A randomized, multicenter phase III Study of once-per-cycle administration of efbemalenograstim alfa (F-627), a novel long-acting rhG-CSF, for prophylaxis of chemotherapy-induced neutropenia in patients with breast cancer.

BACKGROUND: F-627 (efbemalenograstim alfa) is a novel long acting granulocyte colony-stimulating factor (G-CSF) that contains two human G-CSF fused to a human immunoglobulin G2 (hIgG2) -Fc fragment with a peptide linker. This studyevaluated the efficacy and safety of F-627, also known as efbemalenograstim alfa (Ryzneuta®) in reducing neutropenia compared with filgrastim (GRAN®). METHODS: This was a multicenter, randomized, open-label, active-controlled non-inferiority study. Two hundred thirty nine (239) patients were enrolled in thirteen centers and received the chemotherapy with epirubicin (100 mg/m2) and cyclophosphamide (600 mg/m2) on day 1 of each cycle for a maximum of four cycles. Patients were randomized to receive either a single 20 mg subcutaneous (s.c.) injection of F-627 on day 3 of each cycle or daily s.c. injection of filgrastim 5 µg/kg/d starting from day 3 of each cycle. The primary endpoint was the duration of grade 3 or 4 neutropenia in cycle 1. The safety profile was also evaluated. RESULTS: The mean (SD) duration of grade 3 or 4 neutropenia in cycle 1 was 0.68 (1.10) and 0.71 (0.95) days for the F-627 and the filgrastim groups, respectively. The Hodges-Lehmann estimate of the between-group median difference (F-627 vs filgrastim) in the duration of grade 3 or 4 neutropenia in cycle 1 was 0 day and the upper limit of the one-sided 97.5% CI was 0 day, which was within the prespecified non-inferiority margin of 1-day. Results for all efficacy endpoints in cycles 2 - 4 were consistent with the results in cycle 1, however a trend towards a lower incidence and a shorter duration of grade 3 or 4 neutropenia and grade 4 neutropenia was observed in the F-627 group compared with the filgrastim group. The ANC nadir in the F-627 group was significantly higher than that in the filgrastim group in each cycle. A single fixed dose of F-627 was well tolerated and as safe as standard daily filgrastim. CONCLUSIONS: A single fixed dose of 20 mg of F-627 in each cycle was as safe and effective as a daily dose of filgrastim 5 µg/kg/d in reducing neutropenia and its complications in patients who received four cycles of EC. TRIAL REGISTRATION: ClinicalTrials.gov: NCT04174599, on 22/11/2019.

Case report: CSF hypotension secondary to a free syringo-subarachnoid-peritoneal shunt.

INTRODUCTION: Syringomyelia is present in 40% of pediatric patients with Chiari malformation. Typically treated with posterior fossa decompression, some cases require further intervention such as syrinx shunting. CASE REPORT: We report a 16-year-old female with Chiari type 1 malformation and syringomyelia who underwent posterior fossa decompression and subsequent free syringo-subarachnoid-peritoneal shunting. The patient developed symptoms of CSF overdrainage, and imaging indicated CSF hypotension. A distal catheter ligation temporarily improved symptoms, but eventually, a programmable ventricular shunt was necessary due to shunt dependence. CONCLUSION: This case highlights the rare complication of CSF overdrainage from syrinx shunting and the importance of shunt selection considerations.

Functional MRI Study with Conductivity Signal Changes During Visual Stimulation.

