RNA Interactions Are Essential for CTCF-Mediated Genome Organization.

The function of the CCCTC-binding factor (CTCF) in the organization of the genome has become an important area of investigation, but the mechanisms by which CTCF dynamically contributes to genome organization are not clear. We previously discovered that CTCF binds to large numbers of endogenous RNAs, promoting its self-association. In this regard, we now report two independent features that disrupt CTCF association with chromatin: inhibition of transcription and disruption of CTCF-RNA interactions through mutations of 2 of its 11 zinc fingers that are not required for CTCF binding to its cognate DNA site: zinc finger 1 (ZF1) or zinc finger 10 (ZF10). These mutations alter gene expression profiles as CTCF mutants lose their ability to form chromatin loops and thus the ability to insulate chromatin domains and to mediate CTCF long-range genomic interactions. Our results point to the importance of CTCF-mediated RNA interactions as a structural component of genome organization.

Differential DNA methylation and CTCF binding between the ESR1 promoter a of MCF-7 and MDA-MB-231 breast cancer cells.

BACKGROUND: ESR1 is expressed by 60-70% of breast tumours. it's a good prognosis factor and the target of hormone therapy. Optimization of ESR1 reactivation therapy is currently ongoing. Here we probe if the transcription factor CTCF plays a role in the differential expression of ESR1 in the breast cancer cell lines MCF-7 (ESR1+) and MDA-MB-231 (ESR1-). METHODS AND RESULTS: Knockdown of CTCF in MCF-7 resulted in decreased ESR1 gene expression. CTCF binds to the promoter of ESR1 in MCF-7 but not in MDA-MB-231 cells. CTCF ESR1 binding sites are unmethylated in MCF7 but methylated in MDA-MB-231 cells. CONCLUSION: ESR1 expression in MCF7 cells is dependent on CTCF expression. CTCF can bind to specific regions of the promotor of ESR1 gene in MCF-7 cells but not in MDA-MB-231 cells, this correlates with the methylation status of these regions and could be involved in the transcriptional regulation of ESR1.

A Driver in Health Outcomes: Developing Discrete Categories of Transportation Insecurity.

Research suggests that transportation is an important social determinant of health, because the ability to get around is consequential for accessing health care and nutritious food and for making social connections. We used an inductive mixed-methods approach and a quantitative k-means clustering approach to identify 5 categories of transportation insecurity using the validated 16-item Transportation Security Index. The resulting 5-category measure distinguished among respondents with qualitatively different experiences of transportation insecurity. Analyzing data from 2018 that were representative of the US adult population aged 25 years or older, we demonstrated a nonparametric association between transportation insecurity and 2 different health measures (self-rated health and depressive symptoms). There was a threshold relationship between self-rated health and any level of transportation insecurity. High transportation insecurity had a very strong relationship with depressive symptoms. The categorical Transportation Security Index will be useful for clinicians who wish to screen for transportation-related barriers to health care. It will also facilitate research investigating the influence of transportation insecurity on health outcomes and provide the basis for interventions designed to address health disparities.

Lonely Individuals Process the World in Idiosyncratic Ways.

Loneliness is detrimental to well-being and is often accompanied by self-reported feelings of not being understood by other people. What contributes to such feelings in lonely people? We used functional MRI of 66 first-year university students to unobtrusively measure the relative alignment of people's mental processing of naturalistic stimuli and tested whether lonely people actually process the world in idiosyncratic ways. We found evidence for such idiosyncrasy: Lonely individuals' neural responses were dissimilar to those of their peers, particularly in regions of the default-mode network in which similar responses have been associated with shared perspectives and subjective understanding. These relationships persisted when we controlled for demographic similarities, objective social isolation, and individuals' friendships with each other. Our findings raise the possibility that being surrounded by people who see the world differently from oneself, even if one is friends with them, may be a risk factor for loneliness.

Urea Reduction in Acute Kidney Injury and Mortality Risk.

