Prospective Cohort Pilot Study Research Articles

Epidemiology and outcomes of critically ill patients in the emergency department of a tertiary teaching hospital in Rwanda.

The introduction of Emergency Medicine in Rwanda in 2015 has been associated with a mortality reduction in patients presenting to Kigali University Teaching Hospital (KUTH). In the context of increasing numbers of critically ill patients presenting to Emergency Departments (ED) globally, the aim of this study was to describe the characteristics of critically ill patients, the critical care interventions performed, and the outcomes of critically ill patients presenting to the KUTH ED with the goal of informing future research into the root causes of mortality of critically ill ED patients and of identifying high yield topics for didactic and procedural training. A descriptive observational prospective cohort pilot study analyzed all patients ≥15 years who presented to KUTH between April and June 2022 with modified South African Triage Scores of Red with alarm, Red without alarm, and Orange. Of 320 patients, 66.9% were male and median age was 40 years. Patients were triaged as Orange (65.3%), Red without alarm (22.8%), and Red with alarm (11.9%). Presentations were categorized as: medical emergencies (48.0%), traumatic injury (44.5%), and surgical emergencies (7.6%). Median length of stay was 31h (IQR 28, 56) and boarding was 23h (IQR 8, 48). Overall mortality was 12.2% and highest among medical emergencies (16.5%, p = 0.048) and increased significantly with triage color: Red with alarm (47.4%), Red without alarm (16.4%), and Orange (4.3%, p < 0.0001). Cardiopulmonary resuscitation (CPR) (10.3%), endotracheal intubation (8.8%), and vasopressor administration (3.1%) were the most frequent critical interventions performed. Survival after cardiac arrest was 9.1% and 32.1% after intubation. Mortality was associated with the following interventions: CPR, intubation, and use of vasopressors (p < 0.05). This pilot study identified the most common critical care interventions performed and a high mortality among patients who required these interventions in the ED of a tertiary teaching hospital in Rwanda. These findings will inform didactics and procedural training for emergency care providers. Future research should focus on the root causes of mortality in these specific patient populations and identify areas of system strengthening to reduce mortality.

Respiratory Effects of Maximal Lung Recruitment Maneuvers Using Single-Breath Estimation in ARDS.

Determining which patients with ARDS are most likely to benefit from lung recruitment maneuvers is challenging for physicians. The aim of this study was to assess whether the single-breath simplified decremental PEEP maneuver, which evaluates potential lung recruitment, may predict a subject's response to lung recruitment maneuvers, followed by PEEP titration. We conducted a pilot prospective single-center cohort study with a 3-step protocol that defined sequential measurements. First, potential lung recruitment was assessed by the single-breath maneuver in the volume controlled mode. Second, the lung recruitment maneuver was performed in the pressure controlled mode, with a fixed driving pressure of 15 cm H2O and a maximum PEEP of 30 cm H2O. Third, the lung recruitment maneuver was followed by decremental PEEP titration to determine the optimal PEEP, defined as the lowest driving pressure. Responders to the lung recruitment maneuver were defined by an improvement in PaO2 /FIO2 > 20% between the baseline state and the end of the PEEP titration phase. Forty-two subjects with moderate-to-severe ARDS were included. The mean ± SD lung recruitment was 149 ± 104 mL. A threshold lung recruitment of 195 mL (area under the receiver operator characteristic curve 0.62, 95% CI 0.43-0.80) predicted a positive response to the maximal lung recruitment maneuver. The lung recruitment maneuver, followed by PEEP titration, resulted in a modification of PEEP in 74% of the subjects. PEEP was increased in more than two thirds of the responders and decreased in almost half of the non-responders to the lung recruitment maneuver. In addition, a decrease in driving pressure and an increase in respiratory system compliance were reported in 62% and 67% of the subjects, respectively. The single-breath maneuver for evaluating lung recruitability predicted, with poor accuracy, the subjects who responded to the lung recruitment maneuver based on PaO2 /FIO2 improvement. Nevertheless, the lung recruitment maneuver, followed by PEEP titration, improved ventilator settings and respiratory mechanics in a majority of subjects.

Oral, Vaginal, and Stool Microbial Signatures in Patients With Endometriosis as Potential Diagnostic Non-Invasive Biomarkers: A Prospective Cohort Study.

