Predictors and moderators of treatment outcome in the Pediatric Obsessive Compulsive Treatment Study (POTS I).
To identify predictors and moderators of outcome in the first Pediatric OCD Treatment Study (POTS I) among youth (N = 112) randomly assigned to sertraline, cognitive behavioral therapy (CBT), both sertraline and CBT (COMB), or a pill placebo. Potential baseline predictors and moderators were identified by literature review. The outcome measure was an adjusted week 12 predicted score for the Children's Yale Brown Obsessive Compulsive Scale (CY-BOCS). Main and interactive effects of treatment condition and each candidate predictor or moderator variable were examined using a general linear model on the adjusted predicted week 12 CY-BOCS scores. Youth with lower obsessive-compulsive disorder (OCD) severity, less OCD-related functional impairment, greater insight, fewer comorbid externalizing symptoms, and lower levels of family accommodation showed greater improvement across treatment conditions than their counterparts after acute POTS treatment. Those with a family history of OCD had more than a sixfold decrease in effect size in CBT monotherapy relative to their counterparts in CBT without a family history of OCD. Greater attention is needed to build optimized intervention strategies for more complex youth with OCD. Youth with a family history of OCD are not likely to benefit from CBT unless offered in combination with an SSRI. Treatment of Obsessive Compulsive Disorder (OCD) in Children, http://www.clinicaltrials.gov, NCT00000384.


FEF200-1200, FEF25-75% and FEF75-85% in non-smokers of either sex and in male smokers residing at an altitude of 2150 M above MSL in Himachal Pradesh.
Forced expiratory flow (FEF) rates were determined for 170 (130 males and 40 females) healthy adults of Himachal Pradesh in the age group of 19-26 years for males and 19-22 years for females. The subjects had been staying at an altitude of 2150 M above mean sea level (MSL) for at least three years prior to the conduct of the study. FEF200-1200, FEF25-75% and FEF75-85% were found to be high as compared to those reported from most other parts of the country. The results were found comparable with those reported from certain populations in U.S.A. Further, the flow rates of the non-smoker males were compared with those of the male smokers. FEF75-85% in smokers was significantly less than that of non-smokers, whereas the differences in FEF200-1200 and FEF25-75% were found to be nonsignificant.


Gamma heavy chain disease in a patient with diabetes and chronic renal insufficiency: diagnostic assessment of the heavy chain fragment.
Heavy chain diseases are rare B-cell disorders that are characterized by an overproduction of abnormal and structurally incomplete monoclonal immunoglobulin (Ig) heavy chains and are devoid of light chains. We describe a case of a 62 year-old African-American woman with a long history of poorly controlled type 2 diabetes and subsequent probable diabetic nephropathy, hypertension, and recent onset of peripheral neuropathy involving all extremities. Routine laboratory testing revealed a distinct beta spike by urine protein electrophoresis (UPEP). No serum abnormality was noted on serum protein electrophoresis (SPEP). Serum and urine immunofixation demonstrated an IgG heavy chain protein devoid of any corresponding light chains. IgG subclasses identified IgG1 as the predominant IgG component but when we added all the subclasses, the sum, 683.4 mg/dL, failed to come close to our total IgG of 1,770 mg/dL. Therefore, a urine IgG subclass determination was performed in-house and we identified a subclass 3 gamma chain. In conclusion, we portray a patient with an underlying monoclonal gamma heavy chain disease (HCD) who presented with a complex medical history. The evaluation of IgG subclasses in the context of a HCD may be limited by the capability of the test to recognize the particular IgG fragment.


Subtyping of Strengths and Difficulties in a Spanish Children Sample: A Latent Class Analysis.
Externalizing and internalizing problems are frequent during childhood and are often comorbid. The current study aimed to explore subtypes of strengths and difficulties among Spanish children. The Spanish version of the Strengths and Difficulties Questionnaire was administered to 325 children (47.1% girls), aged 7-12 years (M = 9.64; SD = 1.34). Latent Class Analysis (LCA) was carried out to identify risk profiles in children according to their symptoms. ANOVA and multinomial logistic regression were performed to explore associations between latent classes and SDQ subscales, gender, age and number of siblings. Five latent classes were found: "high difficulties" (34.2%), "internalizing" (5.2%), "externalizing" (26.5%), "hyperactive" (14.5%), and "well-adjusted" (19.7%). Children belonging to "High difficulties" group showed the highest scores for emotional symptoms, conduct problems, and hyperactivity/inattention. The most prevalent pure profiles were the "externalizing" and "hyperactivity" group, which represent children with elevated behavioral problems, restlessness and distraction. Children in the "internalizing" class showed the highest risk of peer problems. These results support the diversity in the patterns of psychological strengths and difficulties and highlight the importance of early detection and intervention, especially in children's behavioral problems, in order to avoid vulnerability to comorbidity and more severe symptoms in the future.


The two authentic methionine aminopeptidase genes are differentially expressed in Bacillus subtilis.
Two putative methionine aminopeptidase genes, map (essential) and yflG (non-essential), were identified in the genome sequence of Bacillus subtilis. We investigated whether they can function as methionine aminopeptidases and further explored possible reasons for their essentiality or dispensability in B. subtilis. In silico analysis of MAP evolution uncovered a coordinated pattern of MAP and deformylase that did not correlate with the pattern of 16S RNA evolution. Biochemical assays showed that both MAP (MAP_Bs) and YflG (YflG_Bs) from B. subtilis overproduced in Escherichia coli and obtained as pure proteins exhibited a methionine aminopeptidase activity in vitro. Compared with MAP_Bs, YflG_Bs was approximately two orders of magnitude more efficient when assayed on synthetic peptide substrates. Both map and yflG genes expressed in multi-copy plasmids could complement the function of a defective map gene in the chromosomes of both E. coli and B. subtilis. In contrast, lacZ gene transcriptional fusions showed that the promoter activity of map was 50 to 100-fold higher than that of yflG. Primer extension analysis detected the transcription start site of the yflG promoter. Further work identified that YvoA acted as a possible weak repressor of yflG expression in B. subtilis in vivo. Both MAP_Bs and YflG_Bs are functional methionine aminopeptidases in vitro and in vivo. The high expression level of map and low expression level of yflG may account for their essentiality and dispensality in B. subtilis, respectively, when cells are grown under laboratory conditions. Their difference in activity on synthetic substrates suggests that they have different protein targets in vivo.


Increase in manganese superoxide dismutase activity in the mouse heart after X-irradiation.
Local X-irradiation of mouse heart caused a large increase in manganese superoxide dismutase activity (MnSOD) in this organ but not in copper and zinc containing superoxide dismutase (Cu-Zn SOD) activity. MnSOD induction was both dose and time dependent. Another mitochondrial enzyme, citrate synthase, was not induced by X-irradiation. The amount of immunoreactive MnSOD also increased after X-irradiation, showing that the amount of MnSOD protein increased after X-irradiation. The response to X-irradiation was found to be biphasic--with one large peak and one smaller peak of manganese superoxide dismutase activity. The effect of various inhibitors of cellular activities on these two peaks of MnSOD activity was examined. Cycloheximide, a cytosolic protein synthesis inhibitor, abolished both peaks of MnSOD activity, while chloramphenicol, a mitochondrial protein synthesis inhibitor, has no effect on either peak. Actinomycin D, a RNA-synthesis inhibitor, lowered both peaks, but had more of an effect on the second peak than on the first. In vivo protein synthesis studies using [3H]arginine showed that an increase in new protein synthesis occurred during the time period of the second peak, but did not occur during the first peak. These results are consistent with the hypothesis that MnSOD induction occurs in two peaks with the first peak due to a preformed MnSOD protein or mRNA for MnSOD and the second peak due to an increase in new protein synthesis.


Ethylene glycol assisted synthesis of hierarchical Fe-ZSM-5 nanorods assembled microsphere for adsorption Fenton degradation of chlorobenzene.
A unique zeolite catalyst, Fe doped ZSM-5 microsphere assembled by uniform nanorod-like crystals with hierarchical pore structure, was successfully synthesized and applied for the adsorption and degradation of trace chlorobenzene (CB) in the presence of H2O2. The organic ferric salts as the precursors, ethylene glycol as a chelating/reducing agent and the dynamic two-stage temperature-varied hydrothermal technique, together made the synthesized hierarchical Fe-ZSM-5 nanorods assembled microspheres (FZ-CA-5EG) to be characterized by abundant highly dispersed and valency-controlled framework Fe3+/2+ species. As a result of these features, the FZ-CA-5EG showed excellent ability of adsorption and degradation efficiency of CB, and enhanced durability due to negligible leaching of framework Fe species. Moreover, the hydroxyl radicals were determined as the main the reactive oxygen species of CB oxidation degradation, and a possible adsorption-oxidation degradation pathway was proposed.


Disparities in access to care among patients with appendiceal or colorectal cancer and peritoneal metastases: A medicare insurance-based study in the United States.
Prior studies attempting to identify disparities in the care of patients with appendiceal (AC) or colorectal cancer (CRC) with peritoneal metastasis (PM) are limited to single-institution, highly selected patient populations. This observational cohort study sought to identify factors associated with specialty care for Medicare beneficiaries with AC/CRC-PM. Patients >65 years old in the United States diagnosed with AC/CRC and isolated PM were identified within the Medicare Standard Analytic File (2013-2017). Mixed-effects analyses assessed patient factors associated with cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS/HIPEC) and outpatient consultation with a peritoneal surface malignancy (PSM) surgeon, and Cox proportional-hazards analysis compared 3-year overall survival (OS) between patients receiving CRS/HIPEC versus systemic therapy alone. Among 7,653 patients, only 250 (3.3%) underwent CRS/HIPEC. Among those individuals who did not undergo CRS/HIPEC (N=7,403), only 475 (6.4%) had outpatient consultation with a PSM surgeon. Patient factors independently associated with lower odds of CRS/HIPEC and PSM surgery consultation included older age, greater comorbidity burden, higher social vulnerability index, and further distance from a PSM center (p<0.05). CRS/HIPEC was independently associated with better 3-year OS compared with systemic therapy alone (HR=0.29, 95%CI=0.21-0.38). An exceedingly small proportion of Medicare beneficiaries with AC/CRC-PM undergo CRS/HIPEC or even have an outpatient consultation with a PSM surgeon. Significant disparities in treatment and access to care exist for patients with higher levels of social vulnerability and those that live further away from a PSM center. Future research and interventions should focus on improving access to care for these at-risk patient populations.