INTRODUCTION: Urea is a toxin present in acute kidney injury (AKI). We hypothesize that reduction in serum urea levels might improve clinical outcomes. We examined the association between the reduction in urea and mortality. METHODS: Patients with AKI admitted to the Hospital Civil de Guadalajara were enrolled in this retrospective cohort study. We create 4 groups of urea reduction ratio (UXR) stratified by their decrease in urea from the highest index value in comparison to the value on day 10 (0%, 1-25%, 26-50%, and >50%), or at the time of death or discharge if prior to 10 days. Our primary endpoint was to observe the association between UXR and mortality. Secondary observations included determination of which types of patients achieved a UXR >50%, whether the modality of kidney replacement therapy (KRT) effected changes in UXR, and if serum creatinine (sCr) value changes were similarly associated with patient mortality. RESULTS: A total of 651 AKI patients were enrolled. The mean age was 54.1 years, and 58.6% were male. AKI 3 was present in 58.5%; the mean admission urea was 154 mg/dL. KRT was started in 32.4%, and 18.9% died. A trend toward decreased risk of death was observed in association with the magnitude of UXR. The best survival (94.3%) was observed in patients with a UXR >50%, and the highest mortality (72.1%) was observed in patients achieving a UXR of 0%. After adjusting for age, sex, diabetes mellitus, CKD, antibiotics, sepsis, hypovolemia, cardio-renal syndrome, shock, and AKI stage, the 10-day mortality was higher in groups that did not achieve a UXR of at least 25% (OR: 1.20). Patients achieving a UXR >50% were most likely initiated on dialysis due to a diagnosis of the uremic syndrome or had a diagnosis of obstructive nephropathy. Percentage change in sCr was also associated with increased mortality risk. CONCLUSIONS: In our retrospective cohort of AKI patients, the percent decrease in UXR from admission was associated with a stratified risk of death. Patients with a UXR >25% had the best associated outcomes. Overall, a greater magnitude in UXR was associated with improved patient survival.

Methemoglobinemia in Hemodialysis Patients due to Acute Chlorine Intoxication: A Case Series Calling Attention on an Old Problem.

INTRODUCTION: Hemodialysis uses municipal water that must be strictly purified and sterilized to be used for that procedure. Large amounts of decontaminants are often used, such as chlorine, and if these compounds are not subsequently removed they can be transferred to the blood of patients causing complications including methemoglobinemia. METHODS: In this case series study, dialysis patients in one unit were evaluated. We reviewed clinical characteristics and laboratory findings obtained on the day when the water supply was disinfected with chlorine, with the aim to quantify methemoglobin concentrations. Our objective was to characterize the clinical presentation and management of patients who presented with methemoglobinemia on a specific index day. We also reviewed reported cases in the literature regarding this underreported complication. RESULTS: Eight patients who presented with chlorine intoxication were evaluated. The methemoglobin concentrations were between 1.3% and 7.9% (reference value 0-1%). We believe this to be caused by water containing 0.78 mg/L of total chlorine. Seven patients presented with cyanosis, 4 with dizziness, 6 with dark brown blood, 4 with dyspnea, and 4 with headache and hemolytic anemia. Subjects were treated with supplemental oxygen, methylene blue, intravenous vitamin C, blood transfusions, and increased doses of erythropoietin. No patient died, and all continued with their usual hemodialysis sessions. CONCLUSION: Acute chlorine intoxication transferred by the water used during hemodialysis sessions can present with methemoglobinemia accompanied by cyanosis, oxygen desaturation, and hemolytic anemia. Chlorine levels should be carefully monitored in the water used for hemodialysis treatment.

Probiotics in septic acute kidney injury, a double blind, randomized control trial.