To identify a microbial signature for endometriosis for use as a diagnostic non-invasive biomarker. Prospective cohort pilot study. Nepean Hospital and UNSW Microbiome Research Centre, Australia. Sixty-four age- and sex-matched subjects (n = 19 healthy control (HC); n = 24 non-endometriosis (N-ENDO) and n = 21 confirmed endometriosis (ENDO)). All study participants, besides healthy controls, underwent laparoscopic surgical assessment for endometriosis, and histology was performed on excised lesions. Oral, stool and, vaginal samples were self-collected at a single time point for healthy controls, and preoperatively for patients undergoing laparoscopy. Samples underwent 16S rRNA amplicon sequencing, followed by bioinformatics analysis. Compositional differences between cohorts as identified by diversity analyses, and differentially abundant microbial taxa, as identified by LEfSE analysis. The composition of the oral (adjusted p = 0.003), and stool (adjusted p = 0.042) microbiota is different between the three cohorts. Differentially abundant taxa are present within each cohort as identified by LEfSE analysis. Particularly, Fusobacterium was enriched in the oral samples of patients with moderate/severe endometriosis. Taxonomic and compositional differences were found between the microbiota in the mouth, gut and, vagina of patients with and without endometriosis and healthy controls. Fusobacterium was enriched in patients with moderate/severe endometriosis. Fusobacterium is noted as a key pathogen in periodontal disease, a common comorbidity in endometriosis. These findings suggest a role for the oral, stool and, vaginal microbiome in endometriosis, and present potential for microbial-based treatments and the design of a diagnostic swab.

Weight changes during the initial phase of orthodontic treatment with clear aligners versus fixed appliances: a pilot prospective cohort study.

The aim was to assess adult patients' weight changes during the initial phase of orthodontic treatment (OT) with clear aligner therapy (CAT) versus fixed bracket systems (FBS). This pilot prospective cohort study included systemically-healthy adults who were treated with CAT or FBS. Participants with eating disorders and/or undergoing dietary programs were excluded. Weight was assessed using a calibrated digital scale at the start of OT (T1), 6-8 weeks (T2), and 12-16 weeks (T3) after starting OT. A questionnaire was used to record demographic variables, diet and exercise habits, self-reported compliance and discomfort during OT, and use of analgesics. Repeated measures analysis of variance was used to assess the association between treatment type (FBS versus CT) and time-period (T1, T2, T3) on participants' weight while adjusting for baseline covariates such as age, gender, height, and weight. Forty-two participants (CAT group: 22 and FBS group: 20) were analysed. In the CAT group, there was a slight initial mean weight gain of 0.2 Kg (~ 0.4 pounds) (T2-T1), and 0.8 Kg (~ 1.7 pounds) overall mean weight loss between T1 and T3. For the FBS group, there was a slight initial weight loss of 0.3 Kg (~ 0.7 pounds) (T2-T1) and no overall change between T1 and T3. Treatment type (CAT versus FBS) was not significantly associated with weight changes during the initial phase of OT. There were no significant differences in the weight changes during the initial phase of OT with CAT compared with FBS. Clear aligner therapy should not be advertised as a weight loss strategy.

Plasma bioactive adrenomedullin predicts mortality and need for dialysis in critical COVID-19

COVID-19 is a severe respiratory disease affecting millions worldwide, causing significant morbidity and mortality. Adrenomedullin (bio-ADM) is a vasoactive hormone regulating the endothelial barrier and has been associated with COVID-19 mortality and other adverse events. This prospective cohort pilot study included 119 consecutive patients with verified SARS-CoV-2 infection admitted to two intensive care units (ICUs) in Southern Sweden. Bio-ADM was retrospectively analysed from plasma on ICU admission, and days 2 and 7. Information on comorbidities, adverse events and mortality was collected. The primary outcome was 90-day mortality, and secondary outcomes were markers of disease severity. The association between bio-ADM and outcomes was analysed using survival analysis and logistic regression. Bio-ADM on admission, day 2, and day 7 only moderately predicted 90-day mortality in univariate and multivariate Cox regression. The relative change in bio-ADM between sample times predicted 90-day mortality better even when adjusting for the SAPS3 score, with an HR of 1.09 (95% CI 1.04–1.15) and a C-index of 0.82 (95% CI 0.72–0.92) for relative change between day 2 and day 7. Bio-ADM had a good prediction of the need for renal replacement therapy in multivariate Cox regression adjusting for creatinine, where day 2 bio-ADM had an HR of 3.18 (95% CI 1.21–8.36) and C-index of 0.91 (95% CI 0.87–0.96). Relative changes did not perform better, possibly due to a small sample size. Admission and day 2 bio-ADM was associated with early acute kidney injury (AKI). Bio-ADM on ICU admission, day 2 and day 7 predicted 90-day mortality and dialysis needs, highlighting bio-ADM’s importance in COVID-19 pathophysiology. Bio-ADM could be used to triage patients with a risk of adverse outcomes and as a potential target for clinical interventions.

Predictive value of methylene blue combined with indocyanine green in sentinel lymph node metastasis in breast cancer: a prospective pilot cohort study.

