Quantitation of rubella virus by competitive enzyme immunosorbent assay.
A simple and rapid technique for quantification of rubella virus is described. The specificity of the competitive enzyme linked immunosorbent assay (ELISA) was quantified by comparing the slopes and the Y intercepts of the curves obtained when viral antigen vs. control antigen was used. The curves were derived by plotting the absorbance ratio against the logarithm of the antigen concentration. The technique was reproducible, its sensitivity depending on the purity of the antigen used. In ELISA, when an antigen precipitated by ammonium sulfate was used, the sensitivity expressed in protein/ml was 7 micrograms and when an antigen purified by sucrose gradient was used, 70 ng, whereas the limit of sensitivity in the standard technique of hemagglutination was only 38 micrograms.


Characterization of β-Glucan-Peanut Protein Isolate/Soy Protein Isolate Conjugates and Their Application on Low-Fat Sausage.
Polysaccharide-protein conjugates can improve the functional properties and expand the application field. The emulsifying, thermal properties of WSG-PPI conjugates and WSG-SPI conjugates were improved, compared to WSG, PPI and SPI. The Maillard reaction was confirmed by Fourier transform infrared spectroscopy (FT-IR). Circular dichroism (CD) exhibited that the structure of the conjugates was more expanded. Cryo-SEM and AFM demonstrated that the WSG, WSG-PPI and WSG-SPI conjugates had a morphology of a chain. When the conjugates were added as fat substitutes to low-fat sausage, the cooking yield, hardness and chewiness increased. The objective of this research was to study the emulsifying property, thermal property and structural changes of β-glucan-peanut protein isolate (WSG-PPI) conjugates and β-glucan-soy protein isolate (WSG-SPI) conjugates prepared through wet-heated Maillard reaction, and their effect on the texture of low-fat sausage.


Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8.
Preclinical studies on gene delivery into mouse lymphocytes are often hampered by insufficient activity of lentiviral (LV) and adeno-associated vectors (AAVs) as well as missing tools for cell type selectivity when considering in vivo gene therapy. Here, we selected designed ankyrin repeat proteins (DARPins) binding to murine CD8. The top-performing DARPin was displayed as targeting ligand on both vector systems. When used on engineered measles virus (MV) glycoproteins, the resulting mCD8-LV transduced CD8+ mouse lymphocytes with near-absolute (>99%) selectivity. Despite its lower functional titer, mCD8-LV achieved 4-fold higher gene delivery to CD8+ cells than conventional VSV-LV when added to whole mouse blood. Addition of mCD8-LV encoding a chimeric antigen receptor (CAR) specific for mouse CD19 to splenocytes resulted in elimination of B lymphocytes and lymphoma cells. For display on AAV, the DARPin was inserted into the GH2-GH3 loop of the AAV2 capsid protein VP1, resulting in a DARPin-targeted AAV we termed DART-AAV. Stocks of mCD8-AAV contained similar genome copies as AAV2 but were >20-fold more active in gene delivery in mouse splenocytes, while exhibiting >99% specificity for CD8+ cells. These results suggest that receptor targeting can overcome blocks in transduction of mouse splenocytes.


Interaction of plasma proteins with negatively charged sites on the pulmonary capillary endothelium of the rat.
The endothelial glycocalyx, a polyanionic structure which may regulate the passage of solutes and water through the endothelium, readily binds cationic ferritin (CF). In normal, nonexchange-transfused rats, however, only 7.5% and 6.0% of the luminal plasma membrane and 7.5% and 5.0% of vesicle diaphragms on the thick and thin side of pulmonary capillaries, respectively, bound cationic ferritin. With the graded removal of circulating proteins by exchange transfusion with fluorocarbon emulsion, up to 89 and 82% of the luminal surface, and 76 and 73% of vesicle diaphragms on the thick and thin sides, respectively, bound CF. Although the extent of binding on the thin side was consistently less than on the thick side, the difference was not statistically significant. The extensive binding of CF to the glycocalyx in totally exchange-transfused rats was completely reversible upon addition of lyophilized rat serum protein to the perfusate. These data suggest that in vivo anionic sites of the endothelial glycocalyx are partially masked by adsorbed plasma proteins.


New Therapies for Lowering Triglyceride-Rich Lipoproteins: JACC Focus Seminar 3/4.
Emerging evidence suggests that elevated concentrations of triglyceride-rich lipoprotein remnants (TRLs) derived from hepatic and intestinal sources contribute to the risk of atherosclerotic cardiovascular events. Natural selection studies support a causal role for elevated concentrations of remnant cholesterol and the pathways contributing to perturbations in metabolic pathways regulating TRLs with an increased risk of atherosclerotic cardiovascular disease events. New therapies targeting select catalytic pathways in TRL metabolism reduce atherosclerosis in experimental models, and concentrations of TRLs in patients with a vast range of triglyceride levels. Clinical trials with inhibitors of angiopoietin-like 3 protein and apolipoprotein C-III will be required to provide further guidance on the potential contribution of these emerging therapies in the paradigm of cardiovascular risk management in patients with elevated remnant cholesterol.


Procollagen III peptide and fibronectin in alcohol-related chronic liver disease: correlations with morphological features and biochemical tests.
In order to clarify the significance of procollagen III peptide (PIIIP) and fibronectin (FN) blood concentration in alcohol related chronic liver disease (ALD), we have investigated their relationships with histological liver features and biochemical liver tests in 44 ALD patients. PIIIP was measured in serum by radioimmunoassay whereas FN was determined in plasma using an immunonephelometric method. In each liver biopsy, steatosis, portal infiltrate, lobular necro-inflammation, portal fibrosis and lobular fibrosis were semiquantitatively assessed by scoring from 0 to 3. A close correlation of PIIIP was found with morphological features of fibrosis (both of lobular and portal type), but not with necro-inflammation or steatosis. PIIIP was also positively correlated with ALP and GGT and exhibited a good diagnostic value in liver fibrosis. On the contrary, FN did not distinguish between normals and patients and was not correlated with any morphological liver feature or biochemical liver test. We also conclude that serum NP3P effectively reflects liver fibrosis, whereas plasma FN seems not related to any of the main histological aspects of liver damage in ALD.


Linking obesity-induced leptin-signaling pathways to common endocrine-related cancers in women.
Obesity is related to many major diseases and cancers. Women have higher rates of obesity and obesity is linked to commonly occurring cancers in women. However, there is a lack of knowledge of the unique mechanism(s) involved in each type of cancer. The objective of this review is to highlight the need for novel experimental approaches and a better understanding of the common and unique pathways to resolve controversies regarding the role of obesity in cancer. In women, there is a link between hormones and obesity-associated genes in cancer development. Leptin is an obesity-associated gene that has been studied extensively in cancers; however, whether the defect is in the leptin gene or in its signaling pathways remains unclear. Both leptin and its receptor have been positively correlated with cancer progression in some endocrine-related cancers in women. This review offers an up-to-date and cohesive review of both upstream and downstream pathways of leptin signaling in cancer and a comprehensive picture of cancer pathogenesis in light of current evidence of leptin effects in several major types of cancer. This work is intended to aid in the design of better therapeutic strategies for obese/overweight women with cancer.


Comparing self-reported reactogenicity between adolescents and adults following the use of BNT162b2 (Pfizer-BioNTech) messenger RNA COVID-19 vaccine: a prospective cohort study.
Although clinical data have shown that the BNT162b2 vaccine, which is widely used in many countries, is safe and effective as a protection against the SARS-CoV-2 infection, extant research in adverse reactions using real-world data of various sociodemographic characteristics is scant. We conducted a prospective cohort study to compare age differences in self-reported reactogenicity of BNT162b2 in Hong Kong. A total of 1,516 participants were intensively followed up for two weeks following both doses of BNT162b2 vaccination, during which their basic demographic, health conditions, and medication information were collected. Results from the generalized mixed model showed that compared with adults aged 18 to 59 years, older adults aged 60 years or above had a lower risk of adverse reactions and adolescents aged 12 to 17 years had a moderately higher risk. Results of this study should be informative to parents considering BNT162b2 vaccination for their children in that moderately increased reactogenicity compared with adults is anticipated.


Characteristics Associated with Feeding Tube Placement: Retrospective Cohort Study of Texas Nursing Home Residents with Advanced Dementia.
To investigate resident-level, provider-type, nursing home (NH), and regional factors associated with feeding tube (FT) placement in advanced dementia. Retrospective cohort study. NH residents in Texas with dementia diagnosis and severe cognitive impairment (N = 20,582). This study used 2011-2016 Texas Medicare data to identify NH residents with a stay of at least 120 days who had a diagnosis of dementia on Long Term Care Minimum Data Set (MDS) evaluation and severe cognitive impairment on clinical score. Multivariable repeated measures analyses were conducted to identify associations between FT placement and resident-level, provider-type, NH, and regional factors. The prevalence of FT placement in advanced dementia in Texas between 2011 and 2016 ranged from 12.5% to 16.1% with a nonlinear trend. At the resident level, the prevalence of FT decreased with age [age > 85 years, prevalence ratio (PR) 0.60, 95% confidence interval (CI) 0.52-0.69] and increased among residents who are black (2.74, 95% CI 2.48-3.03) or Hispanic (PR 1.91, 95% CI 1.71-2.13). Residents cared for by a nurse practitioner or physician assistant were less likely to have an FT (PR 0.90, 95% CI 0.85-0.96). No facility characteristics were associated with prevalence of FT placement in advanced dementia. There were regional differences in FT placement with the highest use areas on the Texas-Mexico border and in South and East Texas (Harlingen border area, PR 4.26, 95% CI 3.69-4.86; San Antonio border area, PR 3.93, 95% CI 3.04-4.93; Houston, PR 2.17, 95% CI 1.87-2.50), and in metro areas (PR 1.36, 95% CI 1.22-1.50). Regional, race, and ethnic variations in prevalence of FT use among NH residents suggest opportunities for clinicians and policy makers to improve the quality of end-of-life care by especially considering other palliative care measures for minorities living in border towns.


Purification, characterization and antibacterial activities of red color-related protein found in the shell of kuruma shrimp, Marsupenaeus japonicus.
The well-known red color change plays a significant role in consumer acceptability of crustacean species. In this study, we described the purification of the red color-related protein named MjRCP75 from the shell of Marsupenaeus japonicus. It was a homogeneous monomer with molecular mass of 75 kDa and rich in α-helix conformation. The α-helix content decreased within the increasing of heating temperature and was transformed dominantly to β types. Identification and structural analysis revealed that MjRCP75 belonged to hemocyanin family. The released pigment from heated MjRCP75 showed a λmax at 483 nm in acetone. MjRCP75 showed clearly antibacterial activity against Escherichia coli, Staphylococcus aureus, and Vibrio parahaemolyticus. These findings identify MjRCP75 as the red color-related protein in M. japonicus shell and reveal its involvement in antibacterial activities.


