Association between effector-type regulatory T cells and immune checkpoint expression on CD8+ T cells in malignant ascites from epithelial ovarian cancer

Background Regulatory T cells (Tregs) play an important role in the antitumor immune response in epithelial ovarian cancer (EOC). To understand the immune-inhibitory networks of EOC, we addressed the association between Tregs and immune checkpoint expression on T cells in the tumor microenvironment of EOC Methods A total of 41 patients with stage IIIC and IV EOC were included in the analysis. We harvested cells from malignant ascites and investigated them using multi-color flow cytometry. We categorized the Tregs into 3 groups: effector-type Tregs, naïve Tregs and non-Tregs, based on the expression patterns of CD45RA and Foxp3 in CD4+ T cells. Furthermore, the relationships between the expression of various immune checkpoint molecules, such as PD-1, on CD8+T cells and each of the Treg subtypes was also evaluated. Results The median frequency of naïve Tregs, effector-type Tregs and non-Tregs were 0.2% (0-0.8), 2.0% (0-11.4) and 1.5% (0.1-6.3) in CD4+ T cells of malignant ascites from EOC patients, respectively. A high frequency of effector-type Tregs was associated with high-grade serous carcinoma compared with the other histotypes. Patients with higher proportions of effector-type Tregs showed a trend towards increased progression-free survival. We also demonstrated a correlation between a higher proportion of effector-type Tregs and increased PD-1 expression on CD8+ T cells. In addition, C-C chemokine receptor 4 expression was also observed in effector-type Tregs. Conclusion These data suggest that multiple immune-inhibitory networks exist in malignant ascites from EOC patients, suggesting an approach towards combinational immunotherapies for advanced EOC patients.

Mosaic deletions in X-linked dystonia-parkinsonism influence repeat stability and disease onset

Background: While multiple genetic causes of movement disorders have been identified in the past decade, modifying factors of disease expression are still largely unknown for most conditions. X-linked dystonia-parkinsonism (XDP) is an inherited neurodegenerative disease caused by a SINE-VNTR-Alu (SVA)-type retrotransposon insertion that contains a hexanucleotide repeat within an intron of the TAF1 gene. To date, four putative genetic modifiers explain about 65% of variance in age at onset in XDP. However, additional genetic modifiers are conceivably at play in XDP and may include mismatches of the SVA hexanucleotide repeat motif. We aim to identify additional genetic modifiers of XDP expressivity and age at onset (AAO). Methods: Third-generation sequencing of PCR amplicons from XDP patients (n=202) was performed to assess potential repeat interruption and instability. Repeat-primed PCR and Cas9-mediated targeted enrichment were used to confirm the presence of identified repeat mismatches. Results: An increased frequency of deletions at the beginning of the hexanucleotide repeat (CCCTCT)n domain was found. Specifically, three deletions at positions 11, 14, and 17 of the TAF1 SVA repeat motif of somatic mosaic origins were detected in different combinations. The most common one was three deletions (1-2-3) at a median frequency 0.425 (IQR:0.42-0.43) and deletions within positions 11 and 14 (1-2-wt) at a median frequency 0.128 (IQR:0.12-0.13). The frequency of deletions at positions 11 and 14 correlated with repeat number (r=-0.48, p=9.5x10-13) and AAO (r=0.34, p=9.5x10-7). The association with AAO still stands when including other modifier genotypes (MSH3 and PMS2) in a regression model. However, the association dissipates when including repeat numbers. Conclusion: We present a novel mosaic repeat motif deletion within the hexanucleotide repeat (CCCTCT)n domain of TAF1 SVA. Our study illustrates: 1) the importance of somatic mosaic genotypes; 2) the biological plausibility of multiple modifiers (both germline and somatic) that can have additive effects on repeat instability; 3) that these variations may remain undetected without assessment of single molecules.