Vitamin D status and physical activity interact to improve bone mass in adolescents. The HELENA Study.
The effects of vitamin D concentrations on bone mineral content in adolescents are still unclear. Vitamin D and physical activity (PA) may interact to determine bone mineral content (BMC) in two possible directions; 25(OH)D sufficiency levels improve BMC only in active adolescents, or PA increases BMC in individuals with replete vitamin D levels. The effects of suboptimal 25-hydroxycholecalciferol (25(OH)D) concentrations on BMC in adolescents are still unclear. The main aim of this study was to evaluate the influence of 25(OH)D on BMC in adolescents, considering the effect of body composition, sex, age, Tanner stage, season, calcium and vitamin D intakes, physical fitness and PA. Serum 25(OH)D concentrations, anthropometric measurements, dual energy X-ray absorptiometry measurements, calcium and vitamin D intakes, PA and physical fitness were obtained in 100 Spanish adolescents (47 males), aged 12.5-17.5 years, within the framework of the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study. Relations were examined using ANCOVA and regression analyses including BMC as dependent variable. Linear regression of BMC suggested that 25(OH)D concentrations independently influenced total and leg BMC after controlling for age, sex, lean mass, seasonality and calcium intake (B = 0.328, p < 0.05, and B = 0.221, p < 0.05, respectively) in the physically active group. No significant influence of 25(OH)D concentrations on BMC was observed in the inactive group. Significant effect was shown between the interaction of 25(OH)D and PA on BMC for the total body and legs (both p < 0.05). Vitamin D and PA may interact to determine BMC. 25(OH)D sufficiency levels improve bone mass only in active adolescents, or PA has a positive influence on BMC in individuals with replete vitamin D levels.


Detecting functional rare variants by collapsing and incorporating functional annotation in Genetic Analysis Workshop 17 mini-exome data.
Association studies using tag SNPs have been successful in detecting disease-associated common variants. However, common variants, with rare exceptions, explain only at most 5-10% of the heritability resulting from genetic factors, which leads to the common disease/rare variants assumption. Indeed, recent studies using sequencing technologies have demonstrated that common diseases can be due to rare variants that could not be systematically studied earlier. Unfortunately, methods for common variants are not optimal if applied to rare variants. To identify rare variants that affect disease risk, several investigators have designed new approaches based on the idea of collapsing different rare variants inside the same genomic block (e.g., the same gene or pathway) to enrich the signal. Here, we consider three different collapsing methods in the multimarker regression model and compared their performance on the Genetic Analysis Workshop 17 data using the consistency of results across different simulations and the cross-validation prediction error rate. The comparison shows that the proportion collapsing method seems to outperform the other two methods and can find both truly associated rare and common variants. Moreover, we explore one way of incorporating the functional annotations for the variants in the data that collapses nonsynonymous and synonymous variants separately to allow for different penalties on them. The incorporation of functional annotations led to higher sensitivity and specificity levels when the detection results were compared with the answer sheet. The initial analysis was performed without knowledge of the simulating model.


Surgical bypass of symptomatic central venous obstruction for arteriovenous fistula salvage in hemodialysis patients.
Venous hypertension due to proximal central venous outflow obstruction coexisting with a functioning arteriovenous fistula in the ipsilateral arm presents with a complex management problem in hemodialysis patients. Ligation of the arteriovenous communication is the simplest procedure to relieve symptoms; however, this sacrifices the patient's hemodialysis access, which may be the only available access in that patient. Surgical bypass of the occlusion is a potential option as it obviates the symptoms of venous hypertension while preserving dialysis access. Our objective was to evaluate our experience and outcome with dialysis patients undergoing surgical bypass for symptomatic central venous obstruction and dialysis access salvage. There were three hemodialysis patients with severe venous hypertension secondary to subclavian vein obstruction who had functioning ipsilateral arteriovenous fistulae. All underwent cephalic vein (n = 2) or axillary vein (n = 1) to internal jugular vein bypass of the obstructed subclavian segment via an 8-mm polytetrafluoroethylene bridge graft. All patients had unsuccessful percutaneous transluminal angioplasty (PTA) attempts prior to surgical bypass. In two patients, a wire could not be passed through the occlusion; in the third, PTA was only transiently successful despite four repeated procedures. All patients had complete resolution of symptoms without operative mortality. The bypass grafts remained patent, allowing the arteriovenous fistulae to provide functional access for the entire duration of follow-up after surgery (3-8 months). Surgical bypass of a central vein obstruction relieves the symptoms of venous hypertension and prolongs the use of the existing hemodialysis access. This surgical option should be well recognized within the dialysis community.


Plasmodium falciparum ookinetes migrate intercellularly through Anopheles stephensi midgut epithelium.
The migration of Plasmodium falciparum and P. berghei ookinetes through the midgut epithelium in Anopheles stephensi was studied by transmission electron microscopy. With ruthenium red (RR) staining, the results of previous studies were confirmed: P. falciparum ookinetes take an intercellular route through the midgut epithelium. In the same mosquito species, the rodent parasite P. berghei appeared to take an intracellular position, as previously suggested by other authors. The intra- or intercellular ookinete migration of P. berghei or P. falciparum, respectively, can perhaps be related to the higher mortality of P. berghei-infected mosquitoes within the first 2 days of infection. Evidence is presented that oocyst capsule formation begins as early as during the migration of the ookinete. After localization between the epithelial cells and the midgut basal lamina, the rapidly expanding oocyst stretches the overlying layer of the latter at the haemocoelic surface while a new basal lamina is generated between the oocyst and epithelial cell.


Potential clinical implication of (18) F-FDG PET/CT in diagnosis of periprosthetic infection and its comparison with (18) F-Fluoride PET/CT.
The differentiation between septic and aseptic loosening of prosthetic hip joint is a major challenge for the clinician. This study assessed and compared the diagnostic accuracy of (18) F- flouro-deoxyglucose positron emission tomography/computed tomography ((18) F-FDG PET/CT) with (18) F-fluoride PET/CT for diagnosis of infection in the painful hip prosthesis. In this prospective study, we included the patients with painful hip prostheses with radiological or clinical suspicion of loosening, those who had given the written consent and scheduled for clinical and diagnostic evaluation before revision arthroplasty. To rule out the nature of loosening (septic vs. aseptic), all the patients underwent (18) F-Fluoride PET/CT and (18) F-FDG PET/CT. The reference standard for periprosthetic infection was based on histopathology and/or microbial culture and/or intraoperative findings. We prospectively evaluated 42 patients of hip prostheses before revision arthroplasty. Visual and semi-quantitative analysis of both the positron emission tomography (PET) images was done and the results were compared with a reference standard. The sensitivity, specificity, positive predictive value (PPV), negative predictive values (NPV) and accuracy of (18) F-FDG PET/CT were 93.7%, 92.3%, 88.2%, 96% and 92.8% respectively. The sensitivity, specificity, PPV, NPV and accuracy of (18) F-fluoride PET/CT were 75%, 96.1%, 92.3%, 86% and 88.1% respectively. FDG PET/CT has higher sensitivity, NPV and accuracy as compared to compared to Fluoride PET/CT and comparable specificity and PPV. Fluoride PET/CT had shown slightly higher specificity than FDG-PET/CT, but the overall diagnostic performance of FDG-PET/CT in periprosthetic infection is optimal in routine clinical practice and better than fluoride PET/CT.


Familial abdominal aortic aneurysms.
The case histories of three brothers, the only siblings of one family, all of whom underwent surgery for the treatment of a previously asymptomatic ruptured abdominal aortic aneurysm, are recorded. The possibility of underlying constitutional and hereditary factors is discussed and the suggestion of a primary familial incidence of atheromatous, nondissecting aortic aneurysm is raised.


Haemostatic profile of full-term, healthy, small for gestational age neonates.
Small for Gestational Age (SGA) neonates often appear with haemostatic alterations, principally due to hepatic dysfunction that results from chronic intrauterine hypoxia. Polycythaemia and thrombocytopenia are common findings in this neonatal population. We performed a comparison of coagulation, natural inhibitors and fibrinolysis between SGA and Appropriate for Gestational Age (AGA) infants born full term [gestational age (G.A.) >37 weeks]. Study population consisted of 188 healthy newborns, 90 of whom were SGA (62 females and 28 males), while the rest were the control group (44 females and 54 males). Blood samples were obtained within 30 minutes following birth and before the administration of vitamin K. Investigation included: PT, INR, APTT, fibrinogen, coagulation factors II, V, VII, VIII, IX, X, XI, XII, vWillebrand factor, protein C and free protein S, antithrombin (AT), APCR, tPA and PAI-1. The independent t-test was used to compare the differences between the values of haemostatic parameters. Statistical analysis revealed a significant prolongation in PT, INR, elevated levels of tPA (p<0.015, 0.01 and 0.002 respectively) and a decrease in the values of XII and free protein S (p<0.045 and 0.007 respectively) in SGA full term neonates. The two groups had similar demographic characteristics (except birth weight), without significant differences in the values of other haemostatic parameters. Despite of statistically significant differences in PT, INR, values of tPA, XII and free protein S, levels of haemostatic factors range within laboratory references for healthy full term newborns. These findings were not accompanied with clinical manifestations of altered haemostasis.


NADPH and oxidized thioredoxin mediate redox interconversion of calf-liver and Escherichia coli thioredoxin reductase.
The activity of pure calf-liver and Escherichia coli thioredoxin reductases decreased drastically in the presence of NADPH or NADH, while NADP+, NAD+ and oxidized E. coli thioredoxin activated both enzymes significantly, particularly the bacterial one. The loss of activity under reducing conditions was time-dependent, thus suggesting an inactivation process: in the presence of 0.24 mM NADPH the half-lives for the E. coli and calf-liver enzymes were 13.5 and 2 min, respectively. Oxidized E. coli thioredoxin fully protected both enzymes from inactivation, and also promoted their complete reactivation after only 30 min incubation at 30 degrees C. Lower but significant protection and reactivation was also observed with NADP+ and NAD+. EDTA protected thioredoxin reductase from NADPH inactivation to a great degree, thus indicating the participation of metals in the process; EGTA did not protect the enzyme from redox inactivation. Thioredoxin reductase was extensively inactivated by NADPH under aerobic and anaerobic conditions, thus excluding the participation of O2 or oxygen active species in redox inactivation. The loss of thioredoxin reductase activity promoted by NADPH was much faster and complete in the presence of NAD+ glycohydrolase, thus suggesting that inactivation was related to full reduction of the redox-active disulfide. Those results indicate that thioredoxin reductase activity can be modulated in bacteria and mammals by the redox status of NADP(H) and thioredoxin pools, in a similar way to glutathione reductase. This would considerably expand the regulatory potential of the thioredoxin-thioredoxin reductase system with the enzyme being self-regulated by its own substrate, a regulatory protein.