Vasopressin and epinephrine are equally effective for CPR in a rat asphyxia model.
Epinephrine has been administered as a drug essential for cardiopulmonary resuscitation (CPR). Recently, vasopressin has been reported to be more effective than epinephrine for CPR in a ventricular fibrillation (VF) model. As a different myocardial pathology is speculated to exist between the VF model and the asphyxia model, we investigated whether vasopressin is also effective in a rat asphyxia model. Twenty-one Sprague-Dawley male rats were divided into three groups: vasopressin 0.8 U/kg (Vaso-Gr), epinephrine 0.05 mg/kg (Epi-Gr), and saline same volume as the other two drugs (Sal-Gr). Five minutes after suffocation induced by obstruction of the tracheal tube, CPR was performed using each drug. Although only one animal survived (17%) in the Sal-Gr, 6/7 (85%) survived in both Vaso-Gr and Epi-Gr (P<0.01). Vasopressin is as effective as epinephrine for CPR in asphyxia-induced rats.


Liverwort bHLH transcription factors and the origin of stomata in plants.
Stomata are distributed in nearly all major groups of land plants, with the only exception being liverworts. Instead of having stomata on sporophytes, many complex thalloid liverworts possess air pores in their gametophytes. At present, whether stomata in land plants are derived from a common origin remains under debate.1,2,3 In Arabidopsis thaliana, a core regulatory module for stomatal development comprises members of the bHLH transcription factor (TF) family, including AtSPCH, AtMUTE, and AtFAMA of subfamily Ia and AtSCRM1/2 of subfamily IIIb. Specifically, AtSPCH, AtMUTE, and AtFAMA each successively form heterodimers with AtSCRM1/2, which in turn regulate the entry, division, and differentiation of stomatal lineages.4,5,6,7 In the moss Physcomitrium patens, two SMF (SPCH, MUTE and FAMA) orthologs have been characterized, one of which is functionally conserved in regulating stomatal development.8,9 We here provide experimental evidence that orthologous bHLH TFs in the liverwort Marchantia polymorpha affect air pore spacing as well as the development of the epidermis and gametangiophores. We found that the bHLH Ia and IIIb heterodimeric module is highly conserved in plants. Genetic complementation experiments showed that liverwort SCRM and SMF genes weakly restored a stomata phenotype in atscrm1, atmute, and atfama mutant backgrounds in A. thaliana. In addition, homologs of stomatal development regulators FLP and MYB88 also exist in liverworts and weakly rescued the stomatal phenotype of atflp/myb88 double mutant. These results provide evidence not only for a common origin of all stomata in extant plants but also for relatively simple stomata in the ancestral plant.


Strategies for surgical remediation of the multi-fundoplication failure patient.
Outcomes are not well studied in patients undergoing remediation for multi-fundoplication failure, that is, two or more prior failed fundoplications. Re-operation must balance reflux control and restoration of the ability to eat with the challenge of reconstructing a distorted hiatus and GE junction. The purpose of this study is to present our experience with surgical remediation for multi-fundoplication failure. Medical records were retrospectively reviewed of 91 patients who underwent third time or more esophagogastric operation for fundoplication failure at a single institution from 2007 to 2016. Dysphagia was present in 56% and heartburn in 51%. Median number of prior operations was 2 with range up to 6. Anatomic failure consisted of slipped wrap in 26 cases, wrap herniation in 23, hiatal stenosis in 24, hiatal mesh complication in 8, and wrap dehiscence in 10. Operative approaches generally followed an institutional algorithm and consisted of hiatal hernia repair with: re-do fundoplication in 55%, takedown of fundoplication alone in 24%, Roux-en-Y gastrojejunostomy in 14%, and GE junction resection in 7%. Laparoscopic approach was successful in 81%. Mean duration of operations was 217 min and median length of stay was 3 days. The complication rate was 13%, with 7% undergoing unplanned early re-operation. Patients were followed for mean 11 months, and recurrent hiatal hernia was detected in 13%. Late re-operation was performed in 6% for recurrent hiatal hernia. Recurrent reflux symptomatology resolved in 93%. Dysphagia resolved in 84%. There were no significant differences in outcomes with regard to number of prior operations, operative approach, BMI, or age. There is no single best approach to remediation in the multi-fundoplication failure patient. Re-do fundoplication is appropriate in over half of patients. Reoperation for multi-fundoplication failure can be performed via minimally invasive approach with excellent remediation of symptoms, low morbidity, and low recurrence rates.


Neonatal Thyroid Disease: Testing and Management.
"Thyroid dysfunction that requires prompt diagnosis and treatment often becomes evident in the newborn period because of testing that is done as part of universal newborn screening. Primary congenital hypothyroidism is the most common treatable cause of mental retardation, requiring immediate treatment to prevent abnormal brain development. However, many of the abnormal thyroid test results are less abnormal and difficult to interpret, with a need for repeat testing and careful follow-up before initiation of treatment. Less often, neonatal hyperthyroidism is encountered. This article reviews and discusses management of thyroid dysfunction that may present in the first month after birth."


Long-term corneal complication of retained anterior chamber-angle foreign body.
A 33-year-old patient referred to the cornea and anterior segment department to evaluate inferior corneal edema related to a retained intraocular foreign body (IOFB) in the anterior chamber-angle. The foreign body, which was a single piece of glass caused by an exploded light bulb twenty years back, was surgically removed; edema resolved and vision improved to 20/30. In the presence of an anterior chamber IOFB; long-term adverse effects should be considered. We would advise removal of the FB regardless of the inertness and location facts, as long as the risk and benefits of the surgical intervention are carefully evaluated.


Association between early childhood sleep difficulties and subsequent psychiatric illness.
Chronic disruptions to sleep in childhood are associated with increased prevalence of psychiatric disease later in development. When sleep disruptions remit before adolescence, the increased prevalence of psychiatric disease is no longer observed, highlighting the importance of early detection and intervention. Clinicians typically rely on caregivers' reports for diagnosis and management of childhood sleep challenges. We examined whether findings on polysomnogram (PSG) can offer similar insight into childhood sleep difficulties and the risk of subsequent psychiatric illness. A cohort was identified of 348 children ages 5 years 11 months and younger with sleep difficulties rising to the level of formal clinical workup. A retrospective review of caregiver-reported sleep concerns, PSG results, and subsequent psychiatric illness was completed. PSG findings were compared to presence of psychiatric illness later in life as well as caregivers' reported concerns. Chi-squared and Fisher's exact tests were completed to evaluate correlations and Cohen's kappa was used to evaluate agreement. With only a few exceptions, comparisons between clinician findings on PSG and subsequent psychiatric diagnoses were statistically nonsignificant. Similarly, the relationship between caregivers' subjective reports about sleep and clinicians' findings on PSG demonstrated only slight to fair agreement, suggesting reported concerns were not predictive of PSG results. Parental reports of subjective sleep concerns are indicative of different sleep pathologies compared to sleep pathologies detected on PSG. The addition of PSG to caregiver-reported data appears to have limited clinical utility in understanding sleep concerns associated with the risk of subsequent psychiatric illness in young children. Pease E, Shekunov J, Savitz ST, et al. Association between early childhood sleep difficulties and subsequent psychiatric illness. J Clin Sleep Med. 2023;19(12):2059-2063.


Spinocerebellar ataxias.
Conventional MRI in patients presenting with progressive ataxia demonstrates the three main patterns of macroscopic damage, namely spinal atrophy, olivopontocerebellar atrophy and cortical cerebellar atrophy. Moreover it contributes to the diagnosis of fragile-X tremor ataxia syndrome and siderosis of the CNS. Non conventional MRI techniques detect nervous tissue abnormalities before development of atrophy which are correlated with the severity of the clinical deficit.


Effects of colchicine treatment on mean platelet volume and the inflammatory markers in recurrent aphthous stomatitis.
Recurrent aphthous stomatitis (RAS) is the most commonly seen inflammatory disease in the oral mucosa affecting 5%-25% of the general population. The etiology of RAS is still not fully understood and its treatment is very challenging. With its anti-inflammatory affects, colchicine is used for systematic treatment of RAS. In this study, we want to examine the effects of colchicine on platelet density, mean platelet volume (MPV), neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and red cell distribution width (RDW) of the patients with RAS. Fifteen male and 45 female RAS patients that were taking colchicine were investigated retrospectively. The whole blood parameters of the patients were observed before starting colchicine treatment and in the third month of colchicine treatment. Significant decrease in the levels of NLR, white blood cell count and RDW of the RAS patients under colchicine treatment was observed. Moreover, no changes were seen on MPVs, PLRs and hemoglobin (Hb) levels. It was determined that colchicine lowers the levels of NLR, white blood cell count and RDW. Furthermore, no changes were seen on MPVs, PLRs and Hb levels.


[Corticosteroids].
Adrenal steroids, particularly glucocorticoids, are used in a variety of conditions ranging from adrenal insufficiency requiring substitution therapy (glucocorticoids and mineral-corticoids), to a wide range of clinical disorders in which undesired inflammatory reactions must be reduced (glucocorticoids). The anti-inflammatory effect, typical of glucocorticoids, is mainly due to the interference with arachidonic acid metabolism, from which both prostaglandins and leukotriens, mediators of inflammation, take origin. ACTH is also employed with the same indications of glucocorticoids, but its use is limited, since its effect depends on the stimulation of cortico-adrenal gland with subsequent release of not only glucocorticoids but also mineral-corticoids and androgens. The therapeutical indications of glucocorticoids are numerous, but dose and duration of treatment vary in relation of the disease that is to be cured. The widespread use of steroids is accompanied by numerous unwanted reactions which depend on their metabolic actions. Therefore corticosteroids must be reserved to well defined clinical conditions and their use should be guided by criteria that are now codified. In this review the physiological and pharmacological effects and the clinical use of corticosteroids have been examined.


Carotid intima-media wall thickness in elderly women with and without atherosclerosis of the abdominal aorta.
In the present study the association was evaluated between non-invasively assessed atherosclerosis of the abdominal aorta and ultrasonographically measured intima-media wall thickness of the common carotid arteries in a population-based study of 41 elderly women. Atherosclerosis of the abdominal aorta was assessed in 1985 using a lateral X-ray of the lumbar spine, on which the presence of calcified deposits was determined. The carotid arteries were ultrasonographically evaluated in 1990 for presence of atherosclerotic plaques and intima-media wall thickness of the distal common carotid was measured off line using dedicated software. The age-adjusted mean intima-media wall thickness of the right common carotid artery was significantly higher in subjects with calcified deposits in the aorta (n = 16) compared with those without deposits (n = 25) with a mean difference of 0.15 mm (95% confidence interval (CI) 0.03, 0.26). For the left side similar results were observed. Mean common carotid intima-media wall thickness, ((left+right)/2), differed significantly across groups with a mean difference of 0.11 mm (95% CI 0.01, 021). Additional adjustment for differences across groups in body mass index, serum cholesterol, hypertension and smoking did not change the magnitude of the observed association: mean difference of 0.12 mm (95% CI -0.01, 0.25). The findings of the present study provide evidence that among subjects with atherosclerotic plaques in the abdominal aorta, the intima-media wall thickness of the distal common carotid arteries is increased.


