Effect of a CIDR insert and flunixin meglumine, administered at the time of embryo transfer, on pregnancy rate and resynchronization of estrus in beef cattle.
The objectives of this study were to evaluate the effects of flunixin meglumine (FM), an inhibitor of PGF(2alpha) synthesis, and insertion of an intravaginal progesterone-releasing device (CIDR), on pregnancy rates in beef cattle embryo transfer (ET) recipients, and to examine the effect of a CIDR after embryo transfer on the synchrony of the return to estrus in non-pregnant recipients. Cows (n=622) and heifers (n=90) at three locations were assigned randomly to one of four groups in a 2x2 factorial arrangement of treatments with FM administration (500 mg i.m.) 2-12 min prior to ET, and insertion of a CIDR (1.38 g progesterone) immediately following ET as main effects. Fresh or frozen embryos (Stage=4 or 5; Grade=1 or 2) were transferred on Days 6-9 of the estrous cycle and CIDR devices were removed 13 days after ET. Recipients at Location 2 only were observed for signs of return to estrus. Recipients that returned to estrus at Location 2 were either bred by AI or received an embryo 7 days after estrus. Following the initial ET, there was an FMxlocation interaction on pregnancy rate (P<0.01; Location 1, 89% versus 57%; Location 2, 69% versus 64%; Location 3, 64% versus 67% for FM versus no FM, respectively). Pregnancy rates of embryo recipients were not affected by CIDR administration (P>0.05; 65% with CIDR, 70% without CIDR), however, the timing of the return to estrus was more synchronous (P<0.01) for recipients given a CIDR. Pregnancy rate of recipients bred following a return to estrus did not differ between cows receiving or not receiving a CIDR for resynchronization (P>0.13). Effects of FM on pregnancy rate were location dependent and CIDR insertion at ET improved synchrony of the return to estrus.


Glycosylation as a regulator of site-specific metastasis.
Metastasis is considered to be responsible for 90% of cancer-related deaths. Although it is clinically evident that metastatic patterns vary by primary tumor type, the molecular mechanisms underlying the site-specific nature of metastasis are an area of active investigation. One mechanism that has emerged as an important player in this process is glycosylation, or the addition of sugar moieties onto protein and lipid substrates. Glycosylation is the most common post-translational modification, occurring on more than 50% of translated proteins. Many of those proteins are either secreted or expressed on the cell membrane, thereby making glycosylation an important mediator of cell-cell interactions, including tumor-microenvironment interactions. It has been recently discovered that alteration of glycosylation patterns influences cancer metastasis, both globally and in a site-specific manner. This review will summarize the current knowledge regarding the role of glycosylation in the tropism of cancer cells for several common metastatic sites, including the bone, lung, brain, and lymph nodes.


Direct prenatal chromosome diagnosis of a malignancy.
A fetal tumor was suspected at 31 weeks of gestation. The occurrence of polyhydramnios led to an ultrasound examination, which revealed deformation of the fetal head, face, eye, and neck. This was confirmed by computerized tomography. Amniocentesis yielded cells with an inverted duplication of chromosome #1. This abnormality of chromosome #1 marked the malignant teratoma cells in the amniotic fluid. Cytogenetic analysis of tumor tissue and of normal tissue obtained postnatally confirmed that the abnormality of chromosome #1 observed in amniotic fluid cells was confined to the tumor. The constitutional karyotype was normal. To our knowledge, this is the first report of the direct chromosomal detection of malignancy before birth.


Examination of the foreign body response to biomaterials by nonlinear intravital microscopy.
Implanted biomaterials often fail because they elicit a foreign body response (FBR) and concomitant fibrotic encapsulation. To design clinically relevant interference approaches, it is crucial to first examine the FBR mechanisms. Here, we report the development and validation of infrared-excited nonlinear microscopy to resolve the three-dimensional (3D) organization and fate of 3D-electrospun scaffolds implanted deep into the skin of mice, and the following step-wise FBR process. We observed that immigrating myeloid cells (predominantly macrophages of the M1 type) engaged and became immobilized along the scaffold/tissue interface, before forming multinucleated giant cells. Both macrophages and giant cells locally produced vascular endothelial growth factor (VEGF), which initiated and maintained an immature neovessel network, followed by formation of a dense collagen capsule 2-4 weeks post-implantation. Elimination of the macrophage/giant-cell compartment by clodronate and/or neutralization of VEGF by VEGF Trap significantly diminished giant-cell accumulation, neovascularization and fibrosis. Our findings identify macrophages and giant cells as incendiaries of the fibrotic encapsulation of engrafted biomaterials via VEGF release and neovascularization, and therefore as targets for therapy.


Endothelium-derived relaxing factor inhibition and the pressor response to norepinephrine in the pregnant rat.
To determine whether endothelium-derived relaxing factor (EDRF) plays a role in the blunted pressor response to norepinephrine that is characteristic of normal pregnancy. Eleven pregnant (mean +/- standard error of the mean 18 +/- 1 days of gestation) and 11 virgin conscious, unrestrained Sprague-Dawley rats with long-term indwelling vascular catheters were studied. Mean arterial pressure (MAP) and heart rate were measured in response to continuous infusions of either vehicle (5% dextrose) or increasing concentrations of norepinephrine (25, 100, and 200 ng/kg/minute) before and after administration of NG-monomethyl-L-arginine (LNMMA), a specific inhibitor of EDRF synthesis. Baseline MAP was lower in pregnant than in virgin rats (96 +/- 3 versus 105 +/- 3 mmHg; P < .05). Before LNMMA administration, the pregnant rats exhibited a significantly blunted pressor response to increasing concentrations of norepinephrine compared to that of virgin rats (P < .005). Given alone, LNMMA produced a greater increase in baseline MAP in virgin rats than in pregnant rats (rise in MAP of 44 +/- 2 versus 31 +/- 2 mmHg; P < .001). However, LNMMA abolished the blunted pressor response to norepinephrine in the pregnant animals and did not significantly affect the pressor response to norepinephrine in virgin rats. Heart rate responses to increasing concentrations of norepinephrine in the presence and absence of LNMMA were not significantly different in the two groups of animals. Stimulated EDRF production may contribute to the blunted pressor response to norepinephrine characteristic of pregnancy in the rat.


Hemodialysis-related induction of beta 2-microglobulin synthesis and release by mononuclear phagocytes.
We have investigated beta 2-microglobulin (beta 2M) synthesis and release by blood leukocytes and isolated mononuclear phagocytes. Since beta 2M was discovered to form amyloid fibrils in patients undergoing long term, chronic hemodialysis and monomeric beta 2M levels in plasma of these patients are highly elevated, and since hemodialysis-related factors that increase beta 2M production are unknown, we evaluated beta 2M production by mononuclear phagocytes under a variety of conditions. We utilized a novel enzyme-lined immunoabsorbant assay to quantitate beta 2M release. Adherence of macrophages onto polystyrene or Cuprophan membranes does not induce beta 2M synthesis or release. In contrast, interaction of macrophages with lipopolysaccharide, gamma-interferon, tumor necrosis factor, or interleukin 1 induces synthesis or release. In contrast, interaction of macrophages with lipopolysaccharide, gamma-interferon, tumor necrosis factor, or interleukin 1 induces synthesis and release of beta 2M, indicating that beta 2M synthesis is increased during macrophage activation. Exposing leukocytes or macrophages to changes in osmotic or oncotic pressure induces a rapid release of beta 2M and interleukin 1 into the cellular medium. These results suggest that during hemodialysis, beta 2M production is more likely to result from endotoxin contamination, or osmotic and oncotic changes, rather than direct interaction of mononuclear phagocytes with cellulosic membranes.


Immunohistochemical localization of neurocalcin in the rat inner ear.
Localization in the rat inner ear of neurocalcin, a three EF-hand calcium-binding protein, was examined immunohistochemically. Neurocalcin-like immunoreactivity was restricted to neurons in neuroepithelial receptor organs, while hair cells and supporting cells showed no such immunoreactivity. In the organ of Corti, both afferent and efferent nerve terminals, which formed synapses on both inner and outer hair cells, showed distinct immunoreactions. Spiral ganglion neurons and cochlear nerves were immunopositive. In the cristae ampullaris, macula utriculi and macula sacculi, afferent nerve terminals forming nerve calices or terminal boutons were strongly immunopositive. Efferent nerve terminals making synapses either on nerve calices or on hair cells showed an intense immunoreactivity. Vestibular ganglion neurons were strongly immunopositive. In electron microscopy, immunoreaction products were diffuse in the cytoplasm of ganglion neurons and nerve terminals. Neurocalcin-like immunoreactivity occurred in association with microtubules, outer mitochondrial membranes, synaptic vesicles and synaptic membranes. It is thus likely that neurocalcin is involved in neural functions in each type of afferent and efferent transmission in the inner ear.


Shrimp thrombospondin (TSP): presence of O-β1,4 N-acetylglucosamine polymers and its function in TSP chain association in egg extracellular matrix.
We characterized the existence of O-β(1,4)-GlcNAc polymers (β1,4GNP) that were anchored on the O-linked glycosylation sites of shrimp thrombospondin (pmTSP-II). There were five putative β1,4GNP linkages on the epithelial growth factor-like domain of pmTSP-II. Antibody against O-β-GlcNAc (CTD110.6) was used to prove the existence of linear and complex β1,4GNP. The antibody well reacted with linear chito-triose, -tetraose and -pentaose conjugated with phosphatidylethanolamine lipid. The immunoreactivity could also be detected with a complex β1,4GNP within pmTSP-II (at MW > 250 kDa). Upon denaturing the protein with SDS-PAGE buffer, the size of pmTSP-II was shifted to be 250 kDa, approximately 2.5 folds larger than the deduced molecular mass of pmTSP-II (110 kDa), suggesting additional association of pmTSP-II apart from its known disulfide bridging. This was confirmed by chitinase digestion on pmTSP-II protein leading to the subsequent smaller protein bands at 110-170 kDa in time- and concentration-dependent manners. These bands well reacted with CTD110.6 antibody and disappeared after extensive chitinase hydrolysis. Together, we believe that β1,4GNP on pmTSP-II serve the function in an inter-chain association to provide structural architecture of egg extracellular matrix, a novel function of pmTSP-II in reproductive biology.


