Immunochemical identification of subgroups of vomeronasal nerve fibers and their segregated terminations in the accessory olfactory bulb.
Vomeronasal nerve (VNN) fibers and their terminations in the accessory olfactory bulb (AOB) were studied immunohistochemically using 3 monoclonal antibodies (MAbs). One MAb (R2D5) labeled all VNN fibers. Another MAb (R4B12) labeled a subgroup of the VNN fibers which terminated in the rostrolateral glomeruli in the AOB. The third MAb (R5A10) recognized a complementary subgroup of the VNN which terminated in the caudomedial portion of the AOB. These results for the first time show occurrence of subtypes in the VNN axons with segregated terminations in the AOB.


Perfluorooctylbromide-loaded fucoidan-chlorin e6 nanoparticles for tumor-targeted photodynamic therapy.
Efficient delivery of a photosensitizer (PS) and oxygen to tumor tissue is critical for successful photodynamic therapy (PDT). For this purpose, we developed a fucoidan (Fu)-chlorin e6 (Ce6) nanoparticle (NP) containing perfluorooctylbromide (PFOB). Fu, a biopolymer derived from seaweed, made up the hydrophilic shell of the NP and provided specific targeting to tumor cells by P-selectin binding. Conjugation with the hydrophobic Ce6 enabled self-assembly and Ce6-generated cytotoxic reactive oxygen species to kill tumor cells upon laser irradiation. PF supplied oxygen to the hypoxic tumor tissue and increased the efficacy of the PDT. The developed Fu-Ce6-PF-NPs bound specifically to SCC7 tumor cells and killed them via a photodynamic effect on laser irradiation. High accumulation of the NPs in tumor tissue and improved tumor suppression by PDT were observed in SCC7 tumor-bearing mice. The overall data demonstrated the potential of Fu-Ce6-PF-NP as a tumor-targeting drug carrier for effective PDT.


Quality-of-life reporting in randomized controlled trials shaping the EAU Renal Cell Carcinoma Guidelines.
Quality-of-life (QoL) metrics' importance in patient care has been increasingly recognized and has led oncological societies to issue clinical guidances on their collection. We aim to describe how current RCTs shaping the EAU RCC Guidelines collect QoL metrics and how long they report them. We searched EAU Renal Cell Carcinoma Guidelines' references for phase 3 clinical trials. Data related to if, for how long and how QoL metrics were collected and/or reported were obtained. Over 35 identified trials, with 16 (46 %) having complete information, being included on our study. Among the analyzed studies, 11 (69 %) were on metastasized/advanced disease setting and 5 (31 %) in adjuvant therapy after surgery; none included QoL as a primary outcome, and 13 (81 %) reported positive QoL results. Regarding timings, all (100 %) studies reported QoL during intervention, and at end of treatment, 9 (56 %) until progression and none (0 %) until death. Median OS was reached in 11 (69 %) studies. Across all studies, the median QoL reporting time was of 7.6 (5.0-12.0) months and the median observation time of 27.1 (22.5-53.1) months. As such, trials reported QoL metrics for a of 28.0 % of the possible time at time of their publications. We found that only 46 % of RCTs reported on QoL metrics and most of these evaluated QoL during an insufficient timeframe. Most studies in this field are not adequately informing on QoL metrics, even when they are reported.


Analysis of the SORT1 gene in familial amyotrophic lateral sclerosis.
Substantial efforts have been deployed in the past decade to identify the genetic causes of amyotrophic lateral sclerosis (ALS), and we hypothesized here that mutations in SORT1 or aberrant SORT1 splicing reduce progranulin level and promote neurodegeneration. We sequenced the coding exons of SORT1 in a cohort of 112 unrelated individuals with familial ALS. We also tested for aberrant SORT1 splicing by RT-PCR using RNA samples from cell lines expressing six different ALS-associated TARDBP mutations. We identified one unique missense and two unique silent mutations in our cohort. None are predicted to have functional effects. No aberrant SORT1 splicing event was observed. SORT1 mutations are not a common cause of familial ALS, and the influence of TARDBP mutations on SORT1 splicing still needs to be clarified.


The Metabolic Profiles in Hematological Malignancies.
Leukemia is one of the most aggressive hematological malignancies. Leukemia stem cells account for the poor prognosis and relapse of the disease. Decades of investigations have been performed to figure out how to eradicate the leukemia stem cells. It has also been known that cancer cells especially solid cancer cells use energy differently than most of the cell types. The same thing happens to leukemia. Since there are metabolic differences between the hematopoietic stem cells and their immediate descendants, we aim at manipulating the energy sources with which that could have an effect on leukemia stem cells while sparing the normal blood cells. In this review we summarize the metabolic characteristics of distinct leukemias such as acute myeloid leukemia, chronic myeloid leukemia, T cell lymphoblastic leukemia, B-cell lymphoblastic leukemia, chronic lymphocytic leukemia and other leukemia associated hematological malignancies such as multiple myeloma and myelodysplastic syndrome. A better understanding of the metabolic profiles in distinct leukemias might provide novel perspectives and shed light on novel metabolic targeting strategies towards the clinical treatment of leukemias.


Variations in apolipoproteins B and A1 during the course of myocardial infarction.
The plasma apolipoproteins B and A1, and plasma lipids and lipoproteins, were studied in fifteen patients with acute myocardial infarction. In the days immediately after acute infarction there was a decrease in total cholesterol, low density lipoprotein-cholesterol, total apolipoprotein-B, low density lipoprotein apolipoprotein-B and high density lipoprotein apolipoprotein A1. High density lipoprotein-cholesterol remained unchanged. In the same period the total triglycerides, very low density lipoprotein-protein, very low density lipoprotein-cholesterol, very low density lipoprotein apolipoprotein-B and very low density lipoprotein apolipoprotein A1 were increased. A reduction of the apolipoprotein ratio CII/CIII occurred after the acute phase. After 25--30 days all these values regained their baseline values.


The Histological Features of a Myocardial Biopsy Specimen in a Patient in the Acute Phase of Reversible Catecholamine-induced Cardiomyopathy due to Pheochromocytoma.
A 63-year-old Japanese woman with an adrenal tumor was transferred to our hospital due to cardiogenic shock. Right and left ventriculography showed severe hypokinesis of the middle segment and the apex in both ventricles, and an endomyocardial biopsy demonstrated a small number of necrotic myocytes and cellular infiltration. She was diagnosed with pheochromocytoma and quickly recovered after treatment with an α-blocker. The functional disability of both the right and left ventricles with less myocardial damage due to an excessive level of catecholamine seemed to be related to the early recovery the present patient with catecholamine-induced cardiomyopathy due to pheochromocytoma.


Global prevalence of congenital heart disease in school-age children: a meta-analysis and systematic review.
Congenital heart disease (CHD) is the commonest birth defect. Studies estimating the prevalence of CHD in school-age children could therefore contribute to quantifying unmet health needs for diagnosis and treatment, particularly in lower-income countries. Data at school age are considerably sparser, and individual studies have generally been of small size. We conducted a literature-based meta-analysis to investigate global trends over a 40-year period. Studies reporting on CHD prevalence in school-age children (4-18 years old) from 1970 to 2017 were identified from PubMed, EMBASE, Web of Science and Google Scholar. According to the inclusion criteria, 42 studies including 2,638,475 children, reporting the prevalence of unrepaired CHDs (both pre-school diagnoses and first-time school-age diagnoses), and nine studies including 395,571 children, specifically reporting the prevalence of CHD first diagnosed at school ages, were included. Data were combined using random-effects models. The prevalence of unrepaired CHD in school children during the entire period of study was 3.809 (95% confidence intervals 3.075-4.621)/1000. A lower proportion of male than female school children had unrepaired CHD (OR = 0.84 [95% CI 0.74-0.95]; p = 0.001). Between 1970-1974 and 1995-1999, there was no significant change in the prevalence of unrepaired CHD at school age; subsequently there was an approximately 2.5-fold increase from 1.985 (95% CI 1.074-3.173)/1000 in 1995-1999 to 4.832 (95% CI 3.425-6.480)/1000 in 2010-2014, (p = 0.009). Among all CHD conditions, atrial septal defects and ventricular septal defects chiefly accounted for this increasing trend. The summarised prevalence (1970-2017) of CHD diagnoses first made in childhood was 1.384 (0.955, 1.891)/1000; during this time there was a fall from 2.050 [1.362, 2.877]/1000 pre-1995 to 0.848 [0.626, 1.104]/1000 in 1995-2014 (p = 0.04). Globally, these data show an increased prevalence of CHD (mainly mild CHD conditions) recognised at birth/infancy or early childhood, but remaining unrepaired at school-age. In parallel there has been a decrease of first-time CHD diagnoses in school-age children. These together imply a favourable shift of CHD recognition time to earlier in the life course. Despite this, substantial inequalities between higher and lower income countries remain. Increased healthcare resources for people born with CHD, particularly in poorer countries, are required.


Calcific tendinopathy of the pronator quadratus muscle: A rare site and cause of ulnar sided wrist pain.
Ulnar -sided wrist pain is a common complaint and can present a diagnostic challenge for a musculoskeletal clinician. Calcific tendinopathy is a well-recognised disorder related to the deposition of calcium hydroxyapatite crystals within tendons. Classically, tendons around the shoulder and the hip joints are the most commonly affected sites. We report a rare site of calcific tendinopathy of the Pronator Quadratus muscle presenting with ulnar-sided wrist pain in a 36-year-old male posing a diagnostic dilemma. This case-report highlights the importance of a thorough clinical evaluation and crucial role of cross-sectional imaging in reaching a diagnosis of calcific tendinopathy of the Pronator Quadratus muscle as unusual cause of ulnar-sided wrist pain. This is the first reported case of calcific tendinopathy in pronator quadratus in the literature we could review.


Mutations in CCNB3 affect its location thus causing a multiplicity of phenotypes in human oocytes maturation by aberrant CDK1 activity and APC/C activity at different stages.
Oocyte maturation arrest results in female infertility and the genetic etiology of this phenotype remains largely unknown. Previous studies have proven that cyclins play a significant role in the cell cycle both in meiosis and mitosis. Cyclin B3 (CCNB3) is one of the members of the cyclin family and its function in human oocyte maturation is poorly understood. 118 infertile patients were recruited and WES was performed for 68 independent females that experienced oocyte maturation arrest. Four mutations in CCNB3 were found and effects of these mutations were validated by Sanger sequencing and in vitro functional analyses. We found these mutations altered the location of cyclin B3 which affected the function of cyclin dependent kinase 1 (CDK1) and led to mouse oocyte arrested at germinal vesicle (GV) stage. And then, low CDK1 activity influenced the degradation of cadherin 1 (CDH1) and the accumulation of cell division cycle 20 (CDC20) which are two types of anaphase-promoting complex/cyclosome (APC/C) activators and act in different stages of the cell cycle. Finally, APC/C activity was downregulated due to insufficient CDC20 level and resulted in oocyte metaphase I (MI) arrest. Moreover, we also found that the addition of PP1 inhibitor Okadic acid and CDK1 inhibitor Roscovitine at corresponding stages during oocyte in vitro maturation (IVM) significantly improved the maturation rates in CCNB3 mutant cRNAs injected oocytes. The above experiments were performed in mouse oocytes. Here, we report five independent patients in which mutations in CCNB3 may be the cause of oocyte maturation arrest. Our findings shed lights on the critical role of CCNB3 in human oocyte maturation.