PATTERN OF SKIN DISEASES IN ARMED FORCES IN PUNE REGION.
A total of 8123 Armed Forces personnel attended dermatological OPD from Jan 1989 to Dec 1994. On analysing the record it is found that 2953 (36.3%) cases had infective dermatosis and 5170 (63.7%) cases had non-infective dermatosis. Common infective dermatosis included fungal infections (17.2%), viral infections (7.7%), bacterial infections (3.6%) and arthropode infestations (7.8%). Common non-infective dermatosis included papulosquamous disorders (13.5%), pigmentary disorders (13.4%), eczemas (11.1%), acne (8.7%), alopecia (6.1%), urticaria (4.2%) and drug eruptions (1.9%).


A Review of the Comparative Anatomy, Histology, Physiology and Pathology of the Nasal Cavity of Rats, Mice, Dogs and Non-human Primates. Relevance to Inhalation Toxicology and Human Health Risk Assessment.
There are many significant differences in the structural and functional anatomy of the nasal cavity of man and laboratory animals. Some of the differences may be responsible for the species-specific nasal lesions that are often observed in response to inhaled toxicants. This paper reviews the comparative anatomy, physiology and pathology of the nasal cavity of the rat, mouse, dog, monkey and man, highlighting factors that may influence the distribution of nasal lesions. Gross anatomical variations such as turbinate structure, folds or grooves on nasal walls, or presence or absence of accessory structures, may influence nasal airflow and species-specific uptake and deposition of inhaled material. In addition, interspecies variations in the morphological and biochemical composition and distribution of the nasal epithelium may affect the local tissue susceptibility and play a role in the development of species-specific nasal lesions. It is concluded that, while the nasal cavity of the monkey might be more similar to that of man, each laboratory animal species provides a model that responds in a characteristic and species-specific manner. Therefore for human risk assessment, careful consideration must be given to the anatomical differences between a given animal model and man.


Schnurri-3 inhibition suppresses bone and joint damage in models of rheumatoid arthritis.
Rheumatoid arthritis (RA) is a chronic inflammatory disease that leads to systemic and articular bone loss by activating bone resorption and suppressing bone formation. Despite current therapeutic agents, inflammation-induced bone loss in RA continues to be a significant clinical problem due to joint deformity and lack of articular and systemic bone repair. Here, we identify the suppressor of bone formation, Schnurri-3 (SHN3), as a potential target to prevent bone loss in RA. SHN3 expression in osteoblast-lineage cells is induced by proinflammatory cytokines. Germline deletion or conditional deletion of Shn3 in osteoblasts limits articular bone erosion and systemic bone loss in mouse models of RA. Similarly, silencing of SHN3 expression in these RA models using systemic delivery of a bone-targeting recombinant adenoassociated virus protects against inflammation-induced bone loss. In osteoblasts, TNF activates SHN3 via ERK MAPK-mediated phosphorylation and, in turn, phosphorylated SHN3 inhibits WNT/β-catenin signaling and up-regulates RANKL expression. Accordingly, knock-in of a mutation in Shn3 that fails to bind ERK MAPK promotes bone formation in mice overexpressing human TNF due to augmented WNT/β-catenin signaling. Remarkably, Shn3-deficient osteoblasts are not only resistant to TNF-induced suppression of osteogenesis, but also down-regulate osteoclast development. Collectively, these findings demonstrate SHN3 inhibition as a promising approach to limit bone loss and promote bone repair in RA.


MPGES-1 derived PGE2 inhibits cell migration by regulating ARP2/3 in the pathogenesis of Hirschsprung disease.
We previously studied the metabolomics, transcriptomics and proteomics of intestinal tissue of Hirschsprung disease (HSCR) patients; the results suggested that the expression of prostaglandin E2(PGE2), prostaglandin E receptor 2(PTGER2) and microsomal prostaglandin E synthase-1 (mPGES-1) notably increased in HSCR colon tissues. We already verified the differential expression of PGE2/EP2 in HSCR patients; therefore we investigate how mPGES-1 derived PGE2 affects the migration and the potential mechanism in cells, revealing the role of mPGES-1 derived PGE2 in the pathogenesis of Hirschsprung disease. SH-SY5Y and SK-N-BE2 cell lines were obtained from American Type Culture Collection (ATCC, USA). Prostaglandin E2 and its synthetase inhibitors were purchased from Med Chem Express (MCE, USA). Migration assays were performed with transwell and scratch assays. Cell proliferation was confirmed by CCK8 method. Flow cytometer was used to detect the cell cycle and cell apoptosis. The expressions of mRNA and protein of EP2, ARP2/3 were determined by qRT-PCR and western blot respectively. Immunofluorescence and confocal laser scanning microscopy were used to observe the morphology and function of cytoskeleton. MPGES-1 derived PGE2 decreased the relative expression of EP2 and ARP2/3 and caused damage to cytoskeleton. As to cell functions, PGE2 inhibited cell migration while having no effects on the proliferation, cell cycle and apoptosis. By adding mPGES-1 inhibitor MK886 the abnormal expression and damaged cell function were reversed. MPGES-1 derived PGE2 inhibits the cell migration by regulating ARP2/3 complex via prostaglandin E2 receptor. Potential mechanisms are the damage of cytoskeleton and related proteins leading to failure of cell polarize and migration. Here we thoroughly inquire the role mPGES-1 derived PGE2 plays in cell migration which might provide a new thinking in the investigation interrelated to the pathogenesis of HSCR.


Decrease of bone mineral density during estrogen substitution therapy.
A decrease in bone mineral density (BMD) in patients treated with hormone replacement therapy (HRT) is sometimes observed in clinical practice. In order to assess the frequency and the characteristics of these cases, we reviewed the data of 102 women treated with HRT for more than 2 years, and who had undergone at least 3 lumbar BMD measurements during that period. For each patient, a linear function was fitted to the BMD data in relation to time. The slope was calculated. There was an overall gain in BMD during treatment, mean (+/- S.E.M.) values of slope 0.007 g/cm2/year (+/- 0.002). Fifty-three patients had a slope higher than 0.005, 28 a slope close to 0 (between 0.005 and -0.005) and 21 a slope lower than -0.005. By dividing the patients in tertiles of slopes (tertile I: slope < 0; tertile II: 0 < slope < 0.011; tertile III: slope > 0.011), significant differences were observed between the three groups of slope for the initial BMD (P < 0.001), hydroxyproline/creatinine ratio (P < 0.01), weight, DHEAS and alkaline phosphatase (P < 0.05). Only 1 of the 15 patients with a low bone mass (lower than mean +/- 1 S.D.) had a negative slope, while 9 of the 16 with a high-bone mass (higher than mean +/- 1 S.D.), had a negative slope. Under HRT, about 21% of postmenopausal women have a slight decrease in BMD as assessed by DPA. Because of the DPA coefficient of variation, however, the exact number of bone losers cannot be determined.(ABSTRACT TRUNCATED AT 250 WORDS)


Fusion of membrane vesicles bearing only the influenza hemagglutinin with erythrocytes, living cultured cells, and liposomes.
Membrane vesicles, bearing only the influenza viral hemagglutinin glycoprotein, were reconstituted following solubilization of intact virions with Triton X-100. The viral hemagglutinin glycoprotein was separated from the neuraminidase glycoprotein by agarose sulfanilic acid column. The hemagglutinin glycoprotein obtained was homogenous in gel electrophoresis and devoid of any neuraminidase activity. A quantitative determination revealed that the hemolytic activity of the hemagglutinin vesicles was comparable to that of intact virions. Incubation of fluorescently labeled hemagglutinin vesicles with human erythrocyte ghosts (HEG) or with liposomes composed of phosphatidylcholine/cholesterol or phosphatidylcholine/cholesterol/gangliosides, at pH 5.0 but not at pH 7.4, resulted in fluorescence dequenching. Very little, if any, fluorescence dequenching was observed upon incubation of fluorescently labeled HA vesicles with neuraminidase or glutaraldehyde-treated HEG or with liposomes composed only of phosphatidylcholine. Hemagglutinin vesicles were rendered non-hemolytic by treatment with NH2OH or glutaraldehyde or by incubation at 85 degrees C or low pH. No fluorescence dequenching was observed following incubation of non-hemolytic hemagglutinin vesicles with HEG or liposomes. These results clearly suggest that the fluorescence dequenching observed is due to fusion between the hemagglutinin vesicles and the recipient membranes. Incubation of hemagglutinin vesicles with living cultured cells, i.e. mouse lymphoma S-49 cells, at pH 5.0 as well as at pH 7.4, also resulted in fluorescence dequenching. The fluorescence dequenching observed at pH 7.4 was inhibited by lysosomotropic agents (methylamine and ammonium chloride) as well as by EDTA and NaN3, indicating that it is due to fusion of hemagglutinin vesicles taken into the cells by endocytosis.


Introducing sense into nonsense in treatments of human genetic diseases.
Approximately one-third of alleles causing genetic diseases carry premature termination codons (PTCs), which lead to the production of truncated proteins. The past decade has seen considerable interest in therapeutic approaches aimed at readthrough of in-frame PTCs to enable synthesis of full-length proteins. However, attempts to readthrough PTCs in many diseases resulted in variable effects. Here, we focus on the efforts of such therapeutic approaches in cystic fibrosis and Duchenne muscular dystrophy and discuss the factors contributing to successful readthrough and how the nonsense-mediated mRNA decay (NMD) pathway regulates this response. A deeper understanding of the molecular basis for variable response to readthrough of PTCs is necessary so that appropriate therapies can be developed to treat many human genetic diseases caused by PTCs.


Isolation of microbial natural products.
In principle, the isolation of secondary metabolites from microbes does not differ from their isolation from other organisms. The extraction procedure may of course be quite different, especially if it is carried out in an industrial scale, but when an extract containing the metabolites of interest is at hand, it is the same palette of adsorbents and chromatographic techniques that provide the major tools for the fractionation and eventual isolation of the pure compounds. Compared to plants, in which one is sure to find secondary metabolites of certain types, e.g., flavonoids, microbes can be expected to produce virtually anything and it is important to go about the fractionation procedure with an open mind. This chapter presents an overview of preparation of extracts from microbial sources, and various methods and strategies involved in the isolation and characterization of microbial natural products.