BACKGROUND: Although blood oxygen level-dependent (BOLD) functional MRI (fMRI) is a standard method, major BOLD signals primarily originate from intravascular sources. Magnetic resonance electrical properties tomography (MREPT)-based fMRI signals may provide additional insights into electrical activity caused by alterations in ion concentrations and mobilities. PURPOSE: This study aimed to investigate the neuronal response of conductivity during visual stimulation and compare it with BOLD. MATERIALS AND METHODS: A total of 30 young, healthy volunteers participated in two independent experiments using BOLD and MREPT techniques with a visual stimulation paradigm at 3T MRI. The first set of MREPT fMRI data was obtained using a multi-echo spin-echo (SE) echo planar imaging (EPI) sequence from 14 participants. The second set of MREPT fMRI data was collected from 16 participants using both a single-echo SE-EPI and a single-echo three-dimensional (3D) balanced fast-field-echo (bFFE) sequence. We reconstructed the time-course Larmor frequency conductivity to evaluate hemodynamics. RESULTS: Conductivity values slightly increased during visual stimulation. Activation strengths were consistently stronger with BOLD than with conductivity for both SE-EPI MREPT and bFFE MREPT. Additionally, the activated areas were always larger with BOLD than MREPT. Some participants also exhibited decreased conductivity values during visual stimulations. In Experiment 1, conductivity showed significant differences between the fixation and visual stimulation blocks in the secondary visual cortex (SVC) and cuneus, with conductivity differences of 0.43% and 0.47%, respectively. No significant differences in conductivity were found in the cerebrospinal fluid (CSF) areas between the two blocks. In Experiment 2, significant conductivity differences were observed between the two blocks in the SVC, cuneus, and lingual gyrus for SE-EPI MREPT, with differences of 0.90%, 0.67%, and 0.24%, respectively. Again, no significant differences were found in the CSF areas. CONCLUSION: Conductivity values increased slightly during visual stimulation in the visual cortex areas but were much weaker than BOLD responses. The conductivity change during visual stimulation was less than 1% compared to the fixation block. No significant differences in conductivity were observed between the primary visual cortex (PVC)-CSF and SVC-CSF during fixation and visual stimulations, suggesting that the observed conductivity changes may not be related to CSF changes in the visual cortex but rather to diffusion changes. Future research should explore the potential of MREPT to detect neuronal electrical activity and hemodynamic changes, with further optimization of the MREPT technique.

An approach to autologous stem cell mobilization: trying to define good mobilizers.

BACKGROUND AND OBJECTIVES: Stem cell mobilization is a well-known procedure to harvest hematopoietic stem cells for autologous stem cell transplantation in certain hematologic diseases. Numerous studies have been conducted to identify risk factors for poor mobilization but there are no studies that identify good mobilizers. In our hospital, we decided to explore good mobilizers, defining them as those with ≥40 CD34+ cells/µL on Day +4 in order to start early apheresis. MATERIAL AND METHODS: A descriptive retrospective study was performed at Hospital Universitari Son Espases. A total of 198 patients mobilized with doses of around 10 µg/kg of granulocyte colony-stimulating factor (G-CSF) every 12 h were analyzed for autologous collection between January 2015 and September 2022. Fifty patients who had ≥40 CD34+ cells/µL on Day +4 started early apheresis; the rest continued mobilization as planned. Success was defined as obtaining over 2.5 × 106 CD34+ cells/kg in a single apheresis. RESULTS: The necessary number of CD34+ cells/kg to perform an autologous stem cell transplantation was reached in a single apheresis session in 62 % of patients with ≥40 CD34+ cells/µL in peripheral blood. A cutoff of 102 CD34+ cells/µL on Day +4 was shown to have the best success rate (94 %). In an analysis of success, age, previously failed mobilization and having one or more adverse factors for bad mobilization were statistically significant. CONCLUSION: Patients considered as good mobilizers were matched with our factors of poor mobilization, revealing that most patients (79 %) had none or only one risk factor for poor mobilization. Apheresis on Day +4 in good mobilizers was shown to be an effective alternative to reduce mobilization duration and decrease the amount of granulocyte-colony stimulating factor administered.

Targeted proteomics of cerebrospinal fluid in treatment naïve multiple sclerosis patients identifies immune biomarkers of clinical phenotypes.