The neuromuscular responses in patients with Parkinson’s disease under different conditions during whole-body vibration training

Background Whole-body vibration (WBV) training can provoke reactive muscle response and thus exert beneficial effects in various neurological patients. This study aimed to investigate the muscles activation and acceleration transmissibility of the lower extremity to try to understand the neuromuscular control in the Parkinson’s disease (PD) patients under different conditions of the WBV training, including position and frequency. Methods Sixteen PD patients and sixteen controls were enrolled. Each of them would receive two WBV training sessions with 3 and 20 Hz mechanical vibration in separated days. In each session, they were asked to stand on the WBV machine with straight and then bended knee joint positions, while the vibration stimulation was delivered or not. The electromyographic (EMG) signals and the segmental acceleration from the lower extremity were recorded and processed. The amplitude, co-contraction indexes (CCI), and normalized median frequency slope (NMFS) from the EMG signals, and the acceleration transmissibility were calculated. Results The results showed larger rectus femoris (RF) amplitudes under 3 Hz vibration than those in 20 Hz and no vibration conditions; larger tibialis anterior (TA) in 20 Hz than in no vibration; larger gastrocnemius (GAS) in 20 Hz than in 3 Hz and no vibration. These results indicated that different vibration frequencies mainly induced reactive responses in different muscles, by showing higher activation of the knee extensors in 3 Hz and of the lower leg muscles in 20 Hz condition, respectively. Comparing between groups, the PD patients reacted to the WBV stimulation by showing larger muscle activations in hamstring (HAM), TA and GAS, and smaller CCI in thigh than those in the controls. In bended knee, it demonstrated a higher RF amplitude and a steeper NMFS but smaller HAM activations than in straight knee position. The higher acceleration transmissibility was found in the control group, in the straight knee position and in the 3 Hz vibration conditions. Conclusion The PD patients demonstrated altered neuromuscular control compared with the controls in responding to the WBV stimulations, with generally higher EMG amplitude of lower extremity muscles. For designing WBV strengthening protocol in the PD population, the 3 Hz with straight or flexed knee protocol was recommended to recruit more thigh muscles; the bended knee position with 20 Hz vibration was for the shank muscles.

Monitoring Dynamic Recrystallisation in Bioresorbable Alloy Mg-1Zn-0.2Ca by Means of an In Situ Acoustic Emission Technique

The tensile behaviour of the biocompatible alloy Mg-1Zn-0.2Ca (in wt.%) in the fine-grained state, obtained by severe plastic deformation via multiaxial isothermal forging, has been investigated in a wide range of temperatures (20 ÷ 300) °C and strain rates (5 × 10−4 ÷ 2 × 10−2) s−1 with the measurements of acoustic emission (AE). The dependences of mechanical properties, including the yield stress, ultimate strength, ductility, and the strain-hardening rate, on the test temperature and strain rate, were obtained and discussed. It is shown for the first time that an acoustic emission method is an effective tool for in situ monitoring of the dynamic recrystallisation (DRX) process. The specific behaviour of the acoustic emission spectral density reflected by its median frequency as a function of strain at various temperatures can serve as an indicator of the DRX process’s completeness.

Effect of housing enrichment and type of flooring on the performance and behaviour of female rabbits