Handheld reflectance confocal microscopy for the diagnosis of molluscum contagiosum: Histopathology and dermoscopy correlation.
Handheld reflectance confocal microscopy may represent an adjunctive, fast, non-invasive tool for the diagnosis of molluscum contagiosum, revealing microscopic details closely related to histopathology, as demonstrated by this study evaluating 19 molluscum lesions in 11 patients. It permits the rapid examination of one or multiple skin lesions in real time and it is perfectly suitable for children.


V-Y Tendon Plasty for Reconstruction of Chronic Achilles Tendon Rupture: A Medium-term and Long-term Follow-up.
To evaluate the surgical results of V-Y tendon plasty in the treatment of chronic Achilles tendon rupture during medium and long-term follow-up. Between June 2005 and September 2017, 20 patients with chronic Achilles tendon rupture underwent V-Y tendon plasty in our hospital. The mean injury-to-surgery time was 20.4 weeks (range, 4-96 weeks). The Matles test and an improved Thompson test was used to examine Achilles tendon rupture. These patients were not able to stand on the tiptoes of the injured lower extremity. X-ray tests ruled out the chance of fracture and were used to examine the Kager triangle. MRI was used to confirm the final diagnosis. The function of the ankle and the foot was assessed using the American Orthopaedic Foot & Ankle Society (AOFAS) score and the Achilles Tendon Total Rupture Score (ATRS). V-Y tendon plasty for Achilles tendon reconstruction was performed. A below-knee cast was used to keep the ankle in plantar flexion (up to 20°) for 4 weeks. Non-weight-bearing exercise was allowed with crutches. After 4 weeks, partial weight-bearing was allowed. Physical exercises were performed under rehabilitation guidelines. At 10-12 weeks postoperation, patients began to return to daily life activity levels without restrictions. Preoperative and postoperative MRI was obtained. The mean follow-up period was 32.8 months. The mean operative gap of the Achilles tendon after debridement was 5 cm (range, 4-9 cm), with 85% of the gap less than or equal to 6 cm. The mean AOFAS score increased from 59.25 ± 12.28 preoperatively to 96.55 ± 3.75 at final follow-up (P < 0.05). The mean ATRS score increased from 39.55 ± 14.21 preoperatively to 94.05 ± 4.89 at final follow-up (P < 0.05). All patients had no recurrent Achilles tendon rupture during the follow up. No patient had developed serious complications, such as sural nerve injury or deep vein thrombosis. Patients were able to return to daily life activity levels without restrictions. At the latest follow-up, all patients were able to perform repetitive single heel rise on the involved limb, and to walk without a visible limp. All of the postoperative MRI showed the continuity of the Achilles tendon with no signs of cysts or inflammation, indicating perfect healing at the final follow-up. V-Y tendon plasty can be used in most cases of chronic Achilles tendon rupture. It yields satisfactory functional results and low complication rates. The advantage of this procedure is that it is an easy and economic method without the need for expensive synthetic implants. V-Y tendon plasty should be considered an acceptable first-choice treatment.


The role of personality traits and leisure activities in predicting wellbeing in young people.
The relationship between wellbeing and personality has been studied extensively, but few studies have examined these in the period of adolescence and emerging adulthood. Moreover, the influence of contextual factors such as engagement in leisure activities are rarely considered. The present study employs a combination of frequentist and Bayesian analyses to evaluate the concurrent impact of personality traits and leisure activities on five conceptions of wellbeing (life satisfaction; positive affect; negative affect; mental health; flourishing) in three cohorts of young people (aged 14-15; 16-17; 18-20 years). Personality traits were the only significant predictors of life satisfaction and negative affect, but leisure activities in the form of socialising or physical activity, in addition to personality traits, predicted positive affect, mental health and flourishing. Neuroticism was the largest predictor of wellbeing overall, whereas conscientiousness was the most consistent. Lower levels of wellbeing were also associated with higher levels of creative potential. The study not only confirms the importance of personality traits as predictors of wellbeing in adolescents and young adults, but also indicates the necessity to consider the impact of leisure activities in different conceptions of wellbeing. The negative relationship between creative potential and wellbeing is in line with the literature which shows a link between mental illness, particularly at subclinical levels, and creativity.


On the causes of geographically heterogeneous parallel evolution in sticklebacks.
The three-spined stickleback (Gasterosteus aculeatus) is an important model system for the study of parallel evolution in the wild, having repeatedly colonized and adapted to freshwater from the sea throughout the northern hemisphere. Previous studies identified numerous genomic regions showing consistent genetic differentiation between freshwater and marine ecotypes but these had typically limited geographic sampling and mostly focused on the Eastern Pacific region. We analysed population genomic data from global samples of the three-spined stickleback marine and freshwater ecotypes to detect loci involved in parallel evolution at different geographic scales. Most signatures of parallel evolution were unique to the Eastern Pacific and trans-oceanic marine-freshwater differentiation was restricted to a limited number of shared genomic regions, including three chromosomal inversions. On the basis of simulations and empirical data, we demonstrate that this could result from the stochastic loss of freshwater-adapted alleles during the invasion of the Atlantic basin and selection against freshwater-adapted variants in the sea, both of which can reduce standing genetic variation available for freshwater adaptation outside the Eastern Pacific region. Moreover, the elevated linkage disequilibrium associated with marine-freshwater differentiation in the Eastern Pacific is consistent with secondary contact between marine and freshwater populations that evolved in isolation from each other during past glacial periods. Thus, contrary to what earlier studies from the Eastern Pacific region have led us to believe, parallel marine-freshwater differentiation in sticklebacks is far less prevalent and pronounced in all other parts of the species global distribution range.


Modification of the wavelet method used in transiently evoked otoacoustic emission pass/fail criterion to increase its accuracy.
Transiently evoked otoacoustic emissions (TEOAEs) are widely used in newborn hearing screening programmes for early detection of hearing losses. To increase the accuracy of a TEOAE pass/fail criterion that uses the wavelet method, it was demonstrated that the large estimation variance is a possible reason for the inaccuracy, and a modified wavelet method is proposed to solve the inaccuracy problem. In the modified wavelet method, N paired buffers, instead of only one, were used to store the total 512 subaveraged responses, and then the average of the calculated N cross-correlation coefficients between N pairs of TEOAE signals was taken in the pass/fail criterion. Four sets of 256 synthesised noise and eight sets of 256 synthesised noisy TEOAE signals were tested, and each set was tested 1,000 times. The results showed that the standard deviation of the correlation estimation was greatly reduced by using this average value with N selected as 4. As a result, the total number of single-scale cross-correlation coefficients below 50% decreased from 1281 to 195 for noisy TEOAE signals, and the total number of single-scale cross-correlation coefficients above 50% decreased from 90 to 0 for synthesised noise.


Impact of therapeutic education on the evolution of social representations of medication in patients with Parkinson's disease: A quantitative and qualitative study (ETPARK REMED).
Among the workshops of our therapeutic patient education (TPE) program, the medication workshop (TPEM workshop) is very frequently proposed to patients in view of the difficulties they encounter related to the complexity of managing antiparkinsonian treatment. Patients' appropriation of their medications could depend on their social representations. To evaluate the effect of our TPEM workshop on the social representations PD patients have of their medications and to compare it with that of another therapeutic intervention such as a talking group defined as the control group. This single-center, prospective, randomized, parallel-group study investigated the social representations of medication through a questionnaire on knowledge about antiparkinsonian medications, a questionnaire on beliefs about medication (BMQ), and a word association task. In the TPEM group (n=16), the workshop induced significant effects over time on the knowledge questionnaire (P=0.01), BMQ specific necessity and concerns scores (P=0.04 and 0.01, respectively), necessity-concerns differential (P=0.04), and BMQ general harm (P=0.04). No significant difference was found in the talking group (n=6). Comparison of the two groups showed a significant difference of the BMQ general harm with a decrease in belief in the harmfulness of the medications in the workshop group (P=0.03). The results of the verbal association task showed a modification in the content and structure of the social representations of medication in the TPEM group. The TPEM workshop helped reduce initial negative aspects of medication representations. Improved knowledge of their medication allowed patients to feel more competent and legitimate in communicating with caregivers, modifying their beliefs about medications. Indeed, the medication was perceived as less restrictive, care becoming central as shown by the emergence of the medical team in the social representations of the medication. All the results show a specific beneficial effect of the TPEM workshop through an evolution of the social representations of medications, which became more positive in our PD patients.


Effect of placing double-lumen irrigation-suction tube on closure of anastomotic defect following rectal cancer surgery.
This study aimed to investigate the effect of placement of double-lumen irrigation-suction tubes (DLIST) on the closure of anastomotic defect (AD) after rectal cancer surgery. The study was carried out at two centers managed by one surgeon, both adopted the same treatments. Patients with postoperative AD after rectal cancer surgery from January 2011 to June 2020 were eligible and were divided into a passive drainage (PD) group and a DLIST group according to whether the PD, placed in the rectal cancer surgery, had been replaced with the DLIST. The effect of DLIST on the AL was evaluated. There distributed 76 patients in the DLIST group and 52 in the PD group. A higher closure rate was reported in the DLIST group (46 patients in DLIST group, for a closure rate of 60.5%, and 21 patients in PD group, for a closure rate of 40.4%. HR = 3.05; 95% CI: 1.79-5.19; P < 0.001). Both length of stay and costs of the treatment in the DLIST group were lower (54 days [interquartile range, IQR: 41-17] days vs. 112 days [IQR: 66-27] days, P = 0.005; and $18,721 [IQR: $14,982-4,960] vs. $40,840 [IQR: $20,932-50,529], P < 0.001). Placement of DLIST might serve as an effective method for treating AD following rectal cancer surgery. In comparison with PD, it costs lower to apply DLIST in the treatment of AD and the length of stay is shorter.