INTRODUCTION: During acute kidney injury (AKI) due to sepsis, the intestinal microbiota changes to dysbiosis, which affects the kidney function recovery (KFR) and amplifies the injury. Therefore, the administration of probiotics could improve dysbiosis and thereby increase the probability of KFR. METHODS: In this double-blind clinical trial, patients with AKI associated with sepsis were randomized (1:1) to receive probiotics or placebo for 7 consecutive days, with the objectives of evaluate the effect on KFR, mortality, kidney replacement therapy (KRT), urea, urine volume, serum electrolytes and adverse events at day 7. RESULTS: From February 2019 to March 2022, a total of 92 patients were randomized, 48 to the Probiotic and 44 to Placebo group. When comparing with placebo, those in the Probiotics did not observe a higher KFR (HR 0.93, 0.52-1.68, p = 0.81), nor was there a benefit in mortality at 6 months (95% CI 0.32-1.04, p = 0.06). With probiotics, urea values decreased significantly, an event not observed with placebo (from 154 to 80 mg/dl, p = 0.04 and from 130 to 109 mg/dl, p = 0.09, respectively). Urinary volume, need for KRT, electrolyte abnormalities, and adverse events were similar between groups. (ClinicalTrial.gov NCT03877081) (registered 03/15/2019). CONCLUSION: In AKI related to sepsis, probiotics for 7 consecutive days did not increase the probability of KFR, nor did other variables related to clinical improvement, although they were safe.

Quantitative magnetization transfer imaging for non-contrast enhanced detection of myocardial fibrosis.

PURPOSE: To develop a novel gadolinium-free model-based quantitative magnetization transfer (qMT) technique to assess macromolecular changes associated with myocardial fibrosis. METHODS: The proposed sequence consists of a two-dimensional breath-held dual shot interleaved acquisition of five MT-weighted (MTw) spoiled gradient echo images, with variable MT flip angles (FAs) and off-resonance frequencies. A two-pool exchange model and dictionary matching were used to quantify the pool size ratio (PSR) and bound pool T2 relaxation ( T2B ). The signal model was developed and validated using 25 MTw images on a bovine serum albumin (BSA) phantom and in vivo human thigh muscle. A protocol with five MTw images was optimized for single breath-hold cardiac qMT imaging. The proposed sequence was tested in 10 healthy subjects and 5 patients with myocardial fibrosis and compared to late gadolinium enhancement (LGE). RESULTS: PSR values in the BSA phantom were within the confidence interval of previously reported values (concentration 10% BSA = 5.9 ± 0.1%, 15% BSA = 9.4 ± 0.2%). PSR and T2B in thigh muscle were also in agreement with literature (PSR = 10.9 ± 0.3%, T2B = 6.4 ± 0.4 us). In 10 healthy subjects, global left ventricular PSR was 4.30 ± 0.65%. In patients, PSR was reduced in areas associated with LGE (remote: 4.68 ± 0.70% vs. fibrotic: 3.12 ± 0.78 %, n = 5, P < .002). CONCLUSION: In vivo model-based qMT mapping of the heart was performed for the first time, with promising results for non-contrast enhanced assessment of myocardial fibrosis.

Rectus abdominis muscle thickness as a predictor of peritoneal catheter dysfunction in emergency-start peritoneal dialysis patients.

BACKGROUND: Percutaneous insertion of a peritoneal dialysis catheter (PDc) is an alternative to open surgical techniques, and the anatomical characteristics of the abdominal wall may predict PDc dysfunction. We evaluated the role of rectus abdominis muscle (RAM) thickness as a predictor of PDc dysfunction. MATERIALS AND METHODS: A prospective cohort of emergency-start PD patients (EmPD) who had their first percutaneous PDc insertion were included. PDc failure was defined as the removal of a PDc due to mechanical failure within the first 30 PD fluid exchanges. Clinical variables were recorded. The skin to parietal peritoneum depth and RAM thickness were determined by abdominal ultrasound. Univariate and multivariate logistic regression models were developed to test associations between clinical parameters and PDc dysfunction. RESULTS: Over 6 months, 119 patients underwent PDc insertion; 73 (61.3%) were males, with a mean age of 46.0 ± 17.8 years. The mean skin-to-peritoneum depth was 2.5 ± 1.0 cm, the RAM thickness was 0.91 ± 0.3 cm, and catheter implantation was successful in 116 (97.4%) patients. Insertion failed in 3 (2.5%) cases, and 30 (25.8%) patients presented with catheter dysfunction. Univariate analysis indicated that RAM thickness ≥ 1.0 cm, skin-to-peritoneum depth > 2.88 cm, abdominal waist > 92.5 cm, and skin-to-RAM fascia distance > 2.3 cm were associated with PDc dysfunction; in multivariate logistic regression analysis, only greater RAM thickness remained a significant predictor (OR 1.6, 95% CI 1.38 - 1.88, p < 0.001). CONCLUSION: In EmPD patients, RAM thickness is associated with PDc dysfunction and could aid in identifying patients at risk for PDc dysfunction in emergency settings. Additional adequately powered studies are needed to confirm our findings.