The evaluation of gallstone formation in patients undergoing Roux-en-Y gastric bypass due to morbid obesity.
This study aimed to evaluate gallstone formation, prophylactic and selective cholecystectomy and the effectiveness of ursodeoxycholic acid treatment following laparoscopic Roux-en-Y gastric bypass (LRYGB) in morbid obese patients. Files of 60 patients who underwent LRYGB between October 2006 and March 2011 were retrospectively reviewed. Patients were evaluated for formation of gallstones. Fifty-three (88.3%) patients were female and seven (11.7%) were male. Eight of the 60 patients (13.3%) had previously undergone cholecystectomy. Six patients (11.5%) underwent cholecystectomy in addition to LRYGB due to preoperatively detected gallstones by ultrasonography. The remaining 46 patients were followed up for a mean duration of 28.57 months (5-56 months). In 10 (21.7%) of these patients, gallstones were detected and five patients with symptomatic gallstones underwent cholecystectomy. Patients who did and did not develop gallstones after LRYGB did not show a significant difference regarding age, gender and the new body mass index (BMI). Three patients were started on ursodeoxycholic acid and the treatment was continued for six months. Gallstones were not detected in these patients. In light of these data, since only a very small portion of patients develops symptomatic gallstones after LRYGB, we recommend cholecystectomy in patients with symptomatic gallstones or the use of ursodeoxycholic acid rather than a prophylactic approach. Prospective randomized controlled studies in larger series are required to support these results.


Ultrasound-guided biopsy of abdominal and pelvic lesions in children. A comparison between fine-needle aspiration and 1.2 mm-needle core biopsy.
Tissue diagnosis is mandatory prior to treatment of an abdominal or pelvic lesion in a child. To compare the diagnostic yield and complications of fine-needle aspiration biopsy (FNAB) and 1.2-mm needle core biopsy (NCB) for abdominal and pelvic lesions in children using US guidance. Ninety consecutive abdominal or pelvic US-guided biopsies in 61 children; 53 FNAB procedures in 43 children and 37 NCB procedures in 34 children were retrospectively analysed. Fifty-seven biopsies were performed on malignant lesions and 33 on benign lesions. In 15 children, both FNAB and NCB were performed at the same occasion. FNAB was diagnostic in 41 (77 %) of 53 biopsies while NCB provided a correct diagnosis in 35 (95 %) of 37 biopsies. Results were similar in both malignant and benign conditions. Clinically important bleeding complicated three biopsies (3.3 %); in two after FNAB and in one after combined FNAB and NCB. If FNAB were to be replaced with 1.2-mm NCB using an automatic gun, the diagnostic yield for abdominal and pelvic focal lesions in children would improve significantly (P < 0.05) while the complication rate would remain low. NCB may reduce the need for diagnostic and staging surgery and repeated procedures.


Clozapine use at a specialised psychiatric hospital in Johannesburg.
Clozapine is the gold standard medication for treatment-resistant psychosis, with robust evidence supporting its efficacy in multiple symptom domains. However, clozapine's side effect profile contributes to its underutilisation and discontinuation. This study aimed to explore the magnitude of clozapine use and describe factors that impact on its effective use among in-patients. Tara Hospital, a specialised psychiatric hospital in Johannesburg. This was a retrospective, cross-sectional file review of clozapine-treated patients admitted over the 2-year study period. Data variables included: demographics, clinical information, discharge prescription, clozapine-related side effects and details of clozapine discontinuation, where applicable. A cohort of 33.2% of patients from Tara's biological wards received a trial of clozapine. Participants experienced anti-cholinergic clozapine-related side effects that included weight gain (79.5%), tachycardia (35.2%) and constipation (35.2%). Clozapine was discontinued in 13.7% of participants, and no life-threatening side effects or deaths occurred. Significantly more use of flupenthixol decanoate (64.3% vs. 30.7%; p = 0.0322) and anticholinergics (35.7% vs. 11.4%; p = 0.0474) occurred in the clozapine-discontinued group. Polypharmacy rates were high for psychiatric and non-psychiatric medications. One-third of patients received clozapine trials, most of whom continued at discharge. Although side effects occurred frequently, life-threatening side effects did not. Clozapine monitoring protocols, side effect rating scales, pre-emptive management of side effects, lifestyle interventions and clinician education may improve outcomes of clozapine use. The use of plasma clozapine levels may be beneficial. This study expands our limited knowledge regarding current clozapine prescribing trends in South Africa.


The influence of complaint symptoms on health care utilisation, medicine use, and sickness absence. A comparison between retrospective and prospective utilisation.
The main aim was to examine the impact of reported individual symptoms on health care utilisation (HCU) in a primary health care practice population and to evaluate the impact of these symptoms on utilisation over time. The study was performed in 1055 out of 1442 consecutive adult patients visiting a Swedish health care centre. Logistic regression analyses were applied to assess the relationships between symptoms and HCU outcomes. Each of the 30 symptoms was related to consultations with GPs and a provider of alternative medicine, use of medications and herbals, and sickness absence. Depression and tension groups were the strongest predictors of utilisation. Multisymptomatics had higher OR for most of the outcome variables than those with no symptoms. The results of this study show a linear correlation between the numbers of symptoms and increasing GP consultations, medicine use and sickness absences days, and this persisted during the years 2001 and 2002.


Anticoagulant effect and action mechanism of sulphated flavonoids from Flaveria bidentis.
Quercetin 3-acetyl-7,3',4'-trisulphate (ATS) and quercetin 3,7,3',4'-tetrasulphate (QTS) obtained from Flaveria bidentis (Asteraceae) were investigated in vitro for anticoagulant activity. Three different concentrations of each flavonoid were assayed at different incubation times, showing at 1 mM significant prolongation on the activated partial thromboplastin time (APTT), less on the prothrombin time (PT), and no effect on the thrombin time (TT). In order to define the action mechanism of the anticoagulant activity, all coagulation factors were evaluated and no important activity decrease was observed, indicating that another mechanism is involved. Thus, thrombin inhibition mediated by antithrombin III (ATIII) and heparin cofactor II (HCII) activation was investigated in comparison to the physiological activators, heparin and dermatan sulphate (DS), respectively. As a conclusion, no activation on ATIII for neither flavonoids was observed. On the contrary, QTS much more than ATS produced an activation on HCII comparable to the one of DS, indicating that these flavonoids act as agonists of this inhibitor. A plausible explanation of the effects of both flavonoids could be due to the different degree of sulphation of these molecules. According to the results obtained, and taking in account the high solubility of these natural products in aqueous media and the nontoxic nature of this family of compounds, further investigation on the antithrombotic effects of these flavonoids are merited.


Credentialing in dermatologic surgery.
Dermatologic surgeons' rights to perform surgery are being and will be challenged more and more. Some methods of peer review and credentialing are suggested and the specialty is urged to move ahead with formal planning for more training in the surgical aspects of dermatology.


Coupling tandem affinity purification and quantitative tyrosine iodination to determine subunit stoichiometry of protein complexes.
Rapid protein purification methodologies, such as strategies involving the tandem affinity purification module, have resulted in the identification of a tremendous number of multisubunit protein complexes. Furthermore, in this modern genomic age, mass spectrometry methods are often coupled with affinity purification to identify the genes that encode each protein subunit. However, simple methodologies to determine the stoichiometry of individual subunits within a multisubunit complex have not received much attention. In this article we describe a procedure to rapidly and efficiently determine the stoichiometry of subunits within multisubunit complexes using a combination of tandem affinity purification and quantitative 125I labeling of subunit tyrosines.


Connective tissue growth factor stimulates renal cortical myofibroblast-like cell proliferation and matrix protein production.
Myofibroblasts primarily contribute to the pathogenesis of renal interstitial fibrosis by unregulated cell proliferation and synthesis of excessive amounts of extracellular matrix (ECM) proteins. We used cultured myofibroblast-like cells obtained by outgrowth from explants of rat kidney cortex to study the effects and relevant signaling pathway of connective tissue growth factor (CTGF) on cell proliferation and ECM production. Exogenous CTGF stimulated proliferation of myofibroblast-like cells in a dose- and time-dependent manner. CTGF also increased the secretion of fibronectin and collagen I protein in the supernatant medium. Nevertheless, CTGF did not affect matrix-degrading metalloproteinases-2 and -9 activities in supernatant medium measured by gelatin zymography. CTGF induced activation of extracellular signal-regulated protein kinase (ERK)1/2 mitogen-activated protein kinase pathway as early as 5 minutes. Inhibition of ERK1/2 activation with PD98059 completely blocked CTGF-induced cell proliferation as well as secretion of fibronectin and collagen I protein. The above results indicate that CTGF triggers cell proliferation and production of ECM proteins in cultured myofibroblast-like cells through the ERK1/2 mitogen-activated protein kinase pathway.


Early Treatment with Vigabatrin Does Not Decrease Focal Seizures or Improve Cognition in Tuberous Sclerosis Complex: The PREVeNT Trial.
This study was undertaken to test the hypothesis that early vigabatrin treatment in tuberous sclerosis complex (TSC) infants improves neurocognitive outcome at 24 months of age. A phase IIb multicenter randomized double-blind placebo-controlled trial was conducted of vigabatrin at first epileptiform electroencephalogram (EEG) versus vigabatrin at seizure onset in infants with TSC. Primary outcome was Bayley Scales of Infant and Toddler Development, Third Edition (Bayley-III) cognitive assessment score at 24 months. Secondary outcomes were prevalence of drug-resistant epilepsy, additional developmental outcomes, and safety of vigabatrin. Of 84 infants enrolled, 12 were screen failures, 4 went straight to open label vigabatrin, and 12 were not randomized (normal EEG throughout). Fifty-six were randomized to early vigabatrin (n = 29) or placebo (n = 27). Nineteen of 27 in the placebo arm transitioned to open label vigabatrin, with a median delay of 44 days after randomization. Bayley-III cognitive composite scores at 24 months were similar for participants randomized to vigabatrin or placebo. Additionally, no significant differences were found between groups in overall epilepsy incidence and drug-resistant epilepsy at 24 months, time to first seizure after randomization, and secondary developmental outcomes. Incidence of infantile spasms was lower and time to spasms after randomization was later in the vigabatrin group. Adverse events were similar across groups. Preventative treatment with vigabatrin based on EEG epileptiform activity prior to seizure onset does not improve neurocognitive outcome at 24 months in TSC children, nor does it delay onset or lower the incidence of focal seizures and drug-resistant epilepsy at 24 months. Preventative vigabatrin was associated with later time to onset and lower incidence of infantile spasms. ANN NEUROL 2023.


The Effect of Multidomain Lifestyle Intervention on Daily Functioning in Older People.
To investigate the effect of a 2-year multidomain lifestyle intervention on daily functioning of older people. A 2-year randomized controlled trial (ClinicalTrials.gov, NCT01041989). Finnish Geriatric Intervention Study to Prevent Cognitive Impairment and Disability. A total of 1260 older adults, with a mean age of 69 years at the baseline, who were at risk of cognitive decline. A multidomain intervention, including simultaneous physical activity intervention, nutritional counseling, vascular risk monitoring and management, and cognitive training and social activity. The ability to perform daily activities (activities of daily living [ADLs] and instrumental ADLs) and physical performance (Short Physical Performance Battery). The mean baseline ADL score was 18.1 (SD = 2.6) points; the scale ranges from 17 (no difficulties) to 85 (total ADL dependence). During the 2-year intervention, the ADL disability score slightly increased in the control group, while in the intervention group, it remained relatively stable. Based on the latent growth curve model, the difference in the change between the intervention and control groups was -0.95 (95% confidence interval [CI] = -1.61 to -0.28) after 1 year and -1.20 (95% CI = -2.02 to -0.38) after 2 years. In terms of physical performance, the intervention group had a slightly higher probability of improvement (from score 3 to score 4; P = .041) and a lower probability of decline (from score 3 to scores 0-2; P = .043) for chair rise compared to the control group. A 2-year lifestyle intervention was able to maintain the daily functioning of the at-risk older population. The clinical significance of these results in this fairly well-functioning population remains uncertain, but the study results hold promise that healthy eating, exercise, and cognitive and social activity may have favorable effects on functional independence in older people.


