed the medical records of 397 consecutive patients who underwent either open or arthroscopic shoulder surgery in the beach chair position by a single surgeon. Patient demographic and surgical data including age, gender, weight, body mass index (BMI), diabetes, procedure duration, and anesthesia type (general, regional, regional/general) were recorded. LFCN palsy symptoms were recorded prospectively at the initial postoperative visit and identified clinically by focal pain, numbness, and/or tingling over the anterolateral thigh. The median patient age was 59.0 years and consisted of 158 males (40%) and 239 (60%) females. Five cases of LFCN palsy were identified for a prevalence of 1.3%. These patients had a higher median weight (108.9 kg vs 80.7 kg, P = .005) and BMI (39.6 vs 29.4, P = .005) than the patients who did not develop LFCN palsy. Median age, gender, diabetes, and surgical time were not significantly different between the groups. All cases resolved completely within 6 months. LFCN palsy after shoulder surgery in the beach chair position in our study has a prevalence of 1.3%, making it an uncommon complication. Patients with elevated BMI should be counseled about its possible occurrence after shoulder surgery in the beach chair position. Level IV, prognostic.


Black tea benefits short-chain fatty acid producers but inhibits genus Lactobacillus in the gut of healthy Sprague-Dawley rats.
The gut microbiota plays important roles in physiological and pathological processes of the host. The effect of black tea on the gut microbiota of healthy individuals remains unclear. Healthy Sprague-Dawley (SD) rats were given black tea for 4 weeks, and cecum content, serum, intestinal, and hepatic samples were collected. The results showed that black tea increased α-diversity and modulated β-diversity of the gut microbiota. Additionally, black tea enriched several short-chain fatty acid (SCFA) producers but suppressed genus Lactobacillus. Further tests revealed that the enrichment of SCFA producers was associated with a decrease in the oxidative stress of cecum content caused by black tea, and related to increased luminal butyric acid levels and enhanced intestinal barrier function. The suppression of genus Lactobacillus was related to the increase in luminal total bile acids caused by black tea. In vitro tests showed that bile acids rather than black tea directly inhibited Lactobacillus strains. The reduction in genus Lactobacillus did not affect the effects of black tea on intestinal barrier function and lipid levels. Our results imply that the effects of black tea on gut microbiota in healthy individuals are complex and provide a new perspective on the associations among black tea, gut microbiota, and health. © 2020 Society of Chemical Industry.


Depression recognition using resting-state and event-related fMRI signals.
This paper aimed to develop a method for depression detection using blood-oxygen-level-dependent (BOLD) response estimated from event-related signals and resting-state functional magnetic resonance imaging (fMRI) signals together. Thirteen patients with unipolar depression and matched healthy subjects were recruited. Resting state data of each subject were collected. Thereafter, event-related paradigm was undertaken using sad facial stimuli. The resting-state fMRI signal was deemed as the baseline of each subject's activity. Coefficient marks were designed to sort and select temporal independent components of event-related signals. Thereafter, stimulus-evoked BOLD response components inside event-related signal were extracted and taken as features to discriminate depressive patients from healthy controls. Accuracy rate for depression recognition was 77.27% with P value of .017 for whole-brain analysis and 81.82% with P value of .009 for region-of-interest analysis. The effectiveness and the superiority of the proposed method for disease recognition were demonstrated via the performance comparison with three other typical methods. The proposed model was effective in depression recognition.


Host-Epstein-Barr virus relationship affected by immunostimulation in HIV-infected patients representing distinct progressor profile groups.
Human immunodeficiency virus (HIV) infection with pronounced immunosuppression disrupts Epstein-Barr virus (EBV)-host balance with increased lymphoma risk. We explored whether different host responses to HIV are reflected in the EBV-host balance. Eleven unvaccinated HIV-positive patients and 16 participants in a vaccine trial were included in the study. Blood samples were collected, B cells extracted, and EBV DNA load was determined using a semiquantitative polymerase chain reaction (PCR) method. Treatment-naïve patients with a history of symptomatic primary HIV infection showed non-significant, but higher EBV load compared to untreated long-term non-progressors. A significant difference in HIV RNA titres between these groups correlated weakly to EBV DNA load. Patients in the vaccine trial with recombinant HIV gp160 and/or adjuvant and with a history of symptomatic primary HIV infection, showed a 1-log increase in EBV load compared to patients with long-lasting HIV disease. Different host responses to HIV infection, especially in combination with vaccination, can be reflected in the EBV-host balance.