Universality of the Nonphononic Vibrational Spectrum across Different Classes of Computer Glasses.

It has been recently established that the low-frequency spectrum of simple computer glass models is populated by soft, quasilocalized nonphononic vibrational modes whose frequencies ω follow a gapless, universal distribution D(ω)∼ω^{4}. While this universal nonphononic spectrum has been shown to be robust to varying the glass history and spatial dimension, it has so far only been observed in simple computer glasses featuring radially symmetric, pairwise interaction potentials. Consequently, the relevance of the universality of nonphononic spectra seen in simple computer glasses to realistic laboratory glasses remains unclear. Here, we demonstrate the emergence of the universal ω^{4} nonphononic spectrum in a broad variety of realistic computer glass models, ranging from tetrahedral network glasses with three-body interactions, through molecular glasses and glassy polymers, to bulk metallic glasses. Taken together with previous observations, our results indicate that the low-frequency nonphononic vibrational spectrum of any glassy solid quenched from a melt features the universal ω^{4} law, independently of the nature of its microscopic interactions.

An energy-landscape-based crossover temperature in glass-forming liquids.

The systematic identification of temperature scales in supercooled liquids that are key to understanding those liquids' underlying glass properties, and their formation-history dependence, is a challenging task. Here, we study the statistics of particles' squared displacements δr2 between equilibrium liquid configurations at temperature T and their underlying inherent states, using computer simulations of 11 different computer glass formers. We show that the relative fluctuations of δr2 are nonmonotonic in T, exhibiting a maximum whose location defines the crossover temperature TX. Therefore, TX marks the point of maximal heterogeneity during the process of tumbling down the energy landscape, starting from an equilibrium liquid state at temperature T down to its underlying inherent state. We extract TX for the 11 employed computer glasses, ranging from tetrahedral glasses to packs of soft elastic spheres, and demonstrate its usefulness in putting the elastic properties of different glasses on the same footing. Interestingly, we further show that TX marks the crossover between two distinct regimes of the mean ⟨δr2⟩: a high temperature regime in which ⟨δr2⟩ scales approximately as T0.5 and a deeply supercooled regime in which ⟨δr2⟩ scales approximately as T1.3. Further research directions are discussed.

Contrast-free high-resolution 3D magnetization transfer imaging for simultaneous myocardial scar and cardiac vein visualization.

OBJECTIVE: To develop a three-dimensional (3D) high-resolution free-breathing magnetization transfer ratio (MTR) sequence for contrast-free assessment of myocardial infarct and coronary vein anatomy. MATERIALS AND METHODS: Two datasets with and without off-resonance magnetization transfer preparation were sequentially acquired to compute MTR. 2D image navigators enabled beat-to-beat translational and bin-to-bin non-rigid motion correction. Two different imaging sequences were explored. MTR scar localization was compared against 3D late gadolinium enhancement (LGE) in a porcine model of myocardial infarction. MTR variability across the left ventricle and vessel sharpness in the coronary veins were evaluated in healthy human subjects. RESULTS: A decrease in MTR was observed in areas with LGE in all pigs (non-infarct: 25.1 ± 1.7% vs infarct: 16.8 ± 1.9%). The average infarct volume overlap on MTR and LGE was 62.5 ± 19.2%. In humans, mean MTR in myocardium was between 37 and 40%. Spatial variability was between 15 and 20% of the mean value. 3D whole heart MT-prepared datasets enabled coronary vein visualization with up to 8% improved vessel sharpness for non-rigid compared to translational motion correction. DISCUSSION: MTR and LGE showed agreement in infarct detection and localization in a swine model. Free-breathing 3D MTR maps are feasible in humans but high spatial variability was observed. Further clinical studies are warranted.