Meeting the mental health needs of people with multiple sclerosis: a qualitative study of patients and professionals.
To explore perspectives and experiences of people with multiple sclerosis (MS) and health care professionals of mental health support for MS in the UK. 24 people with MS, 13 practice nurses, 12 general practitioners (GPs) and 9 MS specialist nurses were recruited through community groups and primary care practices across North West England. Semi-structured interviews were conducted, and data analyzed thematically using constant comparative analysis within and across the data sets. The theoretical framework of candidacy was used to interrogate data. Four themes were identified: candidates for care, management choices, defining roles, and permeability and responsiveness. Candidacy for care, and symptom management, depended on the framing of symptoms through a social or medical model of depression. Normalizing symptoms could prevent help-seeking by patients. Reported referral behavior varied by professional group, based on perceived remit, competency and training needs. GPs were perceived by patients and other professionals as central for management of mental health needs in MS, but may not perceive this role themselves, suggesting a need for increased knowledge, training, and improved access to specialist care. Implications for Rehabilitation Anxiety and depression are common in people with MS. Management of mental health needs in people with MS relies on complex decisions made by both people with MS and health care professionals. General practitioners may play a key role in the ongoing management of mental health needs of people with MS.


Significance of Sonographic Subcentimeter, Subpleural Consolidations in Pediatric Patients Evaluated for Pneumonia.
To investigate the rates of radiographic pneumonia and clinical outcomes of children with suspected pneumonia and subcentimeter, subpleural consolidations on point-of-care lung ultrasound. We enrolled a prospective convenience sample of children aged 6 months to 18 years undergoing chest radiography (CXR) for pneumonia evaluation in a single tertiary-care pediatric emergency department. Point-of-care lung ultrasound was performed by an emergency medicine physician with subsequent expert review. We determined rates of radiographic pneumonia and clinical outcomes in the children with subcentimeter, subpleural consolidations, stratified by the presence of larger (>1 cm) sonographic consolidations. The children were followed prospectively for 2 weeks to identify a delayed diagnosis of pneumonia. A total of 188 patients, with a median age of 5.8 years (IQR, 3.5-11.0 years), were evaluated. Of these patients, 62 (33%) had subcentimeter, subpleural consolidations on lung ultrasound, and 23 (37%) also had larger (>1 cm) consolidations. Patients with subcentimeter, subpleural consolidations and larger consolidations had the highest rates of definite radiographic pneumonia (61%), compared with 21% among children with isolated subcentimeter, subpleural consolidations. Overall, 23 children with isolated subcentimeter, subpleural consolidations (59%) had no evidence of pneumonia on CXR. Among 16 children with isolated subcentimeter, subpleural consolidations and not treated with antibiotics, none had a subsequent pneumonia diagnosis within the 2-week follow-up period. Children with subcentimeter, subpleural consolidations often had radiographic pneumonia; however, this occurred most frequently when subcentimeter, subpleural consolidations were identified in combination with larger consolidations. Isolated subcentimeter, subpleural consolidations in the absence of larger consolidations should not be viewed as synonymous with pneumonia; CXR may provide adjunctive information in these cases.


Uromucoid (Tamm-Horsfall glycoprotein) forms different polymeric arrangements on a filter surface under different physicochemical conditions.
Normal human urine cannot be forced through a 0.2 micron filter. To investigate the reason for this phenomenon, uromucoid (Tamm-Horsfall protein) was purified from human urine and its capacity to block a 0.2 micron Millipore filter was measured under different conditions. In the presence of cations (H+, Na+, Ca2+) uromucoid blocked the filter. The blocking varied with cation concentration. Scanning electron microscopy of the filter surface revealed different arrangements of polymerized uromucoid coating the filter surface depending on ionic conditions. In the presence of 100 mmol/l NaCl or 1 mmol/l CaCl2 uromucoid polymers were present in a fibrous arrangement. In the presence of both NaCl and CaCl2 a dense mat of uromucoid polymers was present together with clumps of aggregated polymer. In the absence of ions uromucoid formed a homogeneous coat on the filter surface (as demonstrated by scanning electron microscopy, Western blotting and 125I-uromucoid binding studies) but did not block the filter. Similar fibrous and highly aggregated arrangements of uromucoid polymer were seen in hyaline casts from urine. These data are consistent with the concept that the uromucoid glycoprotein can exist in several different polymeric forms under different ionic conditions.


Collective Approach to Advancing C-H Functionalization.
C-H functionalization is a very active research field that has attracted the interest of scientists from many disciplines. This Outlook describes the collaborative efforts within the NSF CCI Center for Selective C-H Functionalization (CCHF) to develop catalyst-controlled selective methods to enhance the synthetic potential of C-H functionalization.


A split prime editor with untethered reverse transcriptase and circular RNA template.
Delivery and optimization of prime editors (PEs) have been hampered by their large size and complexity. Although split versions of genome-editing tools can reduce construct size, they require special engineering to tether the binding and catalytic domains. Here we report a split PE (sPE) in which the Cas9 nickase (nCas9) remains untethered from the reverse transcriptase (RT). The sPE showed similar efficiencies in installing precise edits as the parental unsplit PE3 and no increase in insertion-deletion (indel) byproducts. Delivery of sPE to the mouse liver with hydrodynamic injection to modify β-catenin drove tumor formation with similar efficiency as PE3. Delivery with two adeno-associated virus (AAV) vectors corrected the disease-causing mutation in a mouse model of type I tyrosinemia. Similarly, prime editing guide RNAs (pegRNAs) can be split into a single guide RNA (sgRNA) and a circular RNA RT template to increase flexibility and stability. Compared to previous sPEs, ours lacks inteins, protein-protein affinity modules and nuclease-sensitive pegRNA extensions, which increase construct complexity and might reduce efficiency. Our modular system will facilitate the delivery and optimization of PEs.


Post-discharge neonatal hyperbilirubinemia surveillance.
To assess implementation of the Israel Neonatal Society's 2008 guidelines for universal community assessment of jaundice within 72 hours of discharge from birth hospitalisation. Mothers of newborns were interviewed at Maternal Child Health Clinics in the Jerusalem District, Israel, and asked whether their newborn had been evaluated for jaundice within the recommended time frame. Newborn discharge letters from Israeli hospitals were assessed for appropriate inclusion of instructions for early follow-up for jaundice. Out of 659, 217 (32.9%) mothers whose newborns were at low risk for neonatal hyperbilirubinemia reported an examination within 72 hours of discharge. Eighteen (8.3%) were referred for a bilirubin test. In contrast, 99.1% (109/110) of high-risk newborns who were specifically invited for a bilirubin test the day following discharge complied. Out of 26, 12 (46.2%) hospital discharge letters specified both a time limit of 72 hours post-discharge and jaundice as a reason for early follow-up. The early community surveillance rate for jaundice was low, contrasting with near universal compliance in those who received a specific instruction for a post-discharge bilirubin blood test. Inclusion of specific written instructions in hospital discharge summaries was also low and may contribute to poor implementation of guidelines.


Survival after recovery from acute myocardial infarction. Two and five year prognostic indices.
A prognostic index for two year survival following recovery from acute myocardial infarction has been verified in an independent group of 105 patients. Five variables comprise the index: systolic blood pressure level on admission, highest blood urea nitrogen level in the cardiac care unit; atrial arrhythmias in the cardiac care unit; angina pectoris for more than three months or a previous myocardial infarction; and more than one ventricular ectopic beat per hour recorded on an 8 hour dynamic electrocardiogram during convalescence just prior to hospital discharge. One hundred twenty-six patients have also been followed for five or more years, and we now report a five year prognostic index. Discriminant analysis indicates that the same five variables, although weighted differently, continue to be significant for prognostic assessment and may be utilized in the identification of patients at high and lower risk.


Association of Pharmacological Treatments and Hospitalization and Death in Individuals With Amphetamine Use Disorders in a Swedish Nationwide Cohort of 13 965 Patients.
There are no medications approved by authorities for the treatment of amphetamine or methamphetamine dependence, and studies investigating the effectiveness of pharmacological treatments in hard outcomes, such as hospitalization and death, are lacking. To investigate the association between pharmacotherapies and hospitalization and mortality outcomes in persons with amphetamine or methamphetamine use disorder. This nationwide register-based cohort study was conducted from July 2006 to December 2018 with a median (IQR) follow-up time of 3.9 (1.0-6.1) years. Data were analyzed from December 1, 2021, to May 24, 2022. All residents aged 16 to 64 years living in Sweden with a registered first-time diagnosis of amphetamine or methamphetamine use disorder and without previous diagnoses of schizophrenia or bipolar disorder were identified from nationwide registers of inpatient care, specialized outpatient care, sickness absence, and disability pension. Medications for substance use disorders (SUDs) or for attention-deficit/hyperactive disorder, mood stabilizers, antidepressants, benzodiazepines and related drugs, and antipsychotics. Medication use vs nonuse was modeled with the PRE2DUP (from prescription drug purchases to drug use periods) method. Primary outcomes were hospitalization due to SUD and any hospitalization or death, which were analyzed using within-individual models by comparing use and nonuse periods of 17 specific medications or medication classes in the same individual to minimize selection bias. The secondary outcome was all-cause mortality, studied using between-individual analysis as traditional Cox models. There were 13 965 individuals in the cohort (9671 [69.3%] male; mean [SD] age, 34.4 [13.0] years). During follow-up, 7543 individuals (54.0%) were taking antidepressants, 6101 (43.7%) benzodiazepines, 5067 (36.3%) antipsychotics, 3941 (28.2%) ADHD medications (1511 [10.8%] were taking lisdexamphetamine), 2856 (20.5%) SUD medications, and 1706 (12.2%) mood stabilizers. A total of 10 341 patients (74.0%) were hospitalized due to SUDs, 11 492 patients (82.3%) were hospitalized due to any cause or died, and 1321 patients (9.5%) died of any cause. Lisdexamphetamine was the only medication in this study that was significantly associated with a decrease in risk of 3 outcomes (adjusted hazard ratio [aHR], 0.82; 95% CI, 0.72-0.94 for SUD hospitalization; aHR, 0.86; 95% CI, 0.78-0.95 for any hospitalization or death; aHR, 0.43; 95% CI, 0.24-0.77 for all-cause mortality). Methylphenidate use also was associated with lower all-cause mortality (aHR, 0.56; 95% CI, 0.43-0.74). Use of benzodiazepines was associated with a significantly higher risk of SUD hospitalization (aHR, 1.17; 95% CI, 1.12-1.22), any hospitalization or death (aHR, 1.20; 95% CI, 1.17-1.24), and all-cause mortality (aHR, 1.39; 95% CI, 1.20-1.60). Use of antidepressants or antipsychotics was associated with a slight increase in risk of SUD hospitalization (aHR, 1.07; 95% CI, 1.03-1.11 and aHR, 1.05; 95% CI, 1.01-1.09) as well as any hospitalization or death (aHR, 1.10; 95% CI, 1.06-1.14 and aHR, 1.06; 95% CI, 1.03-1.10, respectively). In this study, use of lisdexamphetamine was associated with improved outcomes in persons with amphetamine or methamphetamine use disorders, encouraging the conduct of randomized clinical trials. Prescription benzodiazepine use was associated with poor outcomes.