Global estimates and determinants of antituberculosis drug pharmacokinetics in children and adolescents: a systematic review and individual patient data meta-analysis.
Suboptimal exposure to antituberculosis (anti-TB) drugs has been associated with unfavourable treatment outcomes. We aimed to investigate estimates and determinants of first-line anti-TB drug pharmacokinetics in children and adolescents at a global level. We systematically searched MEDLINE, Embase and Web of Science (1990-2021) for pharmacokinetic studies of first-line anti-TB drugs in children and adolescents. Individual patient data were obtained from authors of eligible studies. Summary estimates of total/extrapolated area under the plasma concentration-time curve from 0 to 24 h post-dose (AUC0-24) and peak plasma concentration (C max) were assessed with random-effects models, normalised with current World Health Organization-recommended paediatric doses. Determinants of AUC0-24 and C max were assessed with linear mixed-effects models. Of 55 eligible studies, individual patient data were available for 39 (71%), including 1628 participants from 12 countries. Geometric means of steady-state AUC0-24 were summarised for isoniazid (18.7 (95% CI 15.5-22.6) h·mg·L-1), rifampicin (34.4 (95% CI 29.4-40.3) h·mg·L-1), pyrazinamide (375.0 (95% CI 339.9-413.7) h·mg·L-1) and ethambutol (8.0 (95% CI 6.4-10.0) h·mg·L-1). Our multivariate models indicated that younger age (especially <2 years) and HIV-positive status were associated with lower AUC0-24 for all first-line anti-TB drugs, while severe malnutrition was associated with lower AUC0-24 for isoniazid and pyrazinamide. N-acetyltransferase 2 rapid acetylators had lower isoniazid AUC0-24 and slow acetylators had higher isoniazid AUC0-24 than intermediate acetylators. Determinants of C max were generally similar to those for AUC0-24. This study provides the most comprehensive estimates of plasma exposures to first-line anti-TB drugs in children and adolescents. Key determinants of drug exposures were identified. These may be relevant for population-specific dose adjustment or individualised therapeutic drug monitoring.


Sleep medicine in otolaryngology units: an international survey.
No study to date has described the overall landscape of sleep disorders management and training in otolaryngology departments of different countries. The aim of our study was to investigate and compare settings, diagnostic and therapeutic approaches and training programmes. An international online survey was developed with the collaboration of the YO-IFOS (Young Otolaryngologists-International Federation of Otorhinolaryngological Societies) to assess the current practice of otolaryngologists in the management of sleep disorders. The survey also included a session dedicated to training. A total of 126 otolaryngologists completed the survey. The larger part of responses was collected from Central/South America and Europe. The majority of responders from South/Central America (97%) declared to be certified as sleep specialist while 49% of Europeans stated the opposite. Of responders 83% perform a drug-induced sleep endoscopy (DISE) before planning a possible surgical intervention. Soft palate and base of tongue interventions were the most common procedure, respectively performed in 94% and 79% of the cases. Residents were allowed to perform soft palate surgery in 77% of the cases. Upper airway stimulation (26% vs 10%), trans-oral robotic surgery (36% vs 11%) and radiofrequency of the base of the tongue (58% vs 25%) were preferred more frequently by European responders. The highest caseloads of soft palate surgery and bi-maxillary advancement were registered in the academic institutions. Significant concordance and few interesting divergences in diagnosis and treatment of sleep disorders were observed between nationalities and types of institution. Economic resources might have played a significant role in the therapeutic choice. Trainees' lack of exposure to certain interventions and to a sufficient caseload appeared to be the main burden to overcome.


Experimental manipulation of transforming growth factor-beta isoforms significantly affects adhesion formation in a murine surgical model.
Transforming growth factor-beta (TGF-beta), a multifunctional growth factor, represents three mammalian isoforms, TGF-beta1, TGF-beta2, and TGF-beta3. In cutaneous wound healing, combined neutralization of TGF-beta1 and -beta2 or addition of TGF-beta3 reduces scar formation. Here, we investigated whether experimental manipulation of TGF-beta isoforms reduced adhesion formation after injury to the peritoneum. Adhesions were produced in mice by surgical abrasion of adjacent serosa followed by close apposition. In the first part of this study, a detailed analysis of TGF-beta isoform distribution was performed through immunolocalization. TGF-beta isoforms clearly showed a unique temporal and spatial pattern of expression after peritoneal wounding. Based on this pharmacokinetic data, we next administered neutralizing antibodies to TGF-beta1 and -beta2 or exogenous TGF-beta3 peptide by local application and intraperitoneal injection at various times before and after surgery. At day 7 after surgery, addition of neutralizing antibodies to both TGF-beta1 and -beta2 significantly reduced the number and size of adhesions (P < 0.05) compared with the vehicle control. By contrast, exogenous addition of TGF-beta3 either had no effect or increased adhesion formation compared to the vehicle control. In conclusion, these results show that by blocking both TGF-beta1 and TGF-beta2 using neutralizing antibodies, it is possible to prevent abdominal adhesion formation.


A high-density linkage map and QTL mapping of fruit-related traits in pumpkin (Cucurbita moschata Duch.).
Pumpkin (Cucurbita moschata) is an economically worldwide crop. Few quantitative trait loci (QTLs) were reported previously due to the lack of genomic and genetic resources. In this study, a high-density linkage map of C. moschata was structured by double-digest restriction site-associated DNA sequencing, using 200 F2 individuals of CMO-1 × CMO-97. By filtering 74,899 SNPs, a total of 3,470 high quality SNP markers were assigned to the map spanning a total genetic distance of 3087.03 cM on 20 linkage groups (LGs) with an average genetic distance of 0.89 cM. Based on this map, both pericarp color and strip were fined mapped to a novel single locus on LG8 in the same region of 0.31 cM with phenotypic variance explained (PVE) of 93.6% and 90.2%, respectively. QTL analysis was also performed on carotenoids, sugars, tuberculate fruit, fruit diameter, thickness and chamber width with a total of 12 traits. 29 QTLs distributed in 9 LGs were detected with PVE from 9.6% to 28.6%. It was the first high-density linkage SNP map for C. moschata which was proved to be a valuable tool for gene or QTL mapping. This information will serve as significant basis for map-based gene cloning, draft genome assembling and molecular breeding.


Does percutaneous transluminal angioplasty improve quality of life?
To prospectively assess the impact of peripheral percutaneous transluminal angioplasty (PTA) on quality of life (QOL) in patients with intermittent claudication or critical limb ischemia (CLI). The secondary purpose was to determine if periprocedural medication altered patient QOL. A total of 72 patients underwent lower-limb PTA (86 lesions) over a period of 12 months. QOL data were obtained before PTA and at 1 and 6 months after PTA with use of the Short Form-36 (SF-36) questionnaire. The SF-36 questionnaire was completed in 100% of cases before PTA, in 89% at 1 month after PTA, and in 78% at 6 months after PTA. Baseline QOL was worse than the QOL quoted for the normal population. Overall, there was an improvement in QOL in seven of the nine health domains at 1 month after PTA, and the improvements were significant (P < .01) in five. Improvement was demonstrated at 6 months in all nine QOL domains measured, which was significant (P < .01) in five. Patients with intermittent claudication (n = 40) exhibited an improvement in all nine health domains at 6 months, which was significant (P < .05) in seven. The CLI group (n = 32) showed an improvement in five domains at 6 months, which was significant (P < .05) in two. Subjects with intermittent claudication experienced a more significant improvement than those with CLI. SF-36 pain scores at 1 month after PTA were significantly better in patients taking aspirin than in those who were not (P < .001). PTA results in improved QOL in patients with intermittent claudication and CLI as early as 1 month, which was shown to be maintained at 6 months in our small series. Aspirin significantly improves pain after PTA. QOL assessment is useful after peripheral angioplasty.


Spirometric reference values for white European children and adolescents: Polgar revisited.
We analyzed six spirometric data sets collected in the Netherlands, Austria, the United Kingdom, Spain, and Italy. The objectives were to establish whether (1) it was possible to describe spirometric indices from childhood to adulthood, taking into account the adolescent growth spurt, and (2) there are systematic differences in ventilatory function between children and adolescents in different parts of Western Europe. The study comprised 2,269 girls and 3,592 boys, aged 6-21 years. The range in standing height was 110-185 in girls, 110-205 in boys. The model applicable to all data sets was ln FVC or ln FEV1 = a + (b + c x A) x H, where H = standing height and A = age; this model prevents the phase shift between the adolescent growth spurt in length and lung volume from leading to an age-dependent bias in predicted values. There was surprising agreement between most of the data sets; systematic differences are probably due to technical factors arising from ATPS-BTPS corrections and from defining the end of breath with pneumotachometer systems. Taking those into account, prediction equations for FVC, FEV1, and FEV1%FVC were developed with "lower limits of normal" which should be applicable to children and adolescents of European descent. It is proposed that the approach of analyzing available data sets should also be applied to other ventilatory indices, data collected in adults and elderly subjects, or in other ethnic groups, and that an international data base be set up to that end.


Interactions between light intensity and phosphorus nutrition affect the phosphate-mining capacity of white lupin (Lupinus albus L.).
Light intensity affects photosynthetic carbon (C) fixation and the supply of carbon to roots. To evaluate interactions between carbon supply and phosphorus (P) supply, effects of light intensity on sucrose accumulation, root growth, cluster root formation, carboxylate exudation, and P uptake capacity were studied in white lupin (Lupinus albus L.) grown hydroponically with either 200 µmol m(-2) s(-1) or 600 µmol m(-2) s(-1) light and a sufficient (50 µM P) or deficient (1 µM P) P supply. Plant biomass and root:shoot ratio increased with increasing light intensity, particularly when plants were supplied with sufficient P. Both low P supply and increasing light intensity increased the production of cluster roots and citrate exudation. Transcripts of a phosphoenol pyruvate carboxylase gene (LaPEPC3) in cluster roots (which is related to the exudation of citrate), transcripts of a phosphate transporter gene (LaPT1), and P uptake all increased with increasing light intensity, under both P-sufficient and P-deficient conditions. Across all four experimental treatments, increased cluster root formation and carboxylate exudation were associated with lower P concentration in the shoot and greater sucrose concentration in the roots. It is suggested that C in excess of shoot growth capabilities is translocated to the roots as sucrose, which serves as both a nutritional signal and a C-substrate for carboxylate exudation and cluster root formation.


Arterial Thromboembolism in Patients With AF and CHA2DS2-VASc Score 0-2 With and Without Cancer.
It is unclear whether newly diagnosed cancer adds to the risk of arterial thromboembolism (ATE) in patients with atrial fibrillation/flutter (AF). This is especially relevant for AF patients with low to intermediate CHA2DS2-VASc scores in whom the risk-benefit ratios between ATE and bleeding are delicately balanced. The objectives were to evaluate the ATE risk in AF patients with a CHA2DS2-VASc score of 0 to 2 with and without cancer. A population-based retrospective cohort study was performed. Patients with a CHA2DS2-VASc score of 0 to 2 not receiving anticoagulation at cancer diagnosis (or the matched index date) were included. Patients with embolic ATE or cancer before study index were excluded. AF patients were categorized into AF and cancer and AF and no cancer cohorts. Cohorts were matched for multinomial distribution of age, sex, index year, AF duration, CHA2DS2-VASc score, and low/high/undefined ATE risk cancer. Patients were followed from study index until the primary outcome or death. The primary outcome was acute ATE (ischemic stroke, transient ischemic attack, or systemic ATE) at 12 months using International Classification of Diseases-Ninth Revision codes from hospitalization. The Fine-Gray competing risk model was used to estimate the HR for ATE with death as a competing risk. The 12-month cumulative incidence of ATE was 2.13% (95% CI: 1.47-2.99) in 1,411 AF patients with cancer and 0.8% (95% CI: 0.56-1.10) in 4,233 AF patients without cancer (HR: 2.70; 95% CI: 1.65-4.41). The risk was highest in men with CHA2DS2-VASc = 1 and women with CHA2DS2-VASc = 2 (HR: 6.07; 95% CI: 2.45-15.01). In AF patients with CHA2DS2-VASc scores of 0 to 2, newly diagnosed cancer is associated with an increased incidence of stroke, transient ischemic attack, or systemic ATE compared with matched controls without cancer.