The status of sentinel lymph nodes is crucial for prognosis and treatment decisions in breast cancer patients. This study aimed to evaluate the predictive value of combined methylene blue and indocyanine green for sentinel lymph node metastasis in breast cancer. This prospective cohort study enrolled 90 clinically node-negative breast cancer patients. Methylene blue and indocyanine green were injected locally before surgery. Sentinel lymph nodes were grouped based on fluorescence intensity and methylene blue staining. A binary logistic regression model was established using 285 lymph node groups to predict metastatic risk. A total of 475 lymph nodes were identified, with 33 being metastatic. The metastatic risk reached 70% for partially blue-stained and weakly fluorescent lymph nodes between 1-2 cm. The model revealed associations between lymph node size, dye staining patterns, and metastatic risks (P<0.05). The AUC of the ROC curve was 0.855. The staining pattern of combined methylene blue and indocyanine green could predict risks of sentinel lymph node metastasis and facilitate rapid intraoperative identification of high-risk lymph nodes.

Serum Carnitine Concentrations and Cardiac Function in Pediatric, Adolescent and Young Adult Oncology Patients Receiving High-Dose Anthracyclines

OBJECTIVE Anthracycline chemotherapy agents have significant dose-dependent cardiotoxic effects. ­Carnitine, a non-essential amino acid, is involved in long chain fatty acid oxidation, and carnitine deficiency can result in cardiomyopathy and cardiac arrhythmias. If administered concurrently with chemotherapy, carnitine supplementation could be a potential strategy to prevent cardiotoxicity. However, the association between serum carnitine concentrations and anthracycline cardiotoxicity during cancer treatment in the childhood, adolescent, and young adult (CAYA) age range has not been established. METHODS This prospective pilot cohort study characterized changes in serum carnitine concentrations and cardiac function before, during, and approximately 1 year after large-dose anthracycline therapy in newly diagnosed CAYA cancer patients. RESULTS Among 21 patients with a mean cumulative anthracycline dose exposure of 409 mg/m2 of ­doxorubicin equivalents, left ventricular ejection fraction and relative wall thickness decreased, indicating an overall decline in cardiac function. A reversible decrease in serum carnitine concentrations was also observed. A non-statistically significant positive correlation was observed; for every 1 mmol/L decrease in serum carnitine concentration, there was a 0.09% decrease in LVEF (p = 0.2). CONCLUSIONS These findings from this small pilot study suggest that there may be a relationship between serum carnitine concentrations and cardiac function after anthracycline therapy that should be evaluated in larger studies.

Potential financial loss and risk factors for hamstring muscle injuries in elite male Brazilian soccer players: a season-long prospective cohort pilot study.

The aim of this pilot study was to analyze the potential financial loss and a range of potential risk factors for hamstring muscle injuries in elite Brazilian soccer. Thirty-four male players (age: 25 ± 6 years; stature: 180 ± 8 cm; body mass: 78 ± 9 kg; minutes played in matches: 2243 ± 1423 min) from an elite professional soccer club were monitored during a 12-month season. Muscle injury was identified by magnetic resonance imaging and the severity was defined according to the number of days away: minimal (1-3 days), mild (4-7 days), moderate (8-28 days), severe (>28 days). Potential financial loss due to the team's under achievements was determined. Dorsiflexion range of motion, eccentric knee flexor strength and isokinetic tests were performed during the pre-season. Association between dependent variables and the occurrence of injury was evaluated. Nine hamstring muscle injuries with moderate severity were found in 8 athletes. Recovery time was 22 days off the field on average. Potential financial loss was $-43.2 million USD and earnings on merit money was 21%. Previous injury, increased flexor deficit 60° /sec and increased flexor fatigue index 300°/sec were all associated with a greater chance of hamstring muscle injury. Ankle dorsiflexion range of motion was significantly lower in the injured group (35.6 ± 3° vs. 39.1 ± 4.9°; p = 0.017, effect size = -0.74). High financial burden was found in elite Brazilian soccer during one full season. Injured athletes had high hamstring fatigue index, knee flexor strength deficit, ankle range of motion restriction and previous hamstring muscle injury when compared to non-injured athletes. Therefore, preventive approaches in professional soccer players with previous hamstring injuries should be a priority.

Neuropsychological performance labels: do patients understand?