Poly[μ2-aqua-aqua-[μ3-N-butyl-N-(2-hy-droxy-ethyl)di-thio-carbamato-κ(3) O,O':S]sodium].
In the title compound, [Na(C7H14NOS2)(H2O)2] n , the Na(I) cation is coordinated by five O atoms [Na-O = 2.3142 (11)-2.4677 (10) Å] from three aqua and two N-butyl-N-(2-hy-droxy-eth-yl)di-thio-carbamate (L) ligands and one S atom [Na-S = 3.0074 (6) Å] from a third L ligand in a highly distorted octa-hedral geometry. Two aqua ligands related by an inversion center bridge two Na(I) cations, and each L ligand coordinates three Na(I) cations, leading to a layered arrangement aligned parallel to the bc plane. Inter-molecular O-H⋯S hydrogen bonds are observed in the inner part of each polymeric layer; these are packed along the a axis and held together by weak van der Waals forces.


Morphine three-dimensional T1 gadoxetate MR cholangiography of potential living related liver donors.
To assess low-dose morphine for distension and improved visualization of intrahepatic bile ducts on T1 MR cholangiography (MRC) in preoperative imaging of potential liver donors. Sixty-nine consecutive potential living related liver donors (mean age, 39 years; age range, 20 to 59 years) referred for pre-transplant MRI evaluation were evaluated without (n=30) or with (n=39) intravenous morphine injection (0.04 mg/kg). Morphine was injected pre-MRI while establishing intravenous access to allow ∼1 h for biliary distension before T1 MRC. Three radiologists reviewed intrahepatic biliary branch order visualization, common bile duct (CBD) diameter, and overall image quality. In 25 patients undergoing liver donation surgery, T1 MRC findings were correlated with intraoperative findings. This retrospective study was approved by the institutional review board. Biliary visualization was improved post-morphine administration with biliary duct branch order visualization score of 3.2 and 3.3 at 45 and 60 min, respectively, compared with 2.7 without morphine (P<0.002); CBD diameter measured 5.3 and 5.5 versus 4.1 mm (P<0.005), and overall image quality score was 2.4 and 2.6 versus 1.8 (P<0.0006). Operative notes confirmed T1 MRC findings in 6/11 donors without morphine and 14/14 donors with morphine. Intravenous low-dose morphine distends and improves visualization of bile ducts on T1 gadoxetate MRC.


What do people with lung cancer and stroke expect from patient navigation? A qualitative study in Germany.
This qualitative study investigated patients' needs and wishes in relation to patient navigation. A qualitative interview study was conducted. Participants were invited to take part in three in-depth interviews over a period of 6-12 months. Thematic analysis was used. Interviewees were sought in the Berlin metropolitan area of Germany in academic university hospitals, in rehabilitation clinics and through self-help organisations. The sample consisted of individuals diagnosed with lung cancer (n=20) or stroke (n=20). From the perspective of interviewees, patient navigators should function as consistent contact persons, present during the whole care trajectory. Their role would be to guide patients through an often confusing healthcare landscape, offering practical, advisory and emotional assistance corresponding to patients' needs. The study shows that-independent of the disease-participants had similar expectations and needs regarding support from navigators. For chronic and complex diseases-as is the case with lung cancer and stroke-it appears less important for navigators to fulfil disease-specific tasks. Rather, they should ensure that patients' more general needs, in relation to social, practical and emotional support, are met in a way that suits their individual wishes. Following these results, patient navigation programmes might be designed to include generic elements, which should then be adapted to the infrastructure in a particular healthcare region and to the particularities of a specific healthcare system.


Binding of additional histones to chromatin core particles.
Isolated nucleosome core particles from chick erythrocytes and calf thymus have been used in experiments to show that chromatin subunits can absorb more than an additional equivalent of the histones making up the nucleosome octamer with no significant alteration in external shape and structure.


Rotational deviation of 3-acetyl group from cyclic tetrapyrrole pi-plane in synthetic bacteriochlorophyll-a analogs by 20-substitution.
The 3-acetyl groups of synthetic methyl pyropheophorbides were rotated around the 3-3(1) bond and the rotational conformers were obtained in a dichloromethane solution of 20-bromo- and methyl-substituted compounds, based on their electronic and vibrational absorption spectra. Such a rotational deviation of the 3-acetyl group from the cyclic tetrapyrrole plane induced less pi-conjugation to affect the redmost Q(y) band, which has been observed in natural photosynthetic antenna systems, bacteriochlorophyll-a molecules in oligopeptides.


Cross-reactions of sera from Toxascaris leonina and Ascaris suum infected mice with Toxocara canis, Toxascaris leonina and Ascaris suum antigens.
The ELISA method using larval ES products and homogenized Toxocara canis, Toxascaris leonina and Ascaris suum adult worms extract, was used to determine the possible cross-reactions in BALB/c and C57BL/10 mice inoculated with embryonated eggs or adult worms extract of T. leonina of T. leonina or A. suum in single and multiple doses. When we used sera of mice infected with embryonated eggs of T. leonina against different heterologous Ag, no cross-reactions against T. canis ES and A. suum ES Ag were observed using a single dose. Similarly, in multiple doses no response against T. canis ES Ag was observed. In mice inoculated with adult worms extract of T. leonina cross-reactions with T. canis ES and A. suum ES Ag did not occur. Sera from BALB/c mice infected with embryonated eggs of A. suum, was tested using ES Ag from both A. suum and T. canis and no reactions were observed. This fact confirmed the resistance of this murine strain to A. suum embryonated eggs. When we used sera of susceptible C57BL/10 mice infected against different heterologous Ag, we observed no cross-reactions against T. canis ES Ag. In the case of both BALB/c and C57BL/10 and C57BL/10 mice immunized with a single dose of A. suum adult crude extract no cross-reactions were seen against ES T. canis Ag and with sera from C57BL/10 mice against ES T. leonina. These facts confirmed the specificity of the ES T. canis Ag.


Clinical significance of hematuria in patients on anticoagulant therapy.
We evaluated 29 consecutive patients in whom gross or microscopic hematuria developed while they were on heparin or warfarin anticoagulant therapy. Patients who had bleeding as a result of anticoagulant overdosage and/or from an additional organ system(s) other than the urinary tract were excluded from this review. Significant pathological findings consisting of carcinoma, calculi, renal infarction, infection, benign prostatic hyperplasia and/or adult polycystic renal disease were identified in 17 patients. Insignificant or incidental pathological findings classified as posterior urethritis, simple renal cyst or renal scarring were noted in 6 patients. No pathological condition was found in the remaining 6 patients. We conclude that a thorough and appropriate evaluation of the urinary tract should be conducted in patients on anticoagulant therapy who have gross or microscopic hematuria, since a pathological lesion of variable clinical significance often is discovered.


Measurement of axonal conduction velocity in single mammalian motor axons.
In deeply anesthetized cats, determinations of motor-axonal conduction velocity (CV) were made using extracellular potentials recorded from single, functionally isolated motor axons innervating the muscle tibialis posterior. Axons were activated by suprathreshold electrical stimulation at the ventral-root level. Action potentials were recorded with 3 bipolar electrodes located on the muscle nerve at the level of the popliteal fossa. The most proximal and distal of the bipolar muscle-nerve electrodes were 16.4-22.0 mm apart. Estimates were made of CV from ventral root to muscle nerve (conventional CV) and between the proximal and the distal pairs of muscle-nerve electrodes (muscle-nerve CV). An evaluation was based on comparison of these CVs, estimates of uncertainties in time and distance measurements and simulations of the effects of recording conditions on CV estimates. The analysis indicated that the uncertainty in the conventional CV measurement of mammalian motor axons is at least +/- 2%. However, variability may be as great as 20% between muscle-nerve CV measurements from different experiments, probably due to such factors as regional variation in CV and differences in recording configuration.


Endometrial receptivity and conception outcome among women with light menstrual bleeding of unidentified etiology.
To investigate endometrial receptivity (ER), conception outcome, and other factors among women with light menstrual bleeding of unidentified etiology. A prospective study was undertaken at a center in China. Between January 1, 2011, and December 31, 2014, women meeting the inclusion criteria (aged 20-35 years, without any history of pregnancy, desire to conceive, follicle-stimulating hormone <10 IU on second or third day of cycle, and regular menstrual periods) were enrolled. Participants with a menstrual blood volume of 5-20 mL formed the study group; others were in the control group. ER measures were obtained by transvaginal Doppler ultrasonography. Serum estradiol and progesterone levels were measured and items of prethrombotic state were screened in mid-luteal phase. Participants were followed up for 1 year to establish conception outcome. The ER (Salle) score was significantly lower in the study group (n=110) than in the control group (n=330; 9.41 ± 3.02 vs 15.67 ± 2.63, P<0.001). The natural rate of conception within 1 year was lower (60.0% [63/105] vs 72.1% [230/319], P=0.020), and the spontaneous abortion rate in early pregnancy was higher (34.4% [21/61] vs 18.1% [41/227], P=0.006) in the study group than in the control group. Women with light menstrual bleeding of unidentified etiology had an increased prevalence of low ER and poor conception outcome.