The pathway from cognitive impairment to caries in older adults: A conceptual model.
Caries significantly increases in people with cognitive impairment (PWCI). However, the interrelationships among cognitive impairment, dentally related function (DRF), oral hygiene, and caries remain unclear, increasing difficulties to care appropriately for PWCI. In this study, the researchers tested a conceptual model in which impaired DRF acts as a mediator between cognitive impairment and poor oral hygiene, which then leads to caries. The researchers recruited 68 older adult dentate participants with normal to severely impaired cognition from 3 assisted-living communities in North Carolina. Within 1 week of a calibrated oral examination performed by a geriatric dentist, a trained examiner blinded to the oral examination results conducted cognitive and functional assessments. The researchers conducted statistical analyses to examine the mediating effect of DRF on the relationship between cognitive impairment and oral hygiene or caries. Cognitive impairment (β = 0.05; standard error [SE], 0.02; P = .017) and DRF (β = -0.07; SE, 0.02; P = .005) were significantly associated with oral hygiene. However, the association between cognitive impairment and oral hygiene was greatly reduced and became nonsignificant after controlling for DRF (β = 0.01; SE, 0.03; P = .430), suggesting that the effect of cognitive impairment on oral hygiene was mediated by DRF. DRF accounted for 80% of the total effect of cognitive impairment on oral hygiene. The ratio of the indirect effect to the total effect was .8. Associations between either cognitive impairment or DRF and caries were not significant, but oral hygiene was significantly associated with caries (β = 0.87; SE, 0.28; P < .001). DRF mediated the association between cognitive impairment and oral hygiene. It accounted for 80% of the total impact of cognitive impairment on oral hygiene, which was associated with increased caries in PWCI. As a result of cognitive impairment, impaired DRF plays a critical role in the pathway from cognitive impairment to oral health decline. Therefore, DRF assessment should be a standard component of geriatric dental assessment. A functionally-tailored oral hygiene intervention is essential to improve oral health for persons with cognitive impairment.


The application portfolio empowerment method and ELECTRE-I for optimising the control of ammonia release during the aerobic fermentation process.
Aerobic fermentation composting can transform solid organic waste into biological organic fertiliser, while reducing resource wastage and ecological damage. However, in the composting process, a serious loss of nitrogen occurs, primarily in the form of the release of ammonia gas. The release of ammonia gas not only pollutes the environment, but also diminishes the presence of nutrient elements, resulting in compost products that are lower in quality. Given that many factors influence the release of ammonia gas during the aerobic fermentation process, it is difficult to determine optimal process parameters. In an effort to address this issue, we propose herein a combinational weighting method based on the analytic hierarchy process (AHP) and entropy weighting method to determine the weight of each secondary index. We also establish a parametric optimisation model based on the ammonia release conditions of the ELECTRE-I method that provides a theoretical underpinning and a decision basis for optimising the process parameters that mediate the release of ammonia during the aerobic fermentation process. This method can be widely employed to reduce the release of ammonia gas and may be of significance to the future development of bioengineering-based composting technology.


Perioperative Anuria Resulting in the Diagnosis of Infrasphincteric Ectopic Ureter in an Adult Female.
Ectopic ureter, particularly in females, often presents at a young age as incontinence. Otherwise healthy girls who have continuous urinary wetting during day and night despite adequate toilet training should be held with a high level of suspicion for an extravesical infrasphincteric ectopic ureteral orifice. If such a diagnosis goes undiscovered, frequent reflux or obstruction may cause permanent damage to the renal unit, as in the patient discussed here.


Magnetic fields mapping with the phase reference method.
A simple method for imaging the magnetic field produced by arbitrary current distributions is presented. The method is based on the use of a phase reference image and can be easily adapted to any standard magnetic resonance imaging scanner. Examples are given to application to gradient coil characterization and quantitative field measurements.


Executive function deficit in bipolar offspring: A neurocognitive endophenotype?
Recent studies in bipolar offspring (BO) showed that a low cognitive performance, especially executive function deficit, could be an early marker of bipolar disorder (BD). Nevertheless, these findings have not been replicated (specifically attentional control, flexibility, and working memory). In addition, most studies have focused on children and adolescents, but few studies analyze the executive function performance in BO adults. Our goal was to compare the neurocognitive performance of BO with control parent-offspring (CO) in a sample that included various age groups. We conducted a cohort study, including subjects between six to 30 years old. We evaluated 129 BO and 113 CO subjects using validated psychiatric diagnostic interviews and an extensive neuropsychological battery. Compared to the CO group, the BO group presented a lower performance in several executive functioning domains, mainly in tasks of attentional control, flexibility, and working memory. All age groups exhibited these findings. BO group presents executive function deficits, regardless of the age group: children, adolescents, and adults. This neurocognitive deficit should be accountable as a neurocognitive endophenotype candidate in BD.


PTP1B Inhibitor Claramine Rescues Diabetes-Induced Spatial Learning and Memory Impairment in Mice.
Accumulating clinical and epidemiological studies indicate that learning and memory impairment is more prevalent among people with diabetes mellitus (DM). PTP1B is a member of protein tyrosine phosphatase family and participates in a variety of pathophysiological effects including inflammatory, insulin signaling pathway, and learning and memory. This study was aimed to investigate the effects of CA, a specific inhibitor of PTP1B, on spatial learning and memory impairment in diabetic mice caused by high-fat diet and injection of streptozotocin. We found that the protein expressions of PTP1B increased in hippocampal CA1, CA3, and PFC regions of diabetic mice. Network pharmacology results showed that PTP1B might be one of the key targets between diabetes and cognitive dysfunction, and CA might alleviate DM-induced cognitive dysfunction. Animal experiments showed that CA ameliorated DM-induced spatial learning and memory impairment, and improved glucose and lipid metabolic disorders. Moreover, administration of CA alleviated hippocampal structure damage and enhanced the expressions of synaptic proteins, including PSD-95, SYN-1, and SYP in diabetic mice. Furthermore, CA treatment not only significantly down-regulated the expressions of PTP1B and NLRP3 inflammatory related proteins (NLRP3, ASC, Caspase-1, COX-2, IL-1β, and TNF-α), but also significantly up-regulated the expressions of insulin signaling pathway-related proteins (p-IRS1, p-PI3K, p-AKT, and p-GSK-3β) in diabetic mice. Taken together, these results suggested that PTP1B might be a targeted strategy to rescue learning and memory deficits in DM, possibly through inhibition of NLRP3 inflammasome and regulation of insulin signaling pathway.


Computation of kinetic isotope effects for enzymatic reactions.
We describe a computational approach, incorporating quantum mechanics into enzyme kinetics modeling with a special emphasis on computation of kinetic isotope effects. Two aspects are highlighted: (1) the potential energy surface is represented by a combined quantum mechanical and molecular mechanical (QM/MM) potential in which the bond forming and breaking processes are modeled by electronic structure theory, and (2) a free energy perturbation method in path integral simulation is used to determine both kinetic isotope effects (KIEs). In this approach, which is called the PI-FEP/UM method, a light (heavy) isotope is mutated into a heavy (light) counterpart in centroid path integral simulations. The method is illustrated in the study of primary and secondary KIEs in two enzyme systems. In the case of nitroalkane oxidase, the enzymatic reaction exhibits enhanced quantum tunneling over that of the uncatalyzed process in water. In the dopa delarboxylase reaction, there appears to be distinguishable primary carbon-13 and secondary deuterium KIEs when the internal proton tautomerism is in the N-protonated or in the O-protonated positions. These examples show that the incorporation of quantum mechanical effects in enzyme kinetics modeling offers an opportunity to accurately and reliably model the mechanisms and free energies of enzymatic reactions.


Serum apolipoprotein A-I depletion is causative to silica nanoparticles-induced cardiovascular damage.
The rapid development of nanotechnology has greatly benefited modern science and engineering and also led to an increased environmental exposure to nanoparticles (NPs). While recent research has established a correlation between the exposure of NPs and cardiovascular diseases, the intrinsic mechanisms of such a connection remain unclear. Inhaled NPs can penetrate the air-blood barrier from the lung to systemic circulation, thereby intruding the cardiovascular system and generating cardiotoxic effects. In this study, on-site cardiovascular damage was observed in mice upon respiratory exposure of silica nanoparticles (SiNPs), and the corresponding mechanism was investigated by focusing on the interaction of SiNPs and their encountered biomacromolecules en route. SiNPs were found to collect a significant amount of apolipoprotein A-I (Apo A-I) from the blood, in particular when the SiNPs were preadsorbed with pulmonary surfactants. While the adsorbed Apo A-I ameliorated the cytotoxic and proinflammatory effects of SiNPs, the protein was eliminated from the blood upon clearance of the NPs. However, supplementation of Apo A-I mimic peptide mitigated the atherosclerotic lesion induced by SiNPs. In addition, we found a further declined plasma Apo A-I level in clinical silicosis patients than coronary heart disease patients, suggesting clearance of SiNPs sequestered Apo A-I to compromise the coronal protein's regular biological functions. Together, this study has provided evidence that the protein corona of SiNPs acquired in the blood depletes Apo A-I, a biomarker for prediction of cardiovascular diseases, which gives rise to unexpected toxic effects of the nanoparticles.