Case report. Successful treatment of cutaneous Paecilomyces lilacinus infection with oral itraconazole in an immune competent host.
A 59-year-old woman presented to us with an eight-month history of Paecilomyces lilacinus soft tissue infection. She was treated with a six-month course of oral itraconazole. The lesion resolved fully without the need for adjunctive surgery. Susceptibility testing confirmed susceptibility to itraconazole. This is the second case of confirmed P. lilacinus soft tissue infection in an immunocompetent host treated with oral itraconazole alone.


Retrospective clinical study of mandible fractures.
As society becomes more complex, the incidence of mandibular fractures is increasing. This study aimed to analyze the incidence and type and identify etiological factors of mandibular fractures to use them in future treatments. Data were collected from 224 patients who visited the department of oral and maxillofacial surgery at the Kyung Hee Medical Center dental hospital during a 6-year period (2016 to 2021). A logistic regression model was used for data analysis. In a total of 224 patients, 362 fractures were appeared. The average age of the patients was 34.1 years, with the highest incidence in the 20s. And the ratio between male and female was 4.09:1. Symphysis fractures were the most prevalent of all patients (52.7%), followed by unilateral condyle (37.1%), angle (36.2%), bilateral condyle (9.4%), body (8%), and coronoid (2.2%). The most common cause of fracture was daily-life activity (57.6%), followed by violence (30.4%), traffic accidents (8.5%), and syncope (3.6%). Patients with symphysis fracture were at low risk (OR < 1) of angle, body, and unilateral condyle fractures. Similarly, patients with unilateral fracture were at low risk (OR < 1) of symphysis, angle, body, and others site fractures. In contrast, patient with bilateral condyle fracture were at high risk (OR > 1) of coronoid fractures. And younger patients were high risk of mandibular angle fractures. Through this study, it was confirmed that etiological factors of mandibular fractures were like those of previous studies.


Transabdominal chorionic villus sampling in the second trimester.
Transabdominal chorionic villus sampling under ultrasound guidance was carried out in 19 patients in the second trimester. We found that needle size combination 17/19 provided the best results with consistently adequate samples of tissue. Transabdominal chorionic villus sampling is a potentially useful technique for providing relatively rapid prenatal genetic diagnosis in patients at high risk in the second trimester.


The very-rapid and the ultra-rapid virologic response to two treatment options in patients with chronic hepatitis C: an interim report of a prospective randomized comparative effectiveness study.
We aimed in this interim report to compare two registered generic sofosbuvir products for the degree and speed of virologic response to a dual antiviral treatment protocol within the first 2 weeks of treatment. Data collected during the period of this interim report from the first 25 patients randomized to either one of two generic sofosbuvir products (Grateziano or Gratisovir) at a daily dose of one 400 mg tablet plus a weight-based ribavirin dose were analyzed for both the degree and speed of virus load reduction at the end of 1 and 2 weeks from starting treatment. The baseline Log10 transformed virus load (Log polymerase chain reaction) showed a fairly similar marked and significant reduction in both groups by more than 4 and 5 Logs at the end of week 1 and 2 of starting treatment, respectively. The differences between the two treatment groups at both analysis points were not statistically significant (P>0.05) by repeated measures factorial analysis of variance test. The differences in proportions of patients with ultra-rapid virologic response at the end of week 1 and very-rapid virologic response at the end of week 2 in both groups were also not statistically significant (P>0.05). We can conclude from this interim report that the two generic products Gratisovir and Grateziano are almost equally fast and efficacious in reducing the hepatitis C virus load in our study setting.


The effects of ionizing radiation on the glycosaminoglycan synthesis in embryonic cartilage in vitro.
The effects of ionizing radiation on the glycosaminoglycan synthesis of cartilaginous tibiae of 6.5 day old chick embryos have been studied in vitro over a 3 day period. As a parameter for the glycosaminoglycan synthesis, the incorporation of (3)H-glucosamine is evaluated by liquid scintillation counting. A radiation induced inhibition ranging from 6% at 20 Gy to 68% at 200 Gy is observed. Light microscopical autoradiographs reveal that the secretion of the newly synthesized glycosaminoglycans is not affected significantly by ionizing radiation up to 150 Gy. Electron microscopical pictures confirm this observation and show that the chondrocytes in the irradiated tibiae remain viable. At 200 Gy most cells are necrotic. The results indicate that the radiation effects on the synthesis of the extracellular matrix components are determined by the damage of the RNA transcription.


Analysis of human class II antigens by cloned cytolytic T cell reagents: a study using HLA loss mutant lymphoblastoid cell lines and monoclonal antibodies detecting the HLA-DP product(s).
We have utilized cloned T cell reagents and ionizing radiation-induced mutants of an HLA heterozygous lymphoblastoid cell line (LCL) to investigate the determinants detected by the cell-mediated lympholysis (CML) assay. Cells of an LCL clone, 721.501, an HLA haplotype loss mutant expressing the HLA-A2-Cw1-Bw51-DR1-Dw1-DQw1-DPw2-GLO haplotype were used as sensitizing cells for responder cells in vitro. "Cloned" reagents were generated by single-cell deposition of cells of a bulk reagent primed against 721.501 cells. Those clones were screened for cytolytic activity against HLA loss mutant targets (derived from LCL 721) of four different categories; HLA-A2 loss only, A2-Cw1-Bw51 loss, A2-Cw1-Bw51-DR1-DQw1 loss, and the entire HLA haplotype loss. Of 196 clones tested, 36 were cytolytic, including three anti-A2, five anti-Bw51/Cw1, 12 anti-DR1/DQw1, 13 anti-DP region associated with DPw2, and three of undetermined specificity, based on cytolytic patterns against the HLA loss mutant targets. Of 25 anti-HLA class II lytic clones, 23 (92%) fitted the characteristics of helper cell-independent cytolytic T cells (HITc), whereas only two of eight (25%) anti-class I clones were HITc. The 13 anti-DP region clones were divided into three subgroups defined by blocking by anti-FA and not Tü39 monoclonal antibodies (MoAb), by Tü39 and not anti-FA, and by both MoAbs.


Effect of iron-saccharin injections of Finnish horses.
In four training stables five trotter horses of the Finnish coldblooded breed and seven warmblooded trotters were treated with a ferrissaccharin preparation (Hippiron, Hausmann) intravenously. A short increase in heart frequency after the injection was noted. The cold solution injections, less than + 15 degrees C, may cause dangerous reactions. All horses showed an enhanced appetite, a substantially livier habitus after treatment.


RGFP966 exerts neuroprotective effect via HDAC3/Nrf2 pathway after surgical brain injury in rats.
Histone deacetylase 3 (HDAC3) restores chromatin nucleosomes to a transcriptional repression state, thereby inhibiting gene expression. Studies have found that HDAC3 expression is upregulated in a variety of pathological states of the central nervous system and related to its neurotoxicity. However, the role of HDAC3 in surgical brain injury (SBI) has not been thoroughly explored. To observe the role of HDAC3 in SBI and the outcome of SBI after its suppression. Rat SBI model was used, and intraperitoneal injection of RGFP966 (HDAC3 specific inhibitor) was used to detect the changes of HDAC3 expression and neuronal apoptosis indexes in the surrounding cortex of SBI rats, and the cerebral edema and neurological outcome of rats were observed. The expression of HDAC3 in the peripheral cortex of SBI rats was increased, and RGFP966 inhibited the upregulation of HDAC3 and saved the nerve cells around the damaged area. In addition, RGFP966 increased the expression of anti-oxidative stress proteins such as heme oxygenase-1 (HO-1) and superoxide dismutase 2 (SOD2). At the same time, the expression of apoptotic marker protein cleaved-caspase-3 (cle-caspase-3) was decreased, while the expression level of apoptotic protective marker protein B-cell lymphoma 2 (Bcl-2) was increased. In addition, this research demonstrated that in the RGFP966 rat SBI model, the expression level of antioxidant modifier nuclear factor-erythroid 2-related factor 2 (Nrf2) was increased. RGFP966 might activate HDAC3/Nrf2 signaling pathway by inhibiting HDAC3, regulated oxidative stress and nerve cell apoptosis induced by SBI in rat SBI model, reduced brain edema, and had a protective effect on nerve injury. It might be a potential target of SBI pathology.


Association of malondialdehyde-acetaldehyde (MAA) adducted proteins with atherosclerotic-induced vascular inflammatory injury.
Atherosclerosis is a vascular injury characterized by elevated tissue levels of tumor necrosis factor-alpha (TNF-alpha), increased expression of endothelial cell adhesion molecules, and vascular wall inflammatory cell infiltration. Foam cells are associated with atherosclerotic plaque material, and low density lipoprotein (LDL) is a lipid component of foam cells. Malondialdehyde (MDA) is an oxidative product of unsaturated fatty acids and is also present in atherosclerotic lesions. MDA-modified (adducted) proteins, including MDA-modified LDL, are present in atherosclerotic human vascular tissue. Acetaldehyde (AA) is the major metabolic product of ethanol oxidation. Both MDA and AA are highly reactive aldehydes and will combine with proteins to produce an antigenically distinct protein adduct, termed the MAA adduct. This study demonstrates that proteins modified in the presence of high concentrations of MDA can produce MAA-modified proteins in vitro. In addition, MAA adducted proteins are capable of inducing rat heart endothelial cell cultures (rHEC) to produce and release TNF-alpha, and cause rHEC upregulation of endothelial adhesion molecule expression, including ICAM-1. These adhesion molecules are required for circulating inflammatory cells to adhere to endothelium which allows inflammatory cell tissue infiltration. Additionally, MAA modified proteins were defected in human atherosclerotic aortic vascular tissue but not in normal aortic tissue. Since atherosclerosis is associated with an inflammatory vascular injury characterized by elevated tissue TNF-alpha concentrations and inflammatory cell infiltration, these data suggest that MAA-adducted proteins may be formed in atherosclerotic plaque material and may be involved in the inflammatory reaction that occurs in atherosclerosis. These data further suggest that previous studies demonstrating MDA modified protein in atherosclerotic plaque may in fact have MAA modified proteins associated with them.


An interferon-induced mouse protein involved in the mechanism of resistance to influenza viruses. Its purification to homogeneity and characterization by polyclonal antibodies.
Interferon-alpha + beta (IFN-alpha + beta) plays a central role in the specific resistance to influenza virus infection of those mice carrying the gene Mx (for review, see Haller, O. (1981) Curr. Topics Microbiol. Immun. 92, 25). Particularly, mouse IFN-alpha + beta induces a unique protein in cultivated Mx-bearing cells which is associated with a highly efficient and specific antiviral resistance to influenza viruses (Horisberger, M. A., Staeheli, P., and Haller, O. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 1910). In this report, a procedure is described for the induction of this protein in several organs of Mx-bearing mice and a method for its purification from liver tissue. The protein Mx is nucleophilic and has a Mr approaching 78,000. It is not concentrated in nucleoli and it is not tightly bound to chromatin or nuclear matrices. Polyclonal antibodies to the protein Mx were raised in BALB/c mice. They recognized the protein Mx immobilized on nitrocellulose in a dot immunoassay and they immunoprecipitated the IFN-induced protein Mx from cultivated Mx-bearing cells labeled with a radioactive tracer.