Multiple sclerosis (MS) is an inflammatory demyelinating disease with heterogeneous clinical presentations and variable long-term disability accumulation. There are currently no standard criteria to accurately predict disease outcomes. In this study we investigated the cross-sectional relationship between disease phenotype and immune-modulating cytokines and chemokines in cerebrospinal fluid (CSF). We analyzed CSF from 20 DMT-naïve MS patients using Olink Proteomics' Target 96 Inflammation panel and correlated the resulting analytes with respect to (1) disease subtype, (2) patient age and sex, (3) extent of clinical disability, and (4) MRI segmental brain volumes. We found that intrathecal IL-4 correlated with higher Expanded Disability Status Scale (EDSS) scores and longer 25-foot walk times, and CD8A correlated with decreased thalamic volumes and longer 9-hole peg test times. Male sex was associated with higher FGF-19 expression, and Tumefactive MS with elevated CCL4. Several inflammatory markers were correlated with older age at the time of LP. Finally, higher intrathecal IL-33 correlated with increased MS lesion burden and multi-compartment brain atrophy. This study confirms immune heterogeneity underlying CSF profiles in MS, but also identifies several inflammatory protein biomarkers that may be of use for predicting clinical outcomes in future algorithms.

Changes of NLRP3 in serum and cerebrospinal fluid of patients after moderate to severe traumatic brain injury and their predictive values for prognosis.

BACKGROUND: Traumatic brain injury (TBI) remains a major concern for global health. Recent studies have suggested the role of NOD-like receptor pyrin domain-containing protein 3 (NLRP3), an inflammatory marker, in the cerebrospinal fluid (CSF) and serum as potential indicators of TBI prognosis. The objective of the study was to characterize NLRP3 as a clinically applicable tool for predicting the outcomes of TBI patients. METHODS: A total of 270 patients with moderate to severe TBI were included in this retrospective analysis. Serum and CSF samples were collected at 1-, 3-, 7-, and 21-day post-injury to measure NLRP3 levels. The prognosis of patients was evaluated after 3 months using the Glasgow Outcome Scale (GOS). Patients were categorized into good prognosis (GOS score >3) and poor prognosis (GOS score ≤3) groups. The relationship between NLRP3 levels and prognosis was analyzed. RESULTS: Patients with poor prognosis had significantly elevated NLRP3 levels in their serum on days 1 and 3 post-injury compared with those with a good prognosis. The difference was more pronounced during these early days compared with days 7 and 21. However, NLRP3 levels in CSF consistently showed a large difference between the two groups throughout the observation period. Receiver operating characteristic analysis revealed that the level of NLRP3 in the CSF on day 3 post-injury had the highest predictive value for prognosis, with an area under the curve of 0.83, followed by the level of NLRP3 in the serum on day 3 post-injury. CONCLUSIONS: The levels of NLRP3, especially in the CSF on day 3 post-injury, can serve as a potential biomarker for predicting prognosis in moderate to severe TBI patients. Early measurement of NLRP3 levels can provide valuable insights into patient outcomes and guide therapeutic strategies.

Assessing The Feasibility of Selective Piezoelectric Osteotomy in Transorbital Approach to The Middle Cranial Fossa: Anatomical and Quantitative Study and Surgical Implications.

OBJECTIVES: To verify the feasibility and to discuss advantages and disadvantages of a piezoelectric orbitotomy (PO) during Superior Eyelid Endoscopic Transorbital Approach (SETOA). METHODS: Five adult specimens underwent exoscopic/endoscopic SETOA to middle cranial fossa. The surgical corridor was created via piezoelectric orbitotomy by performing three selective and safe micrometric bone cuts providing a one-piece trapezoid bone flap which was repositioned and secured at the end of procedure. A 3D scan of the bone flap allowed us to reconstruct a 3D model and calculate its volume. An illustrative case demonstrating the application of this novel technique was also presented. RESULTS: Anatomical-morphometric quantitative analysis showed a mean bony-volume gain of 1,574.26 mm3 by using PO. PO yielded concrete surgical advantages and theoretical benefits in terms of functional and esthetic outcomes. All osteotomies were micrometric clear-cut and precise, resulting in a very thin bony gap; a complete sparing of soft tissues and neurovascular structures in- and around the orbit was observed. Lateral orbital wall reconstruction by replacing the bone flap aims to mitigate the risk of enophthalmos, proptosis, CSF leakage, pseudomeningocele and pulsatile headache, which represent significant challenges in the relevant literature. CONCLUSION: PO may offer a viable, selective, effective, safe alternative to high-speed drilling during SETOA, especially for patients affected by intra-axial pathologies in which a watertight closure is mandatory. This procedure could prevent/decrease the risk of some of the main postoperative complications associated to the standard SETOA, resulting potentially in better functional and esthetic outcome.