Translation, cross-cultural adaptation and validation of the sickle cell self-efficacy scale (SCSES).
To translate, cross-culturally adapt and validate the Sickle Cell Self-Efficacy Scale for application in the Brazilian cultural context. This is a methodological study performed in 6 steps: 1- Forward translation; 2- Translation synthesis; 3- Back-translation; 4- Assessment by expert committee, with computation of the Content Validity Index (CVI); 5- Cultural adequacy (pre-test); 6- Reproducibility. A pre-test was performed with the participation of 10 adolescents/young adults with sickle cell disease through a telephone call and their responses were recorded on a form in a web platform. The instrument validation step was carried out with 55 adolescents/young adults with sickle cell disease, 43 of them having participated in the retest. The analysis of internal consistency and reproducibility was calculated using the Cronbach's alpha coefficient and the Intraclass Correlation Coefficient (ICC), in the R statistical programming environment. The translated instrument had good acceptance among the experts, reaching an average CVI of 1.0. In assessing reliability, the scale showed acceptable internal consistency, with a Cronbach's alpha of 0.84. In the agreement analysis, the ICC was 0.923 (95% CI: 0.857 to 0.958), which indicates good temporal reproducibility. Following the process of translation, cross-cultural adaptation and validation, we obtained the Brazilian version of the Sickle Cell Self-Efficacy Scale, considered valid and reliable to be applied to adolescents and young adults with sickle cell disease in Brazil.


HLA class I antigen (heavy and light chain) expression by Langerhans cells and keratinocytes of the normal human epidermis: ultrastructural quantitation using immunogold labelling procedure.
Using an immunogold labelling procedure, we quantified the density of major histocompatibility (MHC) class I antigens on the surface of Langerhans cells (LCs) and keratinocytes of the normal human epidermis. According to ultrastructural features, keratinocytes were divided into three subpopulations: stratum basalis (SBK), stratum spinosum (SSK), and stratum granulosum keratinocytes (SGK), and analyzed separately. For this purpose, three monoclonal antibodies (MCAs) were employed: an anti-HLA A,B,C, and anti-B2-microglobulin (B2-m), and a polymorphic anti-HLA A2 Aw69 MCA. Under electron microscopy, quantitative analysis demonstrated: (a) the presence of a high amount of HLA monomorphic determinants on SBK and SSK and moderate but significant labelling of SGK; (b) the very weak density of MHC class I antigens on the surface of epidermal LCs; (c) the expression, at an identical level, of the HLA heavy chain common determinant (HLA A,B,C), B2-m, and the alloantigen HLA A2 by all epidermal cells (ECs) apart from SGKs and LCs that presented far fewer HLA A2 sites than monomorphic determinants (B2-m and HLA A,B,C); (d) the absence of HLA class I on corneocytes and a moderate labelling of melanocytes. A knowledge of the precise quantitative distribution of HLA class I antigens among various cell subpopulations of the normal human epidermis would be very useful for the study and follow-up of cutaneous malignancies that are known to lose these molecules as well as for the understanding of immune responses, especially allospecific, that involve the skin.


Characterization of neuropathology in the HIV-1 transgenic rat at different ages.
The transgenic HIV-1 rat (Tg) is a commonly used neuroHIV model with documented neurologic/behavioral deficits. Using immunofluorescent staining of the Tg brain, we found astrocytic dysfunction/damage, as well as dopaminergic neuronal loss/dysfunction, both of which worsening significantly in the striatum with age. We saw mild microglial activation in young Tg brains, but this decreased with age. There were no differences in neurogenesis potential suggesting a neurodegenerative rather than a neurodevelopmental process. Gp120 CSF levels exceeded serum gp120 levels in some animals, suggesting local viral protein production in the brain. Further probing of the pathophysiology underlying astrocytic injury in this model is warranted.


Effect of combined treatment with sodium hypochlorite and alpha-tocopherol on prooxidant and antioxidant system of the blood during experimental bile peritonitis.
Combined postoperation treatment with sodium hypochlorite and alpha-tocopherol broke the chain of free radical reactions in lipids and promoted normalization of LPO processes in the plasma and erythrocytes of 250 rats with experimental bile peritonitis.


Achromatic contrast effects in infants: adults and 4-month-old infants show similar deviations from Wallach's ratio rule.
When adults view a disk of light embedded in a higher luminance surround, the perceived lightness of the disk is largely determined by the surround to disk luminance ratio (Wallach's ratio rule). In the present study, both adult and infant subjects were tested with multiple discrete trial procedures in which the surround luminance was decreased between the study and test phases of each trial. Tested with sequential lightness matching, adult subjects showed an approximate ratio rule, with a small but consistent deviation in the direction of a luminance match. Tested with a forced-choice novelty preference technique in combination with a cross-familiarization paradigm, 4-month-old infants showed preference minima that fell closer to the mean adult match than to the ratio rule. This finding suggests that, at least for a relatively simple visual display, 4-month-old infants' looking preferences are governed by an adult-like achromatic contrast system.


Standardized Added Metabolic Activity Predicts Survival After Intra-arterial Resin-Based 90Y Radioembolization Therapy in Unresectable Chemorefractory Metastatic Colorectal Cancer to the Liver.
Standardized added metabolic (SAM) activity is a functional objective measurement of the total tumoral metabolic activity that avoids partial volume effect and thresholding, which limit conventional PET parameters. The purpose of this study is to investigate the role of SAM in predicting survival in unresectable, chemorefractory colorectal hepatic metastatic disease treated with resin-based Y radioembolization. This is a prospective correlative study of patients with unresectable, chemorefractory colorectal liver metastasis who underwent F-FDG PET/CT and CT/MRI before and after Y. Target RECIST, PERCIST, change in total glycolytic activity (ΔTGA), and ΔSAM treatment response were assessed. Percentage changes in diameter, SUVpeak, TGA, and SAM were calculated pre- and post-Y therapy and objective response was defined as >30% change (responders). Survival analysis by Kaplan-Meier, log-rank, and Cox proportional hazard models were performed and significance was set at <0.05. Sixteen patients (mean age of 61.6) were enrolled and performed a total of 20 Y therapies. After Y, target ΔSAM showed an objective response rate of 40% vs. 35%, 30%, and 22.2% based on target ΔTGA, PERCIST, and RECIST criteria, respectively. Median overall survival (OS) of the cohort after Y was 9.2 months (CI 95% 2.2-16.2). Patients demonstrating objective response based on ΔSAM had a median OS of 22.7 months (CI 95% 12.4-33.0) vs. 6.7 (CI 95% 4.2-9.2) in non-responders (P = 0.007). On multivariate analysis, hazard ratios for the objective response group based on target ΔSAM were 0.01 (P = 0.03) vs. 0.05 (P = 0.08), 0.20 (P = 0.29), and 0.91 (P = 0.98) based on target ΔTGA, PERCIST, and RECIST criteria, respectively. In unresectable colorectal liver metastatic disease refractory to standard chemotherapy, ΔSAM predicted OS for assessment of response following Y radioembolization therapy, whereas RECIST, PERCIST, and ΔTGA did not.


Differential diagnostic value of tumor markers and contrast-enhanced computed tomography in gastric hepatoid adenocarcinoma and gastric adenocarcinoma.
This study aimed to investigate the effectiveness of tumor markers and contrast-enhanced computed tomography (CE-CT) in differentiating gastric hepatoid adenocarcinoma (GHA) from gastric adenocarcinoma (GA). This retrospective study included 160 patients (44 with GHA vs. 116 with GA) who underwent preoperative CE-CT. Preoperative serum concentrations of tumor biomarkers and CT imaging features were analyzed, including alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 125 (CA125), tumor location, growth pattern, size, enhancement pattern, cystic changes, and mass contrast enhancement. Multivariate logistic regression analyses were performed to evaluate useful tumor markers and CT imaging features for differentiating GHA from GA. When compared to GA, GHA showed a higher serum AFP [13.27 ng/ml (5.2-340.1) vs. 2.7 ng/ml (2.2-3.98), P <0.001] and CEA levels [4.07 ng/ml (2.73-12.53) vs. 2.42 ng/ml (1.38-4.31), P <0.001]. CT imaging showed GHA with a higher frequency of tumor location in the gastric antrum (P <0.001). GHA had significantly lower attenuation values at the portal venous phase [PCA, (82.34 HU ± 8.46 vs. 91.02 HU ± 10.62, P <0.001)] and delayed phase [DCA, (72.89 HU ± 8.83 vs. 78.27 HU ± 9.51, P <0.001)] when compared with GA. Multivariate logistic regression analyses revealed that tumor location, PCA, and serum AFP level were independent predictors of differentiation between GHA and GA. The combination of these three predictors performed well in discriminating GHA from GA, with an AUC of 0.903, a sensitivity of 86.36%, and a specificity of 81.90%. Integrated evaluation of tumor markers and CT features, including tumor location, PCA, and serum AFP, allowed for more accurate differentiation of GHA from GA.


Efficient dynamic kinetic resolution of racemic secondary alcohols by a chemoenzymatic system using bifunctional iridium complexes with C-N chelate amido ligands.
The combined catalyst system of bifunctional amidoiridium complexes derived from benzylic amines with CALB was found to provide a range of chiral acetates from racemic secondary alcohols in excellent yields with nearly perfect enantioselectivities via dynamic kinetic resolution.


Detection and characterization of methicillin-resistant Staphylococcus pseudintermedius in healthy dogs in La Rioja, Spain.
The objective was to identify the methicillin-resistant coagulase-positive staphylococci (MRCoPS) nasal carriage rate of healthy dogs in La Rioja (Spain) and to characterize the recovered isolates by different molecular techniques. Nasal samples from 196 dogs were obtained (98 household-dogs, 98 pound-dogs). Isolates were identified and characterized by spa-, SCCmec- and MLST-typing, SmaI-PFGE, antimicrobial susceptibility, determination of antimicrobial resistance and toxin genes profiling. S. pseudintermedius was the only species recovered. Nine methicillin-resistant S. pseudintermedius (MRSP) were obtained from 9 of 196 sampled dogs (8% pound-dogs, 1% household-dogs). MRSP isolates were typed (MLST/PFGE/spa/SCCmec) as: ST71/A/t02/II-III (7 isolates), ST92/C/t06/V (1 isolate), and ST26/B/non-typable/non-typable (1 isolate). All MRSP were resistant to [resistance gene/number isolates]: β-lactams [mecA+blaZ/9], tetracycline [tet(K)/7, tet(M)/2], macrolides and lincosamides [erm(B)/9], aminoglycosides [aacA-aphD+aadE+aphA-3/9], and co-trimoxazol [dfr(G)/9]. Eight MRSP isolates showed also resistance to fluoroquinolones and amino acid changes in GyrA [Ser84Leu+Glu714Lys, 7 isolates; Ser84Leu, 1 isolate] and GrlA [Ser80Ile, 8 isolates] proteins were detected. The remaining isolate was chloramphenicol resistant and harboured cat(pC221) gene. All MRSP isolates harboured the aadE-sat4-aphA-3 multiresistance-gene-cluster linked to erm(B) gene as well as the siet, si-ent and lukS/F-I toxin genes. MRSP is a moderately common (4.6%) colonizer of healthy dogs in Spain. A major MRSP lineage (ST71) was detected and its future evolution should be tracked.