High-dimensionality undersampled patch-based reconstruction (HD-PROST) for accelerated multi-contrast MRI.

PURPOSE: To develop a new high-dimensionality undersampled patch-based reconstruction (HD-PROST) for highly accelerated 2D and 3D multi-contrast MRI. METHODS: HD-PROST jointly reconstructs multi-contrast MR images by exploiting the highly redundant information, on a local and non-local scale, and the strong correlation shared between the multiple contrast images. This is achieved by enforcing multi-dimensional low-rank in the undersampled images. 2D magnetic resonance fingerprinting (MRF) phantom and in vivo brain acquisitions were performed to evaluate the performance of HD-PROST for highly accelerated simultaneous T1 and T2 mapping. Additional in vivo experiments for reconstructing multiple undersampled 3D magnetization transfer (MT)-weighted images were conducted to illustrate the impact of HD-PROST for high-resolution multi-contrast 3D imaging. RESULTS: In the 2D MRF phantom study, HD-PROST provided accurate and precise estimation of the T1 and T2 values in comparison to gold standard spin echo acquisitions. HD-PROST achieved good quality maps for the in vivo 2D MRF experiments in comparison to conventional low-rank inversion reconstruction. T1 and T2 values of white matter and gray matter were in good agreement with those reported in the literature for MRF acquisitions with reduced number of time point images (500 time point images, ~2.5 s scan time). For in vivo MT-weighted 3D acquisitions (6 different contrasts), HD-PROST achieved similar image quality than the fully sampled reference image for an undersampling factor of 6.5-fold. CONCLUSION: HD-PROST enables multi-contrast 2D and 3D MR images in a short acquisition time without compromising image quality. Ultimately, this technique may increase the potential of conventional parameter mapping.

Non-contrast enhanced simultaneous 3D whole-heart bright-blood pulmonary veins visualization and black-blood quantification of atrial wall thickness.

PURPOSE: Pre-interventional assessment of atrial wall thickness (AWT) and of subject-specific variations in the anatomy of the pulmonary veins may affect the success rate of RF ablation procedures for the treatment of atrial fibrillation (AF). This study introduces a novel non-contrast enhanced 3D whole-heart sequence providing simultaneous information on the cardiac anatomy-including both the arterial and the venous system-(bright-blood volume) and AWT (black-blood volume). METHODS: The proposed MT-prepared bright-blood and black-blood phase sensitive inversion recovery (PSIR) BOOST framework acquires 2 differently weighted bright-blood volumes in an interleaved fashion. The 2 data sets are then combined in a PSIR-like reconstruction to obtain a complementary black-blood volume for atrial wall visualization. Image-based navigation and non-rigid respiratory motion correction are exploited for 100% scan efficiency and predictable acquisition time. The proposed approach was evaluated in 11 healthy subjects and 4 patients with AF scheduled for RF ablation. RESULTS: Improved depiction of the cardiac venous system was obtained in comparison to a T2 -prepared BOOST implementation, and quantified AWT was shown to be in good agreement with previously reported measurements obtained in healthy subjects (right atrium AWT: 2.54 ± 0.87 mm, left atrium AWT: 2.51 ± 0.61 mm). Feasibility for MT-prepared BOOST acquisitions in patients with AF was demonstrated. CONCLUSION: The proposed motion-corrected MT-prepared BOOST sequence provides simultaneous non-contrast pulmonary vein depiction as well as black-blood visualization of atrial walls. The proposed sequence has a large spectrum of potential clinical applications and further validation in patients is warranted.