Bioaccumulation and reproductive toxicity of bisphenol A in male-pregnant seahorse (Hippocampus erectus) at environmentally relevant concentrations.
Seahorses, with brood pouch in adult males, are a bioindicator species that exhibit specialized reproductive strategy of "male pregnancy". Bisphenol A (BPA), one of the most pervasive endocrine-disrupting chemicals (EDCs), is hazardous for reproductive, immune, and neurological systems. However, no evidence of BPA toxicity to the male-pregnant animals is available. Herein, the reproductive toxicity of BPA was evaluated in lined seahorses (Hippocampus erectus) following exposure to environmentally relevant concentrations (10, 100, and 1000 μg/L) through physiological, histological, and transcriptional analyses. Our results indicated BPA bioaccumulation to be positively correlated with exposure doses in both sexes. Ovarian failure was only observed in the high-dose BPA treatment group, accompanied by the apoptosis of follicular cells and up-regulation of pro-apoptotic genes. However, brood pouches maintenance were surprisingly inhibited at low concentration, and transcriptomic analysis revealed disturbed profiles of genes involved in the extracellular matrix and cell-cell adhesion pathways. Interestingly, seahorse testes were less sensitive to BPA exposure than that in other teleosts. Thus, our study suggests that BPA at environmentally relevant concentrations might cause reproductive dysfunction in seahorses, potentially exerting adverse effects on the seahorse population since most of them inhabit shallow coastal areas with prevalent estrogenic contaminants.


Key aspects of modern GPCR drug discovery.
G-protein-coupled receptors (GPCRs) are the largest and most versatile cell surface receptor family with a broad repertoire of ligands and functions. We've learned an enormous amount about discovering drugs of this receptor class since the first GPCR was cloned and expressed in 1986, such that it's now well-recognized that GPCRs are the most successful target class for approved drugs. Here we take the reader through a GPCR drug discovery journey from target to the clinic, highlighting the key learnings, best practices, challenges, trends and insights on discovering drugs that ultimately modulate GPCR function therapeutically in patients. The future of GPCR drug discovery is inspiring, with more desirable drug mechanisms and new technologies enabling the delivery of better and more successful drugs.


Clinical impact of low serum free T4 in patients with non-small cell lung cancer treated with nivolumab.
Nivolumab improves the prognosis of non-small cell lung cancer (NSCLC) but can cause immune-related adverse events (irAEs). Reports have indicated longer progression-free survivals (PFSs) in patients with irAEs than in those without irAEs. We reported associations between programmed death ligand-1 (PD-L1) single nucleotide polymorphisms (SNPs) and PFS after nivolumab treatment. We hypothesized that adverse events might be associated with the SNPs of PD-L1. We analyzed data from 111 patients with NSCLC treated with nivolumab. The response rate was 14%, and the median PFS was 68 days. We found patients with the adverse event of low free tetraiodothyronine (fT4) had significantly longer PFSs than those without low fT4 (not reached vs 67 days; hazard ratio [HR], 0.297; P = 0.010). Moreover, median overall survival was longer in patients with low fT4 than those without low fT4 (not reached vs 556 days, HR, 0.139; P = 0.020). Patients with the T allele of rs1411262 (P = 0.0073) and with the A allele of rs822339 (P = 0.0204) developed low fT4. Patients with the T/T genotype had longer PFSs than with those with the C/T and C/C genotypes of rs1411262 (165 vs. 67 days, HR, 1.65; P = 0.040), and those with the A/A genotype had longer PFSs than those with the A/G and G/G genotypes of rs822339 (182 vs. 67 days, HR, 1.76; P = 0.025). In the patients with advanced NSCLC, low fT4 after nivolumab treatment was associated with significantly longer PFSs. The SNPs of PD-L1 may be associated with the adverse events of nivolumab.


Diagnosis of an interstitial pregnancy with 4-dimensional volume contrast imaging.
The purpose of this study was to investigate the use of a new ultrasound technique, termed 4-dimensional volume contrast imaging in coronal-plane to facilitate the early detection of interstitial pregnancy. A case of interstitial pregnancy was diagnosed accurately at 6 weeks of gestation with 4-dimensional transvaginal volume contrast imaging in coronal plane technology. A scan of the uterus revealed that the eccentric gestational sac was located in the right uterotubal junctional area, lying 1 cm outside the most upper lateral edge of the uterine cavity with thick endometrial echoes. After the appropriate counseling, the patient was treated with an ultrasonographically guided intracardiac injection of 2.5 mEq of potassium chloride followed by aspiration of the gestational sac. A transvaginal scan immediately after the procedure demonstrated a collapsed empty gestational sac. Three months after the conservative treatment, the serum beta-human chorionic gonadotropin level fell to 1.17 mIU/mL, and normal echotexture of the uterus was noted. Four-dimensional volume contrast imaging in coronal-plane technology provides scan planes that are not accessible by conventional 2-dimensional scanning, with enhanced tissue contrast resolution in region of interest. This new ultrasound technique has the potential to provide more accurate, inexpensive and less variable virtual reality image-guided interventional options through more realistic interaction with the virtualized in utero condition, particularly when 2-dimensional transvaginal scan fails to differentiate between interstitial, angular, and cornual pregnancies.


Influence of variants in serotonin modulating genes on the risk, aggressiveness, and prognosis of oropharynx cancer.
Serotonin (5-HT) is involved in regulating tumor growth, as well as psychiatric disorders. It is synthesized by tryptophan hydroxylase (TPH) and acts through 5-HT receptors (HTRs). Single-nucleotide variations (SNVs) in TPH1 rs623580 (T>A), TPH2 rs4570625 (G>T), and HTR1D rs674386 (G>A) may affect 5-HT levels. However, the effect of these SNVs on oropharynx carcinoma (OPC) is unknown. DNA from 251 patients with OPC and 254 controls was analyzed by RT-PCR. Transcriptional activity of TPH1 rs623580 and HTR1D rs674386 was studied by luciferase assays. Multivariate statistical tests were utilized to evaluate group differences and survival outcomes. TPH1 TT was more frequent in patients than in controls (OR: 1.56, p = 0.03). Patients with HTR1D GG/GA showed invasive tumors (p = 0.01) and shorter survival (HR: 1.66, p = 0.04). TPH1 TT (0.79-fold, p = 0.03) and HTR1D GG (0.64-fold, p = 0.008) presented lower transcriptional activity. Our data suggest that SNVs in 5-HT modulating genes can influence OPC.


Therapeutic apheresis in kidney AB0 incompatible transplantation.
Desensitization strategies to safely perform ABO incompatible living donor kidney transplantations are various and still evolving. Given the successful outcome of the majority of the approaches, the current trend is focused on a minimization of treatments. Based on this consideration, the evolution at a single Center of the desensitization protocol is herein described. Starting from 2010, 58 AB0 incompatible living donor kidney transplantations were performed at the University-Hospital of Padua. Over the years, the initial desensitization strategy with rituximab single-dose induction, pre-and post-transplant plasmapheresis and CMV-specific immunoglobulin administration has been shifted to a minimized approach, omitting post-transplant antibody removal in 25 cases. The results of such reduction in post-transplant antibody removal did not affect the outcome of AB0-incompatible kidney transplants, with a reduction in costs and hospitalization.


Steroidal modulation of pituitary gonadotropin-releasing hormone responsiveness in young turtles, Pseudemys scripta.
Steroid-modulated pituitary secretion and glandular content of gonadotropin (Gth: LH and FSH) was studied in young slider turtles. Injection (ip) of both 17 beta-estradiol (E2) and testosterone (T) reduced pituitary content of both Gths and caused significant inhibition of basal LH secretion and gonadotropin-releasing hormone (GnRH)-stimulated LH and FSH secretion measured in vitro. However, gonadectomy did not affect pituitary Gth secretion or response in these juveniles, and anti-estrogen and anti-androgen compounds had some steroid agonistic action on the pituitary gland. Exposure to E2, T, and 5 alpha-dihydrotestosterone (DHT) in vitro for 4, 24, or 48 hr either had no effect or completely inhibited pituitary GnRH responsiveness. Progesterone (P) alone had no effect on pituitary GnRH response and in combination did not alter the typical inhibitory effect of E2. There were several indications of differential effects of steroids on secretion of the two Gths, especially in response to GnRH and tetraethyl chloride (receptor independent) stimulation. The results suggest that steroids may act directly at the pituitary level to alter Gth secretion and that steroidal modulation of pituitary secretion might play a role in differential regulation of LH and FSH in turtles.


A randomized, placebo-controlled phase 3 trial of the PI3Kδ inhibitor leniolisib for activated PI3Kδ syndrome.
Activated phosphoinositide 3-kinase delta (PI3Kδ) syndrome (APDS) is an inborn error of immunity with clinical manifestations including infections, lymphoproliferation, autoimmunity, enteropathy, bronchiectasis, increased risk of lymphoma, and early mortality. Hyperactive PI3Kδ signaling causes APDS and is selectively targeted with leniolisib, an oral, small molecule inhibitor of PI3Kδ. Here, 31 patients with APDS aged ≥12 years were enrolled in a global, phase 3, triple-blinded trial and randomized 2:1 to receive 70 mg leniolisib or placebo twice daily for 12 weeks. Coprimary outcomes were differences from baseline in the index lymph node size and the percentage of naïve B cells in peripheral blood, assessed as proxies for immune dysregulation and deficiency. Both primary outcomes were met: the difference in the adjusted mean change (95% confidence interval [CI]) between leniolisib and placebo for lymph node size was -0.25 (-0.38, -0.12; P = .0006; N = 26) and for percentage of naïve B cells, was 37.30 (24.06, 50.54; P = .0002; N = 13). Leniolisib reduced spleen volume compared with placebo (adjusted mean difference in 3-dimensional volume [cm3], -186; 95% CI, -297 to -76.2; P = .0020) and improved key immune cell subsets. Fewer patients receiving leniolisib reported study treatment-related adverse events (AEs; mostly grades 1-2) than those receiving placebo (23.8% vs 30.0%). Overall, leniolisib was well tolerated and significant improvement over placebo was notable in the coprimary endpoints, reducing lymphadenopathy and increasing the percentage of naïve B cells, reflecting a favorable impact on the immune dysregulation and deficiency seen in patients with APDS. This trial was registered at www.clinicaltrials.gov as #NCT02435173.