Behavioral characterization of the 6-hydroxidopamine model of Parkinson's disease and pharmacological rescuing of non-motor deficits.
Parkinson's disease (PD) is a chronic neurodegenerative condition that is characterized by motor symptoms as a result of dopaminergic degeneration, particularly in the mesostriatal pathway. However, in recent years, a greater number of clinical studies have focused on the emergence of non-motor symptoms in PD patients, as a consequence of damage on the mesolimbic and mesocortical dopaminergic networks, and on their significant impact on the quality of life of PD patients. Herein, we performed a thorough behavioral analysis including motor, emotional and cognitive dimensions, of the unilateral medial forebrain bundle (MFB) 6-hydroxidopamine (6-OHDA)-lesioned model of PD, and further addressed the impact of pharmacological interventions with levodopa and antidepressants on mood dimensions. Based on apomorphine-induced turning behaviour and degree of dopaminergic degeneration, animals submitted to MFB lesions were subdivided in complete and incomplete lesion groups. Importantly, this division also translated into a different severity of motor and exploratory impairments and depressive-like symptoms; in contrast, no deficits in anxiety-like and cognitive behaviors were found in MFB-lesioned animals. Subsequently, we found that the exploratory and the anhedonic behavioural alterations of MFB-lesioned rats can be partially improved with the administration of both levodopa or the antidepressant bupropion, but not paroxetine. Our results suggest that this model is a relevant tool to study the pathophysiology of motor and non-motor symptoms of PD. In addition, the present data shows that pharmacological interventions modulating dopaminergic transmission are also relevant to revert the non-motor behavioral deficits found in the disease.


Autoanalyzer for continuous fractionation and quantitation of the polyphenols content in wines.
A simple continuous flow autoanalyzer for the on-line fractionation of the polyphenols content in wines is proposed. The target compounds are isolated from the matrix by solid-phase extraction on an RP-C18 sorbent column, using selective solvents for the sequential elution of each polyphenol family. Moreover, evaporative light scattering detection (ELSD) is used for the first time for the on-line monitorization of the three polyphenol fractions present in the wine samples. Thus, a single sample injection is required to determine the global concentration of the three selected polyphenol fractions and the whole analysis is completed within a few minutes. Three calibration graphs were constructed for quantitative analysis of the global compounds concentration in every fraction and covered the range 5-300 mg l(-1) (expressed as gallic acid). Average repeatability, expressed as relative standard deviation, was 4%. The proposed autonalyzer was applied to the analysis of a variety of commercial wine samples. The results obtained were compared with those provided by the Folin-Ciocalteau method, being similar in all instances.


The CNS mononuclear phagocyte system in health and disease.
CNS-resident macrophages-including parenchymal microglia and border-associated macrophages (BAMs)-contribute to neuronal development and health, vascularization, and tissue integrity at steady state. Border-patrolling mononuclear phagocytes such as dendritic cells and monocytes confer important immune functions to the CNS, protecting it from pathogenic threats including aberrant cell growth and brain malignancies. Even though we have learned much about the contribution of lymphocytes to CNS pathologies, a better understanding of differential roles of tissue-resident and -invading phagocytes is slowly emerging. In this perspective, we propose that in CNS neuroinflammatory diseases, tissue-resident macrophages (TRMs) contribute to the clearing of debris and resolution of inflammation, whereas blood-borne phagocytes are drivers of immunopathology. We discuss the remaining challenges to resolve which specialized mononuclear phagocyte populations are driving or suppressing immune effector function, thereby potentially dictating the outcome of autoimmunity or brain cancer.


The role of subtalar motion and ankle contact pressure changes from angular deformities of the tibia.
It is a well known entity that fractures of the tibia heal with some component of angular deformity. Ankle and subtalar joints may compensate for small degrees of angular deformities, but the exact amount of malunion that can be accepted without development of late sequalae has yet to be determined. Two recent studies from this institution have concluded that contact changes at the tibiotalar joint tend to be greater with distal third tibial fracture deformities compared to proximal and middle with the ankle in neutral, 5 degrees dorsiflexion, and 20 degrees of plantar flexion. Anterior and posterior bow deformities produced a greater change in contact area of the tibiotalar joint than with valgus or varus deformities. This phenomena may be possibly explained by the subtalar motion in the horizontal plane which averages 23 degrees. Thus, it was the primary purpose of this paper to determine the exact role, if any, in subtalar motion on tibiotalar contact in angular deformities of the tibia. To achieve this objective the subtalar joint was transfixed thereby eliminating its perceived compensatory movement. Six cadaveric lower extremities were disarticulated at the knee joint and stripped of soft tissue preserving capsular and ligamentous structures. A custom universal joint was used to create various angulatory deformities at proximal, middle, and distal third levels of the tibia.(ABSTRACT TRUNCATED AT 250 WORDS)


Fertilization and development in vitro of bovine oocytes following intracytoplasmic injection of heat-dried sperm heads.
This study investigated the development of bovine oocytes following intracytoplasmic injection of sperm heads from spermatozoa dried by heating. When sperm suspension was heated in a dry oven at 50, 56, 90, and 120 degrees C, the mean amounts of residual water were about 0.3 g water/g dry weight within 8 h, 6 h, 1.5 h, and 20 min of heating, respectively. Oocyte activation, cleavage of oocytes, and development of cleaved embryos to the morula stage were better in oocytes injected with spermatozoa stored at 25 degrees C for 7-10 days following drying at 50 and 56 degrees C than at 90 and 120 degrees C; however, only a small proportion of oocytes developed to the blastocyst stage. When spermatozoa were dried at 50 degrees C for 16 h, activation, male pronucleus (MPN) formation, cleavage, and development to the morula stage were less good than when spermatozoa were dried for 8 and 10 h and no blastocysts were obtained. The development of oocytes was significantly better when spermatozoa were stored for 7-10 days at 4 degrees C than 25 degrees C after drying at 50 degrees C for 8 h. Longer storage (7 days-12 mo) of heat-dried spermatozoa at 4 degrees C did not affect MPN formation in activated oocytes, but blastocyst development was significantly lower when spermatozoa were stored for 3 mo or more. These results demonstrate that bovine oocytes can be fertilized with heat-dried spermatozoa and that the fertilized oocytes can develop at least to the blastocyst stage.


Diagnostic accuracy of MRI with extracellular vs. hepatobiliary contrast material for detection of residual hepatocellular carcinoma after locoregional treatment.
To compare the diagnostic accuracy of extracellular gadolinium-based contrast-enhanced MRI (Gd-MRI) and gadoxetic acid-enhanced MRI (EOB-MRI) for the assessment of hepatocellular carcinoma (HCC) response to locoregional therapy (LRT) using explant correlation as the reference standard. Forty-nine subjects with cirrhosis and HCC treated with LRT who underwent liver MRI using either Gd-MRI (n = 26) or EOB-MRI (n = 23) within 90 days of liver transplantation were included. Four radiologists reviewed the MR images blinded to histology to determine the size and percentage of viable residual HCC using a per-lesion explant reference standard. Sensitivities, specificities, accuracies, and agreement with histology for the detection residual HCC were calculated. Gd-MRI had greater agreement with histology (ICC: 0.98 [0.95-0.99] vs. 0.80 [0.63-0.90]) and greater sensitivity for viable HCC (76% [13/17 50-93%] vs. 58% [7/12; 28-85%]) than EOB-MRI; specificities were similar (84% [16/19; 60-97%] vs. 85% [23/27; 66-96%]). Areas under ROC curves for detecting residual viable tumor were 0.80 (0.64-0.92) for Gd-MRI and 0.72 (0.55-0.85) for EOB-MRI. Gd-MRI had greater inter-rater agreement than EOB-MRI for determining the size of residual viable HCC (ICC: 0.96 [0.92-0.98] vs. 0.85 [0.72-0.92]). Gd-MRI may be more accurate and precise than EOB-MRI for the assessment of viable HCC following LRT.


Overexpression of cystic fibrosis transmembrane conductance regulator (CFTR) is associated with human cervical cancer malignancy, progression and prognosis.
To investigate the correlation of cystic fibrosis transmembrane conductance regulator (CFTR) to cervical cancer progression and prognosis by examining CFTR expression levels in different cervical tissues and cell lines. Paraffin-embedded cervical tissue samples (n=192) were collected for immunohistochemistry (IHC), while fresh cervical tissue samples (n=165) and human cervical cell lines were collected for protein and mRNA detection by quantitative real-time PCR and western blot, respectively. Correlations between CFTR expression levels to cancer clinicopathologic features and prognosis were statistically analyzed. Both CFTR mRNA and protein expression gradually increased from normal to precancerous (LSIL, HSIL) and cervical cancer tissues (p<0.05). Furthermore, CFTR expression level was well-correlated to tumor stage (p<0.001), histological grades (p<0.001), lymphatic metastasis (p<0.001), vascular invasion (p<0.05), interstitial invasive depth (p<0.05), tumor size (p<0.05) and HPV infection (p<0.05). In vitro, CFTR mRNA and protein were expressed strongly both in SiHa and HeLa, but little was seen in Caski and H8 (p<0.05). More importantly, overexpression of CFTR conferred significantly poorer survival in cervical carcinoma (Log rank p=0.028), although it was not an independent predictor for prognosis according to multivariate analysis (p>0.05). These results suggest that higher CFTR expression is closely associated with cervical cancer progression, aggressive behaviors and poorer prognosis, indicating that CFTR may function as a novel tumor marker, a prospective prognostic indicator and a potential therapeutic target for cervical cancer.


Neuroprotective effects of lactation against kainic acid treatment in the dorsal hippocampus of the rat.
Marked hippocampal changes in response to excitatory amino acid agonists occur during pregnancy (e.g. decreased frequency in spontaneous recurrent seizures in rats with KA lesions of the hippocampus) and lactation (e.g. reduced c-Fos expression in response to N-methyl-d,l-aspartic acid but not to kainic acid). In this study, the possibility that lactation protects against the excitotoxic damage induced by KA in hippocampal areas was explored. We compared cell damage induced 24 h after a single systemic administration of KA (5 or 7.5 mg/kg bw) in regions CA1, CA3, and CA4 of the dorsal hippocampus of rats in the final week of lactation to that in diestrus phase. To determine cellular damage in a rostro-caudal segment of the dorsal hippocampus, we used NISSL and Fluorojade staining, immunohistochemistry for active caspase-3 and TUNEL, and we observed that the KA treatment provoked a significant loss of neurons in diestrus rats, principally in the pyramidal cells of CA1 region. In contrast, in lactating rats, pyramidal neurons from CA1, CA3, and CA4 in the dorsal hippocampus were significantly protected against KA-induced neuronal damage, indicating that lactation may be a natural model of neuroprotection.


The propagation of motor unit action potential and the location of neuromuscular junction investigated by surface electrode arrays.
A myoelectric signal was recorded from the biceps brachii muscle using surface electrode arrays which consisted of 15 pieces of stainless steel wire placed parallel to each other with 5 mm interelectrode spacing. The electrode assembly was positioned parallel to the underlying muscle fibers and amplifiers were connected bipolarly to 11 pairs of adjacent electrodes. In the chart of recorded signals a propagation of the action potential was observed, originating from a neuromuscular junction and traveling both in the proximal and distal directions. In one recording under an isotonic and isometric contraction condition, two neuromuscular junctions were found to be active. One was located approximately at the center of the muscle and the other was 15 mm distal from it. It was found that the amplitude of the myoelectric potential originating from the distal neuromuscular junction increased gradually during a 1 min heavy muscular loading (50% maximum voluntary contraction).