PhoP and OxyR transcriptional regulators contribute to Yersinia pestis virulence and survival within Galleria mellonella.
The virulence of Yersinia pestis KIM6+ was compared with multiple isolates of Yersinia pseudotuberculosis and Yersinia enterocolitica toward larvae of the greater wax moth Galleria mellonella. Although Y. pestis and Y. pseudotuberculosis were able to cause lethal infection in G. mellonella, these species appeared less virulent than the majority of Y. enterocolitica strains tested. Y. pestis survived primarily within hemocytes of G. mellonella, and induced a strong antibacterial peptide response that lasted for at least 3 days in surviving larvae. Immunization with dead bacteria to induce an antibacterial response led to increased survival of the larvae following infection. Mutant strains lacking the either phoP or oxyR, which were less resistant to antibacterial peptides and hydrogen peroxide respectively, were attenuated and restoration of the wild-type genes on plasmids restored virulence. Our results indicate that the Y. pseudotuberculosis-Y. pestis lineage is not as virulent toward G. mellonella as are the majority of Y. enterocolitica isolates. Further, we have shown that G. mellonella is a useful infection model for analyzing Y. pestis host-pathogen interactions, and antibacterial peptide resistance mediated by phoP and reactive oxygen defense mediated by oxyR are important for Y. pestis infection of this insect.


An inverse correlation between TNF alpha serum levels and heart rate variability in patients with heart failure.
Recent evidence indicates that chronic heart failure (CHF) is accompanied by both activation of the immune system and autonomic imbalance. There is a growing body of evidence that increased levels of proinflammatory cytokines and other inflammatory markers have important roles as mediators of disease progression and markers of mortality in patients with CHF. The aim of this study was to investigate connection between autonomic imbalance [obtained by analysis of heart rate variability (HRV)] and activation of the immune system [as measured by serum levels of tumor necrosis factor (TNF)-α] in patients with chronic heart failure. This cross-sectional study included 21 patients with CHF and 8 age- and gender-matched healthy control subjects. We assessed HRV by 24-hour electrocardiographic Holter monitoring and measured serum levels of TNF-α using an enzyme-linked immunosorbent assay. Clinical assessment and echocardiography were also performed. There was an inverse correlation between serum level of TNF-α and a time-domain parameter of HRV - SDNN (r=-0.542, p<0.05). A similar result was found for HRV triangular index, a geometric measure of HRV (r=-0.556; p<0.05). The correlation was stronger for subjects with a diabetes mellitus, females, and TNFA2 allele carriers (an "A" at position -308A). The pNN50, indirect marker of cardiac vagal activity, was not significantly associated with serum concentration of TNF-α. In conclusion, the results of the present study indicate that increased serum TNF-α level is significantly associated with reduced HRV indices, suggesting that activation of the immune system in patients with CHF is closely related to autonomic imbalance.


Endovascular Repair of Anomalous Splenic Artery Aneurysm with Systemic Lupus Erythematosus.
Splenic artery aneurysm with systemic lupus erythematosus is rare but serious. We report a case of a 45-year-old female with systemic lupus erythematosus who presented with a splenic artery aneurysm of anomalous origin from the superior mesenteric artery. This is the first time to report about using an endovascular approach to treat splenic artery aneurysm combined with systemic lupus erythematosus. Endovascular repair of the aneurysm was conducted using a covered stent and multiple coils to exclude both the aneurysm and total splenic artery embolization for treating thrombocytopenia, and this approach was determined to be safe and effective for yielding short-term results.