Objective: Descriptive labels to communicate performance are integral in neuropsychological reports. Although the American Academy of Clinical Neuropsychology (AACN) reached consensus on standardizing uniform labels of performance, consumer understanding of these labels is unclear. This pilot prospective cohort study evaluated patient, family member, and medical provider understanding of neuropsychological performance labels. Method: Patients (n = 62), family members (n = 31), and referral sources (n = 34) completed a 13-question survey evaluating the uniform label system and an alternative, patient-centered language system (PCL). Questions assessed label knowledge and associated percentiles and relative relationships between labels. Results: Patients had significantly higher accuracy on knowledge questions using PCL labels (54.8%) versus AACN labels (32.2%) as were families (51.6% versus 35.5%). When ranking labels, providers and patients were more accurate when using PCL labels compared to AACN labels, although no significant difference was seen in the family group. Generally, participants were more accurate in identifying highest/lowest scores using AACN labels and more accurate in identifying relatively higher and lower scores using PCL terminology. Conclusions: Results illustrate that current and alternative score labels may not be understandable for audiences who read and base decisions on neuropsychological reports. The findings highlight the need for further consideration of patient-centered language to improve understanding of neuropsychological score labels.

Cerebral oxygenation in preterm infants at high altitude: a prospective pilot cohort study

Cerebral oxygenation in preterm infants at high altitude: a prospective pilot cohort study

Associations of maternal serum concentration of iron-related indicators with gestational diabetes mellitus in Chinese: A pilot prospective cohort study

BackgroundWe aimed to provide a comprehensive understanding of the associations between iron metabolism and gestational diabetes mellitus (GDM) by examining multiple iron-related indicators. MethodsWe conducted a prospective study involving 907 Chinese pregnant women. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum concentrations of iron-related indicators during the first trimester (≤ 14 weeks of gestation). GDM outcomes were measured through oral glucose tolerance tests (OGTT) conducted between weeks 24 and 28 of gestation. ResultsSubjects with iron-related indicators below the 10th percentile (except for serum iron and soluble transferrin receptor) had a higher risk of GDM compared to normal subjects (10th-90th percentiles). The ORs (95 %CI; p-value) were 1.88 (1.10, 3.20; P=0.020) for ferritin, 1.88 (1.10, 3.19; P=0.020) for hepcidin. Higher levels of ferritin (> 90th percentile) were associated with a higher risk of abnormal fasting blood glucose, while lower levels (< 10th percentile) of ferritin, hepcidin, and transferrin were associated with a higher risk of one-hour postprandial glucose ≥ 8.6 mmol/L in the OGTT. ConclusionsLower levels (< 10th percentiles) of several iron-related indicators (ferritin, hepcidin, and transferrin) were associated with a higher risk of GDM and abnormal blood glucose compared to normal subjects.

The association between exosomal proteins and the efficacy of thermoradiochemotherapy in overweight/obese rectal cancer patients: a pilot prospective cohort study

Background: Overweight and especially obesity are associated with the risk of the development and progression of colorectal cancer. It can be assumed that there are multifaceted interactions between the tumor and adipose tissue during anti-tumor treatment. Cancer cells secrete exosomes, extracellular vesicles affecting the microenvironment of the tumor and promoting its progression or regression. The presence of transcription/translation/folding factors (heat shock proteins (HSPs), matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in exosomes secreted by irradiated cells and cells exposed to hyperthermia, indicates the cell adaptation to the thermal and radiation stress. Aim: To analyze the MMPs, TIMP1, and HSPs on CD9-positive (CD9+) exosomes, as well as on exosomes of adipocytic origin (FABP4+) in rectal cancer patients with overweight/obesity under thermoradiochemotherapy (TRCT) and their association with the immediate treatment efficacy. Methods: Since 2021, 20 patients (of those 8 men; median age 59.0 [52.0; 63.0] years, median body mass index 29.6 [28.5; 33.1] kg/m2) with morphologically verified rectal cancer (T3-4N0M0 and T3-4N1M0, differentiation grade G1–G3) have been participating in the study. They were treated with TRCT: external gamma therapy (2 Gy, 1 fraction/day, 5 days/week, total focal dose 54 Gy), chemotherapy with capecitabine (825 mg/m2 twice daily) combined with local hyperthermia (42–44 °C, 60 min, 3 times/week, 10 sessions). The TRCT efficacy was assessed by RECIST 1.1 and ESGAR criteria. Blood samples for exosomes were taken from the patients at baseline (point 1), in the middle of the treatment course (point 2), at 6 to 10 weeks after the end of TRCT (point 3), and at 6 months after point 1 (point 4). Small extracellular vesicles were isolated from plasma by ultrafiltration with double ultracentrifugation. The isolated exosomes were characterized by transmission electronic microscopy, flow cytometry and nanoparticle trajectory analysis (NTA). Results: TRCT resulted in complete tumor regression in 13/20 of the rectal cancer patients and partial regression or stabilization in 7/20. Four subpopulations of CD9+ and FABP4+ exosomes associated with the TRCT efficacy were identified (CD9+MMP2+, СD9+MMP2+9+TIMP1+, СD9+MMP2+9+TIMP1-, and FABP4+MMP2+9-TIMP1+). Compared to the CD9+ exosomes, the adipocytic vesicles had higher MMP2 expression (p = 0.026); however, the adipocyte vesicles subpopulation were virtually free of vesicles with combined MMP2 and MMP9 gelatinase expression. The HSPs expression by circulating exosomes at various TRCT steps was associated neither with direct treatment efficacy nor with the vesicle type. Conclusion: The expression of MMPs and TIMP1 on CD9+ and FABP4+ exosomes is associated with TRCT efficacy. In the future, vesicular markers could be used to build prognostic models, to identify patient groups with an unfavorable prognosis, and to personalize treatment and follow-up.