Croton golden mosaic virus: a new bipartite begomovirus isolated from Croton hirtus in Colombia.

A new begomovirus infecting Croton hirtus (Euphorbiaceae) from Colombia has been characterized. The complete DNA-A and DNA-B components were determined to be 2613 and 2551 nt in length, respectively, showing the typical genome organization of bipartite New World begomoviruses. DNA-A showed the highest nucleotide sequence identity (81.2%) to sida yellow mottle virus (JN411687), a begomovirus isolated from Sida rhombifolia in Cuba. Based on the current ICTV species demarcation criterion for the genus Begomovirus, we report a new member of this genus infecting C. hirtus. We propose that it be named Croton golden mosaic virus (CroGMV), based on the symptoms observed in the weed. Phylogenetic analysis revealed that CroGMV segregates into a single clade and has some relationship with viruses from Central America and the Caribbean. CroGMV is the first Croton-infecting bipartite begomovirus reported in the world.

Molecular identification of a new begomovirus infecting yellow passion fruit (Passiflora edulis) in Colombia.

The complete genome sequence of a bipartite begomovirus (genus Begomovirus, family Geminiviridae) infecting yellow passion fruit (Passiflora edulis) in the state of Valle del Cauca (Colombia) has been determined. The complete DNA-A and DNA-B components were determined to be 2600 and 2572 nt in length, respectively. The DNA-A showed the highest nucleotide sequence identity (87.2 %) to bean dwarf mosaic virus (M88179), a begomovirus found in common bean crops in Colombia, and only 77.4 % identity to passion fruit severe leaf distortion virus (FJ972767), a begomovirus identified infecting passion fruit in Brazil. Based on its sequence identity to all other begomoviruses known to date and in accordance with the ICTV species demarcation criterion for the genus Begomovirus (≥91 % sequence identity for the complete DNA-A), the name passion fruit leaf distortion virus is proposed for this new begomovirus. To our knowledge, this is the first report of a bipartite begomovirus affecting passion fruit in Colombia and the second report of a geminivirus affecting this crop worldwide.

Experimental and Theoretical Thermochemistry of the Isomers 3- and 4-Nitrophthalimide.

This work presents a thermochemical study of two derivatives of phthalimide: the isomers 3-nitrophthalimide and 4-nitrophthalimide. The enthalpies of formation for these compounds in the solid phase were obtained by combustion calorimetry. Using ths thermogravimetry technique, the enthalpies of vaporization were obtained. The enthalpies of sublimation were calculated from enthalpies of fusion and vaporization. From experimental data and by ab initio methods, the enthalpies of formation in the gas phase were calculated. With these results, it was possible to determine their relative stability, and it was found that 4-nitrophthalimide is more stable than its isomer 3-nitrophthalimide. This tendency is similar to that of 3-nitrophthalic anhydride and 4-nitrophthalic anhydride, as reported in a previous work by our research group. The enthalpy of isomerization was also obtained, and a good correlation with that of phthalic anhydride derivatives was found. Finally, with the values obtained, the enthalpic difference resulting when the imide group is substituted by an anhydride group was determined.

Effect of a symbiotic gel (Lactobacillus acidophilus + Bifidobacterium lactis + inulin) on presence and severity of gastrointestinal symptoms in hemodialysis patients.