Expert testimony in sexual assault cases: Alcohol intoxication and memory.
At court-martial tribunals in the United States military, cases involving alcohol facilitated sexual assault often pivot on the alleged victim's level of intoxication or impairment and ability to consent to the sexual act. These cases frequently arise following a night of partying and heavy drinking among a group of friends and acquaintances, military and civilian. The determination of whether a sexual act was consensual may rest on estimates of the alleged victim's blood alcohol concentration and related behavioral indicia of impairment. Expert testimony may be presented by the prosecution and/or the defense, from forensic toxicologists and psychiatrists or psychologists regarding the potential involvement of alcohol and its impact on the participants relevant to the charges at court-martial. A review of the state of the science is offered to bring such testimony into perspective. Appellate cases illustrate that the experts' testimony may sometimes elucidate, sometimes obfuscate, and sometimes exceed professional expertise and invade the province of the factfinder.


Dynorphin in pro-opiomelanocortin neurons of the hypothalamic arcuate nucleus.
Considerable evidence suggests that dynorphin participates in the regulation of energy balance. In this study, we have used immunohistochemistry to investigate in detail the cellular localization of pro-dynorphin (DYN) immunoreactive cell bodies in the mediobasal hypothalamus with special reference to neurons producing orexigenic or anorexigenic transmitters. In colchicine-treated rats, DYN immunoreactivity was demonstrated in many cell bodies of the arcuate nucleus (Arc). Double-labeling revealed that DYN immunoreactivity was present in approximately 30% of pro-opiomelanocortin (POMC) neurons in the ventrolateral Arc as shown by presence of alpha-melanocyte-stimulating hormone (alpha-MSH) and cocaine- and amphetamine-regulated transcript (CART). In contrast, DYN immunoreactivity was not demonstrated in agouti-related peptide (AgRP)- or neuropeptide Y (NPY) -containing neurons in the ventromedial aspect of the Arc. Dynorphin immunoreactivity was also colocalized with the vesicular acetylcholine (ACh) transporter (VAChT; a marker for cholinergic neurons) in the cell soma of Arc POMC neurons. Brainstem POMC neurons in the commissural part of the solitary tract nucleus (NTS) were devoid of DYN immunoreactivity, whereas DYN immunoreactivity was detected in a few NPY-containing NTS neurons and cholinergic DMX neurons. Our results showing presence of DYN together with alpha-MSH in a subpopulation of hypothalamic POMC neurons further point to the neurochemical heterogeneity of hypothalamic POMC neurons. The results suggest a role for DYN in control of energy balance by mediating the effect of peripheral hormones such as leptin and insulin.


The prevalence of posttraumatic stress among women requesting induced abortion.
To describe the prevalence and pattern of traumatic experiences, to assess the prevalence of posttraumatic stress disorder (PTSD) and posttraumatic stress symptoms (PTSS), to identify risk factors for PTSD and PTSS, and to analyse the association of PTSD and PTSS with concomitant anxiety and depressive symptoms in women requesting induced abortion. A Swedish multi-centre study of women requesting an induced abortion. The Screen Questionnaire - Posttraumatic Stress Disorder was used for research diagnoses of PTSD and PTSS. Anxiety and depressive symptoms were evaluated by the Hospital Anxiety and Depression Scale (HADS). Of the 1514 respondents, almost half reported traumatic experiences. Lifetime- and point prevalence of PTSD were 7% (95% confidence interval [CI]: 5.8-8.5) and 4% (95% CI: 3.1-5.2), respectively. The prevalence of PTSS was 23% (95% CI: 21.1-25.4). Women who reported symptoms of anxiety or depression when requesting abortion were more likely to have ongoing PTSD or PTSS. Also single-living women and smokers displayed higher rates of ongoing PTSD. Although PTSD is rare among women who request an induced abortion, a relatively high proportion suffers from PTSS. Abortion seeking women with trauma experiences and existing or preexisting mental disorders need more consideration and alertness when counselled for termination.


Use of laser knife in thoracic surgery.
CO2 laser knife was applied in 18 thoracic operations. The laser knife can be used for the sterile and ablastic cutting of skin, soft parts, ribs, sternum, bronchi and lung parenchyma. By its use intraoperative bleeding is substantially diminished.