Morphological and biochemical properties of a Sphaerotilus sp. Isolated from paper mill slimes.
Four strains of filamentous bacteria were isolated from slimes collected in different paper mill factories. Morphological and physiological characterization of the isolates indicated an affiliation with the genus Sphaerotilus. However, while the physiological properties of the isolates were almost identical, pronounced physiological differences between the isolates and Sphaerotilus natans DSM 6575(T), DSM 565, and DSM 566 with respect to their ability to metabolize complex polysaccharides, sugars, polyalcohols, or organic acids as carbon sources were detected. In contrast to the analyzed culture collection strains of S. natans, all paper mill isolates were able to grow at elevated temperatures of up to 40 degrees C. Comparative sequence analysis of nearly complete 16S ribosomal DNA (rDNA) sequences from the four new isolates demonstrated that the retrieved sequences were highly similar to each other (99.6 to 99.8% similarity) and to previously published partial 16S rDNA sequences of S. natans DSM 6575(T) and ATCC 15291. Polyphasic characterization of the isolated Sphaerotilus strains revealed interesting adaptations of the strains to the environmental paper mill conditions with regard to temperature tolerance and utilization of cellulose and starch.


[Melanosis of the urinary bladder].
Melanosis of the bladder is rare. Only 10 cases have been described in the literature. We present the case of an 80-year-old woman with neurogenic lower urinary tract dysfunction due to spinal paralysis. During the diagnostic work-up which included cystoscopy, black spots in the bladder wall were observed. Histopathological evaluation revealed a benign suburothelial melanosis. Thus, with cystoscopic suspicion of a malignancy (melanoma), a biopsy is mandatory and regular cystoscopic follow-up is recommended.


Hirmi Valley liver disease: a disease associated with exposure to pyrrolizidine alkaloids and DDT.
Hirmi Valley liver disease was first reported in 2001 in Tigray, Ethiopia. 591 cases, including 228 deaths, were reported up to December 2009. The pyrrolizidine alkaloid acetyllycopsamine was detected in stored grain and residents reported adding the pesticide DDT (dichlorodiphenyldichloroethylene) directly to their food stores. We aimed to characterise the clinical features of the disease, and explore the role of these chemicals in its aetiology. 32 cases were examined and full clinical histories taken. Nine cases underwent liver biopsy in hospitals. Serum and urine samples were collected from cases and controls. Urine was analysed for acetyllycopsamine by UPLC-MS. Total DDT in serum was measured by ELISA. Hepatotoxicity of DDT and acetyllycopsamine alone or in combination was explored in C57BL/6J mice. Clinical presentation included epigastric pain, abdominal swelling, bloody diarrhoea, hepatomegaly, splenomegaly, and ascites. Histology revealed acute injury characterised by centrilobular necrosis or chronic injury with bile ductular reaction, cytomegaly and fibrosis but no hepatic vein occlusion. Acetyllycopsamine was detected in urine samples taken in the affected area with significantly greater concentrations in 45 cases than in 43 controls (p=0.02). High levels of DDT (>125 ppb) were detected in 78% of serum samples. In mice, DDT (3 × 75 mg/kg) significantly increased the hepatotoxicity (plasma ALT, p=0.0065) of acetyllycopsamine (750 mg/kg), and in combination induced liver pathology similar to Hirmi Valley liver disease including centrilobular necrosis and cytomegaly. This novel form of disease appears to be caused by co-exposure to acetyllycopsamine and DDT.


MRTX1719 Is an MTA-Cooperative PRMT5 Inhibitor That Exhibits Synthetic Lethality in Preclinical Models and Patients with MTAP-Deleted Cancer.
Previous studies implicated protein arginine methyltransferase 5 (PRMT5) as a synthetic lethal target for MTAP-deleted (MTAP del) cancers; however, the pharmacologic characterization of small-molecule inhibitors that recapitulate the synthetic lethal phenotype has not been described. MRTX1719 selectively inhibited PRMT5 in the presence of MTA, which is elevated in MTAP del cancers, and inhibited PRMT5-dependent activity and cell viability with >70-fold selecti-vity in HCT116 MTAP del compared with HCT116 MTAP wild-type (WT) cells. MRTX1719 demonstrated dose-dependent antitumor activity and inhibition of PRMT5-dependent SDMA modification in MTAP del tumors. In contrast, MRTX1719 demonstrated minimal effects on SDMA and viability in MTAP WT tumor xenografts or hematopoietic cells. MRTX1719 demonstrated marked antitumor activity across a panel of xenograft models at well-tolerated doses. Early signs of clinical activity were observed including objective responses in patients with MTAP del melanoma, gallbladder adenocarcinoma, mesothelioma, non-small cell lung cancer, and malignant peripheral nerve sheath tumors from the phase I/II study. PRMT5 was identified as a synthetic lethal target for MTAP del cancers; however, previous PRMT5 inhibitors do not selectively target this genotype. The differentiated binding mode of MRTX1719 leverages the elevated MTA in MTAP del cancers and represents a promising therapy for the ∼10% of patients with cancer with this biomarker. See related commentary by Mulvaney, p. 2310. This article is featured in Selected Articles from This Issue, p. 2293.


Impact of personal protective equipment on prehospital endotracheal intubation performance in simulated manikin.
Tracheal intubation in COVID-19 patients is a potentially high-risk procedure for healthcare professionals. Personal protective equipment (PPE) is recommended to minimize contact with critical patients with COVID-19 infection. This study aimed to primarily examine the effect of PPE use on intubation time and success rate among prehospital healthcare professionals; additionally, we compared intubation times among prehospital health care professionals using PPE with direct laryngoscopy and video laryngoscopy assistance. In this prospective simulation study, we compared the intubation times and success rates among prehospital healthcare professionals who were or were not using PPE. Furthermore, demographic data, previous intubation experience, and previous intubation experience with PPE were recorded. Overall time to intubation with PPE use was 51.28±3.89s, which was significantly higher than that without PPE use (33.03±2.65s; p<0.001). In addition, the overall success rate with PPE use was 74.4%, which was significantly lower than that without PPE use (93%;p<0.001). PPE use increased the average intubation time by 19.73±2.59s with direct laryngoscopy and by 16.81±2.86s with video laryngoscopy (p<0.001). PPE use is associated with increased intubation time and decreased success rate. Video laryngoscopy assistance in cases where PPE use is required facilitates faster endotracheal intubation than does direct laryngoscopy assistance.


Independent protective effects for tumor necrosis factor and lymphotoxin alpha in the host response to Listeria monocytogenes infection.
Although the essential role of tumor necrosis factor (TNF) in resistance to Listeria monocytogenes infection is well established, the roles of the related cytokines lymphotoxin alpha (LTalpha) and lymphotoxin beta (LTbeta) are unknown. Using C57BL/6 mice in which the genes for these cytokines were disrupted, we examined the contributions of TNF, LTalpha, and LTbeta in the host response to Listeria. To overcome the lack of peripheral lymph nodes in LTalpha(-/-) and LTbeta(-/-) mice, bone marrow chimeras were constructed. TNF(-/-) and LTalpha(-/-) chimeras that lacked both secreted LTalpha(3) and membrane-bound LTalpha(1)beta(2) and LTalpha(2)beta(1) were highly susceptible and succumbed 4.5 and 6 days, respectively, after a low-dose infection (200 CFU). LTbeta(-/-) chimeras, which lacked only membrane-bound LT, controlled the infection in a manner comparable to wild-type (WT) chimeras. The Listeria-specific proliferative and gamma interferon T-cell responses were equivalent in all five groups of infected mice (LTalpha(-/-) and LTbeta(-/-) chimeras, WT chimeras, and TNF(-/-) and WT mice). TNF(-/-) mice and LTalpha(-/-) chimeras, however, failed to generate the discrete foci of lymphocytes and macrophages that are essential for bacterial elimination. Rather, aberrant necrotic lesions comprised predominantly of neutrophils with relatively few lymphocytes and macrophages were observed in the livers and spleens of TNF(-/-) and LTalpha(-/-) chimeras. Therefore, in addition to TNF, soluble LTalpha(3) plays a separate essential role in control of listerial infection through control of leukocyte accumulation and organization in infected organs.


Exploring the impacts of street-level greenspace on stroke and cardiovascular diseases in Chinese adults.
In recent years, cardiovascular diseases (CVDs) have become the primary cause of death in the world. Existing studies have found that greenspace is important for the prevention of CVDs and stroke. However, since they only focus on large green infrastructure (e.g., urban parks) or the general greenspace (usually being evaluated through normalized difference vegetation index), little information exists regarding the association between street-level greenspace and CVDs (stroke). In this study, the CVDs and stroke data of participants were retrieved from the 33 Chinese Community Health Study. We measured participants' exposure to street-level greenspace exposure using street view images and machine learning technique. Multilevel logistic regressions were applied. While controlling for confounders, we found that higher level of street-level greenspace exposure was associated with lower CVDs prevalence. However, street-level greenspace exposure was associated with stroke prevalence only for females. The associations were stronger among females, younger adults, participants with educational attainment above high school, physically active participants and participants who were not overweight. None of the mediators (air pollution, physical exercise, and BMI) can explain the associations between street-level greenspace exposure and CVDs (stroke) prevalence. Our findings suggest that street-level vegetation should be increased to cope with the rapid growth of the CVDs burdens. Also, the differences between the effect of street-level trees and grasses should be noted before formulating specific urban planning policies.


Impaired lymphocyte blastogenic response in patients with colon adenocarcinoma: effects of disease and age.
The in vitro blastogenic response of 76 patients with colon adenocarcinoma and 29 age-matched normal healthy volunteers was compared. An age-related decline in cellular immunocompetence was observed in both the patient and control group. Within each age interval, the patients' blastogenic responses were lower than the controls' and the rate of decline of the blastogenic response in the patient group was greater in magnitude, suggesting that their disease state also contributed to loss of cellular immunocompetence. Thus, both increasing age and presence of tumor are factors that contribute to the decline the immunocompetence in patients with colon adenocarcinoma.


Severe combined respiratory and myocardial failure treated with high-frequency ventilation.
High levels of positive end-expiratory pressure (PEEP) impair cardiac output. The subsequent lowering of mixed venous oxygenation, when coupled with a significant intrapulmonary shunt, may dramatically depress PaO2. We present a patient whose severe myocardial and respiratory insufficiency was unmanageable on conventional ventilation with high levels of PEEP and maximal inotropic support. High-frequency ventilation superimposed on conventional ventilation lowered peak airway pressure and dramatically improved both cardiac and pulmonary function.