Comparison of outcomes of the patients with acute cholecystitis treated in the COVID-19 pandemic and pre-pandemic period.
The coronavirus disease 2019 (COVID-19) pandemic has affected the health care system in an unpredictable way. In this study, we aimed to analyze the effects of the pandemic process on the disease severity on admission, management strategies, and outcomes of patients. The medical records of the patients who applied to the emergency department and consulted to the general surgery clinic from March 2020 until January 2021 were retrospectively reviewed as the pandemic period. For the control group, patients' medical records in the same time interval of 2019 were evaluated as the pre-pandemic period. A total of 88 patients in the pre-pandemic period and 89 patients in the pandemic period were treated for acute cholecystitis. There was no statistically significant difference between the two groups in terms of the treatment strategies and length of hospital stay between the two periods (p=0.087 and p=0.587, respectively). In the pandemic period, it is thought that postponing and bridging treatments may replace surgery for reducing the risk of contamination of both patients and health care workers.


Goal engagement and goal attainment in adolescents with and without visual impairment.
The present longitudinal study analyzed the effects of domain-specific goal engagement on the attainment of four developmental goals in 133 adolescents with visual impairment and in 449 sighted peers. Goal engagement predicted stronger progress in goal attainment with regard to getting access to a peer group, career choice and development of romantic relationships at the 1-year-follow-up. However, goal engagement at T(1) did not predict progress in the achievement of autonomy from ones' parents. Furthermore, we found an interaction effect between vision status and goal engagement on progress in career choice. Higher initial levels of goal engagement predicted stronger progress in goal attainment of sighted adolescents. No such effect was found in adolescents with visual impairment. It is concluded that effects of goal engagement vary by goal contents and contexts.


Design and synthesis of novel leucomycin analogues modified at the C-3 position. Part II: 3-O-(3-Aryl-2-propenyl)leucomycin analogues.
The design and synthesis of 16-membered macrolides modified at the C-3 position are described. Starting from fully protected intermediate (5), appropriate modifications including Heck reaction were performed to furnish 3-O-(3-aryl-2-propenyl)leucomycin A(7) analogues (9a-9m). These leucomycin A(7) derivatives showed improved in vitro antibacterial activities against clinically important pathogens including erythromycin-resistant Streptococcus pneumoniae (ERSP). SAR analysis of derivatives modified at the C-3 and C-3'' positions suggested that single modification at C-3 or C-3'' was effective for in vitro antibacterial activity.


Long-term complications of diabetes mellitus, Part I: Retinopathy, nephropathy, neuropathy.
Diabetic retinopathy, nephropathy, and neuropathy occur infrequently in small animals, but are capable of causing significant disease. The clinical and histopathologic findings seen with these late complications of diabetes are discussed. The pathogenesis of these disorders is most likely multifactorial; metabolic alterations secondary to the hyperglycemic state, and microvascular changes seen with diabetes have both been implicated. Current treatment consists of aggressive control of the hyperglycemia, as research continues into the pathology of the late complications in attempts to find a definitive therapy.


Effects of peritoneal ropivacaine nebulization for pain control after laparoscopic gynecologic surgery.
To evaluate the effects of peritoneal cold nebulization of ropivacaine on pain control after gynecologic laparoscopy. Evidence obtained from a properly designed, randomized, double-blind, placebo-controlled trial (Canadian Task Force classification I). Tertiary care center. One hundred thirty-five women with American Society of Anesthesiologists disease classified as ASA I-III who were scheduled to undergo operative laparoscopy. Patients were randomized to receive either nebulization of 30 mg ropivacaine before surgery (preoperative group), nebulization of 30 mg ropivacaine after surgery (postoperative group), instillation of 100 mg ropivacaine before surgery (instillation group), or instillation of saline solution (control group). Nebulization was performed using the Aeroneb Pro device. Pain scores, morphine consumption, and ambulation time were collected in the post-anesthesia care unit and at 4, 6, and 24 hours postoperatively. One hundred eighteen patients completed the study. Patients in the preoperative group reported lower pain Numeric Ranking Scale values compared with those in the control group (net difference 2 points; 95% confidence interval [CI], 0.3-3.1 at 4 hours, 1-3 at 6 hours, and 0.7-3 at 24 hours; p = .01) Patients in the preoperative group consumed significantly less morphine than did those in the control group (net difference 7 mg; 95% CI, 0.7-13; p = .02). More patients who received nebulization walked without assistance within 12 hours after awakening than did those in the instillation and control groups (net difference 15%; 95% CI, 6%-24%; p = .001). Cold nebulization of ropivacaine before surgery reduced postoperative pain and morphine consumption and was associated with earlier walking without assistance.


Survivorship of the native hip joint after percutaneous repair of acetabular fractures in the elderly.
Our purpose was to examine survivorship of the native hip joint in patients ages 60 and over who underwent percutaneous reduction and fixation of acetabular fractures. A retrospective review at a University Level I Trauma Center was performed. Our institutional trauma database was reviewed. Patients aged 60 or older treated with percutaneous reduction and fixation of acetabular fractures between 1994 and 2007 were selected. 79 consecutive patients with 80 fractures were identified. Rate of conversion to total hip arthroplasty were used to construct a Kaplan-Meier curve showing survivorship of the native hip joint after treatment. 75 fractures had adequate clinical follow-up with a mean of 3.9 years (range 0.5-11.9 years). Average blood loss was 69 cc and there were no postoperative infections. 19/75 (25%) were converted to total hip arthroplasty at a mean time of 1.4 years after the index procedure. Survivorship analysis demonstrated a cumulative survival of 65% at 11.9 years of follow-up. There were no conversions to arthroplasty beyond 4.7 years postoperatively. There were no statistically significant associations between conversion to arthroplasty and age, sex, closed vs. limited open reduction, and simple vs. complex fracture pattern. Percutaneous fixation is a viable treatment option for patients age 60 or greater with acetabular fractures. Rates of conversion to total hip arthroplasty are comparable to open treatment methods and if conversion is required, soft tissues are preserved for future surgery.


Intra and interrater reliability for lower limb flexibility assessment using photogrammetry.
Photogrammetry represents an advancement in the flexibility evaluation, and although it was highly explored for postural assessment, there is a scarcity of studies analyzing lower limb angular measurements using it. The purpose of this study is to verify the reliability of intrarater and interrarater photogrammetry in assessing lower limb flexibility. This was a randomized cross-sectional observational study with test-retest design and a two-day interval. Thirty healthy, physically active adults were included. Three novice raters assessed the participants through flexibility tests of iliopsoas, hamstring, quadriceps and gastrocnemius on two occasions, and independently analyzed the captured images to establish reliability. Intraclass correlation coefficient (ICC), standard error of measurement (SEM), and minimal detectable change (MDC) were calculated. Intrarater reliability was excellent for iliopsoas (ICC = 0.96; SEM = 1.4; MDC = 3.8), hamstring (ICC = 0.99; SEM = 1.1; MDC = 3.1), quadriceps (ICC = 0.99; SEM = 0.8; MDC = 2.3) and gastrocnemius (ICC = 0.98; SEM = 0.9; MDC = 2.5). Interrater reliability was excellent for iliopsoas (ICC = 0.94; SEM = 1.7; MDC = 4.6) and gastrocnemius (ICC = 0.91; SEM = 2.1; MDC = 5.8), but good for hamstring (ICC = 0.90; SEM = 2.8; MDC = 7.9) and quadriceps (ICC = 0.85; SEM = 3.0; MDC = 8.3). The excellent intrarater and good to excellent interrater reliability suggest that photogrammetry assessment of lower limb flexibility by novice raters is reliable. However, clinicians should consider the higher threshold of range of motion change necessary to outweigh measurement error due to interrater variability.


[A case of an effective application of probiotics in the complex therapy of severe type 1 diabetes mellitus and intestinal disbacteriosis].
The article presents a case illustrating an association between the severity of type 1 diabetes mellitus and disorder in the intestinal microbiocenosis. The authors stress that clinical signs of disbacteriosis are mostly manifested by disturbances in the metabolic processes. Clinical effectiveness of probioticotherapy in the complex treatment of the disease is shown.


Mt. Apo Biotic Index (MABI): a macroinvertebrate-based multimetric index for assessing stream biotic integrity of wadeable streams within a geothermal production field in Mindanao, Philippines.
Monitoring the ecological integrity of streams is a challenge, especially in the tropics, which experience high rates of degradation. Multimetric scoring systems have been widely used in other countries in evaluating current stream conditions; however, it has never been done in the Philippines. This study focuses on the development of a benthic macroinvertebrate-based multimetric index for the overall assessment of streams in Mt. Apo, Mindanao, Philippines. The index was used to develop existing physicochemical and biological data obtained during 2010 to 2015 surveys from 15 monitoring sites within the Mt. Apo Geothermal Project (MAGP). Metrics related to benthic macroinvertebrate abundance, richness, composition, functional habit groups, functional feeding groups, and pollution tolerance were screened for their range, temporal stability, sensitivity, discrimination efficiency (DE), redundancy, and responsiveness to anthropogenic impacts. The resulting multimetric index, the Mt. Apo Biotic Index (MABI), is computed as the sum of the individual metric scores after metric transformation using the discrete scoring method DRQ1 (D = discrete, R = reference, Q1 = 25th percentile) of the six core metrics: (1) number of Coleoptera individuals (abundance), (2) number of taxa (richness); (3) [%] Coleoptera taxa (composition), (4) number of sprawler individuals (functional habit group), (5) [%] collector-filterer taxa (functional feeding group), and (6) the Biological Monitoring Working Party Thai version (BMWP-Thai; pollution tolerance). MABI scores were classified into five condition ratings of stream biotic integrity: very poor (6 to 10), poor (11 to 15), fair (16 to 20), good (21 to 25), and excellent (26 to 30). The study demonstrated that the resulting pilot index may provide useful information that will benefit policymakers and resource managers in formulating more comprehensive stream management approaches and conservation plans for priority sites in the region.


Erratum in the Article on Wounds from Fire Arms without Ball, Contained in the Eclectic Department of This No.
[This corrects the article on p. 668 in vol. 2.].