National survey of pediatric services available in US emergency departments.
Children account for nearly 20% of all US emergency department (ED) visits, yet previous national surveys found that many EDs lack specialized pediatric care. In response, a 2001 joint policy statement recommended resources needed by EDs for effective pediatric emergency care delivery. We sought to update and enhance previous estimates of pediatric services available in US EDs. We administered a telephone survey to a 5% random sample (n = 279) of all US EDs from the 2007 National Emergency Department Inventory-USA. The survey collected data on local capabilities (including typical management of three clinical scenarios) and prevalence of a coordinator for pediatric emergency care. We used descriptive statistics to summarize data. Multivariable logistic regression was used to examine the association between survey respondent and ED characteristics as well as the presence of a coordinator for pediatric emergency medicine. Data were collected from 238 hospitals (85% response rate). A minority of hospitals had pediatric departments (36%) or intensive care units (12%). The median annual number of ED visits by children was 3,870 (interquartile range 1,500-8,800). Ten percent of hospitals had a separate pediatric ED; only 17% had a designated pediatric emergency care coordinator. Significant positive predictors of a coordinator were an ED pediatric visit volume of ≥1 patient per hour and urban location. Most EDs treated only mild-to-moderate cases of childhood bronchiolitis and asthma exacerbation (77% and 65%, respectively). Less than half (48%) of the hospitals reported the ability to surgically manage a child with acute appendicitis. We found little change in pediatric emergency services compared to earlier estimates. Our study results suggest a continued need for improvements to ensure access to emergency care for children.


Application of wastewater-based epidemiology to estimate the usage of beta-agonists in 31 cities in China.
The illegal use of beta-agonists could cause severe problems to human health. In this study, the usage of beta-agonists in 31 cities across China was estimated using wastewater-based epidemiology (WBE). The proposed method is based on solid-phase extraction (SPE) and LC-MS/MS and was developed and validated to determine the concentration of seven beta-agonists in wastewater. A population model based on cotinine (COT), NH4-N and the flow volume was constructed to estimate the population equivalents for different wastewater treatment plants (WWTPs). Clenbuterol and ractopamine are banned in China for both animal husbandry and medical use, but were nevertheless detected in some wastewater samples at rates of 6.2 % and 4.7 %, respectively (n = 339). The WBE-based consumption of clenbuterol and ractopamine were compared with the acceptable daily intake (ADI) and the health risks were assessed by their hazard quotients (0.26-6.62 for clenbuterol and 9.27 × 10-4-0.05 for ractopamine). Salbutamol, clorprenaline and terbutaline were observed in practically all wastewater samples at concentrations of up to several ng/L, whereas the formoterol and bambuterol concentrations were below the detection limit in all samples. Salbutamol consumption (7.35 ± 4.14 mg/1000 inh/day) was highest among the examined beta-agonists and varied regionally. Beta-agonist consumption based on WBE was higher in some cities than that based on medical survey data, indicating potential illegal use. These results show that WBE can be a straightforward and supplementary method for monitoring beta-agonist usage at the population level and spatially.


Pediatric update #8. Acute hematogenous osteomyelitis in children. Long-term results of surgical treatment.
A retrospective review (1970-1982) of the clinical experience of 68 children with acute hematogenous osteomyelitis (AHO) is presented. Analysis of the epidemiologic, clinical, laboratory, radiologic and bacteriologic findings is made. Our series was treated mainly by an aggressive primary surgical protocol which is described in detail. Seventeen percent developed chronic osteomyelitis. The role of surgery in the treatment of AHO is discussed. It is our belief that surgical treatment has definite advantages over the conservative regime and it remains our treatment of choice.


Toxic environmental exposures and kidney health in children.
High-level exposures to a number of agents are known to have direct nephrotoxic effects in children. A growing body of literature supports the hypothesis that chronic, relatively low-level exposure to various nephrotoxicants may also increase the risk for chronic kidney disease (CKD) or accelerate its progression. In this review we highlight several environmental nephrotoxicants and their association with CKD in children and adolescents. We also discuss unique epidemiological challenges in the use of kidney biomarkers in environmental nephrotoxicology.