Injectable silk fibroin/polyurethane composite hydrogel for nucleus pulposus replacement.
In degenerative disc disease, an injectable hydrogel can fill a degenerate area completely, reduce the risk of implant migration and subsequent loss of height of the intervertebral disc, and minimise surgical defects. Here, we propose a method of preparing an injectable silk fibroin/polyurethane (SF/PU) composite hydrogel by chemical cross-linking under physiological conditions. Mechanical testing was used to determine the mechanical strength of the hydrogel. The impact of hydrogel height on the biomechanical properties was discussed to estimate the working capacity of the hydrogel for further clinical application. Rheological properties were also examined to assess the practical ability of the hydrogel for clinical application. Hydrogel injection and cell assessment is also of interest for clinical application. An SF/PU composite hydrogel can be injected through a small incision. A cell proliferation assay using bone marrow stromal cells showed positive cell viability and increased proliferation over a seven-day period in culture. Importantly, the hydrogel can be monitored in real-time using X-ray fluoroscopy during and after surgery according to the results of X-ray fluoroscopy examination, and shows good visibility based on X-ray assays. In particular, the hydrogel offers the clinically important advantage of visibility in CT and T2-weighted magnetic resonance imaging. Based on the results of the current study, the SF/AU composite hydrogel may offer several advantages for future application in nucleus pulposus replacement.


Merkel Cell Carcinoma of the Maxillary Sinus: An Unusual Presentation of a Common Tumor.
Merkel cell carcinoma is most commonly seen in the skin of sun exposed areas, particularly the head and neck and is associated with Merkel cell polyomavirus. Merkel cell carcinoma at an extracutaneous mucosal site of the head and neck is rare. We report a case of a 74-year-old women who presented with an enlarging thyroid mass found to be neuroendocrine carcinoma consistent with Merkel cell carcinoma (positive for synaptophysin, chromogranin, CK20). Subsequent work up revealed a maxillary sinus mass with extension into the nasal cavity. Biopsy was diagnostic for Merkel cell carcinoma (positive for synaptophysin, chromogranin, CK20 and Merkel cell polyomavirus). There are only case reports and small case series of Merkel cell carcinoma arising in the mucosal sites of the head and neck most commonly in the oral cavity, rarely the sinonasal mucosa. Merkel cell carcinoma metastasizing to the thyroid has only been reported in three other case reports, all from skin primaries. In addition to our case, we review the literature of extracutaneous sinonasal Merkel cell carcinoma and metastases to the thyroid.


Formation of a new O-polysaccharide in Escherichia coli O86 via disruption of a glycosyltransferase gene involved in O-unit assembly.
The majority of hetero-polysaccharide biosynthesis in Gram-negative bacteria utilizes the wzy-dependent pathway, in which repeating O-units are first synthesized in the cytosol and then subsequently translocated to the periplasmic face of the inner membrane where polymerization is initiated by the Wzy polymerase. Wzy proteins share little primary sequence homology and are specific for their cognate O-unit structures. Our previous studies on O-polysaccharide biosynthesis in Escherichia coli O86 identified the wbnI gene, which encodes a galactosyltransferase responsible for the introduction of alpha-(1-->3)-Galp residues as side chains of the polysaccharide. In this work, we functionally inactivated the wbnI gene and showed that the mutant strain produced a different polysaccharide without the side chain Galp residue. The yield of the polysaccharide was substantially lower than the one produced by the wild-type strain. This study indicated that the complete O-unit structure is the preferred substrate for the polymerization, thus further confirming the specificity of Wzy. On the other hand, these studies also suggest that the Wzy polymerase might have moderate tolerance of side-chain truncated O-unit substrates.


Automated on-line in-tube solid-phase microextraction-assisted derivatization coupled to liquid chromatography for quantifying residual dimethylamine in cationic polymers.
A method for the analysis of dimethylamine (DMA) by automated in-tube solid-phase microextraction (IT-SPME)-supported chemical derivatization coupled with high-performance liquid chromatography was developed. Extraction, derivatization and desorption were studied by using a capillary coated with 95% polydimethylsiloxane and 5% polydiphenylsiloxane. Solution derivatization and automated IT-SPME derivatization using 9-fluorenylmethyl chloroformate (FMOC) were compared. The proposed procedure provided adequate linearity, accuracy and precision in the 0.2-2.0 microg/mL concentration interval, and the limit of detection (LOD) was 50 ng/mL. The main advantages of the proposed procedure are: (i) no off-line sample manipulation, (ii) rapidity, as the total analysis time is about 10 min, (iii) specificity for the samples assayed, (iv) minimal consumption of FMOC reagent and (v) minimal residues. Therefore, the proposed method is an environmental-friendly and cost-effective alternative for the control of residual DMA in polymeric cationic surfactants used like flocculants in water treatment.


Working memory performance and thalamus microstructure in healthy subjects.
Research on the neural basis of working memory (WM) has generally focused on cortical regions, specifically frontal and parietal areas. Comparatively, evidence of a possible involvement of deep gray matter structures, that are parts of cortico-cortical circuits linking anterior and posterior cortical areas, is far less clear. The goal of the present study is to test the hypothesis that individual structural variations within deep gray matter structures may affect the cortical networks involved in WM. To this aim, a large sample (n=181) of healthy subjects underwent a high-resolution structural magnetic resonance imaging (MRI) and a diffusion tensor imaging (DTI) scan protocol. Data of micro- (mean diffusivity, MD) and macro- (volume) structural variations of six bilateral deep gray matter structures (thalamus, caudate nucleus, putamen, hippocampus, amygdala and pallidum) and lateral ventriculi volume were analyzed in association with score in a WM (the so-called n-back task) and other neuropsychological tasks. Results showed that increased MD of bilateral thalami was the only structural parameter that significantly correlated with reduced WM performance. In particular, a voxel-by-voxel analysis revealed that the greater percentage of voxels showing significant anticorrelation between WM score and MD values were localized in those thalamic nuclei projecting to prefrontal and posterior parietal cortices. Results highlight the specific involvement of thalamus microstructure, not volume, in modulating WM performance, possibly by regulating the connections among cortical areas that are recruited during WM tasks.


Long-Term Blood Pressure Variability Across the Clinical and Biomarker Spectrum of Alzheimer's Disease.
Elevated blood pressure is linked to cognitive impairment and Alzheimer's disease (AD) biomarker abnormality. However, blood pressure levels vary over time. Less is known about the role of long-term blood pressure variability in cognitive impairment and AD pathophysiology. Determine whether long-term blood pressure variability is elevated across the clinical and biomarker spectrum of AD. Alzheimer's Disease Neuroimaging Initiative participants (cognitively normal, mild cognitive impairment, AD [n = 1,421]) underwent baseline exam, including blood pressure measurement at 0, 6, and 12 months. A subset (n = 318) underwent baseline lumbar puncture to determine cerebrospinal fluid amyloid-β and phosphorylated tau levels. Clinical groups and biomarker-confirmed AD groups were compared on blood pressure variability over 12 months. Systolic blood pressure variability was elevated in clinically diagnosed AD dementia (VIM: F2,1195 = 6.657, p = 0.001, η2 = 0.01) compared to cognitively normal participants (p = 0.001), and in mild cognitive impairment relative to cognitively normal participants (p = 0.01). Findings were maintained in biomarker-confirmed AD (VIM: F2,850 = 5.216, p = 0.006, η2 = 0.01), such that systolic blood pressure variability was elevated in biomarker-confirmed dementia due to AD relative to cognitively normal participants (p = 0.005) and in biomarker-confirmed mild cognitive impairment due to AD compared to cognitively normal participants (p = 0.04). Long-term systolic blood pressure variability is elevated in cognitive impairment due to AD. Blood pressure variability may represent an understudied aspect of vascular dysfunction in AD with potential clinical implications.


A modal theory of discrimination.
Discrimination is a central epistemic capacity but typically, theories of discrimination only use discrimination as a vehicle for analyzing knowledge. This paper aims at developing a self-contained theory of discrimination. Internalist theories of discrimination fail since there is no compelling correlation between discriminatory capacities and experiences. Moreover, statistical reliabilist theories are also flawed. Only a modal theory of discrimination is promising. Versions of sensitivity and adherence that take particular alternatives into account provide necessary and sufficient conditions on discrimination. Safety in contrast is not sufficient for discrimination as there are cases of safety that are clearly instances of discrimination failure. The developed account of discrimination between objects will be extended to discrimination between kinds and between types.


Substructure and higher structure of chicken smooth muscle alpha-actinin molecule.
Substructure of chicken gizzard smooth muscle alpha-actinin molecule was deduced by domainal mapping of the proteolytic fragments with alpha-chymotrypsin. There were three chymotryptic cleavage sites (Sites I, II, and III, from the amino terminus). Cleavage at Site I generated two fragments, i.e. an NH2-terminal 36-kDa fragment and a COOH-terminal 70-kDa fragment. The 70-kDa fragment generated either a 55-kDa fragment by cleavage at Site II or a 65-kDa fragment by cleavage at Site III. Purified NH2-terminal 36-kDa fragment bound to F-actin, whereas the 55-kDa fragment formed a dimeric molecule. Circular dichroism and electron microscopic experiments demonstrated that the alpha-helical content of the 55-kDa fragment was 14% higher than that of native gizzard alpha-actinin, coinciding with the apparently rod-shaped configuration of this fragment. A 110-kDa product was generated from two 55-kDa fragments in a cross-linking study with the zero-length cross-linker 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. Two cross-linkable sites in the 55-kDa, A- and B-site, were shown to be involved in this reaction. Further, it was demonstrated by using N-(7-dimethylamino-4-methyl-3-coumarinyl)maleimide labeling and immunoblotting analyses that the A-site on one 55-kDa fragment was cross-linked to the B-site on the other. These results suggest that smooth muscle alpha-actinin formed an antiparallel dimeric molecule in which the 55-kDa fragments connected the two actin-binding domains composed of the 36-kDa fragments.


Recurrent cystitis in post-menopausal patients more or less than 65 years of age.
We conducted a prospective monocentric study to confirm the difference in clinical, biological, bacteriological, and radiological presentations of patients with recurrent cystitis in patients more or less than 65 years of age. We analyzed the characteristics of post-menopausal patients with recurrent cystitis. We compared patients less than 65 years of age to patients more than 65 years of age. Two hundred and sixty-eight patients were included. No statistical difference was found between the two groups apart from sexual activity that was superior in younger patients. Clinical and bacteriological data were similar in both groups. The analysis of a series of post-menopausal patients with recurrent cystitis did not reveal any difference in clinical, radiological, or biological presentation whether patients were more or less than 65 years of age. Taking this age threshold for the treatment of recurrent cystitis does not seem validated according to our results.


Rate of adjudication of radiological progression in rheumatoid arthritis randomized controlled trials depending on preset limits of agreement: a pooled analysis from 15 randomized trials.
The aim of this study is to provide data on the adjudication rate for a predetermined threshold of difference in change score between two readers in randomized controlled trials (RCTs). Fifteen datasets from RCTs in RA were scored by 13 experienced readers as pairs according to the modified Sharp-van der Heijde method. The theoretical adjudication rates for thresholds of between 3 and 20 units were calculated. We investigated the influence of the number of time points within the same session, the length of the interval and disease duration on the adjudication rates. A total of 21,295 time points from 7643 patients from 15 databases were included in the analysis. The adjudication rate was inversely related to the threshold. Higher adjudication rates were observed with a higher number of time points, longer time intervals and in early versus established RA. The adjudication rates ranged from 0% to 22% depending on the scenario. With trained and experienced readers, the adjudication rate in RA RCTs is low even with very conservative adjudication thresholds.