Site of tubulin synthesis in Tetrahymena pyriformis.
Free and membrane-bound polysomes were isolated from the protozoa Tetrahymena pyriformis, and the contribution of these two types of polysomes to tubulin synthesis were studied using immunoprecipitation of the 35S-translational products in a rabbit reticulocyte lysate. One-dimensional electrophoretic analysis shows that tubulin is synthesized by polyadenylated RNA isolated from free and membrane-bound polysomes. Non-polyadenylated RNAs of free polysomes are also able to direct tubulin synthesis. Two-dimensional electrophoretic analysis using O'Farrell's system confirms these results and also reveals the existence of the alpha- and beta-tubulin subunits.


PAX-8 expression in primary and metastatic Merkel cell carcinoma: an immunohistochemical analysis.
PAX-8, a nephric cell lineage transcription factor initially characterized in renal cell carcinomas, is also well recognized as a marker of Müllerian tract and thyroid tumors. From a previous tissue microarray study of nonrenal neoplasms including a variety of skin tumors, we identified PAX-8 staining in a small set of Merkel cell carcinomas, a finding not previously described. Herein, we explore PAX-8 immunoreactivity in 34 whole-section Merkel cell carcinomas (24 primary, 10 metastatic) using polyclonal PAX-8 (prediluted) and 2 varieties of monoclonal PAX-8 (prediluted clone MRQ-50 and 1:100 dilution clone BC12). Nuclear staining intensity and extent was semiquantitatively analyzed with a comparison of staining thresholds required for a "positive" result (≥ 2+ vs. 1+). Thirty-three of 34 (97%) cases showed positive Cell Marque polyclonal PAX-8 staining, whereas 31 of 34 (91%) cases showed positive Cell Marque monoclonal PAX-8 staining. There was no significant difference in staining between primary versus metastatic tumors. The Cell Marque polyclonal PAX-8 was superior to their monoclonal PAX-8, maintaining strong sensitivity using a ≥ 2+ versus 1+ staining cut point for positive results (79% vs. 18%, respectively), which may be important in cases with scant tissue or background staining. The Biocare monoclonal PAX-8 was negative in all cases. PAX-8 staining in Merkel cell carcinoma expands the spectrum of tumors showing immunoreactivity and may prove to be a useful addition to a diagnostic panel. Awareness of this immunoreactivity and recognition of the antibody source and clone are important to preclude diagnostic pitfalls with tumors in the differential diagnosis.


Microsatellite instability detection in breast cancer using drop-off droplet digital PCR.
The use of conventional methods (immunohistochemistry, pentaplex PCR) for detecting microsatellite instability (MSI), a predictive biomarker of immunotherapy efficacy, is debated for cancers with low MSI prevalence, such as breast cancer (BC). We developed two multiplex drop-off droplet digital PCR (ddPCR) assays targeting four microsatellites, initially identified from public BC whole-genome sequencing dataset. Performances of the assays were investigated and 352 tumor DNA and 28 circulating cell-free DNA from BC patients, with unknown MSI status were blindly screened. Cross-validation of ddPCR MSI status with other MSI detection methods was performed. We then monitored circulating tumor DNA (ctDNA) dynamics before and during pembrolizumab immunotherapy in one patient with MSI-high (MSI-H) metastatic BC. The assays showed high analytical specificity and sensitivity (limit of detection = 0.16%). Among N = 380 samples, seven (1.8%) were found as MSI-H by ddPCR with six of them confirmed by next-generation sequencing (NGS). Specificity was 100% in N = 133 microsatellite stable BC submitted to NGS. In the patient with MSI-H metastatic BC, ctDNA monitoring revealed an early decrease of microsatellite mutant allelic frequencies during immunotherapy. These results demonstrated MSI detection by ddPCR, a non-invasive, fast and cost-effective approach, allowing for large pre-screening of BC patients who may benefit from immunotherapy.


The role of electrically stimulated endocytosis in gene electrotransfer.
Gene electrotransfer is an established method for transfer of genes into cells, however, the mechanism of transfer of DNA across the cell membrane is still not known. Some studies suggest that DNA is translocated through membrane pores while others propose that DNA enters the cell via electro-endocytosis, but no direct observation was performed. In this paper we investigated the second hypothesis. Cells were stained with membrane dye FM 1-43FX, which is used for observation of endocytosis, and then exposed to electric pulses. We analyzed if endocytosis was stimulated by applying electric pulses with intensities below and above the threshold value for gene electrotransfer. No increase in endocytosis from 20 min or even up to 2h after the pulse delivery was observed, regardless of the electric field strength. These observations do not correlate with electrotransfer efficiency, which increases with field strength and is observed only above the threshold value. Our results suggest that electro-endocytosis is not a crucial mechanism for gene electrotransfer and that the hypothesis of DNA entry by translocation through permeabilized membrane is more plausible. The presented results are important for better understanding of the mechanisms of gene electrotransfer and for its optimization for clinical applications.


Pornography Consumption and Extramarital Sex Attitudes Among Married U.S. Adults: Longitudinal Replication.
Social scientific interest in pornography use and effects dates back to at least the mid-twentieth century. Despite this, recent meta-analyses reveal a need for additional longitudinal studies, in general; a need for attitudinal studies, specifically; and a need for studies of U.S. consumers, in particular. In response to these needs and recent calls for the fields of communication and psychological science to prioritize replication, the present study probed whether Wright et al. (Psychol Pop Media 3(2):97-109, 2014) novel longitudinal findings on pornography consumption and extramarital sex attitudes among married U.S. adults were replicable. As in Wright et al., a distal assessment of extramarital sex attitudes did not predict interindividual increases in the likelihood of pornography consumption. Contrary to Wright et al., a distal assessment of pornography consumption also failed to predict interindividual increases in positive attitudes toward extramarital sex. However, more proximal measures of extramarital sex attitudes and pornography consumption did predict over time interindividual change in pornography use and attitudinal positivity, respectively, even after adjusting for participants' age, divorce history, education, race, sex, general unhappiness, martial unhappiness, liberal-conservative political orientation, and religiosity. These results are consistent with prior panel studies in the pornography literature in the macro, but also highlight a need for theoretical development (and testing) on the duration and time-course of selection and socialization effects in the context of pornography use and sexual attitudes.


The feasibility and acceptability of a population-level antenatal risk factor survey: Cross-sectional pilot study.
Australian data on the prevalence and distribution of antenatal risk factors are scarce. This study aimed to investigate (i) the feasibility and acceptability of an antenatal risk factor survey collected in public settings and (ii) whether the survey risk factors co-occur with more sensitive risk factors that are privately asked by clinicians. Design and setting: study-designed survey linked with clinician-collected risk factors. pregnant women attending antenatal clinics at two Victorian hospitals. (i) study-designed survey: young pregnancy, no support, poor/fair/good general health, anxious mood, not finishing high school, no income, long-term illness, living without another adult, not employed, never had a job and proxy poverty measures; (ii) Clinician-collected data: smoking, alcohol/marijuana/drug use, domestic violence, social issues, history of mental health problems and depression symptoms. (i) feasibility and acceptability were assessed through survey completion. Each item was assessed for whether they discriminated risk; dichotomised into risk versus no risk; with a total count calculated. (ii) Co-occurrence was assessed by examining how the risk factor count agreed with clinician-collected items. One hundred and sixty-six of 186 (89%) eligible women completed the survey; 139 of 166 (84%) consented to linking clinician-collected data. The high response and zero missing data demonstrated feasibility and acceptability. Of women with linked data, 92 of 139 (66%) had ≥1 survey risk factor and 30 of 139 (22%) had ≥3; 36 of 139 (26%) had at least one co-occurring clinician-collected risk factor. This survey provides a simple, rapid approach to gathering antenatal risk data publicly. It may be a helpful addition to clinicians' standard history collection.


Ovarian hyperstimulation presenting as acute hydrothorax after in vitro fertilization.
Ovarian hyperstimulation that occurs with an isolated hydrothorax as the only symptom has previously been reported. We describe the second such case, the first to occur after in vitro fertilization and embryo transfer.


Maternal aromatase inhibition via letrozole altered RFamide-related peptide-3 and gonadotropin-releasing hormone expression in pubertal female rats.
Despite prevalence of polycystic ovary syndrome (PCOS) among childbearing women and development of many animal models for this syndrome, information on its etiology is still scarce. The intrauterine hyperandrogenic environment may underlie changes at the level of hypothalamus, pituitary, ovary organization in female offspring, and PCOS later in life. Letrozole has been shown to mimic reproductive and metabolic characteristics of PCOS in adult rodent models. Therefore, this research aimed to assess the condition in a prenatal letrozole-treated rat model. Twenty-eight female rats dams receiving letrozole at certain doses during late pregnancy were used in the trial. Pregnant Sprague-Dawley rats (n=21) received letrozole treatment on gestation days 16-18 at doses of 1.25, 1.0, 0.75, 0.5, and 0.25 mg/kg body weight (BW). Prenatal letrozole treatment delayed parturition time and reduced the litter size in pregnant dams (P<0.0001). Late puberty onset, irregular ovarian cyclicity, increased anogenital distance (AGD), body weight gain, serum testosterone concentration, and reduced estradiol levels (P<0.0001) were observed in the female offspring of dams receiving 1.25 and 1 mg/kg BW letrozole. Furthermore, letrozole at 1.25 and 1 mg/kg BW showed increased RFRP and decreased GnRH mRNA expression (P<0.0001). Letrozole treatment at doses of 1 mg/kg BW and lower was not fetotoxic. It was concluded that 1 mg/kg BW letrozole may be suggested for prenatal PCOS induction.


Gliomatosis cerebri, report of a clinically diagnosed and histologically confirmed case.
Pathological criteria for diffuse gliomatosis are clearly demarcated but its clinical diagnosis has always been hampered by the imaging techniques used. Although with the advent of CT scanning clinical possibilities were improved, isodense brain lesions are difficult to detect and in these cases the diagnosis gliomatosis cerebri can only be suspected from the clinical signs and the absence of radiological signs. We have studied a young patient with a peculiar clinical history, in whom comparison of the CT-scan and NMR scan yielded remarkable results. Our clinical diagnosis of (diffuse) gliomatosis was confirmed by biopsy.


Edge effects and intraguild predation in native and introduced centipedes: evidence from the field and from laboratory microcosms.
Human alteration of habitat has increased the proportion of forest edge in areas of previously continuous forest. This edge habitat facilitates invasion of exotic species into remaining fragments. The ability of native species to resist invasion varies and may depend on intrinsic variables such as dispersal and reproductive rates as well as external factors such as rate of habitat change and the density of populations of introduced species in edge habitat. We examined the distributional and competitive relationships of two members of the class Chilopoda, Scolopocryptops sexspinosus, a centipede native to the eastern US, and Lithobius forficatus, an exotic centipede introduced from Europe. We found that L. forficatus was most abundant in edge habitat and S. sexspinosus was most abundant in the interior habitat at our field sites. Although L. forficatus was present in habitat interiors at 11 of 12 sites, there was no correlation between fragment size and numbers of L. forficatus in interior habitat. The native centipede was rarely found occupying fragment edges. We used laboratory microcosms to examine potential competitive interactions and to indirectly assess prey preferences of the two species. In microcosms both species consumed similar prey, but the native centipede, S. sexspinosus, acted as an intraguild predator on the introduced centipede. Native centipedes were competitively superior in both intraspecific and interspecific pairings. Our results suggest that intraguild predation may aid native centipedes in resisting invasion of introduced centipedes from edge habitat.