Increases in amyloid-ß42 slow cognitive and clinical decline in Alzheimer's disease trials.

Positive effects of new anti-amyloid-ß (Aß) monoclonal antibodies in Alzheimer's disease (AD) have been attributed to brain amyloid reduction. However, most anti-Aß antibodies also increase the CSF levels of the 42-amino acid isoform (Aß42). We evaluated the associations of changes in CSF Aß42 and brain Aß-PET with cognitive and clinical end points in randomized trials of anti-Aß drugs that lowered (ß- and γ-secretase inhibitors) or increased CSF Aß42 levels (anti-Aß monoclonal antibodies) to test the hypothesis that post-treatment increases in CSF Aß42 levels are independently associated with cognitive and clinical outcomes. From long-term (≥12 months) randomized placebo-controlled clinical trials of anti-Aß drugs published until November 2023, we calculated the post-treatment versus baseline difference in ADAS-Cog (cognitive subscale of the Alzheimer's Disease Assessment Scale) and CDR-SB (Clinical Dementia Rate-Sum of Boxes) and z-standardized changes in CSF Aß42 and Aß-PET Centiloids (CL). We estimated the effect size [regression coefficients (RCs) and confidence intervals (CIs)] and the heterogeneity (I2) of the associations between AD biomarkers and cognitive and clinical end points using random-effects meta-regression models. We included 25 966 subjects with AD from 24 trials. In random-effects analysis, increases in CSF Aß42 were associated with slower decline in ADAS-Cog (RC: -0.55; 95% CI: -0.89, -0.21, P = 0.003, I2 = 61.4%) and CDR-SB (RC: -0.16; 95% CI: -0.26, -0.06, P = 0.002, I2 = 34.5%). Similarly, decreases in Aß-PET were associated with slower decline in ADAS-Cog (RC: 0.69; 95% CI: 0.48, 0.89, P < 0.001, I2 = 0%) and CDR-SB (RC: 0.26; 95% CI: 0.18, 0.33, P < 0.001, I2 = 0%). Sensitivity analyses yielded similar results. Higher CSF Aß42 levels after exposure to anti-Aß drugs are independently associated with slowing cognitive impairment and clinical decline. Increases in Aß42 may represent a mechanism of potential benefit of anti-Aß monoclonal antibodies in AD.

Granulocyte colony stimulating factor promotes scarless tissue regeneration.

Mammals typically heal with fibrotic scars, and treatments to regenerate human skin and hair without a scar remain elusive. We discovered that mice lacking C-X-C motif chemokine receptor 2 (CXCR2 knockout [KO]) displayed robust and complete tissue regeneration across three different injury models: skin, hair follicle, and cartilage. Remarkably, wild-type mice receiving plasma from CXCR2 KO mice through parabiosis or injections healed wounds scarlessly. A comparison of circulating proteins using multiplex ELISA revealed a 24-fold higher plasma level of granulocyte colony stimulating factor (G-CSF) in CXCR2 KO blood. Local injections of G-CSF into wild-type (WT) mouse wound beds reduced scar formation and increased scarless tissue regeneration. G-CSF directly polarized macrophages into an anti-inflammatory phenotype, and both CXCR2 KO and G-CSF-treated mice recruited more anti-inflammatory macrophages into injured areas. Modulating macrophage activation states at early time points after injury promotes scarless tissue regeneration and may offer a therapeutic approach to improve healing of human skin wounds.