Volume-based enteral feeding for ward patients with acute neurological conditions: a pilot prospective cohort study.

Patients requiring enteral nutrition (EN) after neurological insults experience feeding interruptions, contributing to inadequate nutrition delivery. This prospective cohort study investigated if volume-based enteral feeding (VBF) improved the delivery of prescribed EN volume in ward patients with acute neurological conditions. Over two sequential periods, the usual care group received standard continuous rate-based feeding, and the intervention group received VBF with bi-daily EN rate adjustments to achieve target daily volume. The primary outcome was percentage of prescribed daily EN formula volume delivered. Differences in energy and protein provision, weight, malnutrition and safety were explored. An evaluation survey captured nurse acceptability of the protocol. The intervention group (n = 32) achieved greater median interquartile range (IQR) EN adequacy of prescribed volume at 92% (88-97) compared to 67% (54-78) for usual care (n = 35) (p < 0.001). VBF compared to rate-based feeding resulted in patients receiving more kilojoules (131 [121-138] kJ/kg vs. 84 [64-99] kJ/kg; p < 0.001) and protein (1.3 [1.2-1.5] g/kg vs. 0.9 [0.6-1.1] g/kg; p < 0.001). There were no differences in gastrointestinal intolerance between groups. Compliance to the VBF protocol was 90%, and 78% of staff reported high confidence using the protocol. The intervention group had less median weight loss at discharge (-1.4 [0.1 to -4.3] kg) than usual care (-3.6 [-1.3 to 8.4] kg; p < 0.011), but no differences in malnutrition status were observed. A VBF protocol delivered greater EN volume, energy and protein following neurological injury. The VBF protocol was feasible with high acceptability from nursing staff.

P-701 Impact of vaginal microbiome on vaginal absorption of exogenous progesterone: a pilot study

Abstract Study question Does vaginal microbiome have an impact on vaginal absorption of exogenous progesterone (P intake) in artificial cycles? Summary answer Results suggest that the presence of lactobacillus in the vagina may positively impact the uptake of P. What is known already Low serum P levels on ET day decrease significantly live birth rates. About 20% of patients receiving micronized vaginal P show inadequate levels. It is of interest to find out which intrinsic factors might influence on the capacity of absorption (P intake) of vaginal P, and if they could be treated to prevent this situation. Vaginal microbiome hast been suggested as a possible factor, although this has not been addressed yet. The aim of this study was to explore if there is a correlation between vaginal microbiome and pH, and P intake. Study design, size, duration A prospective single-centre cohort pilot study including 92 ART patients was conducted from February 2022 to January 2023 in IVI-RMA Valencia. Embryo transfer was conducted in the context of a hormonal replacement therapy cycle with use of vaginal P (400mg/12h) for luteal phase support. “Progesterone intake” capacity was evaluated according to the vaginal microbiome status (lactobacillus dominant (LDM, &amp;gt;90%) or non-lactobacillus dominant (NLDM) and pH levels determinations (considered normal when moderately acidic ( &amp;lt; =4.8). Participants/materials, setting, methods Samples were taken twice: A) proliferative phase (day of initiation of exogenous P) and B) mid-luteal phase (day of ET, P + 5). Serum E2 and P levels as well as vaginal microbiome and pH were analysed. A validated qPCR methodology was used to accurately assess the presence and relative abundance of the four major lactobacillus species associated with eubiosis (crispatus, gasseri, iners, and jensenii) and the most common bacterial species implicated in dysbiosis. Main results and the role of chance Distribution of LDM and NLDM profiles were comparable between Samples A and B. No significant difference between any of the potential confounding factors (age, BMI,baseline P) between the lactobacillus CST groups (CST-I,-II,-III,-V &amp; lactobacillus) and the dysbiotic CST group (CST-IV) in Samples A and B were observed. Serum P levels on the ET day were significantly higher in LDM profiles both in Sample A (15.2 vs. 12.9ng/mL in NLDM;p=0.009) and B (15.0 vs. 12.8ng/mL;p=0.014). A positive significant correlation was found between serum P levels and Lactobacillus abundance in Samples A (r = 0.28; p = 0.008) and B (r = 0.30; p = 0.004). The relative abundance of Gardnerella vaginalis in Sample A was negatively related to serum P levels on the ET day (r=-0.24;p=0.02). Vaginal pH was significantly lower in LDM profiles both in Sample A (4.9 vs. 5.6 in NLDM; p = 0.002) and B (4.7 vs. 5.1;p&amp;lt;0.001). pH levels were negatively correlated with Lactobacillus abundance in Samples A (r=-0.34;p=0.0012) and B (r=-0.41;p&amp;lt;0.001). Additionally, serum P levels were negatively correlated with pH levels in Sample A (r=-0.14;p=0.191) and B (r=-0.27; p = 0.010). Indeed, when patients are grouped based on a cut-off of P = 8.8ng/mL, patients with a pH ≤ 4.8 have significantly higher P levels than patients with a pH &amp;gt; 4.8. Limitations, reasons for caution This is a pilot study, thus further trials with larger sample sizes should be performed in order to confirm these results. Additionally, vaginal microbiome alone does not explain the total variability in serum P levels measured in artificial cycles when using MVP. Wider implications of the findings We might have found one of the causes of the high heterogeneity on vaginal absorption of MVP in artificial cycles. This finding may help us with patient management when programming an ET in this type of cycles, improving personalised patient care and progressing on luteal phase support individualisation. Trial registration number 2110-VLC-095-EL