Discrimination between subtypes of apamin-sensitive Ca(2+)-activated K+ channels by gallamine and a novel bis-quaternary quinolinium cyclophane, UCL 1530.
1. Gallamine, dequalinium and a novel bis-quaternary cyclophane, UCL 1530 (8,19-diaza-3(1,4),5(1,4)-dibenzena-1 (1,4),7(1,4)-diquinolina-cyclononadecanephanedium) were tested for their ability to block actions mediated by the small conductance, apamin-sensitive Ca(2+)-activated K+ (SKCa) channels in rat cultured sympathetic neurones and guinea-pig isolated hepatocytes. 2. SKCa channel block was assessed in sympathetic neurones by the reduction in the slow afterhyperpolarization (AHP) that follows an action potential, and in hepatocytes by the inhibition of the SKCa mediated net loss of K+ that results from the application of angiotensin II. 3. The order of potency for inhibition of the AHP in sympathetic neurones was UCL 1530 > dequalinium > gallamine, with IC50 values of 0.08 +/- 0.02, 0.60 +/- 0.05 and 68.0 +/- 8.4 microM respectively, giving an equi-effective molar ratio between gallamine and UCL 1530 of 850. 4. The same three compounds inhibited angiotensin II-evoked K+ loss from guinea-pig hepatocytes in the order dequalinium > UCL 1530 > gallamine, with an equi-effective molar ratio for gallamine to UCL 1530 of 5.8, 150 fold less than in sympathetic neurones. 5. Dequalinium and UCL 1530 were as effective on guinea-pig as on rat sympathetic neurones. 6. UCL 1530 at 1 microM had no effect on the voltage-activated Ca2+ current in rat sympathetic neurones, but inhibited the hyperpolarization produced by direct elevation of cytosolic Ca2+. 7. Direct activation of SKCa channels by raising cytosolic Ca2+ in hepatocytes evoked an outward current which was reduced by the three blockers, with dequalinium being the most potent. 8. These results provide evidence that the SKCa channels present in guinea-pig hepatocytes and rat cultured sympathetic neurones are different, and that this is not attributable to species variation. UCL 1530 and gallamine should be useful tools for the investigation of subtypes of apamin-sensitive K+ channels.


The parathyroid glands in the rat as seen by ultrathin step and serial sectioning.
Parathyroid cells exhibit ultrastructural heterogeneity. This has been interpreted as an indication that the cells are undergoing secretory cycles, varying in synthetic activity. By electron microscopy we examined step sections at 1 and 2 micron intervals and serial sections of parathyroids from normocalcemic rats of three different age groups. The secretory cells in all three age groups exhibit structural polarity: nuclei and secondary lysosomes are generally located towards the interstitial tissue, whereas the Golgi is confined mainly to regions in the interior of the cell sheets. Cell surfaces facing the interstitial tissue are smooth, whereas plasma membranes in the interior of the cell sheets form tortuous areas. Consequently, an individual cell profile may show quite different appearance depending on section level. Our results do not support the concept that the observed heterogeneity is necessarily due to differences in organelle contents between the cells. The present investigation thus offers an alternative explanation to the ultrastructural variability among secretory parathyroid cells in the rat.


Clinical Determinants Affecting Indications for Surgery and Chemotherapy in Recurrent Ovarian Granulosa Cell Tumor.
Because reports on the management of recurrent granulosa cell tumor have been sparse, a consensus as to which patients should undergo surgical resection and which patients should be considered for chemotherapy has not been established. A total of 21 tumor recurrences in eight patients with granulosa cell tumor were reviewed. Surgery was performed as the main treatment for 13 recurrences, while chemotherapy was chosen as the main treatment for eight recurrences. Complete tumor resection could be accomplished in 13 of 16 surgeries (81.3%), which include all the ten recurrences without involvement of liver or diaphragm and without ascites. The number of recurrent masses was significantly higher in the early recurrence group (progression free survival < 2 years) than in the late recurrence (progression free survival > 2 years). All cases with a solitary recurrent tumor at an extra-peritoneal site presented a significantly longer progression free survival. For patients with recurrent granulosa cell tumor, surgery may provide the best disease control. In cases with complete resection, the number of recurrent masses was the predictive factor for the next recurrence, and adjuvant chemotherapy might be considered in such cases.


Mini Safe Havens for population recovery and reintroductions 'beyond-the-fence'.
In response to the ongoing decline of fauna worldwide, there has been growing interest in the rewilding of whole ecosystems outside of fenced sanctuaries or offshore islands. This interest will inevitably result in attempts to restore species where eliminating threats from predators and competitors is extremely challenging or impossible, or reintroductions of predators that will increase predation risk for extant prey (i.e., coexistence conservation). We propose 'Mini Safe Havens' (MSHs) as a potential tool for managing these threats. Mini Safe Havens are refuges that are permanently permeable to the focal species; allowing the emigration of individuals while maintaining gene flow through the boundary. Crucial to the effectiveness of the approach is the ongoing maintenance and monitoring required to preserve a low-to-zero risk of key threats within the MSH; facilitating in-situ learning and adaptation by focal species to these threats, at a rate and intensity of exposure determined by the animals themselves. We trialled the MSH approach for a pilot reintroduction of the Australian native New Holland mouse (Pseudomys novaehollandiae), in the context of a trophic rewilding project to address potential naïveté to a reintroduced native mammalian predator. We found that mice released into a MSH maintained their weight and continued to use the release site beyond 17 months (525 days) post-release. In contrast, individuals in temporary soft-release enclosures tended to lose weight and became undetectable approximately 1-month post-release. We discuss the broad applicability of MSHs for population recovery and reintroductions 'beyond-the-fence' and recommend avenues for further refinement of the approach. The online version contains supplementary material available at 10.1007/s10531-022-02495-6.


[Gender patterns in Spanish otolaryngologic doctoral theses].
In last decades women in Spain have a greater access to postgraduate education. The objective of this study was to perform a gender analysis on the Otolaryngology doctoral theses presented in a 25 year-period. The TESEO data base on doctoral theses was searched for theses on Otorhinolaryngology written between 1981 and 2005. As strategy for the research we employed the terms: 1) Otorrinolaringología (Otorhinolaryngology); 2) Cirugía de garganta, nariz y oídos (Ear, nose and throat surgery); 3) Fisiología de la audición (Physiology of hearing); 4) Fisiología del equilibrio (Physiology of balance); 5) Física de la audición (Physics of hearing); and 6) Bioacústica (Bioacoustics). A total of 450 theses (18.0±8.3 theses/year) were found, of which 129 were written by females (28.6%). There was a gender imbalance among authors, with 5.2±3.4 theses/year for women vs. 12.9±6.6 theses/year for men (p=0.0002). Nevertheless, there was a tendency toward equality in the last 10 years (p=0.001). On the other hand, the PhD student's gender was clearly related to the supervisor's gender (p=0.0001). With respect to the main topics in our area (otology, audiology/vestibular diseases, rhinology and pathology of paranasal sinuses and neck diseases), there were no significant differences between males and females (p=0.231). These results indicate a tendency towards equality in the number of men and women successfully completing doctoral studies in Otolaryngology. However, the PhD student's gender is clearly related to the supervisor's gender.


Structural and optical properties of thermal decomposition assisted Gd2O3:Ho(3+)/Yb(3+) upconversion phosphor annealed at different temperatures.
The infrared to visible upconversion fluorescent nanoparticles of Ho(3+)/Yb(3+) codoped Gd2O3 phosphor is synthesized via thermal decomposition route. The as-synthesized sample was annealed at 800, 1000 and 1200°C for 3h and then structural and optical properties were studied. The Rietveld refinement of X-ray diffraction (XRD) data was analyzed to probe the effect of Ho(3+)/Yb(3+) dopant on the structural parameters of Gd2O3 host. The upconversion emission spectra of as-synthesized and annealed samples are compared using 980nm diode laser excitation and five emission bands noticed at 490, 539, 550, 667 and 757nm corresponding to the (5)F3→(5)I8, (5)F4→(5)I8, (5)S2→(5)I8,(5)F5→(5)I8 and (5)I4→(5)I8 manifolds, respectively. The local temperature induced by laser light is also calculated. The fluorescence intensity ratio (FIR) of two thermally coupled transitions (5)F4→(5)I8 and (5)S2→(5)I8 is plotted against the sample temperature and sensor sensitivity of sample is calculated.


Problems in the investigation of urine from patients suffering from primary hyperoxaluria type 1.
Regular calculation of urinary crystallization risk indices in patients suffering from urolithiasis is a recommended measure for treatment adjustment. The more the patient experiences either extensive stone formation or an enhanced recurrence rate, the more important risk index calculations. In patients suffering from primary hyperoxaluria type 1 (PH1), both criteria are met. Different methods of risk determination are known. All strategies for measuring the calcium oxalate (CaOx) crystallization risk of a given urine principally determine this parameter from voided urine. This "bladder urine", however, has possibly passed stone material located in the urinary tract and thus may be depleted in lithogenic components. This is commonly the case for patients with PH1, who mostly experience a massive stone burden or severe nephrocalcinosis. Hence, the question arises as to whether we can adequately determine the crystallization risk in the urine of stone-bearing PH1-patients or not. Based on model calculations, we show that the determination of CaOx formation risk in PH1-patients requires knowledge of the restrictions in risk index interpretation: risk indices calculated from urinalysis (e.g. EQUIL) still indicate, even after strong in vivo stone formation, an enhanced but in fact strongly underestimated risk value. However, the outcome "enhanced" masks the patient's true risk situation. The BONN Risk Index (BRI), in contrast, discloses the process of extreme in vivo crystal formation. As determined, inter alia, from the urinary concentration of free ionized calcium ([Ca(2+)]), BRI approaches abnormally low values, as, in consequence of CaOx - formation, [Ca(2+)] tends to values close to zero. Thus, calculations of urinalysis-based risk indices alone are insufficient strategies for the quantification of a PH1 patient's CaOx crystallization risk.


Amphibians: lungs' lift lost.
The reduction or loss of lungs in some amphibian species is a rare evolutionary occurrence. The discovery of the first lungless frog sheds further light on the physiological, morphological and behavioral changes that allowed sufficient external respiration in the absence of lungs.