Retroviral gene transfer of interferon-inducible protein 10 inhibits growth of human melanoma xenografts.
Interferon-inducible protein 10 (IP-10) is an immunomodulatory chemokine recently recognized to have potent antiangiogenic activity in vivo. Due to difficulties in the stability, manufacture and chronic administration of recombinant forms of endogenous antiangiogenic proteins, antiangiogenic gene therapy has emerged as a promising new form of cancer treatment. We retrovirally transduced A375 human melanoma cells with the human IP-10 gene and injected cells subcutaneously into nude mice. IP-10-transduced cells also were mixed with null-transduced cells in varying proportions before injection. In vivo growth of IP-10-transduced melanoma cells was markedly diminished compared to parental or null-transduced cells (p = 0.0002, Kruskal-Wallis test). This growth inhibition was associated with a marked reduction in microvessel density. The degree of growth inhibition of tumors following injection of a mixed population of null- and IP-10-transduced cells was directly associated with the fraction of IP-10-transduced cells present. We conclude that retroviral transduction of human melanoma cells with the IP-10 gene leads to sufficient protein secretion to inhibit angiogenesis and tumor growth. These findings suggest that IP-10 gene therapy might be an effective therapy in patients with cancer.


Is partial nephrectomy worth performing compared to radical nephrectomy for small, localised renal cortical tumours in geriatric patients?
This study aimed to evaluate the likelihood of progression to chronic kidney disease (CKD) in a cohort of geriatric Asian patients who underwent either partial nephrectomy (PN) or radical nephrectomy (RN) for localised pT1 kidney tumours. Geriatric patients aged > 65 years who underwent PN or RN from 2005 to 2014 for localised kidney masses < 7 cm at a single institution were reviewed retrospectively. Preoperative and postoperative estimated glomerular filtration rates (eGFRs) were calculated using the Modification of Diet in Renal Disease Study equation. Clinicopathological data was analysed for new-onset CKD after at least five years of follow-up. Postoperative complications were measured using the modified Clavien classification system. Among 67 patients, 36.4% had diabetes mellitus and 80.3% had hypertension. Pre-existing CKD with eGFR ≤ 60 mL/min/1.73 m2 was present in 23.3% of PN and 43.2% of RN patients. RN and PN patients had similar postoperative morbidity. Those who underwent PN had immediate decline followed by gradual improvement of kidney function at six months after surgery. This improvement was sustained even for patients with four years of clinical follow-up. Among patients with normal preoperative kidney function, new-onset CKD after surgery was found in 20.0% and 52.9% of PN and RN patients, respectively (p = 0.007). In our study, geriatric patients who underwent PN for localised kidney mass < 7 cm had immediate decline but gradual and sustained improvement in kidney function. PN also resulted in less new-onset CKD through a five-year follow-up period.


A comparison of empiric to physician-tailored programming of implantable cardioverter-defibrillators: results from the prospective randomized multicenter EMPIRIC trial.
The purpose of this randomized study was to determine whether a strategically chosen standardized set of programmable settings is at least as effective as physician-tailored choices, as measured by the shock-related morbidity of implantable cardioverter-defibrillator (ICD) therapy. Programming of ventricular tachyarrhythmia (ventricular tachycardia [VT] or ventricular fibrillation [VF]) detection and therapy for ICDs is complex, requires many choices by highly trained physicians, and directly influences the frequency of shocks and patient morbidity. A total of 900 ICD patients were randomly assigned to standardized (EMPIRIC, n = 445) or physician-tailored (TAILORED, n = 455) VT/VF programming and followed for 1 year. The primary end point was met: the adjusted percentages of both VT/VF (22.3% vs. 28.7%) and supraventricular tachycardia or other non-VT/VF event episodes (11.9% vs. 26.1%) that resulted in a shock were non-inferior and lower in the EMPIRIC arm compared to the TAILORED arm. The time to first all-cause shock was non-inferior in the EMPIRIC arm (hazard ratio = 0.95, 90% confidence interval 0.74 to 1.23, non-inferiority p = 0.0016). The EMPIRIC trial had a significant reduction of patients with 5 or more shocks for all-cause (3.8% vs. 7.0%, p = 0.039) and true VT/VF (0.9% vs. 3.3%, p = 0.018). There were no significant differences in total mortality, syncope, emergency room visits, or unscheduled outpatient visits. Unscheduled hospitalizations occurred significantly less often (p = 0.001) in the EMPIRIC arm. Standardized empiric ICD programming for VT/VF settings is at least as effective as patient-specific, physician-tailored programming, as measured by many clinical outcomes. Simplified and pre-specified ICD programming is possible without an increase in shock-related morbidity.


Proliferative effects of eosinophil lysates on cultured human airway smooth muscle cells.
The hypertrophy/hyperplasia of airway smooth muscle (ASM) cells is one of the characteristic features of bronchial asthma. This structural change leads to the thickening of airway walls resulting in the amplification of airway narrowing. However, the pathogenesis of this structural change has not yet been determined. Eosinophils, which play a pathogenic role in asthma, have been demonstrated to have proliferative effects on fibroblasts and vascular smooth muscle cells. We attempted to investigate the potential of eosinophils to induce the proliferation of ASM cells. We examined the effect of lysates of eosinophils purified from peripheral blood of healthy donors on cultured human ASM cell proliferation. Eosinophil lysates significantly induced ASM cell proliferation in time- and dose-dependent manners, reaching a maximum on day 6 at 50% of eosinophil lysates (6.0 +/- 0.7 x 104 [mean +/- SD] /well, n = 5 vs. 4.5 +/- 1.1 x 104/well, n = 5; P < 0.05). This proliferative activity was heat-sensitive and recovered in the soluble fraction of the eosinophil lysates. Furthermore, the molecular weight of the mitogenic activity in the soluble fraction was identified as lower than 10 kDa. The inhibitory activity to ASM cell proliferation was also found in the insoluble fraction of the lysates. These results indicate that circulating eosinophils store mitogenic activity for ASM cells, suggesting that eosinophils might contribute to the development of the hyperplasia of ASM cells in asthmatics through the release of the stored mitogenic activity upon stimulation at the site of inflammation.


Retrograde BDNF to TrkB signaling promotes synapse elimination in the developing cerebellum.
Elimination of early-formed redundant synapses during postnatal development is essential for functional neural circuit formation. Purkinje cells (PCs) in the neonatal cerebellum are innervated by multiple climbing fibers (CFs). A single CF is strengthened whereas the other CFs are eliminated in each PC dependent on postsynaptic activity in PC, but the underlying mechanisms are largely unknown. Here, we report that brain-derived neurotrophic factor (BDNF) from PC facilitates CF synapse elimination. By PC-specific deletion of BDNF combined with knockdown of BDNF receptors in CF, we show that BDNF acts retrogradely on TrkB in CFs, and facilitates elimination of CF synapses from PC somata during the third postnatal week. We also show that BDNF shares signaling pathway with metabotropic glutamate receptor 1, a key molecule that triggers a canonical pathway for CF synapse elimination. These results indicate that unlike other synapses, BDNF mediates punishment signal for synapse elimination in the developing cerebellum.During development, synapses are selectively strengthened or eliminated by activity-dependent competition. Here, the authors show that BDNF-TrkB retrograde signaling is a "punishment" signal that leads to elimination of climbing fiber-onto-Purkinje cell synapses in the developing cerebellum.


Free radical scavenger and antioxidant capacity correlation of alpha-tocopherol and Trolox measured by three in vitro methodologies.
The objective of this study was to correlate the free radical scavenging and antioxidant activity of two known substances (Trolox and alpha-tocopherol), using three in vitro methods (linoleic acid emulsion, brain homogenate and 2,2-diphenyl-1-picryl-hydrazyl [DPPH]). At steady state, alpha-tocopherol showed a greater inhibition of spontaneous oxidation of brain homogenate (59.42%+/-1.91) than Trolox (38.50%+/-2.38), while the latter showed a better antioxidant activity performance regarding inhibition of linoleic acid peroxidation (100% versus 84.02%+/-1.98) and free radical scavenging activity (93.56%+/-5.71 versus 66.72%+/-6.28). When the IC50 value was used as a parameter, alpha-tocopherol presented greater antioxidant activity than Trolox evaluated in brain homogenate and DPPH, without a significant difference when using linoleic acid emulsion. Both compounds showed the same antioxidant efficiency measured by DPPH kinetics (0.37 mM). Antioxidant activity significantly changed according to the substrate, the parameter adopted to compare the substances in the same method and the form used to express antioxidant concentration.


Clinical utility of near-infrared perfusion assessment of the gastric tube during Ivor Lewis esophagectomy.
Anastomotic leakage (AL) after Ivor Lewis esophagectomy with intrathoracic anastomosis carries a significant morbidity. Adequate perfusion of the gastric tube (GT) is an important predictor of anastomotic integrity. Recently, near infrared fluorescent (NIRF) imaging using indocyanine green (ICG) was introduced in clinical practice to evaluate tissue perfusion. We evaluated the feasibility and efficacy of GT indocyanine green angiography (ICGA) after Ivor Lewis esophagectomy. This retrospective analysis used data from a prospectively kept database of consecutive patients who underwent Ivor Lewis (IL) esophagectomy with GT construction for cancer between January 2016 and December 2020. Relevant outcomes were feasibility, ICGA complications and the impact of ICGA on AL. 266 consecutive IL patients were identified who matched the inclusion criteria. The 115 patients operated with perioperative ICGA were compared to a control group in whom surgery was performed according to the standard of care. ICGA perfusion assessment was feasible and safe in all 115 procedures and suggested a poorly perfused tip in 56/115 (48.7%) cases, for which additional resection was performed. The overall AL rate was 16% (43/266), with 12% (33/266) needing an endoscopic our surgical intervention and 6% (17/266) needing ICU support. In univariable and multivariable analyses, ICGA was not correlated with the risk of AL (ICGA:14.8% vs non-ICGA:17.2%, p = 0.62). However, poor ICGA perfusion of the GT predicted a higher AL rate, despite additional resection of the tip (ICGA poorly perfused: 19.6% vs ICG well perfused: 10.2%, p = 0.19). ICGA is safe and feasible, but did not result in a reduction of AL. The interpretation and necessary action in case of perioperative presence of ischemia on ICGA have yet to be determined. Prospective randomized trials are warranted to analyze its benefit on AL in esophageal surgery. Trial registration Ethical approval for a prospective esophageal surgery database was granted by the Ethical committee of the Ghent University Hospital. Belgian registration number: B670201111232. Ethical approval for this retrospective data analysis was granted by our institutional EC. BC-09216.