O-273 A transcriptomic approach for embryo selection after preimplantation genetic diagnosis for aneuploidy

Abstract Study question Is it possible to predict an euploid chromosomal constitution from RNA-Seq data and identify a transcriptomic profile compatible with extended embryonic development? Summary answer It has been possible to obtain a karyotype comparable to PGT-A, in addition to acquiring a transcriptomic signature of embryos suggestive of improved implantation capacity What is known already Conventional assessment of embryo competence, based on morphology and morphokinetic, lacks knowledge of molecular aspects and faces controversy in predicting ploidy status. Understanding the embryonic transcriptome is crucial, as gene expression influences development and implantation. Preimplantation genetic testing (PGT) has improved pregnancy rates, but problems persist when high-quality euploid embryos do not reach term. In fact, around 50 to 60% implant, of which 10% miscarry. Comprehensive approaches, including RNA-Seq, offer potential to discover molecular markers of reproductive competence. Extended-embryo culture platforms up to day 14 can be utilized as a proxy to study embryo development at post-implantation stages Study design, size, duration Prospective pilot cohort study conducted from March 2023 to August 2023. A total of 30 vitrified human blastocysts with previous PGT-A diagnosis on day 5 (D5) or day 6 (D6) of development were analysed: n = 15 euploid and n = 15 aneuploid embryos. Finally, 21 embryo samples were included in the study (n = 10 euploid and n = 11 aneuploid), the rest (n = 9) were excluded due to poor quality after sequencing. All embryos were donated for research purposes Participants/materials, setting, methods Following warming and re-expansion, embryos underwent a second trophectoderm biopsy. The remaining embryo was cultured until day 11 to assess its development. After biopsy analysis by RNA-Seq, karyotypes were studied, as well as differential expression gene (DEG) of the comparison: unattached embryos (n = 12) vs embryos attached (n = 9) to the plate (padj&amp;lt;0.05 and FC &amp;gt; 1). In addition, we obtained a transcriptomic signature specific to embryos with a “theoretical” capacity for sustained implantation (raw p-value&amp;lt;0.05 and shrunk LFC&amp;gt;1.1) Main results and the role of chance Digital karyotype concordance was obtained with a sensitivity of 81%, a specificity of 83%, a Kappa index of 65.5% and an area under the ROC curve of 90%. At the gene level, 76 DEGs (56 upregulated and 20 downregulated) were found in the comparison of embryos unattached to the plate in extended culture versus those that were attached. To address the functional implications of these differences, significantly deregulated pathways according to GO and KEGG categories were identified. The mural trophectoderm of the analysed blastocysts showed 63 significantly deregulated terms (30 GO-Biological Process, 13 GO-Cellular Component, 12 GO-Molecular Function and 8 KEGG pathways), displaying upregulation of autophagy, apoptosis, protein kinase and ubiquitin-like protein ligase activity, and downregulation of ribosome, spliceosome, kinetochore, segregation, and chromosome condensation processes. The first described transcriptomic signature specific to embryos with theoretical enhanced implantation capacity consists of 501 genes, including: EMP2, AURKB, FOLR1, NOTCH3, LRP2, FZD5, MDH1, APOD, GPX8, COLEC12, HSPA1A, CMTM7, BEX3, which are related to implantation and embryonic development. These findings indicate that it would be possible to identify euploid embryos with a greater capacity for implantation and development, excluding those embryos that, although they could be implanted and viable, present chromosomal alterations Limitations, reasons for caution This study included a small sample size and a remarkable variability between samples. Structural chromosomal abnormalities were not included. It is not possible to diagnose mosaic embryos. Trophectoderm biopsy does not assure the chromosomal status of the whole embryo. Maximum number of days for in vitro development was 11 Wider implications of the findings Gene expression profiles of embryos with theoretical increased implantation capacity have been obtained. Clinical implementation of this technique, which assesses chromosomal content and transcriptomic profile, could be used to significantly increase clinical outcomes in assisted reproduction, allowing informed decisions to be made about future treatments and the prospects of success Trial registration number Not applicable