GP laughter in lifestyle behaviour consultations: A conversation analytical study of general practice.
This study investigates laughter by General Practitioners (GPs) in response to patient laughter in lifestyle behaviour consultations. We examined video-recorded consultations involving 44 patients of four GPs in Australia. After identifying 33 cases of patient laughter, we examined whether GPs laughed in response. We used Conversation Analysis to explore the appropriateness of GP laughter and non-laughter by investigating the talk before and after the occurrence of patient laughter. GP reciprocal laughter was found in thirteen occasions when patients unsolicitedly mentioned their behaviours, laughed and displayed their evaluative stances (whether the behaviours were positive or negative). On twenty occasions, patients laughed in response to GP enquiries, which worked to problematise particular behaviours. In this context, patient laughter was not usually reciprocated (19/20 cases) because reciprocal laughter may risk being interpreted as laughing at the patient, as evidenced by one deviant case. GP reciprocal laughter may be problematic when the behaviour issues are raised by GPs and patients' evaluative stances regarding their behaviour have not yet been revealed. To decide when it is appropriate to reciprocate laughter, GPs should consider the contexts that lead to patient laughter and patients' evaluative stances.


The Heritage Harbour Health Group: doing it our way.
The Heritage Harbour Health Group is a privately run, nonprofit health care organization founded by the residents of a retirement community for their own use. Such a facility is an original concept, and this article describes how home health care is being provided with a unique blend of professional and volunteer cooperation. Health care professionals may find information and inspiration to conduct their own research into similar programs.


Navigation-guided osteotomies improve margin delineation in tumors involving the sinonasal area: A preclinical study.
To demonstrate and quantify, in a preclinical setting, the benefit of three-dimensional (3D) navigation guidance for margin delineation during ablative open surgery for advanced sinonasal cancer. Seven tumor models were created. 3D images were acquired with cone beam computed tomography, and 3D tumor segmentations were contoured. Eight surgeons with variable experience were recruited for the simulation of osteotomies. Three simulations were performed: 1) Unguided, 2) Guided using real-time tool tracking with 3D tumor segmentation (tumor-guided), and 3) Guided by 3D visualization of both the tumor and 1-cm margin segmentations (margin-guided). Analysis of cutting planes was performed and distance from the tumor surface was classified as follows: "intratumoral" when 0 mm or negative, "close" when greater than 0 mm and less than or equal to 5 mm, "adequate" when greater than 5 mm and less than or equal to 15 mm, and "excessive" over 15 mm. The three techniques (unguided, tumor-guided, margin-guided) were statistically compared. The use of 3D navigation for margin delineation significantly improved control of margins: unguided cuts had 18.1% intratumoral cuts compared to 0% intratumoral cuts with 3D navigation (p < 0.0001). This preclinical study has demonstrated the significant benefit of navigation-guided osteotomies for sinonasal tumors. Translation into the clinical setting - with rigorous assessment of oncological outcomes - would be the proposed next step.


Carbon dioxide exchange characteristics of C4 Hawaiian Euphorbia species native to diverse habitats.
The characteristics of the photosynthetic apparatus of 11 Hawaiian Euphorbia species, all of which possess C4 photosynthesis but range from arid habitat, drought-deciduous shrubs to mesic or wet forest evergreen trees and shrubs, were investigated under uniform greenhouse conditions. Nine species exhibited CO2 response curves typical of C4 plants, but differed markedly in photosynthetic capacity. Light-saturated CO2 uptake rates ranged from 48 to 52 μmol m-2 s-1 in arid habitat species to 18 to 20 μmol m-2 s-1 in mesic and wet forest species. Two possessed unusual CO2 response curves in which photosynthesis was not saturated above intercellular CO2 pressures [p(CO2)] of 10 to 15 Pa, as typically occurs in C4 plants.Both leaf (g'1) and mesophyll (g'm) conductances to CO2 varied widely between species. At an atmospheric p(CO2) of 32 Pa, g'1 regulated intercellular p(CO2) at 12-15 Pa in most species, which supported nearly maximum CO2 uptake rates, but did not result in excessive transpiration. Intercellular p(CO2) was higher in the two species with unusual CO2 response curves. This was especially apparent in E. remyi, which is native to a bog habitat. The regulation of g'1 and intercellular p(CO2) yielded high photosynthetic water use efficiencies (P/E) in the species with typical CO2 response curves, whereas P/E was much lower in E. remyi.Photosynthetic capacity was closely related to leaf nitrogen content, whereas correlations with leaf morphological characteristics and leaf cell surface area were not significant. Thus, differences in photosynthetic capacity may be determined primarily by investment in the biochemical components of the photosynthetic apparatus rather than by differences in diffusion limitations. The lower photosynthetic capacities in the wet habitat species may reflect the lower light availability. However, other factors, such as reduced nutrient availability, may also be important.


A multivariate approach to analyzing the relation between occlusion and craniofacial morphology.
This study examined the association between occlusion and craniofacial morphology using univariate and multivariate statistical methods. Data were obtained from study casts and lateral cephalometric radiographs of 164 children in the early permanent dentition. The following multiple features of occlusion were assessed: molar relation, overjet, overbite, and anterior crowding. Angular skeletal measures assessed cranial base flexure, maxillary horizontal and vertical positions, mandibular horizontal and vertical positions, horizontal and vertical maxillary-mandibular relations, and positions of the incisors. The relation between the Occlusal Index, which is a malocclusion severity index, and skeletal morphology was also investigated. Associations were examined by use of linear correlation, stepwise multiple regression, and canonical correlation analyses. Individually and in combination, occlusal features were poorly associated with individual skeletal measures (r2 less than or equal to 0.35). The strongest association occurred between a linear combination of occlusal features and a linear combination of skeletal measures (R2 = 0.66, p = 0.0001). A malocclusion severity index did not aid in the identification of craniofacial morphology. The results suggested that combinations of certain occlusal characteristics may be associated with specific skeletal types; however, a generalized statement of this concept could not be supported.


Silencing of PMEL attenuates melanization via activating lysosomes and degradation of tyrosinase by lysosomes.
The functionally specialized melanosome is a membrane-enclosed lysosome-related organelle, which coexists with lysosomes in melanocytes. Pre-melanosomal protein (PMEL) initiates pre-melanosome morphogenesis and is the only cell-specific pigment protein required for the formation of fibrils on which melanin is deposited in melanosomes. But the effects of PMEL on melanin synthesis and lysosome activity remain unclear. In the study, PMEL was silenced in human epidermal melanocytes by siRNA transfection. Compared to the non-treated group, melanin content in the transfected cells was greatly reduced. Real-time qPCR, Western blotting and immunofluorescence analyses all showed that PMEL-siRNA transfection reduced protein level of tyrosinase, a key enzyme in melanogenesis, but it does not affect tyrosinase gene expression. Moreover, in the absence of PMEL, lysosomal activation was manifested by an increase in the number of lysosomes and activity of hydrolysis enzymes. The lysosome inhibitors restored tyrosinase expression after PMEL silencing, indicating that tyrosinase was degradated by lysosomes. The data collectively showed that silencing of PMEL suppressed melanization through activating lysosomes and degradation of tyrosinase by lysosomes. Our findings provide novel insight into the interaction between the melanosome and its related organelle, the lysosome, supplying a new idea for the pathogenesis and clinical treatment of pigmented diseases.


[The importance of silicone oil removal in prophylaxis of the late complications of internal tamponade].
To present the functional and anatomical results after silicone oil removal. Covers 32 patients (34 eyes), age 29-77 years. Silicone oil removal was performed between February 1993 and December 1994. The indication for silicone oil tamponade was: PDVR (17/34), rhegmatogenous retinal detachment (12/34), trauma (4/34) and vitreous haemorrhage (1/34). The conditions of silicone oil removal were complete retinal attachment within at least 3 months, visual acuity not worse than 1/50, no proliferations, laser photocoagulation of peripheral retina. In postoperative period retina was attached in 30 cases (88%). Retinal detachment was observed in 4 cases (12%). Visual acuity remains unchanged in 27 cases. We found that silicone oil removal leads in the majority of eyes to a stabilisation of visual acuity and reduces the progress of silicone oil-related complications.


Scalp and depth recordings of induced deep cerebral potentials.
A balanced square wave was introduced between two adjacent depth electrodes implanted in the course of studying patients with intractable epilepsy and who were being considered for surgery. The stimulus current was designed so that charge density loading was well within limits of safety to avoid tissue damage. No neuronal activation was seen, and the stimulus intensity was significantly less than that used in subsequent stimulation session for the purpose of eliciting a clinical response and after-discharges. Averaging techniques were used to record the stimulus at distant electrodes both within the cerebrum and on the scalp. The recorded voltage decrement from the source was nearly identical with the theoretical voltage decrement predicted using principles of electric field theory in which the brain was assumed to be a homogeneous conductive medium. When the voltage recorded on the scalp was compared with the voltages recorded from depth electrodes, it was found that the effect of the highly resistive skull on voltage decrement was relatively less the more centric the source. This result also confirmed predictions based on electric field theory. Most significantly, voltages well within the physiologic range introduced in deep mesial temporal lobe structures were recorded from the scalp.


Individual-level modifiers of the acute effects of air pollution on mortality in Wuhan, China.
Ambient air pollution has posed negative effects on human health. Individual-level factors may modify this effect, but previous studies have controversial conclusions, and evidence is lacking especially in developing countries. This study aims to examine the modifying effects of sex, age, and education level of individuals on the associated between daily mortality and air pollutants, including particulate matter < 10 μm in aerodynamic diameter (PM10), sulfur dioxide (SO2), and nitrogen dioxide (NO2). Time-series analysis was conducted to investigate the acute effects of the air pollution on daily mortality from January 2002 to December 2010 in Wuhan, China. Generalized Additive Models (GAM) were used to examine the association stratified by sex for non-accidental, cardiovascular, and respiratory mortality. For non-accidental mortality, stratified analysis was also conducted by age and educational level. Outdoor air pollution was associated with daily non-accidental and cardiovascular mortality. An increase of 10 μg/m3 in a 2-day average concentration of PM10, SO2, and NO2 was corresponding to the increase in non-accidental mortality of 0.29% (95%CI: 0.06-0.53%), 1.22% (95%CI: 0.77-1.67%) and 1.60% (95%CI: 1.00-2.19%), respectively. The effects of air pollution were faster in females than males. The magnitude of the estimates was higher for females with low education, aged 65-75 years for PM10 and < 65 years for SO2. To be more specific, we observed that per 10 μg/m3 increase in SO2 was association with increases in non-accidental mortality of 2.03% (95%CI: 1.38-2.67) for all females and 3.10% (95%CI: 2.05-4.16) for females with low education. Females and people with low-education are more susceptible to the effect of air pollution, which would provide a sound scientific basis for determination of air pollution standards.