Super-Resolving and Denoising 4D flow MRI of Neurofluids Using Physics-Guided Neural Networks.

PURPOSE: To obtain high-resolution velocity fields of cerebrospinal fluid (CSF) and cerebral blood flow by applying a physics-guided neural network (div-mDCSRN-Flow) to 4D flow MRI. METHODS: The div-mDCSRN-Flow network was developed to improve spatial resolution and denoise 4D flow MRI. The network was trained with patches of paired high-resolution and low-resolution synthetic 4D flow MRI data derived from computational fluid dynamic simulations of CSF flow within the cerebral ventricles of five healthy cases and five Alzheimer's disease cases. The loss function combined mean squared error with a binary cross-entropy term for segmentation and a divergence-based regularization term for the conservation of mass. Performance was assessed using synthetic 4D flow MRI in one healthy and one Alzheimer' disease cases, an in vitro study of healthy cerebral ventricles, and in vivo 4D flow imaging of CSF as well as flow in arterial and venous blood vessels. Comparison was performed to trilinear interpolation, divergence-free radial basis functions, divergence-free wavelets, 4DFlowNet, and our network without divergence constraints. RESULTS: The proposed network div-mDCSRN-Flow outperformed other methods in reconstructing high-resolution velocity fields from synthetic 4D flow MRI in healthy and AD cases. The div-mDCSRN-Flow network reduced error by 22.5% relative to linear interpolation for in vitro core voxels and by 49.5% in edge voxels. CONCLUSION: The results demonstrate generalizability of our 4D flow MRI super-resolution and denoising approach due to network training using flow patches and physics-based constraints. The mDCSRN-Flow network can facilitate MRI studies involving CSF flow measurements in cerebral ventricles and association of MRI-based flow metrics with cerebrovascular health.

Assessing the effects of spontaneous intracranial hypotension on quality of life, work ability and disability.

BACKGROUND: Spontaneous intracranial hypotension (SIH), characterized by headaches due to cerebrospinal fluid leaks or low pressure, is a challenging condition to diagnose and treat and affects the quality of life. METHODS: An 8­week online survey was conducted to assess the impact of SIH on symptoms, sociodemographics and quality of life. The cohort was comprised of patients who had a self-reported diagnosis of SIH and were divided into two groups: those with radiological evidence of SIH and those with clinical suspicion but no radiological evidence. Mental health and disability were evaluated using the Depression, Anxiety and Stress Scale-21 (DASS-21) and the Henry Ford Hospital Headache Disability Inventory (HDI). RESULTS: A total of 86 participants were included in the study, 59 with radiological evidence and 27 without. Most participants were female (84.9%) with a mean age of 44.8 years. Orthostatic headache was more common in participants without radiological evidence (74.1% vs. 42.4%). The severity in those with radiological evidence was 27.1% mild, 27.1% moderate, 30.5% severe and 15.3% extremely severe, while those without had 7.4% mild, 18.5% moderate, 63.0% severe and 11.1% extremely severe headaches. Mental health assessment using the DASS-21 scale showed that 77.9% of all participants reported signs of depression, 96.5% reported anxiety and 89.5% reported stress. The HDI showed 2.3% total disability, 40.7% severe, 19.8% moderate and 37.2% mild. The impact on employment was significant: 15.1% were able to work full-time, 48.8% part-time, 30.2% were unable to work and 5.8% retired early due to SIH. CONCLUSION: The study demonstrates the broad impact of SIH affecting physical health, mental well-being, and socioeconomic status, and calls for multifaceted and robust management approaches to address its complex effects on patients.

Reuse of Nasoseptal Flaps for Endoscopic Endonasal Skull Base Reconstruction.