Blood lactate, pH, base excess and pCO2 as prognostic indicators in caesarean-born kids from goats with pregnancy toxaemia.
The objective of this study was to identify the prognostic value for survival of blood parameters in the immediate post-caesarean surgery period in kids born from pregnancy toxaemia (PT) goats. This study involved 10 PT goats, in which a caesarean surgery was performed. Twenty-five kids were born after caesarean surgery of which 16 survived. A blood sample was collected from the jugular vein of the 10 goats and from the kids immediately after caesarean surgery (within 15 min). There were differences between the kids that survived and the kids that did not survive concerning the blood levels of pH (7.22 vs 7.00), base excess (- 9 vs - 18 mmol/L), pCO2 (46 vs 62 mmHg) and L-lactate (5.6 vs 16 mmol/L). Maternal ketoacidosis due to PT has a negative impact on the survival rate of the offspring. This appears to be associated to a metabolic acidosis of the offspring. However, the only blood parameter in which there was a strong association between the maternal and newborn kids was blood urea nitrogen (r = 0.97).


Lanthanum chloride suppresses hydrogen peroxide-enhanced calcification in rat calcifying vascular cells.
Lanthanum chloride (LaCl(3)) has been shown to retard the progression of established atherosclerotic lesions in animal models, and used as a calcium channel blocker in various cellular experiments. In this study, we assessed the role of lanthanum chloride (LaCl(3)) in H(2)O(2)-enhanced calcification in rat calcifying vascular cells (CVCs) and examined the involvement of MAPK signaling pathways. H(2)O(2) induced growth inhibition of CVCs, as well as increases in intracellular levels of calcium and reactive oxygen species, ALP activity, apoptosis and calcium deposition. These effects of H(2)O(2) were suppressed by pretreatment of the cells with 1 muM of LaCl(3) for 2 h. In addition, H(2)O(2) activated the phosphorylation of ERK1/2, JNK and p38 MAPK, but only the last two were associated with the ALP activity. Our findings demonstrate that H(2)O(2)-enhanced osteoblastic differentiation and apoptosis are responsible for the increased calcification in rat CVCs, and LaCl(3) can counteract these effects by suppressing the activation of JNK (JNK2, but not JNK1) and p38 MAPK signaling pathway.


Liquid-liquid microextraction methods based on ultrasound-assisted emulsification and single-drop coupled to gas chromatography-mass spectrometry for determining strobilurin and oxazole fungicides in juices and fruits.
Two procedures are proposed based on ultrasound-assisted emulsification and single-drop liquid-liquid microextraction for the sensitive determination of seven strobilurin and six oxazole fungicides in fruits and juice samples. Both miniaturized techniques are coupled to gas chromatography with mass spectrometry in the selected ion monitoring mode, GC-MS(SIM). The procedures use low density organic solvents, and several factors influencing the emulsification, extraction and collection efficiency are optimized. The detection limits obtained at a signal-to-noise ratio of 3 are below the MRLs set by the European Commission. Enrichment factors are between 140-1140 for the first technique used and 80-1600 for the latter. The recoveries obtained for spiked samples are satisfactory for all compounds. The methods are validated according to the Commission Decision 2002/657/EC. Different fruit and juices are analyzed by the proposed method and none of the samples contained fungicide residues above the detection limits.


Enzymatic synthesis of trisialoganglioside by particulate fractions of rat liver.
Rat liver contains a particulate (membrane-bound) glycosyl transferase, concentrated in Golgi apparatus fractions, which catalyzes the synthesis of a trisialoganglioside from the ganglioside precursor disialohematoside (GD3). Sialic acid was not incorporated into exogenous GD1a from CMP-N-acetylneuraminic acid suggesting that GD1a is an endproduct of the monosialoganglioside pathway. Thus, the disialoganglioside pathway may be a primary source of trisialoganglioside and higher ganglioside homologs in adult rat liver.