Blocking Triggering Receptor Expressed on Myeloid Cells-1-Positive Tumor-Associated Macrophages Induced by Hypoxia Reverses Immunosuppression and Anti-Programmed Cell Death Ligand 1 Resistance in Liver Cancer.
Tumor-associated macrophages (TAMs) are recognized as antitumor suppressors, but how TAMs behave in the hypoxic environment of hepatocellular carcinoma (HCC) remains unclear. Here, we demonstrated that hypoxia inducible factor 1α induced increased expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in TAMs, resulting in immunosuppression. Specifically, TREM-1-positive (TREM-1+ ) TAMs abundant at advanced stages of HCC progression indirectly impaired the cytotoxic functions of CD8+ T cells and induced CD8+ T-cells apoptosis. Biological and functional assays showed that TREM-1+ TAMs had higher expression of programmed cell death ligand 1 (PD-L1) under hypoxic environment. However, TREM-1+ TAMs could abrogate spontaneous and PD-L1-blockade-mediated antitumor effects in vivo, suggesting that TREM-1+ TAM-induced immunosuppression was dependent on a pathway separate from PD-L1/programmed cell death 1 axis. Moreover, TREM-1+ TAM-associated regulatory T cells (Tregs) were crucial for HCC resistance to anti-PD-L1 therapy. Mechanistically, TREM-1+ TAMs elevated chemokine (C-C motif) ligand 20 expression through the extracellular signal-regulated kinase/NF-κβ pathway in response to hypoxia and tumor metabolites leading to CCR6+ Foxp3+ Treg accumulation. Blocking the TREM-1 pathway could significantly inhibit tumor progression, reduce CCR6+ Foxp3+ Treg recruitment, and improve the therapeutic efficacy of PD-L1 blockade. Thus, these data demonstrated that CCR6+ Foxp3+ Treg recruitment was crucial for TREM-1+ TAM-mediated anti-PD-L1 resistance and immunosuppression in hypoxic tumor environment. Conclusion: This study highlighted that the hypoxic environment initiated the onset of tumor immunosuppression through TREM-1+ TAMs attracting CCR6+ Foxp3+ Tregs, and TREM-1+ TAMs endowed HCC with anti-PD-L1 therapy resistance.


Optical nanoinjection of macromolecules into vital cells.
A tuneable near infrared 90MHz mode locked femtosecond laser was applied for targeted multiphoton optoporation of vital cells. Here, we demonstrate the laser assisted transfer of large amounts of exogenous materials and even macromolecules into living cells via a transient opening of the membrane and quantify the influx. The use of near infrared lasers also allow the optoporation of cells deep in three dimensional biological structures without photodestructive collateral effects.


Expression of the gene encoding transcription factor PaVP1 differs in Picea abies embryogenic lines depending on their ability to develop somatic embryos.
Maturation of Norway spruce (Picea abies L.) somatic embryos is induced by abscisic acid (ABA). Several proteins were proven to be involved in ABA sensing including ABI3/VP1 transcription factors and their orthologue PaVP1 was characterized in spruce. To evaluate the role of PaVP1 both in embryogenic potential and in the process of embryo maturation, we studied PaVP1 expression in lines with contrast embryogenic capacities in parallel with detailed anatomical characterization. PaVP1 expression was determined by northern blot hybridisation, which revealed presence of two differentially regulated VP1-like B3-domain transcripts. Full-length PaVP1 transcript level was negligible in all lines on the proliferation media, but it differed strongly on the maturation media containing ABA. In non-embryogenic line, lacking any differentiated structures, the transcript remained undetectable. In contrast, in embryogenic lines with meristematic centres attached to suspensor cells, PaVP1 expression increased strongly after transition onto the maturation media. In highly embryogenic lines, it kept on a high level until the embryos reached cotyledonary stage, while in developmentally arrested line incapable to form mature embryos, the expression dropped down in connection with advanced disintegration of the meristematic centres. Removal of ABA from the maturation media after 2 weeks of maturation resulted in aberrant embryo development and rapid decrease in PaVP1 expression, indicating the impact of exogenously supplemented ABA on both initiation and maintenance of PaVP1 expression and proper embryo development. Since permanently high or increasing PaVP1 transcript levels accompanied proper embryo development in all experiments, it could be regarded as a good marker of this process.


Programmed appearance of translatable flagellar tubulin mRNA during cell differentiation in Naegleria.
The programmed de novo synthesis of flagellar tubulin during the hour-long differentiation of Naegleria gruberi from amoebae to flagellates is our paradigm for the study of gene expression during cell differentiation. This paper reports the efficient translation of flagellar tubulin mRNA in the wheat germ cell-free system directed by total or polyadenylated RNA extracted from differentiating cells. The tubulin in the in vitro product has a subunit molecular weight of 55,000, separates into alpha and beta subunits under suitable conditions of polyacrylamide gel electrophoreis and co-polymerizes with calf brain tubulin. At least half of the tubulin synthesized in vitro is precipitated by antibodies specific to flagellar tubulin, and the immunoprecipitated tubulin subunits yield peptide maps similar to those of outer doublet tublin. Flagellar tubulin is the predominant protein synthesized in the cell-free system, and amounts to about 5% of the polypeptides whose synthesis is directed by total RNA from differentiating cells. In contrast, little or no flagellar tubulin is synthesized when the cell-free system is directed by RNA extracted from amoebae prior to differentiation. Translation assays show that at least 92% of the flagellar tubulin mRNA appears during differentiation. The time course of appearance of this mRNA was measured by quantitative immunoprecipitation of the cell-free products. Under conditions where cells from flagella 60 min after initiation of differentiation, translatable flagellar tubulin mRNA was first detected at 20 min, reached a maximum at about 60 min and then declined. An excellent correlation was observed between the amount of translatable flagellar tubulin mRNA and the previously measured rates of flagellar tubulin synthesis in vivo. These results indicate that synthesis of flagellar tubulin is a direct reflection of the abundance of its mRNA, and provide the molecular techniques for dissection of the factors that regulate the rapid appearance of this structural protein during differentiation.


Alterations in nuclear anatomy by chemical modification of proteins in isolated rat liver nuclei.
Whole rat liver nuclei were treated with citraconic anhydride, a reagent specific for primary amines. Dramatic changes were observed in nuclear morphology and light scattering properties. An analysis for DNA and RNA content suggested that DNA was released from the nuclei with a short half-time, approximately 2-4s demonstrating a biphasic release profile. RNA was similarly released but with a monophasic profile. Analysis of SDS-PAGE gels of modified nuclei demonstrated a progressive enrichment of nuclear matrix (lamins) polypeptides with extent of modification. H1 histone was quantitatively lost as a function of modification reagent concentration, while approx. 50% of the nucleosomal histones cosedimented with DNA- and RNA-free nuclei. Modification in the presence of 2 mM EGTA released all the DNA and RNA [less than or equal to 1% remaining) while retaining structures characteristic of nuclear matrix, nucleoli, and ribonucleoprotein (predominantly hnRNA group A and B). These nucleic acid-deficient structures have been termed nuclear fossils to differentiate them from high salt detergent-prepared empty nuclear sacks, nuclear remnants, or nuclear scaffolds. Modification in the presence of 2% Triton X-100 results in structures similar to the nuclear fossils (EGTA treatment), but missing the double bilayer and a 51K polypeptide that is a major component of the other structures. The use of chemical modification on the nucleus provides an experimental approach for examining the role of ionic interactions in controlling nuclear structure. Citraconylation may thus serve two functions: (a) as a protein-specific perturbant of nuclei capable of simply and rapidly preparing a range of structural variants for the analysis of nuclear interactions; (b) offer a paradigm for control of nucleic acid-polypeptide interactions based on post-translational alterations in protein charge.


Genetic diversity of the ATAQ gene in Rhipicephalus microplus collected in Mexico and implications as anti-tick vaccine.
The cattle tick Rhipicephalus microplus has a large impact on cattle production due to its bloodsucking habit and transmission of pathogens that cause babesiosis and anaplasmosis. Application of acaricides constitutes the major control method but is often accompanied by serious drawbacks, including environmental contamination and an increase in acaricide resistance by ticks. The recent development of anti-tick vaccines has provided positive results in the post-genomic era, owing to the rise of reverse vaccinological and bioinformatics approaches to analyze and identify candidate protective antigens for use against ticks. The ATAQ protein is considered a novel antigen for the control of the cattle tick R. microplus; it is expressed in midguts and Malpighian tubules of all ticks from the Rhipicephalus genus. However, genetic diversity studies are required. Here, the ATAQ gene was sequenced of seven R. microplus tick isolates from different regions in Mexico to understand the genetic diversity. The results showed that sequence identity among the Mexican isolates ranged between 98 and 100% and 97.8-100% at the nucleotide and protein levels, respectively. Alignments of deduced amino acid sequences from different R. microplus ATAQ isolates in Mexico revealed a high degree of conservation. However, the Mexican isolates differed from the R. microplus "Mozambique" strain, at 20 amino acid residues. Finally, the analysis of more R. microplus isolates, and possibly of other Rhipicephalus species, to determine the genetic diversity in the ATAQ locus is essential to suggest this antigen as a vaccine candidate that might control tick infestations.


Nicotinic cholinergic activation of magnocellular neurons of the hypothalamic paraventricular nucleus.
The aim of the present work was to determine whether paraventricular neurons possess functional acetylcholine nicotinic receptors. Using infrared videomicroscopy and differential interference contrast optics, we performed whole-cell recordings in hypothalamic slices containing the paraventricular nucleus. Acetylcholine, locally applied by pressure microejection in the presence of the muscarinic antagonist atropine, evoked a rapidly rising inward current in paraventricular magnocellular endocrine neurons. This current persisted in the presence of blockers of synaptic transmission. It could be reversibly suppressed by nanomolar concentrations of methyllycaconitine, a selective antagonist of alpha 7-containing nicotinic receptors, but was insensitive to micromolar concentrations of dihydro-beta-erythroidine, an antagonist acting preferentially on non-alpha 7 nicotinic receptors. In addition, the effect of acetylcholine could be mimicked by exo-2-(2-pyridyl)-7-azabicyclo[2.2.1]heptane, a recently synthesized nicotinic agonist specific for alpha 7 receptors. Acetylcholine also desensitized paraventricular nicotinic receptors. Desensitization was pronounced and recovery from desensitization was rapid, consistent with the notion that paraventricular nicotinic receptors contain the alpha 7 subunit. Nicotinic currents could not be evoked in paraventricular parvocellular neurons, suggesting that these neurons are devoid of functional nicotinic receptors. The electrophysiological data were corroborated by light microscopic autoradiography, showing that [(125)I]alpha-bungarotoxin binding sites are present in all the magnocellular divisions of the paraventricular nucleus but are undetectable in other areas of this nucleus. Immunohistochemistry, performed using antibodies directed against vasopressin and oxytocin, indicated that responsiveness to nicotinic agonists was a property of vasopressin as well as of oxytocin magnocellular endocrine neurons, in both the paraventricular and the supraoptic nucleus. We conclude that nicotinic agonists can influence the magnocellular neurosecretory system by directly increasing the excitability of magnocellular neurons. By contrast, they are probably without direct effects on paraventricular parvocellular neurons.