Introduction Pedicled nasoseptal flap (NSF) placement is a critical component of skull base reconstruction after endoscopic endonasal approaches (EEAs). The effectiveness of NSF reuse has not been thoroughly studied. Prior reports using flaps harvested at one center and reused at another may have technical variability bias. Methods We identified patients who underwent both their initial and NSF-reused surgeries at Weill Cornell Medical College from 2004 to 2022 using a prospective database of all EEAs. Surgical pathology, intraoperative leak grade, use of cerebrospinal fluid (CSF) diversion and skull base coverage were examined. The primary outcome measure was occurrence of CSF leak. Results Fourteen patients (six women, eight men) underwent 14 first time and 14 revision operations with median age of 36.6 years (interquartile range [IQR]: 23.9-61.3) at the time of the NSF reuse. The median interval between the first NSF use and reuse was 70.6 months (IQR: 16.6-87). Eight patients were operated on for pituitary adenoma. Nonadenomas included three craniopharyngiomas and one case each of epidermoid, ependymoma, and chordoma. There were 16 high-flow, 8 low-flow intraoperative leaks, and 4 with no leak. CSF diversion was used in 24 operations. There were three postoperative leaks, one after a first operation and two after NSF reuse. All postoperative CSF leaks, whether first or second operations, occurred in cases with both high-flow intraoperative CSF leak and incomplete NSF coverage ( p = 0.006). Conclusion NSF reuse is effective at preventing postoperative CSF leak. The primary predictors of leak are high-flow intraoperative leak and inadequate defect coverage with NSF, regardless of the operation number.

Anatomical-Radiological Aspects and Their Influence on the Results of Pituitary Adenomas Endoscopic Endonasal Surgery.

Introduction Endoscopic endonasal surgery has globally improved postoperative results in pituitary adenomas. Material and Methods We retrospectively analyzed 101 patients who underwent endonasal endoscopic surgery for pituitary adenomas in the period from 2016 to 2021. Data on epidemiological variables, preoperative radiological factors including tumor volume, tumor appearance, cavernous sinus invasion (modified Knosp scale), degree of extension according to the SIPAP (stands for the five directions in which a pituitary adenoma can extend: suprasellar, infrasellar, parasellar, anterior, and posterior) classification, and preoperative visualization of the healthy gland on magnetic resonance imaging (MRI) were collected as well as intra- and postoperative cerebrospinal fluid (CSF) leak. As variables of interest, data on the degree of tumoral resection and preservation of hormonal function were collected. Results Among the preoperative factors related to greater tumoral resection, we found a lesser tumoral extension according to the SIPAP scale, and the absence of a postoperative CSF leak had a statistically significant relation with greater hormonal preservation. Conclusion The SIPAP classification is a simple-to-measure preoperative radiological variable that could predict the extent of resection, and, conversely, the occurrence of a postoperative CSF leak has been associated with an inferior endocrinological outcome in this type of surgery.

Giant metastatic mixed-type spinal ependymoma presenting with hydrocephalus.

Ependymomas are rare nervous system tumors that can arise anywhere in the neuraxis. While having a high propensity for leptomeningeal dissemination, retrograde dissemination (from the spine to the CNS) remains infrequent. We describe the case of a 31-year-old female who presented with hydrocephalus secondary to an intracranial leptomeningeal metastasis of a giant spinal ependymoma with mixed (classic and myxopapillary) histopathologic features, successfully treated with surgical resection and radiotherapy of the entire neuraxis. This case highlights the importance of including ependymomas in the differential diagnosis for lesions in atypical extra-axial locations, of systematically obtaining imaging of the entire neuraxis when suspecting it, and of considering retrograde dissemination when both intracranial and spinal lesions are present.

Thrombospondin-4 as potential cerebrospinal fluid biomarker for therapy response in pediatric spinal muscular atrophy.