This study investigated the effect of housing enrichments (scratching card, gnawing material and a platform), of a change in height and in the type of flooring on the live weight, reproductive performance and behaviour of female rabbits, as well as on the feed intake and spatial distribution of females and their kits. A total of 40 multiparous female rabbits were monitored in three consecutive reproductive cycles (48-d intervals). Four days before parturition in each reproductive cycle, the females were randomly assigned to one of the five types of housing: Control (CNT: 102×47×30 cm, L×W× H); Scratching card (SCT: containing a scratching card); Gnawing materials (GNW: CNT dimensions plus a compressed lucerne hay block and a wooden stick); Platform (PLT: 102×47×60 cm, including a platform with a plastic floor) and Combination (CBN: PLT dimensions with the scratching card, the gnawing materials and a platform). Data were only recorded during the first and third reproductive cycles. The living conditions did not significantly alter the females’ live weight (4889 g at housing; 4890 g at mid-lactation; 4867 g at weaning), reproductive performance (9.0 kits born alive), survival of the kits (90%), nor the feed intake of females and their litters (542 g/day). Providing animals with a gnawing block stimulated Gnawing behaviour (median frequency per group: CTL=0.00, SCT=0.00, GNW=4.69, PLT=0.00, and CBN=2.34; PRearing up behaviour (median frequencies per group: CTL=0.00, SCT=0.00, GNW=0.00, PLT=2.08, and CBN=3.12; P=0.06), and when a platform was present, the rabbits used it (mean values per group: CTL=0.00, SCT=0.00, GNW=0.00, PLT=1.79, and CBN=4.91; P=0.003). Regarding the type of floor, females appeared to prefer the plastic mesh flooring (31.2%) to the wire mesh flooring (18.8%). To sum up, providing female rabbits with simple enrichments appears to stimulate specific behaviours like Gnawing and Rearing up and may contribute to their wellbeing.

Cognitive Effects of Lacosamide in Patients with Drug Refractory Focal Epilepsy: A Prospective Observational Study

Objectives: This study was conducted to obtain data on the cognitive effects of lacosamide in Indian population. Methodology: An open labelled prospective observational study in 22 patients who suffered from focal epilepsy. Results: All the pre and post lacosamide cognition scores showed statistically significant positive correlation in this study. Average initial seizure frequency per month was 3.56 (SD 2.58) and median frequency 2.5 seizures per month. Range being 1-8 per month. At the final follow-up at 6months, 87.5% of the study subjects had no seizures. In the remaining12.5% of patients, reduction in seizure frequency was observed. The difference in frequency is statistically significant (Wilcoxon Signed Ranks TestP <0.001). Conclusion: Excellent seizure control is observed in patients with refractory focal epilepsy treated with lacosamide. Also, lacosamide has no serious adverse effects or drug interactions. In this study, it is observed that unlike many AEDs, lacosamide contributed to significant improvement in cognition.

Frequency and burden of potentially treatable symptoms in glioma patients with stable disease: a classical and a network approach

Background: Glioma patients experience a multitude of symptoms, impacting health-related quality of life. We aimed to assess frequency and burden of patient-reported symptoms in glioma patients with stable disease and whether patients would consider treatment. We also explored how symptoms co-occur and interact within a network.Methods: Patients rated frequency and burden and whether they would consider treatment of seventeen symptoms. Correlations between frequency, burden, and considering treatment were evaluated with Kendall’s Tau correlation coefficients. Partial correlations between symptom frequency scores were visualized as a symptom network.Results: Fifty-two glioma patients with stable disease were included (31 grade II/III, 21 grade IV tumors). The top five symptoms were fatigue, memory problems, reduced physical fitness, concentration problems, and drowsiness. Fatigue had the highest median frequency, 4.5 with an interquartile range of 2.5, on a seven-point Likert scale. More than 50% of patients experienced three or more symptoms simultaneously. In a network, symptoms seem to cluster together in a Fatigue, Cognition and Anxiety cluster. Overall, about one-third of patients would consider treatment for at least one symptom. Considering treatment correlated only moderately with frequency and burden (range of correlations 0.24-0.57 and 0.28-0.61, respectively).Conclusion: Glioma patients with stable disease often experience multiple co-occurring symptoms with a high symptom burden. Despite the high prevalence of symptoms, the inclination to undergo treatment was relatively low. The most frequent and burdensome symptoms and the way they are interrelated and cluster together could serve as a roadmap for future research on symptom management and treatment.