HER2 targeting near-infrared photoimmunotherapy for a CDDP-resistant small-cell lung cancer.
Human epidermal growth factor receptor 2 (HER2) is tyrosine kinase receptor that belongs to the ErbB family and is overexpressed on the membrane surface of various cancer cells, including small cell lung cancer (SCLC); however, no HER2 targeted therapy for SCLC have yet been established. Near-infrared photoimmunotherapy (NIR-PIT) is a novel cancer therapy based on photo-absorber, IRDye-700DX (IR700), -antibody conjugates, and near-infrared (NIR) light. We used HER2-positive SCLC parental cell lines (SBC-3) and its chemoresistant cell lines, and examined therapeutic efficacy of HER2 targeting NIR-PIT using anti HER2 antibody trastuzumab. We found that HER2 expression was upregulated on chemoresistant cell lines, especially cisplatin-resistance (SBC-3/CDDP). In vitro, the rate of cell death increased with the amount of NIR-light irradiation, and it was significantly higher in SBC-3/CDDP than in SBC-3. In vivo, tumor growth was more suppressed in SBC-3/CDDP group than in SBC-3 group, and survival period tended to be prolonged. In this study, we demonstrated that HER2 targeting NIR-PIT using trastuzumab is promising therapy for HER2-positive SCLC, and is more effective when HER2 expression is upregulated due to CDDP resistance, suggesting that the HER2 expression level positively corelated with the efficacy of NIR-PIT.


Promoter region variant C-824T in the TH gene modulates the subjective effects of cocaine.
This study examined how a promoter variant of TH (rs10770141) affects subjective effects of cocaine in 65 nontreatment-seeking individuals with cocaine dependence. Participants received cocaine/saline intravenously, and TH genotypes were evaluated. Homozygous individuals for the minor T allele reported greater "good" and "bad" subjective effects to cocaine than those with the major C allele. TH rs10770141 modulates subjective effects of cocaine in participants with cocaine dependence. These results are among the first to indicate that homozygosity of the T allele of rs10770141 modulates greater sensitivity to cocaine.


Unanswered questions in the regulation and function of the duplicated α7 nicotinic receptor gene CHRFAM7A.
The α7 nicotinic receptor (α7 nAChR) is an important entry point for Ca2+ into the cell, which has broad and important effects on gene expression and function. The gene (CHRNA7), mapping to chromosome (15q14), has been genetically linked to a large number of diseases, many of which involve defects in cognition. While numerous mutations in CHRNA7 are associated with mental illness and inflammation, an important control point may be the function of a recently discovered partial duplication CHRNA7, CHRFAM7A, that negatively regulates the function of the α7 receptor, through the formation of heteropentamers; other functions cannot be excluded. The deregulation of this human specific gene (CHRFAM7A) has been linked to neurodevelopmental, neurodegenerative, and inflammatory disorders and has important copy number variations. Much effort is being made to understand its function and regulation both in healthy and pathological conditions. However, many questions remain to be answered regarding its functional role, its regulation, and its role in the etiogenesis of neurological and inflammatory disorders. Missing knowledge on the pharmacology of the heteroreceptor has limited the discovery of new molecules capable of modulating its activity. Here we review the state of the art on the role of CHRFAM7A, highlighting unanswered questions to be addressed. A possible therapeutic approach based on genome editing protocols is also discussed.


Association between gastrointestinal events and compliance with osteoporosis therapy.
The aim of this study was to estimate the rate of gastrointestinal (GI) events, and association between GI events and compliance with osteoporosis therapy among osteoporotic women. A retrospective cohort study using a large administrative claims database in the United States from 2001 through 2010 was conducted. We studied women ≥ 55 years old who were continuously enrolled in a health plan for at least 2 years, a baseline year before and a follow-up year after the date of the first prescription of oral bisphosphonate as the first oral osteoporosis treatment. Compliance with osteoporosis therapy was measured using the medication possession ratio (MPR), with compliance defined as MPR ≥ 0.8. Multivariate logistic regression was used to assess the association between occurrence of GI events and compliance with osteoporosis therapy after controlling for demographic and clinical characteristics. A sample consisting of 75,593 women taking at least one oral bisphosphonate with mean (SD) age of 64 (8) years was identified. A total of 21,142 (28%) patients experienced at least one GI event during the follow-up period. Only 31,306 (41%) patients were compliant with osteoporosis therapy. Patients who experienced GI events after initiation of oral bisphosphonates were 29% less likely to adhere to osteoporosis therapy as compared to patients who did not experience GI events (odds ratio [95% CI], 0.71 [0.69-0.74]; P < .001). Less than half of the patients were compliant with osteoporosis therapy within one year after initiating oral bisphosphonates, and the likelihood of compliance was significantly lower by 29% among women with GI events.


Emerging organic contaminants in springs of the highly karstified Dinaric region.
Emerging organic contaminants (EOCs) have become of increasing interest due to concerns about their impact on humans and the wider environment. Karst aquifers are globally widespread, providing critical water supplies and sustaining rivers and ecosystems, and are particularly susceptible to pollution. However, EOC distributions in karst remain quite poorly understood. This study looks at the occurrence of EOCs in the Croatian karst, which is an example of the "classical" karst, a highly developed type of karst that occurs throughout the Dinaric region of Europe. Samples were collected from 17 karst springs and one karst lake used for water supply in Croatia during two sampling campaigns. From a screen of 740 compounds, a total of 65 compounds were detected. EOC compounds from the pharmaceutical (n = 26) and agrochemical groups (n = 26) were the most frequently detected, while industrials and artificial sweeteners had the highest concentrations (range 8-440 ng/L). The number of detected compounds and the frequency of detection demonstrate the vulnerability of karst to EOC pollution. Concentrations of 5 compounds (acesulfame, sucralose, perfluorobutane sulfonate, emamectin B1b, and triphenyl phosphate) exceeded EU standards and occurred at concentrations that are likely to be harmful to ecosystems. Overall, most detections were at low concentrations (50 % <1 ng/L). This may be due to high dilution within the exceptionally large springs of the Classical karst, or due to relatively few pollution sources within the catchments. Nevertheless, EOC fluxes are considerable (10 to 106 ng/s) due to the high discharge of the springs. Temporal differences were observed, but without a clear pattern, reflecting the highly variable nature of karst springs that occurs over both seasonal and short-term timescales. This research is one of a handful of regional EOC investigations in karst groundwater, and the first regional study in the Dinaric karst. It demonstrates the need for more frequent and extensive sampling of EOCs in karst to protect human health and the environment.


Bactericidal efficacy of carbon dioxide laser against bacteria-contaminated titanium implant and subsequent cellular adhesion to irradiated area.
The aim of this study was to assess CO2 laser ability to eliminate bacteria from titanium implant surfaces. The changes of the surface structure, the rise in temperature, and the damage of connective tissue cells after laser irradiation were also considered. Streptococcus sanguis and Porphyromonas gingivalis on titanium discs were irradiated by an expanded beam of CO2 laser. Surface alteration was observed by a light, and a scanning electron, microscope. Temperature was measured with a thermograph. Damage of fibroblastic (L-929) and osteoblastic (MC3T3-E1) cells outside the irradiation spot and adhesion of the cells to the irradiated area were also estimated. All the organisms (10(8)) of S. sanguis and P. gingivalis were killed by the irradiation at 286 J/cm2 and 245 J/cm2, respectively. Furthermore, laser irradiation did not cause surface alteration, rise of temperature, serious damage of connective tissue cells located outside the irradiation spot, or inhibition of cell adhesion to the irradiated area. CO2 laser irradiation with expanded beam may be useful in removing bacterial contaminants from implant surface.


Levels and uptake of taurine in various brain regions after drug-induced generalized convulsions.
In a study of the role of taurine in the genesis of epilepsy the effects of metrazol-induced convulsions on the uptake and distribution of taurine in the brain were measured. In vivo we found no significant uptake of taurine in the mouse brain; in rabbit brain in most areas significant taurine uptake was found. The physiological levels of taurine were much higher in mouse brain than in rabbit brain. In vivo the regional levels and the uptake of taurine were not significantly changed after generalized convulsions. Uptake in vivo was lowered in slices obtained from mice treated with metrazol. The lack of effect of metrazol convulsions on cerebral taurine in vivo indicates that further studies are needed to clarify the relationship between taurine, a putative inhibitory transmitter, and epilepsy.


A general procedure for screening inhibitory antibodies: application for identifying anti-protein kinase C antibodies.
In this communication we describe a microfiltration assay to identify monoclonal antibodies that interfere with the activity of enzymes. This method is quick and sensitive to small changes in the activity of the enzyme and does not require highly purified enzyme or large quantities of antibodies. It has been applied to identify anti-protein kinase C antibodies which would have been impossible to identify by classical assays such as enzyme-linked immunosorbent assay.


Effects of exposure to PCBs and related compounds on growth and activity in children.
Two hundred thirty-six children from two established cohorts at risk for exposure to polychlorinated biphenyls (PCBs) and related contaminants were evaluated at age 4 years. Prenatal exposure (assessed by cord serum PCB level) was associated with lower weight, an effect consistent with reports of growth retardation in laboratory rats and in children exposed at high levels in Taiwan and at general population levels in Japan. The highest exposed children weighed 1.8 kg less on the average than the least exposed. Contemporary body burden (assessed by 4-year serum PCB level) was associated with reduced activity based on composite ratings provided by the child's mother and two independent examiners. This effect, attributable to lactation exposure, was strongest among the offspring of women with above average milk PCB levels who breast fed for at least 1 year. While the weight deficit is consistent with previous data linking developmental effects of low-dose human PCB exposures specifically to the prenatal period, activity is the first domain found to be affected by lactation at contemporary levels of exposure.


The effect of ovariectomy on the healing tooth socket of the rat.
Under general anaesthesia, 35-day-old female rats were ovariectomized and the right maxillary molar teeth removed. Dynamic measures of alveolar bone formation were determined at 10 days after surgery, using the fluorochrome labelling technique, and compared with control animals. Ovariectomy significantly increased buccal resorption and palatal bone formation. In a second experiment, ovariectomized rats had the right maxillary molar teeth extracted and were killed at either 5 or 14 days after surgery. The mean mineralizing surface of the alveolar bone (percentage of surfaces occupied by a double fluorescent label) was significantly lower in rats killed at either 5 or 10 days than at 14 days after ovariectomy and tooth extraction. The mean appositional rate was significantly greater at 5 days after ovariectomy and tooth extraction than at 10 or 14 days. Oestrogen deficiency can therefore affect alveolar bone turnover following tooth extraction.