BACKGROUND AND PURPOSE: Spinal muscular atrophy (SMA) as the second most common neurodegenerative disorder in childhood is characterized by the deficiency of survival of motor neuron (SMN) protein leading predominantly to degeneration of alpha motor neurons and consequently to progressive muscle weakness and atrophy. Besides some biomarkers like SMN2 copy number therapeutic biomarkers for SMA with known relevance for neuromuscular transmission are lacking. Here, we examined the potential of Thrombospondin-4 (TSP4) to serve as a cerebrospinal fluid (CSF) biomarker, which may also indicate treatment response. METHODS: We used untargeted proteomic analyses to determine biomarkers in CSF samples derived from pediatric pre-symptomatic (n = 6) and symptomatic (n = 4) SMA patients. The identified biomarker TSP4 was then validated in additional 68 CSF samples (9 adult and 24 pediatric SMA patients, 5 adult and 13 pediatric non-disease controls in addition to 17 pediatric disease controls) by enzyme-linked immunosorbent assay (ELISA) as an additional analytical approach. RESULTS: Untargeted proteomic analyses of CSF identified a dysregulation of TSP4 and revealed a difference between pre-symptomatic SMA patients and patients identified after the onset of first symptoms. Subsequent ELISA-analyses showed that TSP4 is decreased in pediatric but not adult SMA patients. CSF of pediatric patients with other neurological disorders demonstrated no alteration of TSP4 levels. Furthermore, CSF TSP4 levels of pediatric SMA patients increased after first dose of Nusinersen. CONCLUSIONS: We found that TSP4 levels are exclusively reduced in CSF of pediatric SMA patients and increase after treatment, leading us to the hypothesis that TSP4 could serve as a CSF biomarker with the potential to monitor treatment response in pediatric SMA patients. Moreover, TSP4 enable to distinguish pre-symptomatic and symptomatic patients suggesting a potential to serve as a stratification marker.

Heat shock on bovine embryos from day 2.5-3 selects the most competent for progression to the blastocyst stage.

Heat shock can impair embryo formation, while growth factors, such as colony-stimulating factor 2 (CSF2), modulate embryonic development. This study evaluated the effect of heat shock between days 2.5 and 3, as well as the impact of CSF2 at day 5 on bovine embryos cultured in a serum-free in vitro medium. The focus was on blastocyst development, the number of blastomeres, DNA fragmentation (TUNEL-positive cells), and mitochondrial activity. Heat shock reduced the proportion of cleaved embryos that developed into blastocysts (P = 0.0603). The resultant blastocysts exhibited a reduced number and proportion of TUNEL-positive cells in the trophectoderm (P = 0.0270 and P = 0.0240, respectively) and in the entire embryo (P = 0.0029 and P = 0.0031, respectively). Additionally, mitochondrial activity was lower in blastocysts derived from heat-shocked embryos (P = 0.0150) and further reduced in embryos exposed to both heat shock and CSF2 (P = 0.0415). In conclusion, the exposure of cleaved embryos to heat shock reduced their development to the blastocyst stage. However, the resulting blastocysts showed decreased DNA fragmentation and mitochondrial activity.

Combined Lactate Glucose Ratio (CLGR) as a novel marker for rapid diagnosis of CSF bacterial infection in neurosurgical patients: diagnostic accuracy study and benchtop analyzer correlation.

OBJECTIVE: This study aimed to assess the diagnostic accuracy of a novel marker, the combined lactate-to-glucose ratio (CLGR), in identifying cerebrospinal fluid (CSF) bacterial infection (CBI) in neurosurgical patients. Additionally, it seeks to establish cut-off values for CLGR and evaluate the reliability of measurement using blood gas analyzer (BGA). METHODS: CSF samples were collected from two neurosurgical centers in Kuala Lumpur, Malaysia, between January 2022 and October 2023. Conventional markers and CLGR were quantified using standard laboratory methods, with BGA utilized for measurement when feasible. Samples were categorized into confirmed CBI-positive (CBI+) and CBI-negative (CBI-) groups. Marker performance was compared, and receiver operating characteristic analysis conducted. Pearson's correlation assessed the agreement between BGA and laboratory measurements. RESULTS: Among the 130 CSF samples, 11 were CBI+. Both cLac and CLGR were significantly elevated in the CBI+ group (p<0.001). The area under the curve (AUC) for cLac and CLGR was 0.990 and 0.994, respectively. Using a cut-off of 6.0mmol/L, cLac demonstrated sensitivity of 100%, specificity of 93.3%, positive predictive value (PPV) of 57.9%, negative predictive value (NPV) of 100%, and diagnostic accuracy of 93.9%. CLGR ≥20.0 showed even higher accuracy: 100.0% sensitivity, 98.6% specificity, 84.6% PPV, 100% NPV, and overall accuracy of 98.5%. Both markers maintained excellent performance in blood-stained CSF. BGA measurements correlated well with laboratory results (r=0.980 & 0.999, respectively, p<0.001). CONCLUSIONS: CLac levels ≥6.0mmol/L and CLGR ≥20.0 accurately identified CBI in neurosurgical patients, with CLGR exhibiting superior efficacy. The potential for instant BGA measurement suggests promise for point-of-care testing.