Assay of a ribonuclease that preferentially hydrolyses mRNAs containing cytokine-derived UA-rich instability sequences.
mRNA molecules encoding a number of inflammatory cytokines, as well as certain proto-oncogenes, contain a conserved UA-exclusive sequence in the 3' untranslated region that confers message instability in vivo. This sequence may comprise a critical regulatory element, governing the level of these mRNA molecules, and determining the efficiency with which they are translated. Through the use of a double-label RNAse assay, we have determined that lysates prepared from mouse macrophages selectively degrade mRNA molecules containing the 3' untranslated UA sequence found in the mRNA encoding human cachectin/TNF. The degree of instability is dependent upon the number of copies of inserted UA sequence present in the target mRNA molecule (a Xenopus beta-globin mRNA). mRNAs containing randomly generated UA sequences are more labile than unmodified globin mRNA, but less susceptible to degradation than mRNAs containing the authentic cachectin-derived sequence. mRNA molecules containing synthetic UG-exclusive sequences are normally stable or protected in vitro. The destruction of UA-containing mRNA is inhibited by random adenylate/uridilate copolymers, but not by guanylate/uridilate copolymers. Boiling or proteinase K treatment destroys the selective nucleolytic activity of macrophage lysates. We propose that the nuclease measured here may serve to regulate cellular levels of mRNA molecules encoding cachectin, other inflammatory cytokines, and certain proto-oncogene products.


Raman spectroscopy reveals distinct differences between two closely related bacterial strains, Mycobacterium indicus pranii and Mycobacterium intracellulare.
A common technique used to differentiate bacterial species and to determine evolutionary relationships is sequencing their 16S ribosomal RNA genes. However, this method fails when organisms exhibit high similarity in these sequences. Two such strains that have identical 16S rRNA sequences are Mycobacterium indicus pranii (MIP) and Mycobacterium intracellulare. MIP is of significance as it is used as an adjuvant for protection against tuberculosis and leprosy; in addition, it shows potent anti-cancer activity. On the other hand, M. intracellulare is an opportunistic pathogen and causes severe respiratory infections in AIDS patients. It is important to differentiate these two bacterial species as they co-exist in immuno-compromised individuals. To unambiguously distinguish these two closely related bacterial strains, we employed Raman and resonance Raman spectroscopy in conjunction with multivariate statistical tools. Phenotypic profiling for these bacterial species was performed in a kinetic manner. Differences were observed in the mycolic acid profile and carotenoid pigments to show that MIP is biochemically distinct from M. intracellulare. Resonance Raman studies confirmed that carotenoids were produced by both MIP as well as M. intracellulare, though the latter produced higher amounts. Overall, this study demonstrates the potential of Raman spectroscopy in differentiating two closely related mycobacterial strains. Graphical abstract.


Lessons learned from a pandemic: implications for a combined exercise and educational programme for medical students.
The 'MED-WELL' programme is a combined exercise and educational intervention designed to promote well-being among medical students and educate students about prescribing exercise as medicine in clinical practice. Due to COVID-19 public health restrictions of social distancing the 'MED-WELL' programme was offered online instead of in-person in 2021. The aim of this study is to compare the experiences of participants in the 'MED-WELL' programme online to those that previously participated in the same programme in-person to understand the student experience and optimize programme delivery. Purposive sampling was used to recruit 20 participants to a qualitative study using semi-structured interviews. Ten study participants took part in the 'MED-WELL' programme when it was offered in-person, and the other ten study participants took part in the programme when it was offered online. All interviews were audio-recorded and transcribed using Microsoft Teams. A combined inductive and deductive approach was used for analysis. An inductive thematic analysis was utilized to categorize data into higher order codes, themes, and overarching themes. The theory of online learning provided the theoretical framework for a deductive approach. Analysis of the data produced five overarching themes: 'student-student', 'student-teacher', 'student-content', 'student-environment', and 'effects of a pandemic'. The first four themes detail distinct types of interaction that participants had with various entities of the 'MED-WELL' programme and the effects that these interactions had on participant experiences. 'Effects of a pandemic' refers to the context of delivering the 'MED-WELL' programme online during a pandemic and how this mode of delivery influenced participants and the programme. Optimizing the 'MED-WELL' programme relies on an understanding of how participants interact with different entities of the programme and are motivated to attend and engage. Participants tended to favour an in-person mode of delivery, however certain advantages of delivering the programme online were also identified. The findings from this study can be used to inform similar experiential and educational exercise interventions, and may help plan for potential future restrictions on in-person educational and exercise-based programmes.


The circular RNAs differential expression profiles in the metastasis of salivary adenoid cystic carcinoma cells.
In order to reveal circular RNAs (circRNAs) differential expression profiles and investigate the function and mechanism of circRNAs in the metastasis of salivary adenoid cystic carcinoma (SACC), microarray was used to detect differentially expressed circRNAs in SACC-83 and SACC-lung metastasis (LM) cell lines. Up-regulated circRNAs were analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses to further predict their function. Expression of candidate circRNA and microRNA (miRNA) was determined using quantitative real-time polymerase chain reaction (qRT-PCR). Constructed circRNA-miRNA-mRNA co-expression network was based on TargetScan, miRanda databases. Wound healing and transwell assays were completed to examine the effects of hsa_circRNA_001982 and miR-181a-5p on cell migration and invasion. qRT-PCR confirmed hsa_circRNA_092556, hsa_circRNA_101379, and hsa_circRNA_001982 up-regulation in SACC-LM. miR-181a-5p was down-regulated in SACC-LM and correlated with up-regulated hsa_circRNA_001982. Wound healing and transwell assays indicated that silencing hsa_circRNA_001982 inhibited the migration and invasion of the SACC-LM cells. Furthermore, over-expression of hsa_circRNA_001982 promoted the migration and invasion of SACC-83 cells. Interestingly, up-regulation or down-regulation of miR-181a-5p led to the opposite result in wound healing and transwell assays. Overall, differential expression circRNA profiles in SACC-83 and SACC-LM cells may reveal potential targets and a novel mechanism of circRNAs in the metastasis of SACC. Moreover, the interaction of hsa_circRNA_001982/miR-181a-5p is closely related to the metastasis of SACC cells.


Nerve growth factor (NGF) levels in follicular fluid of infertile patients undergoing to in vitro fertilization (IVF) cycle.
The aim of this study was to determine the levels of NGF in follicular fluid and serum samples, and to correlate them with some characteristics of the patients (age) and of the IVF protocol (E2 levels, number of collected oocytes, used FSHr IU) to investigate its rule in the folliculogenesis. This study examined a sample of 78 women undergoing to FIVET/ICSI cycles since October 2011 to March 2013. NGF levels were determinated in follicular fluid (FF) and serum sample using enzyme immunoassay ELISA kit for NGF. The NGF level was significantly higher in FF (213.76 pg/ml) than in basal serum (46.47 pg/ml (p < 0.001) and in serum sample of the pick-up day (60.75 pg/ml (p < 0.001). In FF, the levels of NGF were positively correlated to age of women (corr.coeff. r = 0.44) and units of FSHr used during stimulation protocol (corr.coeff r = 0.34). Our results demonstrate that the FF levels are higher than the blood; so there is a local production of NGF in the ovarian follicle, important for the follicle's growth and oocyte quality. We could also say that the increase of NGF levels is correlated to a lower ovary response, that is obviously less in the older women.


Premature changes in surface activity in lung fluid of fetal lambs.
The present study was conducted to monitor changes in surface activity of lung and amniotic fluid of premature lambs. Lung fluid was continuously obtained from 10 tracheotomized fetal lambs (gestational age 98-112 days) until (premature) parturition. Amniotic fluid was sampled twice a week from 3 other lambs by catheterization, and during cesarean section at the end of the gestational period from another 3 lambs. Surface activity in lung fluid increased on the first postoperative day, the more so when this occurred at a later fetal age in the period of 98-112 days. These findings show that even before day 110 'stress' can result in a release of surface active material. A return to low surface activity (about 26 mN/m) was observed within 3 to 6 days. Thereafter, between days 110 and 120, surface activity increased steeply within the individual animals but with large variability between the fetuses. The steep and premature increased was followed by a further but slow increase during the mature period and Ymin values of approximately 2 mN/m were reached. Amniotic fluid surface activity changed slowly with gestational age, but did not reach Ymin values of less than 15 mN/m. Lung fluid total phospholipid phosphorus values were low (about 6 micro M) up to 125 days, when they increased rapidly (greater than 16 micro M). Lung fluid production increased from 4.9 ml/h at the beginning of the experiment to 12.1 ml/h after day 130 but decreased dramatically 1-3 days before (premature) parturition. Experiments on fetal lung maturation should be started before day 105 of gestation and we suggest that measurement of surface activity of fetal lung fluid may be useful in monitoring changes in the premature alveolar type II cells of lambs.


Female gender is an independent predictor of operative mortality after coronary artery bypass graft surgery: contemporary analysis of 31 Midwestern hospitals.
Women have a higher operative mortality (OM) after coronary artery bypass graft (CABG) surgery than men. Suggested contributing factors have included women's increased age, advanced disease, comorbidities, and smaller body surface area (BSA). It is unclear whether women's increased risk factors fully account for this difference or whether female gender within itself is associated with increased OM. We attempted to determine whether, all other factors being equal, there is a significant difference in OM between men and women undergoing CABG. We retrospectively reviewed a clinical database of 15,440 patients who underwent CABG at 31 Midwestern hospitals in 1999-2000. Each patient record consisted of >400 data elements. Risk-adjusted mortality rates were computed using a predictive equation derived by stepwise logistic regression. Overall, women were older, had a higher incidence of diabetes and valvular disease, and were more likely to be presenting in shock. The OM for the entire population was 2.88% (women 4.24% versus men 2.23%, P<0.0001). Lower BSA was found to be an independent predictor of increased mortality, and a direct inverse relationship between BSA and OM was noted. After adjusting for all comorbidities including BSA, female gender remained an independent predictor of increased mortality (risk-adjusted OM was 3.81% for women and 2.43% for men). Thus, whereas risk adjustment reduced women's OM from 90% higher than men's to 22% higher, a significant difference remained. In this contemporary data set from 31 Midwestern hospitals, female gender was an independent predictor of perioperative mortality, even after accounting for all comorbidities, including low BSA.