Efficacy and Safety of Platelet-Rich Plasma on Bronchopleural Fistula: A Pilot Prospective Cohort Study

IntroductionBronchopleural fistula (BPF) is associated with high morbidity and mortality rates in patients undergoing pulmonary resections. Surgery, bronchoscopy, and conservative management have their limitations for small fistulas. Platelet-rich plasma (PRP) has regenerative properties, which might be efficient in enhancing tissue recovery and repairing small BPF. This study aimed to investigate efficacy and safety of PRP on BPF. MethodsThis is a pilot prospective cohort study. Patients whose fistulas smaller than 4mm were enrolled in this study, treated with PRP under bronchoscopy and followed up at 2 weeks and 4–6 weeks after the last PRP treatment. The cure rate, improvement rate and ineffectiveness rate were investigated. The severity of respiratory symptoms was evaluated by modified Medical Research Council dyspnea scale (mMRC) and COPD Assessment Test (CAT). The recurrence of fistula, new infection and mortality rate were examined. Adverse events were documented to explore the safety profile of PRP therapy. ResultsA total of 16 patients (mean age, 50.1 years) met the eligibility criteria. The median time from the first PRP treatment to the closure of the fistula was 12.0 (IQR 6.0, 21.5) days. Our findings indicate an effectiveness rate of 87.6%, with 68.8% of cure and 18.8% of improvement, along with significant improvement of respiratory symptoms evaluated by mMRC (P<0.001) and CAT (P<0.001). No recurrent of fistulas, newly developed infection, or death was observed. Adverse events of the procedure were most mild (82.6%) and temporary. ConclusionsPRP is a potential treatment for small BPF and is well tolerated.

Empowerment, Pain Control, and Quality of Life Improvement in Early Triple-Negative Breast Cancer Patients through Pain Neuroscience Education: A Prospective Cohort Pilot Study Protocol (EMPOWER Trial).

The treatment of early triple-negative breast cancer (eTNBC) has improved patients' prognosis but often leads to adverse events and sequelae affecting quality of life (QoL). Pain Neuroscience Education (PNE) is a promising non-pharmacological intervention in this field. Preliminary data have shown the beneficial effect of PNE in BC survivors. However, there are still gaps in knowledge regarding its optimal use in eTNBC. To address this issue, a prospective pilot study will enroll 30 consecutive patients diagnosed with eTNBC at IRCCS Humanitas Research Hospital. The PNE program will consist of 10 weekly sessions to be started within 4 weeks of the onset or worsening of a pain syndrome (PS). QoL, pain, and disability will be assessed before, during, at the end of, and 6 months after PNE using validated questionnaires. Peripheral venous blood samples will be taken before and at the end of PNE to evaluate inflammatory serum biomarker levels. The primary objective is to evaluate whether PNE leads to clinical improvement in QoL and pain. If successful, it will be validated in a larger multi-centric cohort, potentially leading to its widespread implementation as a standard pain management tool for eTNBC patients.

Association Between Dietary Intake After Roux-en-Y Gastric Bypass Surgery and Antioxidant/inflammatory Status: A 6-Month Pilot Prospective Cohort Study.