Carotid artery vascular stenosis causes the blood-CSF barrier damage and neuroinflammation.

BACKGROUND: The choroid plexus (ChP) helps maintain the homeostasis of the brain by forming the blood-CSF barrier via tight junctions (TJ) at the choroid plexus epithelial cells, and subsequently preventing neuroinflammation by restricting immune cells infiltration into the central nervous system. However, whether chronic cerebral hypoperfusion causes ChP structural damage and blood-CSF barrier impairment remains understudied. METHODS: The bilateral carotid stenosis (BCAS) model in adult male C57BL/6 J mice was used to induce cerebral hypoperfusion, a model for vascular contributions to cognitive impairment and dementia (VCID). BCAS-mediated changes of the blood-CSF barrier TJ proteins, apical secretory Na+-K+-Cl- cotransporter isoform 1 (NKCC1) protein and regulatory serine-threonine kinases SPAK, and brain infiltration of myeloid-derived immune cells were assessed. RESULTS: BCAS triggered dynamic changes of TJ proteins (claudin 1, claudin 5) accompanied with stimulation of SPAK-NKCC1 complex and NF-κB in the ChP epithelial cells. These changes impacted the integrity of the blood-CSF barrier, as evidenced by ChP infiltration of macrophages/microglia, neutrophils and T cells. Importantly, pharmacological blockade of SPAK with its potent inhibitor ZT1a in BCAS mice attenuated brain immune cell infiltration and improved cognitive neurological function. CONCLUSIONS: BCAS causes chronic ChP blood-CSF damage and immune cell infiltration. Our study sheds light on the SPAK-NKCC1 complex as a therapeutic target in neuroinflammation.

Pan-cancer analysis reveals CCL5/CSF2 as potential predictive biomarkers for immune checkpoint inhibitors.

BACKGROUND: Currently, there are no optimal biomarkers available for distinguishing patients who will respond to immune checkpoint inhibitors (ICIs) therapies. Consequently, the exploration of novel biomarkers that can predict responsiveness to ICIs is crucial in the field of immunotherapy. METHODS: We estimated the proportions of 22 immune cell components in 10 cancer types (6,128 tumors) using the CIBERSORT algorithm, and further classified patients based on their tumor immune cell proportions in a pan-cancer setting using k-means clustering. Differentially expressed immune genes between the patient subgroups were identified, and potential predictive biomarkers for ICIs were explored. Finally, the predictive value of the identified biomarkers was verified in patients with urothelial carcinoma (UC) and esophageal squamous cell carcinoma (ESCC) who received ICIs. RESULTS: Our study identified two subgroups of patients with distinct immune infiltrating phenotypes and differing clinical outcomes. The patient subgroup with improved outcomes displayed tumors enriched with genes related to immune response regulation and pathway activation. Furthermore, CCL5 and CSF2 were identified as immune-related hub-genes and were found to be prognostic in a pan-cancer setting. Importantly, UC and ESCC patients with high expression of CCL5 and low expression of CSF2 responded better to ICIs. CONCLUSION: We demonstrated CCL5 and CSF2 as potential novel biomarkers for predicting the response to ICIs in patients with UC and ESCC. The predictive value of these biomarkers in other cancer types warrants further evaluation in future studies.