Presence of an expressed beta-tubulin gene (TUBB) in the HLA class I region may provide the genetic basis for HLA-linked microtubule dysfunction.
An expressed beta-tubulin gene (TUBB) has previously been localized to chromosome region 6pter-p21 in man. By using a panel of deletion mutant cell lines and radiation-reduced hybrids containing fragments of chromosome 6, the TUBB locus could be mapped to the HLA class I region at 6p21.3. A long range restriction map including TUBB and several HLA class I genes was then generated by rotating field gel electrophoresis. The results show that TUBB maps to a segment 170-370 kb telomeric of HLA-C. This location suggests that a mutation at the TUBB locus could be the cause for certain forms of HLA-linked microtubule dysfunction, including immotile cilia syndrome.


Maxwelleus, a new genus of Curculionidae from the Himalayas, with two new species (Coleoptera: Curculionidae: Molytinae: Euderini).
The new genus Maxwelleus, with two new species, M. tibetanus (China, Tibet) and M. birmanus (Myanmar [Burma]), is described and illustrated. The new genus is immediately recognized by large body size (12.5-13.8 mm), pronotum with a distinct transverse groove or constriction behind apex and elytra with narrow but distinct transverse patches of yellow scales. A key to the species is given.


Identification of the components of the murine T cell antigen receptor complex.
In addition to the alpha and beta chains of the MHC class II restricted antigen receptor, monoclonal anti-receptor antibodies coprecipitate four polypeptides that appear to be noncovalently associated with the alpha-beta dimer of murine T cells. Included in the murine T cell antigen receptor complex are two glycoproteins of 25 kd (gamma) and 21 kd (delta) and two nonglycosylated polypeptides of 26 kd (epsilon) and 16 kd (zeta). The epsilon chain appears to possess an intrachain disulfide bond and zeta exists in the complex as a disulfide-linked homodimer. The delta chain is phosphorylated on a serine residue in response to T cell activation with antigen. In contrast, both delta and epsilon are phosphorylated in response to treatment of the T cells with phorbol 12-myristate 13-acetate. These polypeptides may play a role in the transduction of the signal(s) in T cell activation.


Analysis of the glycosylation and phosphorylation of the lysosomal enzyme, beta-hexosaminidase B, by site-directed mutagenesis.
Lysosomal enzymes require a mannose 6-phosphate recognition marker, constructed on asparagine-linked oligosaccharide chains, for targeting to lysosomes. We have identified the glycosylation sites of human beta-hexosaminidase B and have determined the influence of individual oligosaccharides on the phosphorylation, lysosomal targeting, and catalytic activity of the enzyme. The five potential glycosylation sites of the hexosaminidase beta-chain were modified individually by site-directed mutagenesis, and the constructs were expressed in COS 1 cells. By this analysis, we determined that four of the five potential sites were glycosylated. Two of the four oligosaccharides were preferentially phosphorylated. The absence of these two preferentially phosphorylated oligosaccharides resulted in greatly reduced amounts of the lysosomal form of the enzyme with increased secretion into the medium. The catalytic activity of beta-hexosaminidase B was not significantly altered by the absence of individual oligosaccharides suggesting the folding and assembly of the enzyme was not disrupted.


Antibody response in elderly vaccinated four times with an mRNA anti-COVID-19 vaccine.
The humoral response after the fourth dose of a mRNA vaccine against COVID-19 has not been adequately described in elderly recipients, particularly those not exposed previously to SARS-CoV-2. Serum anti-RBD IgG levels (Abbott SARS-CoV-2 IgG II Quant assay) and neutralizing capacities (spike SARS-CoV-2 pseudovirus Wuhan and Omicron BA.1 variant) were measured after the third and fourth doses of a COVID-19 mRNA vaccine among 46 elderly residents (median age 85 years [IQR 81; 89]) of an assisted living facility. Among participants never infected by SARS-CoV-2, the mean serum IgG levels against RBD (2025 BAU/ml), 99 days after the fourth vaccine, was as high as 76 days after the third vaccine (1987 BAU/ml), and significantly higher (p = 0.030) when the latter were corrected for elapsed time. Neutralizing antibody levels against the historical Wuhan strain were significantly higher (Mean 1046 vs 1573; p = 0.002) and broader (against Omicron) (Mean 170 vs 375; p = 0.018), following the fourth vaccine. The six individuals with an Omicron breakthrough infection mounted strong immune responses for anti-RBD and neutralizing antibodies against the Omicron variant indicating that the fourth vaccine dose did not prevent a specific adaptation of the immune response. These findings point out the value of continued vaccine boosting in the elderly population.


Molecular Imaging of the GABAergic System in Parkinson's Disease and Atypical Parkinsonisms.
During recent years, there has been a growing interest in GABAergic alterations in parkinsonian disorders. This paper aims to review the latest literature published, focusing on in vivo neuroimaging, and to suggest potential future avenues of research in the field. A growing number of neuroimaging studies have focused on the association with different symptoms of Parkinson's disease, thereby suggesting a GABAergic role in motor symptoms, gait disturbances, frontal cognition, somatic symptom disorder, and hallucinations. However, there are a number of conflicting results, and further investigations in larger, clinically well-defined cohorts are needed to elucidate possible correlations. In progressive supranuclear palsy, recent evidence suggests a decrease of GABA in the frontal lobe. In this narrative review, we discuss the possible GABAergic role in the symptoms of PD and atypical parkinsonisms and outline possible research strategies for future neuroimaging of GABAergic changes in parkinsonian disorders.


Decreased insulin sensitivity is associated with the extent of coronary artery disease in patients with angina.
Insulin resistance has been proposed as an important risk factor in the development of atherosclerosis. To evaluate the association of insulin resistance and coronary atherosclerosis, we investigated the correlation between insulin sensitivity and the degree of coronary stenosis in patients with angina pectoris. The study population consisted of 74 subjects with angina (54 men and 20 women) aged from 31 to 73 years. Coronary angiograms were evaluated by three semiquantitative scoring systems (vessel score, stenosis score and extent score) to estimate the extent of focal and diffuse coronary artery disease (CAD). Insulin sensitivity (K(ITT)) was determined by the insulin tolerance test. There were significant correlations existed between K(ITT) and all three coronary scores. Multivariate analysis revealed significant and independent correlations of all three coronary scores with K(ITT) (vessel score: beta = -0.349, p = 0.004; stenosis score: beta = -0.487, p < 0.001; extent score: beta = -0.481, p < 0.001), even in patients without diabetes mellitus (vessel score: beta = -0.387, p = 0.008; stenosis score: beta = -0.469, p < 0.001; extent score: beta = -0.559, p < 0.001). K(ITT) was significantly lower in patient with diffuse CAD than without diffuse CAD (2.13 +/- 0.66 vs. 2.57 +/- 0.79%/min, p < 0.05). However, K(ITT) was not different between patients with and without focal CAD. Insulin sensitivity has statistically significant and independent associations with the extent of coronary stenosis. These results suggest that insulin resistance may play a major role in the development of diffuse coronary artery stenosis.


Interferon-induced changes in expression of antigens defined by monoclonal antibodies on malignant and nonmalignant mononuclear hematopoietic cells.
The effect of alpha interferon (alpha IFN) on the expression of histocompatibility--as well as differentiation antigens on normal and malignant hematopoietic mononuclear cells--were investigated by cell cytofluorometry using a panel of monoclonal antibodies (MoAbs). An increase in the expression of HLA-antigens detected by beta 2-Microglobulin (beta 2-M) could be demonstrated for peripheral blood mononuclear cells, non-T cells, Null cells, activated T cells, fetal thymocytes, adherent cells, and on four malignant non-T lymphoblastoid cell lines. In contrast, no significant differences were observed in the expression of antigens specific for B-lymphocytes (B1), T-lymphocytes (T3, T4, T6, T8, T11), NK-cells (901) and adherent cells (Mo1-4). Likewise, the expression of Ia-antigens remained unaltered on non-T cells, Null cells, and monocytes. The only other effect of IFN was an increase in the number as well as the amount of lymphocytes expressing the T10 antigen. It thus seems that the enhancing effect of IFN on resting cells of the immune system is highly selective. On the four lymphoblastoid cell lines, the expression of the common acute lymphoblastic leukemia antigen (CALLA) was significantly decreased concomitantly with the increase in MHC-antigens. On the other hand, the density of both a HLA-D related Ia antigen (I2) and a B-lymphocyte differentiation antigen (B1) remained unaltered following IFN treatment. The implications of these findings are discussed.


Depressed prostaglandin release from peritoneal cells induced by a T cell adjuvant, lentinan.
PGE and PGF release from peritoneal exudate cells was studied in mice after injection with two beta (1-3) glucans, the antitumor active lentinan and the inactive pachyman, 4 days after injection of both polysaccharides, the spontaneous and phagocytosis-induced PGE and PGF release was markedly suppressed. However, only the immunopotentiator lentinan induced peritoneal exudate cells which exhibited a longer lasting diminished PG release. The data suggest that the T cell adjuvant lentinan may potentiate cellular immune responses by reducing synthesis of immune suppressive prostaglandins from peritoneal exudate cells.


Determinants of increased risk for recurrence in patients undergoing radical hysterectomy for stage IB and IIA carcinoma of the cervix.
From 1939 to 1977, 431 patients underwent radical hysterectomy as primary therapy for Stage IB or IIA carcinoma of the cervix at Memorial Sloan-Kettering Cancer Center. Only 11 patients were lost to follow-up at intervals of from 1 to 94 months. Assessment of gross and histologic extent of disease was correlated with the prevalence of nodal metastases and survival. Increasing tumor size, depth of invasion, and histologic grade were covariable and predictive of both lymph node metastases and recurrence. After stratifying for nodal metastases, adenocarcinoma cell type, the size of the primary tumor, depth of invasion into the cervix, and histologic grade were associated with decreased survival. For the 85 patients with documented recurrence of their carcinoma, the time to recurrence varied inversely with primary tumor size. Of 56 patients with documented recurrence and negative nodes at the time of their initial therapy, 10 patients (18%) were ultimately salvaged. None of the 29 patients with recurrent carcinoma and positive nodes at the time of their initial lymphadenectomy was successfully treated. Analysis of prognostic factors identifies a group of patients at high risk for recurrence and decreased survival for whom prospective trials of adjunctive treatment should be considered.