Adipose tissue can increase levels of inflammation and oxidative stress, which explains the relationship between obesity and many chronic diseases. Weight loss, changes in adipose tissue metabolism, and dietary nutrient intake changes following bariatric surgery could affect a number of oxidative- and inflammation-related factors. Therefore, this study aimed to assess the potential relationship between dietary intake and inflammatory/antioxidant markers in the 6months following Roux-en-Y gastric bypass surgery (RYGB). This pilot prospective cohort study included 45 patients with severe obesity who underwent RYGB. The patients were assessed at three different time points: baseline, 3 months, and 6 months post-surgery. Throughout the study, dietary intake data, levels of total antioxidant capacity (TAC), NF-κB, and serum levels of certain micronutrients were measured three times. Dietary macro- and micronutrient intake data were obtained three times throughout the study using the 24-h food recall questionnaire. The analysis of dietary indices in the present study found a significant positive correlation between the dietary intake of zinc, copper, MUFA, and serum TAC levels. It also revealed a significant inverse correlation between serum levels of NF-κB with vitamin E and PUFA intake. Additionally, there was a significant positive association between the amount of dietary carbohydrates and saturated fatty acids intake and the levels of NF-κB. Furthermore, within 3 to 6 months after the surgery, patients experienced an increase in serum levels of TAC, ferritin, vitamin D3, vitamin B12, and folate. However, there was a decrease in serum levels of NF-κB, zinc, and copper. Weight loss and nutritional status may potentially impact oxidative stress and inflammation levels within 6months following RYGB surgery. Further research is necessary to comprehensively investigate the different facets of this correlation and elucidate the precise underlying mechanism.

Early effect of onabotulinumtoxinA on EEG-based functional connectivity in patients with chronic migraine: A pilot study.

In this pilot prospective cohort study, we aimed to evaluate, using high-density electroencephalography (HD-EEG), the longitudinal changes in functional connectivity (FC) in patients with chronic migraine (CM) treated with onabotulinumtoxinA (OBTA). OBTA is a treatment for CM. Several studies have shown the modulatory action of OBTA on the central nervous system; however, research on migraine is limited. This study was conducted at the Neurology Unit of "Policlinico Tor Vergata," Rome, Italy, and included 12 adult patients with CM treated with OBTA and 15 healthy controls (HC). Patients underwent clinical scales at enrollment (T0) and 3 months (T1) from the start of treatment. HD-EEG was recorded using a 64-channel system in patients with CM at T0 and T1. A source reconstruction method was used to identify brain activity. FC in δ-θ-α-β-low-γ bands was analyzed using the weighted phase-lag index. FC changes between HCs and CM at T0 and T1 were assessed using cross-validation methods to estimate the results' reliability. Compared to HCs at T0, patients with CM showed hyperconnected networks in δ (p = 0.046, area under the receiver operating characteristic curve [AUC: 0.76-0.98], Cohen's κ [0.65-0.93]) and β (p = 0.031, AUC [0.68-0.95], Cohen's κ [0.51-0.84]), mainly involving orbitofrontal, occipital, temporal pole and orbitofrontal, superior temporal, occipital, cingulate areas, and hypoconnected networks in α band (p = 0.029, AUC [0.80-0.99], Cohen's κ [0.42-0.77]), predominantly involving cingulate, temporal pole, and precuneus. Patients with CM at T1, compared to T0, showed hypoconnected networks in δ band (p = 0.032, AUC [0.73-0.99], Cohen's κ [0.53-0.90]) and hyperconnected networks in α band (p = 0.048, AUC [0.58-0.93], Cohen's κ [0.37-0.78]), involving the sensorimotor, orbitofrontal, cingulate, and temporal cortex. These preliminary results showed that patients with CM presented disrupted EEG-FC compared to controls restored by a single session of OBTA treatment, suggesting a primary central modulatory action of OBTA.

Investigation of the pathogen-specific antibody response in periprosthetic joint infection.

Periprosthetic joint infections (PJIs) are a very demanding complication of arthroplasty. Diagnosis of PJI and pathogen identification pose considerable challenges in clinical practice. We hypothesized that the pathogen-specific immune response to PJI reflects the infection process, provides clinically relevant information on disease course, and has the potential to further optimize antimicrobial therapy. We conducted a prospective matched cohort pilot study with 13 patients undergoing two-stage septic revision arthroplasty (PJI patients) between 06/2020 and 06/2021, as well as 11 control patients undergoing one-stage aseptic revision arthroplasty (Non-PJI patients). Pre-, intra- and postoperative serum samples were collected at standardized time points. We developed a custom Luminex®-based quantitative bead-based suspension array (Infection Array; IA), and used it for simultaneous measurement of antibody specificities against 32 pathogens commonly associated with PJI in 267 serum samples. The IA was able to trace the dynamics of the pathogen-specific humoral immune response in all patients against PJI-related pathogens, prominently coagulase-negative staphylococci and streptococci. Pathogen-specific serum antibody titers declined in 62% of PJI patients over the course of treatment, while no changes in antibody titers were observed in 82% of Non-PJI patients during this study. Our serological data strongly suggested that antibody signatures reflect an immune response to microbial invasion. Our results provide insights into the pathophysiology of PJI and information on the individual disease courses. The IA is therefore a promising and novel serological tool of high resolution for monitoring the immunoproteomic footprints of infectious pathogens in the course of PJI.