Estrogen and androgen receptors in the liver of cynomolgus monkeys (Macaca fascicularis).
Estrogen receptors (ER) and androgen receptors (AR) were evaluated in the hepatic cytosol from cynomolgus macaques to determine if there were differences associated with gender and endogenous hormone secretion. Saturable, high affinity binding (Kd = 0.2-0.8 nM) was demonstrated for both ER and AR from either male or female monkeys. Displacement of tritiated estradiol from the ER was estrogen specific (including ethinyl estradiol). Both androgens and the synthetic progestins (levonorgestrel and norethindrone) displaced tritiated mibolerone from the AR. Both 8S and 4S molecular forms of ER and AR were demonstrated on 5-20% sucrose density gradients. The ER levels were higher in females in the follicular phase of the menstrual cycle (40.5 +/- 1.9 fmol/mg protein) than levels in males (26.4 +/- 4.8 fmol/mg protein; P less than 0.01) or levels in luteal phase females (31.8 +/- 2.4 fmol/mg protein; P less than 0.05). AR levels were not different between females during different phases of the menstrual cycle (65.8 +/- 4.6 and 69.5 +/- 4.3 fmol/mg protein, follicular and luteal, respectively), but there was a tendency (P less than 0.10) for the levels in males (54.4 +/- 6.6 fmol/mg protein) to be lower than female levels. The demonstration of saturable, high affinity binding of androgens and estrogens in liver tissue of these primates, along with differences associated with gender and the stage of the menstrual cycle, suggests that hepatic receptors are functional and may play an important role in hepatic protein secretion.


Towards Improving Embryo Prioritization: Parallel Next Generation Sequencing of DNA and RNA from a Single Trophectoderm Biopsy.
Improved embryo prioritization is crucial in optimizing the results in assisted reproduction, especially in light of increasing utilization of elective single embryo transfers. Embryo prioritization is currently based on morphological criteria and in some cases incorporates preimplantation genetic testing for aneuploidy (PGT-A). Recent technological advances have enabled parallel genomic and transcriptomic assessment of a single cell. Adding transcriptomic analysis to PGT-A holds promise for better understanding early embryonic development and implantation, and for enhancing available embryo prioritization tools. Our aim was to develop a platform for parallel genomic and transcriptomic sequencing of a single trophectoderm (TE) biopsy, that could later be correlated with clinical outcomes. Twenty-five embryos donated for research were utilized; eight for initial development and optimization of our method, and seventeen to demonstrate clinical safety and reproducibility of this method. Our method achieved 100% concordance for ploidy status with that achieved by the classic PGT-A. All sequencing data exceeded quality control metrics. Transcriptomic sequencing data was sufficient for performing differential expression (DE) analysis. All biopsies expressed specific TE markers, further validating the accuracy of our method. Using PCA, samples clustered in euploid and aneuploid aggregates, highlighting the importance of controlling for ploidy in every transcriptomic assessment.


Alterations in composition of immune cells and impairment of anti-tumor immune response in aged oral cancer-bearing mice.
Aging has been suggested to be associated with immune dysregulation. An understanding of alterations in the host immunity with advancing age is, therefore, important for designing immune therapy for elderly cancer patients. In this context, not much is known about age-associated alterations in the immune system in oral cancer. To evaluate age-associated alterations in the immune system, which might affect anti-tumor immune responses in oral cancer, we performed a comparative analysis of the proportion of different immune cells, the proliferative capacity of T cell compartment, and the response against immune therapies targeting immune check point molecules between young and aged oral cancer-bearing mice. The proportion of immune regulatory cells, such as regulatory T cells and myeloid derived suppressor cells, was significantly increased in aged mice compared to that in young mice. Moreover, the expression of PD-1 and CTLA-4 on both CD4+ and CD8+ T cells was elevated in aged mice compared to that in young mice, and the proliferative abilities of CD4+ and CD8+ T cells derived from aged mice were significantly reduced following stimulation of T-cell receptors. Moreover, tumor growth was significantly enhanced in aged mice compared to that in young mice. However, immunotherapies targeting PD-1, CTLA-4, and PD-L1 resulted in faster tumor regression in aged mice than in young mice. Together, our results indicate that age-associated alterations in the immune system are directly associated with the impairment of anti-tumor immunity in aged mice bearing oral cancer, and might facilitate the progression of the tumor.


New and old markers of progression of diabetic nephropathy.
The onset of diabetic nephropathy is characterised by a rise in albumin excretion rate (AER) and/or a transient rise in glomerular filtration rate (GFR) (hyperfiltration). Without intervention AER increases exponentially and there is a linear decrease in GFR after onset of overt nephropathy. In overt nephropathy, AER is a predictor of decline in GFR and the early AER response to antihypertensive therapy correlates with long-term decline in GFR. AER can be measured by immunoassay or by other techniques including HPLC. However, HPLC assays result in higher levels of AER in normal subjects compared with immunoassayable AER. Recent data suggest that there are distinct albuminuric and non-albuminuric pathways to renal impairment in type 1 and type 2 diabetes. In type 2 diabetes, the non-albuminuric pathway may explain a decline in GFR to <60 ml/min/1.73 m(2) in approximately one in four subjects after accounting for the use of renin angiotensin system inhibitors. In established nephropathy (chronic kidney disease (CKD) stages 3 and 4), plasma cystatin C based estimates of GFR are marginally superior to creatinine based estimates. However, cystatin C clearly outperforms creatinine based estimates of GFR decline at GFR levels >60 ml/min/1.73 m(2) (CKD stages 1 and 2). Other potential markers of progression of diabetic nephropathy include transforming growth factor beta (TGFbeta) and connective tissue growth factor (CTGF). However, long-term studies are needed to define their roles as markers of progression. Diabetic nephropathy is likely to be more susceptible to intervention at an early stage and accurate estimation of GFR is already possible with cystatin C. However, improved formulas for estimating GFR based on using creatinine or other markers are still required, because this may still provide the most cost effective approach applicable to existing clinical practice.


The observation of excited-state dynamical evolution in light-harvesting complex LH2 from Rhodobacter sphaeroides 601.
With selective excitation around BChl-B800 and BChl-B850 absorption bands, we observed the evolution of excited-state dynamics in LH2 from Rhodobacter sphaeroides 601. The dynamical traces demonstrate a dominant excited-state absorption (ESA) followed concomitantly by an ultrafast transmission increase and decay with pulse-width limited time scale at 818 nm and 828 nm excitation. The ESA occurring prior to excitonic thermalization or ground-state bleach was observed at 840 nm as well. These experimental results indicate the competition between the transition from excitonic states to higher-lying excited states and interexciton relaxation, which are of physical significance for understanding excitation transfer and related mechanisms in LH2.


High circulating levels of RBP4 and mRNA levels of aP2, PGC-1alpha and UCP-2 predict improvement in insulin sensitivity following pioglitazone treatment of drug-naïve type 2 diabetic subjects.
High levels of circulating retinol-binding protein 4 (RBP4) and baseline expression of adipogenic genes correlate with subsequent improvement in insulin sensitivity following Thiazolidinedione (TZD) treatment. The aim was to identify baseline characteristics and early changes related to TZD treatment that could predict a good treatment response. Subjects were examined with oral glucose tolerance test, intravenous glucose tolerance test, hyperinsulinaemic euglycaemic clamp, body composition and standard blood sampling at baseline and after 4 and 12 weeks treatment. Subcutaneous adipose tissue biopsies were taken from the abdominal region at baseline, after 3 days and 4 weeks treatment to examine the gene expression profile. Research laboratory in a University hospital. Ten newly diagnosed and previously untreated type 2 diabetic subjects were treated with pioglitazone for 3 months. Baseline characteristics and early changes related to TZD treatment that could predict the response after 3 months. Pioglitazone improved insulin sensitivity after 4 weeks combined with lower glucose and insulin levels without any change in BMI. It was accompanied by lower circulating resistin and plasminogen activator inhibitor-1 levels rapidly increased levels of circulating total and high molecular weight adiponectin as well as adiponectin and adipocyte fatty acid-binding protein (aP2) mRNA expression in the adipose tissue. High levels of circulating RBP4 at baseline and adipose tissue expression of aP2, proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1alpha) and uncoupling protein 2 (UCP-2) predicted a good treatment response measured as improvement in insulin-stimulated whole-body glucose uptake after 3 months. Circulating levels of RBP4 as an index of insulin sensitivity and mRNA levels of adipogenic genes correlate with the subsequent improvement in insulin sensitivity following TZD treatment.


Tribological Characterization of the Heat-Assisted Single Point Incremental Forming Process Applied to the Ti6Al4V Alloy with the Definition of an Adhesion Parameter for the Tool Surface.
Heat-assisted single point incremental forming or HA-SPIF has a great potential for producing one-piece batches of hard-to-form materials such as Ti6Al4V alloy for medical and aeronautical applications. One of the limitations of the process is the difficulty in achieving a reasonable surface finish, which makes essential the characterization of the tribological process in the tool-sheet contact. In fact, not much work can be found at this point in literature. In this research, a novel procedure for evaluating the adhesion on the tool surface is proposed and the influence of the temperature is determined. The surface finish of parts is analyzed, and the changes promoted by HA-SPIF appearing in the morphology of the external surface layer are characterized by SEM.


A simple radiochemical assay of thromboxane B2, 12-hydroxyeicosatetraenoic acid (HETE) and 12-hydroxyheptadecatrienoic acid (HHT) synthetized by human platelets.
A simple radiochemical assay of thromboxane B2, 12-hydroxyeicosatetraenoic acid (HETE) and 12-hydroxyheptadecatrienoic acid (HHT) produced by platelets from arachidonic acid has been described. Moreover, this assay is suitable for measuring thromboxane A2 trapped as 11-mono-O-methylthromboxane B2 and minor products such as prostaglandins E2, F2alpha and D2. The advantage of this technique is discussed as to its ease and specificity.


Analysis of the Healthy Subject Response to Prolonged Contact with Tuberculosis Patients.
Prolonged contact of healthy subjects with Mycobacterium tuberculosis can change their blood formula and immune status, thus reflecting adaptive reactions to constant antigenic load. The peripheral blood analysis of health care workers in a tuberculosis hospital demonstrates changes in cell populations which prevent development of tuberculosis, in particular, CD4+ Т cells and CD3+ Т cells. It is shown that the number of the memory CD4+ Т cells specific to M.tuberculosis antigens which produce interferon gamma depends on the duration of work contact with tuberculosis patients. The use of health care workers' blood characteristics as a control for tuberculosis patients is discussed.