Lymphovascular space invasion and estrogen receptor status in high-grade serous ovarian cancer - A multicenter study by the FRANCOGYN group.
The aim of this study was to evaluate the impact of Lymphovascular Space Invasion (LVSI) on Overall Survival (OS) and Recurrence-Free Survival (RFS) in patients managed for high-grade serous epithelial ovarian cancer (HGSOC). Retrospective multicenter study by the FRANCOGYN research group between January 2001 and December 2018. All patients managed for HGSOC and for whom histological slides for the review of LVSI were available, were included. The characteristics of patients with LVSI (LVSI group) were compared to those without LVSI (No LVSI group). A Cox analysis for OS and RFS analysis was performed in all populations. Over the study period, 410 patients were included in the thirteen institutions. Among them, 289 patients had LVSI (33.9%). LVSI was an independent predictive factor for poorer Overall and Recurrence-Free Survival. LVSI affected OS (p<0.001) and RFS (p<0.001), Association of LVSI status and estrogen receptor status (ER) also affected OS and RFS (p = 0.04; p = 0.04 respectively). The presence of LVSI in HGSOC has an impact on OS and RFS and should be routinely included in the pathology examination along with ER status.


Increased macrophages and changed brain endothelial cell gene expression in the frontal cortex of people with schizophrenia displaying inflammation.
Elevated pro-inflammatory cytokines exist in both blood and brain of people with schizophrenia but how this affects molecular indices of the blood-brain barrier (BBB) is unclear. Eight mRNAs relating to BBB function, a microglia and three immune cell markers were measured by qPCR in the prefrontal cortex from 37 people with schizophrenia/schizoaffective disorder and 37 matched controls. This cohort was previously grouped into "high inflammation" and "low inflammation" subgroups based on cortical inflammatory-related transcripts. Soluble intercellular adhesion molecule-1 (sICAM1) was measured in the plasma of 78 patients with schizophrenia/schizoaffective disorder and 73 healthy controls. We found that sICAM1 was significantly elevated in schizophrenia. An efflux transporter, ABCG2, was lower, while mRNAs encoding VE-cadherin and ICAM1 were higher in schizophrenia brain. The "high inflammation" schizophrenia subgroup had lower ABCG2 and higher ICAM1, VE-cadherin, occludin and interferon-induced transmembrane protein mRNAs compared to both "low inflammation" schizophrenia and "low inflammation" control subgroups. ICAM1 immunohistochemistry showed enrichment in brain endothelium regardless of diagnosis and was localised to astrocytes in some brains. Microglia mRNA was not altered in schizophrenia nor did it correlate with ICAM1 expression. Immune cell mRNAs were elevated in "high inflammation" schizophrenia compared to both "low inflammation" schizophrenia and controls. CD163+ perivascular macrophages were identified by immunohistochemistry in brain parenchyma in over 40% of "high inflammation" schizophrenia brains. People with high levels of cytokine expression and schizophrenia display changes consistent with greater immune cell transmigration into brain via increased ICAM1, which could contribute to other neuropathological changes found in this subgroup of people.


An atlas of O-linked glycosylation on peptide hormones reveals diverse biological roles.
Peptide hormones and neuropeptides encompass a large class of bioactive peptides that regulate physiological processes like anxiety, blood glucose, appetite, inflammation and blood pressure. Here, we execute a focused discovery strategy to provide an extensive map of O-glycans on peptide hormones. We find that almost one third of the 279 classified peptide hormones carry O-glycans. Many of the identified O-glycosites are conserved and are predicted to serve roles in proprotein processing, receptor interaction, biodistribution and biostability. We demonstrate that O-glycans positioned within the receptor binding motifs of members of the neuropeptide Y and glucagon families modulate receptor activation properties and substantially extend peptide half-lives. Our study highlights the importance of O-glycosylation in the biology of peptide hormones, and our map of O-glycosites in this large class of biomolecules serves as a discovery platform for an important class of molecules with potential opportunities for drug designs.


Frailty Is Associated with an Increased Risk of Incident Type 2 Diabetes in the Elderly.
To investigate whether frailty is associated with an increased risk of incident type 2 diabetes mellitus (T2DM) in a prospective cohort of community-dwelling older people. Longitudinal study, mean follow-up of 4.4 years. Progetto Veneto Anziani (Pro.V.A.) study that involved older community-dwellers. 1754 men and women older than 65 years without T2DM at baseline. Frailty status was defined according to Fried criteria and categorized as frailty (≥3 criteria), prefrailty (1-2 criteria), or no frailty (0 criterion). Incident T2DM was defined as fasting plasma glucose (FPG) ≥7.0 mmol/L, or glycosylated hemoglobin ≥6.5%, the use of glucose-lowering drugs, or FPG ≥11.1 mmol/L on a 2-hour oral glucose tolerance test during the follow-up. All T2DM diagnoses were confirmed by endocrinologists. At baseline, frail participants (n = 174) were significantly (a) more obese and had higher waist circumference, (b) experienced a higher rate of cardiovascular disease (including hypertension), and (c) presented with higher, but not pathologic, values of glycosylated hemoglobin and FPG than prefrail (n = 830) and nonfrail participants (n = 750). Over a 4.4-year follow-up, 265 individuals developed T2DM. In a logistic regression analysis, adjusted for potential baseline confounders, frailty [odds ratio (OR) = 1.87, 95% confidence interval (CI) = 1.31-2.13, P < .0001] and prefrailty (OR = 1.60, 95% CI = 1.27-2.00, P < .0001) were associated with a significantly higher incidence of T2DM than in nonfrail individuals. Among community-dwelling older people, frailty and prefrailty were significant and independent predictors of T2DM, which is a major and potentially preventable risk factor for multiple comorbidities.


Cross-striated banded structures in cutaneous leishmaniasis.
Banded structures were observed in 4 cases of cutaneous leishmaniasis. These structures were found in proximity to macrophages containing leishmania and were intermingled with native collagen, elastic fibres and ground substance fibrils. The possibility that banded structures are an indication of active, accelerated collagen production is discussed.


Girls' recurrent and concurrent body dissatisfaction: correlates and consequences over 8 years.
The present study examines the patterns of body dissatisfaction during adolescence in order to determine whether girls enter onto trajectories of body dissatisfaction that are associated with eating and depressive symptoms during adolescence and in young adulthood. Body dissatisfaction was studied in 120 adolescent girls drawn from a normal population of students enrolled in private schools in a major metropolitan area. They were seen at three times over an 8-year period (mean ages = 14.3, 16.0, and 22.3). Recurrent body dissatisfaction during adolescence was associated with earlier pubertal maturation and elevated depressive and eating symptoms in young adulthood compared with girls who maintained positive body images during adolescence. Concurrent body dissatisfaction was also a correlate of greater depressed affect and eating symptoms at the time these girls experienced body dissatisfaction. Findings suggest that adolescent body dissatisfaction has consequences for affect in adulthood.


[Evading the catastrophy].
A 21-year-old athletic woman had been suffering from flank pain during fluid intake after sports for some time. Urological work-up revealed hydronephrosis with a "fishhook" shape and medial displacement of the ureter. This ureteral narrowing was studied ureterorenoscopically, bioptically and endoscopic-radiologically without the correct diagnosis of a retrocaval ureter being made. Therapeutically, a DJ stent was inserted several times, a balloon dilatation was performed under anaesthesia three times and finally a permanent DJ catheter was inserted. Due to the patient's dissatisfaction, an endoscopic endopyelotomy using Acucise was offered.


Recipients' perspective on breaking bad news: how you put it really makes a difference.
The goal of this study was to show that physician communication style of breaking bad news affects how the physician is perceived, how satisfied recipients of bad news are with the consultation, and how they feel after the consultation. Female participants (students, N=159) were asked to put themselves in the shoes of a patient receiving the bad news of a breast cancer diagnosis. Participants were randomly assigned to watch one of three prototypical physician communication styles of breaking bad news on videotape: patient-, disease-, or emotion-centered communication. Results showed that these three prototypical communication styles were perceived very differently and they determined how satisfied participants were with the consultation and how they felt after the consultation. Participants exposed to the patient-centered communication perceived the physician as most emotional, least dominant, most appropriate when it comes to conveying information, most available and most expressive of hope. Also, they reported to be most satisfied with the visit and they showed the least increase in negative emotions. A patient-centered communication style has the most positive outcome for recipients of bad news on a cognitive, evaluative, and emotional level. Results of this study provide guidelines to physicians on how to convey bad news.


Sensory cortical control of movement.
Walking in our complex environment requires continual higher order integrated spatiotemporal information. This information is processed in the somatosensory cortex, and it has long been presumed that it influences movement via descending tracts originating from the motor cortex. Here we show that neuronal activity in the primary somatosensory cortex tightly correlates with the onset and speed of locomotion in freely moving mice. Using optogenetics and pharmacogenetics in combination with in vivo and in vitro electrophysiology, we provide evidence for a direct corticospinal pathway from the primary somatosensory cortex that synapses with cervical excitatory neurons and modulates the lumbar locomotor network independently of the motor cortex and other supraspinal locomotor centers. Stimulation of this pathway enhances speed of locomotion, while inhibition decreases locomotor speed and ultimately terminates stepping. Our findings reveal a novel pathway for neural control of movement whereby the somatosensory cortex directly influences motor behavior, possibly in response to environmental cues.


Reducing the availability of food to control feral pigeons: changes in population size and composition.
As feeding by humans is one of the main food resources to pigeons (Columba livia), there is general agreement that public education that aims to reduce the food base may be the most feasible way to reduce pigeon abundance. However, except for the classic example of Basel, the method has rarely been tested or implemented. We provide results from a 1 year study in the city of Barcelona where we tested the effect of public education on pigeon population abundance and composition. The quantity of food provided by people to pigeons was significantly reduced during the study. Feral pigeon density was reduced by 40% in the two experimental districts, but no variation was detected in the control district. Detailed analyses in one of the districts showed that the reduction was mainly related to the reduction in food availability but not to culling. Pigeons captured at the end of the experiment were larger than at the start of the study, but body condition was reduced. Results show the effectiveness of public information to manage feral pigeon populations in a large city, and that control operations can exert important selection pressure on the population, leading to changes in population composition. © 2016 Society of Chemical Industry.