Neuroanatomia: Assiduidade na Monitoria e Desempenho Acadêmico

ResumoA Neuroanatomia é uma vertente complexa do estudo da Anatomia Humana, sendo a monitoria acadêmica (MA) uma forma dinâmica de habituar os discentes às estruturas e contribuir com a assimilação dos assuntos. O objetivo do presente estudo foi analisar a assiduidade nas atividades de MA com o desempenho dos estudantes. Trata-se de um estudo descritivo, transversal e quantitativo, realizado com 91 discentes vinculados aos cursos de Psicologia e Fisioterapia. Os estudantes foram distribuídos em três grupos: indivíduos que não participaram da MA (G1), participaram poucas vezes da MA (G2) e aqueles com participação mais assídua na MA (G3), com base na mediana da frequência dos participantes na MA. A análise estatística foi realizada pelo programa GraphPad Prism 6. Observou-se que, em ambas as avaliações parciais, os indivíduos que apresentaram maior assiduidade (G3) na MA tiveram melhor desempenho em relação àqueles que não participaram da MA (G1). Quando realizada a média final, foi observado que os indivíduos com maior assiduidade (G3) apresentaram melhor desempenho, quando comparados tanto ao G1 como G2. Além disso, o teste de Spearman mostrou correlação positiva entre presença e nota. Os discentes que participam das atividades da monitoria encontraram vantagens pedagógicas, resultando em um melhor domínio do processo de ensino e aprendizagem. Palavras-chave: Anatomia. Ensino. Educação Superior. AbstractNeuroanatomy is a complex aspect of the study of Human Anatomy, and academic monitoring (AM) is a dynamic way to familiarize the students with structures and contribute to the subjects assimilation. The objective of the study was to analyze the assiduity in AM activities with the students' performance. It is a descriptive, transversal, and quantitative study, carried out with 91 students linked to the Psychology and Physiotherapy courses. The students were divided into three groups: individuals who did not participate in AM (G1), participated in AM less often (G2) and those with more frequent participation in AM (G3), based on the participants’ median frequency in AM. Statistical analysis was performed using the GraphPad Prism 6 program. It was observed that in both partial assessments, the individuals who showed greater attendance (G3) in AM had a better performance in relation to those who did not participate in AM (G1). When the final average was performed, it was observed that individuals with greater attendance (G3) performed better when compared to both G1 and G2. In addition, the Spearman test showed a positive correlation between presence and grade. The students who participate in the monitoring activities found pedagogical advantages, resulting in a better mastery of the teaching-learning process. Keywords: Anatomy. Teaching. Education.

Underutilization and Quality Gaps in Blood Culture Processing in Public Hospitals of Peru

Correct processing of blood cultures may impact individual patient management, antibiotic stewardship, and scaling up of antimicrobial resistance surveillance. To assess the quality of blood culture processing, we conducted four assessments at 16 public hospitals across different regions of Peru. We assessed the following standardized quality indicators: 1) positivity and contamination rates, 2) compliance with recommended number of bottles/sets and volume of blood sampled, 3) blood culture utilization, and 4) possible barriers for compliance with recommendations. Suboptimal performance was found, with a median contamination rate of 4.2% (range 0–15.1%), with only one third of the participating hospitals meeting the target value of < 3%; and a median positivity rate of 4.9% (range 1–8.1%), with only 6 out of the 15 surveilled hospitals meeting the target of 6–12%. None of the assessed hospitals met both targets. The median frequency of solitary blood cultures was 71.9% and only 8.9% (N = 59) of the surveyed adult bottles met the target blood volume of 8 – 12 mL, whereas 90.5% (N = 602) were underfilled. A high frequency of missed opportunities for ordering blood cultures was found (30.1%, 95/316) among patients with clinical indications for blood culture sampling. This multicenter study demonstrates important shortcomings in the quality of blood culture processing in public hospitals of Peru. It provides a national benchmark of blood culture utilization and quality indicators that can be used to monitor future quality improvement studies and diagnostic stewardship policies.