Mutations and polymorphisms in hemoglobin genes and the risk of pulmonary hypertension and death in sickle cell disease.
Pulmonary hypertension is a common complication of sickle cell disease (SCD) and a risk factor for early death. Hemolysis may participate in its pathogenesis by limiting nitric oxide (NO) bioavailability and producing vasculopathy. We hypothesized that hemoglobin mutations that diminish hemolysis in SCD would influence pulmonary hypertension susceptibility. Surprisingly, coincident alpha-thalassemia (Odds Ratio [OR]=0.95, 95% CI=0.46-1.94, P=NS) was not associated with pulmonary hypertension susceptibility in homozygous SCD. However, pulmonary hypertension cases were less likely to have hemoglobin SC (OR=0.18, 95% confidence interval [CI]=0.06-0.51, P=0.0005) or Sbeta(+) thalassemia (OR=0.25, 95% CI=0.06-1.16, P=0.10). These compound heterozygotes may be protected from pulmonary hypertension because of reduced levels of intravascular hemolysis, but develop this complication at a lower rate possibly due to the presence of non-hemolytic risk factors such as renal dysfunction, iron overload and advancing age. Despite this protective association, patients with SC who did develop pulmonary hypertension remained at significant risk for death during 49 months of follow-up (Hazard Ratio=8.20, P=0.0057).


Reducing socioeconomic health inequalities? A questionnaire study of majorization and invariance conditions.
We study the appeal of basic preference conditions that underpin health inequality indices, including the widely used concentration index. We did a lab experiment in which 349 respondents had to choose repeatedly between two policies that generated a distribution of income and health among five groups in society. We found stronger support for preference conditions that focus on inequality in the marginal distribution of health (and income) than for preference conditions that favor reduced correlation between both dimensions. Respondents' choices were more in line with the principle of income related health transfers when policies did not affect the ranking of groups in terms of health. Respondents also expressed more concern about the correlation between income and health when health was expressed as a shortfall rather than an attainment. Support for the preference conditions was unaffected when all groups in society experienced the same absolute or relative health change.


Prevalence of new-onset atrial fibrillation in hospitalized patients with community-acquired pneumonia: a systematic review and meta-analysis.
Community-acquired pneumonia (CAP) is a common lower respiratory tract infection, often complicated by cardiovascular events, including cardiac arrhythmias. New-onset atrial fibrillation (newAF) has been associated with increased mortality in CAP patients, especially in those critically ill; however, limited data on the prevalence of newAF in patients with CAP are available. We aim to estimate the pooled prevalence of newAF and its impact on adverse outcomes in patients with CAP, through a systematic review and meta-analysis. MEDLINE and EMBASE were systematically searched from inception to 27 January 2022. All studies reporting the prevalence of newAF in CAP patients were included and all-cause mortality was extracted when available. The pooled prevalence of newAF, 95% Confidence Intervals (CI), and 95% Prediction Intervals (PI) were computed. The inconsistency index (I2) was calculated to measure heterogeneity. Subgroup analyses were also performed. A protocol for this study was registered on PROSPERO (CRD42022307422). Among 7,655 records retrieved, 10 studies were included, with a total of 280,589 CAP patients. Pooled prevalence of newAF in CAP patients was 7.6% (95% CI 6.4-9.0%, 95% PI 4.3-13.1%, I2 = 95%). Subgroup analyses showed no significant differences according to geographical location or study design. Patients with newAF had a higher risk of mortality among the studies included in the systematic review. NewAF is a common complication, occurring in 7.6% of CAP patients, with prediction intervals suggesting an even higher burden. CAP patients who develop newAF during hospitalization may be at higher risk of mortality in both short- and long-term follow-up.


SARS-CoV-2 transmitters have more than three times higher viral loads than non-transmitters - Practical use of viral load for disease control.
Quantitative results of SARS-CoV-2 testing reported as viral load copies/mL can provide valuable information, but are rarely used in practice. We analyze whether viral load in the upper respiratory tract is correlated with transmission and disease course and how this information can be used in practice. Municipal Health Service (MHS) and clinical patients ≥18 years tested positive for SARS-CoV-2 with RT-PCR between June 1 and September 25, 2020 were included. Transmission was defined as an index having at least one contact tested positive. Test delay was defined as the time between symptom onset and SARS-CoV-2 testing. 683 patients were included (656 MHS and 27 clinical patients). The viral load was considerably lower among clinical patients compared to MHS patients: median log10 copies/mL 2.51 (IQR -1.52 - 6.46) vs 4.92 (IQR -0.54 - 8.26), p < 0.0001. However, the test delay was higher for clinical patients (median 7 [IQR 2 - 19] vs 3 [IQR 0 - 26] days, p < 0.0001). SARS-CoV-2 transmitters showed much higher viral loads than non-transmitters (log10 copies/mL 5.23 [IQR -0.52 - 8.26] vs 4.65 [IQR -0.72 - 8.00], p < 0.0001), but not for those with a test delay > 7 days. Higher viral loads were significantly correlated with older age and with more (severe) COVID-19 related symptoms. Indexes that transmitted SARS-CoV-2 had more than three times higher viral loads than non-transmitters. Viral load information can be useful during source and contact tracing to prioritize indexes with highest risk of transmission, taking into account the test delay.


Bacterial biofilm formation on indwelling urethral catheters.
Urethral catheters are the most commonly deployed medical devices and used to manage a wide range of conditions in both hospital and community care settings. The use of long-term catheterization, where the catheter remains in place for a period >28 days remains common, and the care of these patients is often undermined by the acquisition of infections and formation of biofilms on catheter surfaces. Particular problems arise from colonization with urease-producing species such as Proteus mirabilis, which form unusual crystalline biofilms that encrust catheter surfaces and block urine flow. Encrustation and blockage often lead to a range of serious clinical complications and emergency hospital referrals in long-term catheterized patients. Here we review current understanding of bacterial biofilm formation on urethral catheters, with a focus on crystalline biofilm formation by P. mirabilis, as well as approaches that may be used to control biofilm formation on these devices. SIGNIFICANCE AND IMPACT OF THE STUDY: Urinary catheters are the most commonly used medical devices in many healthcare systems, but their use predisposes to infection and provide ideal conditions for bacterial biofilm formation. Patients managed by long-term urethral catheterization are particularly vulnerable to biofilm-related infections, with crystalline biofilm formation by urease producing species frequently leading to catheter blockage and other serious clinical complications. This review considers current knowledge regarding biofilm formation on urethral catheters, and possible strategies for their control.


A novel medium for the isolation of N-acylhomoserine lactone-degrading bacteria.
A novel chemically defined medium, named KG medium, supplemented with N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL), an acylhomoserine lactone (AHL) used as signalling molecules in Gram-negative bacterial cell-to-cell communication, as the sole source of carbon and nitrogen, was designed and successfully used for the enrichment and isolation of AHL-degrading bacteria. A 3-oxo-C6-HSL-degrading bacterium, 13sw7, was isolated from sewage after six enrichment transfers in the 3-oxo-C6-HSL-containing KG medium. On the basis of the almost complete 16S ribosomal DNA sequence, isolate 13sw7 was clustered with unculturable beta-proteobacteria. This study indicates that the AHL-containing KG medium is effective in isolating AHL-degrading bacteria, including those previously considered unculturable, from environmental sources. To the best of our knowledge, this is the first documentation of the isolation of an AHL-degrading proteobacterium from sewage.


Serum oncostatin M predicts mucosal healing in patients with inflammatory bowel diseases treated with anti-TNF, but not vedolizumab.
Oncostatin M was recently highlighted as a promising biomarker for therapeutic effectiveness in inflammatory bowel diseases (IBD), with particular regard for infliximab. The primary aim was to evaluate the ability of serum oncostatin M to predict endoscopic response to different drugs in IBD. We selected two different cohorts of patients with IBD, treated with anti-TNF (infliximab and adalimumab) or with vedolizumab. Therapeutic response was evaluated at week 54 in terms of mucosal healing. Serum oncostatin M and C-reactive protein were measured at baseline; fecal calprotectin was measured at baseline and after 14 weeks of treatment. We evaluated the association of these biomarkers with mucosal healing at week 54. Among 66 patients treated with anti-TNFs and 68 treated with vedolizumab, 35 and 31 attained mucosal healing, respectively. Mucosal healing at 54 weeks was significantly associated with low oncostatin M levels at baseline in the anti-TNF cohort; the diagnostic accuracy of oncostatin M at baseline in predicting mucosal healing was 0.91 (95% CI 0.84 to 0.99) in the anti-TNF cohort and 0.56 (95% CI 0.43 to 0.70, P < 0.001) in the vedolizumab cohort. Mucosal healing was also associated with low fecal calprotectin levels at week 14 in both cohorts. Our study suggests that serum oncostatin M is a drug-specific biomarker, since it could be used to predict therapeutic effectiveness to anti-TNFs but not to vedolizumab. Moreover, these results emphasize the utility of serum oncostatin M measurement in patients treated with anti-TNF.


A redescription of the Late Jurassic (Tithonian) turtle Uluops uluops and a new phylogenetic hypothesis of Paracryptodira.
We study the Late Jurassic (Tithonian) turtle Uluops uluops using micro-computed tomography scans to investigate the cranial anatomy of paracryptodires, and provide new insights into the evolution of the internal carotid artery and facial nerve systems, as well as the phylogenetic relationships of this group. We demonstrate the presence of a canalis caroticus lateralis in Uluops uluops, the only pleurosternid for which a palatine artery canal can be confidently identified. Our phylogenetic analysis retrieves Uluops uluops as the earliest branching pleurosternid, Helochelydridae within Pleurosternidae, and Compsemydidae including Kallokibotion bajazidi within Baenidae, which suggests at least two independent losses of the palatine artery within paracryptodires. We expect future studies will provide additional insights into the evolution of the circulation system of paracryptodires, as well as clarifying relationships along the turtle stem. The online version contains supplementary material available at 10.1186/s13358-021-00234-y.


Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness.
Mitochondrial respiration is an essential component of cellular metabolism. It is a process of energy conversion through enzymatically mediated reactions, the energy of taken-up substrates transformed to the ATP production. Seahorse equipment allows to measure oxygen consumption in living cells and estimate key parameters of mitochondrial respiration in real-time mode. Four key mitochondrial respiration parameters could be measured: basal respiration, ATP-production coupled respiration, maximal respiration, and proton leak. This approach demands the application of mitochondrial inhibitors-oligomycin to inhibit ATP synthase, FCCP-to uncouple the inner mitochondrial membrane and allow maximum electron flux through the electron transport chain, rotenone, and antimycin A to inhibit complexes I and III, respectively. This chapter describes two protocols of seahorse measurements performed on iPSC-derived cardiomyocytes and TAZ knock-out C2C12 cell line.