[Efficacy and safety of treatment with infliximab in Crohn's disease-the experience of single center in Korea].
Infliximab has been shown to be effective and safe for treating refractory luminal and fistulizing Crohn's disease (CD). The aim of this study was to report the efficacy and adverse effect of infliximab therapy in patients with CD at our center. Medical records of thirteen patients who were treated with infliximab for refractory luminal or fistulizing CD were reviewed. Clinical response was classified as complete response, partial response and nonresponse. Seven patients were treated for fistulizing CD, four patients for luminal CD, and two for both. The mean time of follow-up was 13.1 months (3.3-28.1 months). Clinical response was seen in 10/13 (77%); complete response 7/13 (54%), partial response 3/13 (23%), nonresponse 3/13 (23%). Mean time to response was 27.1 days (10-41 days). 4 of 10 responders (40%) maintained remission over 30 weeks. Those who started on immunosuppressive treatment more than 3 months before infliximab infusion achieved lower early recurrence rate (14%) compared with those less than 3 months (67%) (p=0.039). Steroid tapering was successful in 7/12 (58%). Five patients required surgical therapy; three nonresponders, one partial responder and one who recurred after initial complete response. Initial responders required less surgery than nonresponders (p=0.035). Acute infusion reactions were seen in 2/40 infusions (5%). One patient developed herpes zoster 20 weeks after infliximab infusion. During follow-up peried, no patient developed serious infection, tuberculosis or malignancy. Infliximab is effective and safe in clinical practice. Concurrent immunosuppressive use is associated with lower rate of early recurrence.


Mitochondrial DNA 3243A>T mutation in a patient with MELAS syndrome.
Approximately 80% of cases of mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) harbor a heteroplasmic m.3243A>G transition in the tRNALeu (UUR) (MTTL1) gene. We report a MELAS case with a rare heteroplasmic m.3243A>T mutation found by direct sequencing of MTTL1. This mutation has been previously reported in 5 cases, of which 2 cases had the MELAS phenotype. Our case also strengthens the hypothesis that the m.3243A>T mutation can cause the MELAS phenotype.


The sites of encephalic photoreception in phosoperiodic induction of the growth of the testes in the white-crowned sparrow, Zonotrichia leucophrys gambelii.
In bilaterally enucleated White-crowned Sparrows, Zonotrichia leucophrys gambelii, testicular growth and development at rates similar to those of intact birds subjected to long days, can be induced by illumination of sites within and near the ventromedial hypothalamus from the tips of single, chronically implanted, light-conducting fibers. Essentially identical results are obtained with the same mode of encephalic illumination of birds with intact eyes held on short days that are otherwise nonstimulatory. Examination of the distribution of the sites of the fibers from which weak illumination induced testicular growth, increase in plasma level of luteinizing hormone or increase in plasma level of testosterone suggests that most of the photoreceptors must lie either within the ventromedial hypothalamus or in sites ventral thereto, such as the tuberal complex. For the time being, at least, we find it necessary to assume that some photosensitive elements of the system occur outside of the ventral hypothalamus.


Genetic and environmental causes of tracking in explosive strength during adolescence.
To determine whether the observed phenotypic stability in explosive strength during adolescence, as measured by inter-age correlations in vertical jump (VTJ), is mainly caused by genetic and/or environmental factors. Subjects are from the Leuven Longitudinal Twin Study (LLTS) (n = 105 pairs, equally divided over five zygosity groups). VTJ data were aligned on age at peak height velocity (APHV) to attenuate the temporal fluctuations in inter-age correlations caused by differences in timing of the adolescent growth spurt. Simplex models were fitted using structural equation modelling. After aligning the data on APHV, the annual inter-age correlations show a clear simplex structure over a 4 year interval. The best fitting models included additive genetic and unique environmental sources of variation. Heritability estimates ranged between 60.8% (CI 37.7%-77.2%) and 87.3% (CI 74.2%-94.0%) for boys and between 76.5% (CI 56.7%-89.0%) and 88.6% (CI 77.8%-94.1%) for girls. Up to 56.4% and 62.8% of the total variation at the last measurement occasion is explained by additive genetic factors that already explained a significant amount of variation at previous measurement occasions in boys and girls respectively. It thus can be concluded that the observed stability of explosive strength during adolescence is mainly caused by a stable genetic influence in boys and girls. Additive genetic factors seem to be the main cause of the observed phenotypic stability in VTJ performance in boys and girls during adolescence.


Calpain activation is upstream of caspases in radiation-induced apoptosis.
The molecular events involved in apoptosis induced by ionizing radiation remain unresolved. In this paper we show that the cleavage of fodrin to a 150 kDa fragment is an early proteolytic event in radiation-induced apoptosis in the Burkitts' Lymphoma cell line BL30A and requires 100 microM zVAD-fmk for inhibition. Caspases-1, -3, -6 and -7 were shown to cleave fodrin to the 150 kDa fragment in vitro and all were inhibited by 10 microM zVAD-fmk. We also show that the in vitro cleavage of fodrin by calpain is inhibited by 100 microM zVAD-fmk as was the calpain-mediated hydrolysis of casein. We demonstrate that calpain is activated within 15 min after radiation exposure, concomitant with the cleavage of fodrin to the 150 kDa fragment whereas caspase-3 is activated at 2 h correlating with the cleavage of fodrin to the 120 kDa fragment. These results support a role for calpain in the early phases of the radiation-induced apoptosis pathway, upstream of the caspases.


Pharmacodynamic effects of serotonin (5-HT) receptor ligands in pigs: stimulation of 5-HT2 receptors induces malignant hyperthermia.
In pigs, the serotonin-2 (5-HT2) receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), 0.8 mg/kg, induced "psychotic" behaviour (e.g., grimacing, backward locomotion, blank stare) and a muscular syndrome, which is known as malignant hyperthermia (MH) in pigs and humans. This syndrome is characterized by generalized skeletal muscle rigidity, leading to an increase in body temperature, marked acidosis, hyperkaliaemia, cyanosis and elevation of lactate, carbon dioxide and the muscle enzyme creatine kinase (CK) in plasma. In pigs which were selectively bred for susceptibility to MH induction by known triggering agents, such as halothane, the administration of DOI was fatal in 3 out of 5 animals. In genetically susceptible pigs, MH was also induced by 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT), 0.5-1.8 mg/kg, and D-lysergic acid diethylamide (LSD), 60-110 micrograms/kg. Furthermore, 5-MeO-DMT and LSD induced head shakes in the animals, which had not been observed after DOI and could not be blocked by 5-HT2-antagonists, ketanserin (0.5-5 mg/kg) and ritanserin (1-2.5 mg/kg). The psychotomimetic effects of 5-MeO-DMT could be blocked by ketanserin or ritanserin, which, depending on the dose, also reduced or totally prevented the hyperthermia and metabolic changes induced by 5-MeO-DMT in pigs. Administration of 5-MeO-DMT, 1.8 mg/kg, was fatal in 4 of 5 MH-susceptible pigs, whereas pigs injected with this dosage after pretreatment with ketanserin (0.5-5 mg/kg) or ritanserin (1-2.5 mg/kg) did not die. In pigs from MH-resistant littermates, administration of 5-MeO-DMT was not fatal. Comparison of metabolic changes in susceptible and non-susceptible pigs suggested that the marked increase in plasma potassium, which arises principally from damaged muscle cells, is primarily responsible for the fatal effect of DOI and 5-MeO-DMT in genetically susceptible individuals. In MH-susceptible pigs, which were anesthetized, relaxed and artificially ventilated, 5-MeO-DMT did not induce hyperthermia, thus substantiating that the marked hyperthermia observed in conscious pigs was a result of muscle activation and not due to effects on thermoregulation or blood pressure. The results indicate that hallucinogenic drugs with 5-HT2 agonistic effects trigger a life-threatening syndrome, MH, in genetically susceptible pigs. 5-HT2 antagonists, such as ketanserin or ritanserin, are capable of counteracting the fatality of this syndrome.


High binding ability ligand controlled formation of CsPbX3 (X = Cl/Br, Br, I) perovskite nanocrystals with high quantum yields and enhanced stability.
CsPbX3 NCs with both high photoluminescence quantum yields (PLQYs) and enhanced stability have been obtained by using high binding ability ligands. As a result, the CsPbI3 NCs prepared using palmitic acid and oleylamine as ligands have high PLQY (up to 92%), and the PLQYs of CsPbX3 perovskite NCs can be sustained for one month with a slight decrease. Impressively, the presence of palmitic acid and stearic acid can dramatically improve the chemical stability of CsPbX3 NCs. Importantly, 1H NMR measurements indicate that much more palmitic acid remained on the surface of CsPbX3 NCs than oleic acid after purification using solvent, suggesting the higher binding ability of palmitic acid than oleic acid. Finally, the fabricated perovskite NCs were used as luminescent inks and phosphors of white light emitting diodes.


Saksenaea erythrospora infection after medical tourism for esthetic breast augmentation surgery.
Mucormycosis caused by Saksenaea erythrospora is rarely reported in humans. Three previous cases have been reported in the literature, two associated with trauma (a sailing accident in Argentina and a combat trauma in Iraq) and one as a cause of invasive rhinosinusitis (India), all in immunocompetent patients . The first case of mucormycosis following esthetic surgery, associated with medical tourism, is reported herein. A case study of an S. erythrospora infection in an immunocompetent woman after the completion of esthetic surgery (dermolipectomy and breast augmentation) is reported. The infection presented as a rapidly progressive necrotizing infection of the skin and soft tissue, which required a bilateral mastectomy and extensive surgical debridement associated with prolonged antifungal therapy. The organism was identified phenotypically and confirmed biologically after rDNA amplification and sequencing. Two months later, the patient remains hospitalized awaiting the start of reconstructive surgeries. The present case is, to the best of the authors' knowledge, the first report from Colombia. Mucormycosis should be considered in the differential diagnosis of necrotizing infections of the skin and soft tissue that evolve rapidly after cosmetic surgery performed in tropical or subtropical countries.


Investigation of BRAF mutation in a series of papillary thyroid carcinoma and matched-lymph node metastasis with ARMS PCR.
To figure out that if there is a consistency relationship of the BRAFV600E mutation in matched-lymph node metastasis and original papillary thyroid carcinoma (PTC) specimen for the same patient. We collected the specimen of thyroids and matched-lymph node metastases of PTCs and tested the BRAFV600E mutation status with amplification refractory mutation system (ARMS) PCR. 20 patients with PTC and metastasis lymph node were hired. In this cohort, 16 (80%) patients had the same BRAF genetic mutation status in thyroid and metastasis, and the other 4 (20%) had an inconsistent situation. Within our cohort, the data suggested that wild-type BRAFV600E oncogene in thyroid primary tumor does not rule out its mutation in lymph node metastasis, and vice versa.