OBJECTIVE: The study aimed to assess the effect of a symbiotic gel on presence and severity of gastrointestinal symptoms (GIS) in hemodialysis patients. DESIGN: A double-blinded, placebo-controlled, randomized, clinical trial was designed. The study was conducted at 2 public hospitals in Guadalajara, Mexico. SUBJECTS AND INTERVENTION: Twenty-two patients were randomized to the intervention group (nutritional counseling + symbiotic gel) and 20 patients were randomized to the control group (nutritional counseling + placebo), during 2 months of follow-up. MAIN OUTCOME MEASURE: Presence and monthly episodes of GIS were assessed by direct interview and severity by using the self-administered GIS questionnaire. Additionally, biochemical parameters, inflammatory markers, and nutritional status (dietary intake, subjective global assessment, anthropometry, and body composition) were evaluated. RESULTS: After a 2-month treatment, intervention group had a significant reduction in prevalence and monthly episodes of vomit, heartburn, and stomachache, as well as a significant decrease in GIS severity compared with control group. Moreover, intervention group had a greater yet not significant decrease in the prevalence of malnutrition and a trend to reduce their C-reactive protein and tumor necrosis factor α levels compared with control group. No symbiotic-related adverse side effects were shown in these patients. Clinical studies with longer follow-up and sample size are needed to confirm these results. CONCLUSIONS: We concluded that administration of a symbiotic gel is a safe and simple way to improve common GIS in dialysis patients.

Ghrelin decreases motor deficits after traumatic brain injury.

BACKGROUND: Pharmacologic therapy for traumatic brain injury (TBI) has remained relatively unchanged for decades. Ghrelin, an endogenously produced peptide, has been shown to prevent apoptosis and blood-brain barrier dysfunction after TBI. We hypothesize that ghrelin treatment will prevent neuronal degeneration and improve motor coordination after TBI. MATERIALS AND METHODS: A weight drop model created severe TBI in three groups of BALB/c mice: Sham, TBI, and TBI + ghrelin (20 µg intraperitoneal ghrelin). Brain tissue was examined by hematoxylin and eosin and Fluoro-Jade B (FJB) staining to evaluate histologic signs of injury, cortical volume loss, and neuronal degeneration. Additionally, motor coordination was assessed. RESULTS: Ghrelin treatment prevented volume loss after TBI (19.4 ± 9.8 mm(3)versus 71.4 ± 31.4 mm(3); P < 0.05). Similarly, although TBI increased FJB-positive neuronal degeneration, ghrelin treatment decreased FJB staining in TBI resulting in immunohistologic patterns similar to sham. Compared with sham, TBI animals had a significant increase in foot faults at d 1, 3, and 7 (2.75 ± 0.42; 2.67 ± 0.94; 3.33 ± 0.69 versus 0.0 ± 0.0; 0.17 ± 0.19; 0.0 ± 0.0; P < 0.001). TBI + ghrelin animals had significantly decreased foot faults compared with TBI at d 1, 3, and 7 (0.42 ± 0.63; 0.5 ± 0.43; 1.33 ± 0.58; P versus TBI <0.001; P versus sham = NS). CONCLUSIONS: Ghrelin treatment prevented post-TBI cortical volume loss and neurodegeneration. Furthermore, ghrelin improved post-TBI motor deficits. The mechanisms of these effects are unclear; however, a combination of the anti-apoptotic and inflammatory modulatory effects of ghrelin may play a role. Further studies delineating the mechanism of these observed effects are warranted.

Quantitative detection of cassava common mosaic virus for health certification of cassava (Manihot esculenta Crantz) germplasm using qPCR analysis.

Cassava (Manihot esculenta Crantz) is a crop of global economic and food safety importance, used for human consumption and in various industrial applications. The genebank of the Genetic Resources Program of the Alliance of Bioversity International and CIAT currently holds the world's largest cassava collection, with 5965 in vitro accessions from 28 countries. Managing this extensive collection involves indexing quarantine pathogens as a phytosanitary certification requirement for safely distributing cassava germplasm. The study therefore aimed to optimize a quantitative diagnostic protocol to detect cassava common mosaic virus (CsCMV) using quantitative PCR (qPCR) as a better alternative to other molecular techniques. This was done through designing primers and a probe in the RdRP region of CsCMV, and optimizing the qPCR conditions of the diagnostic protocol using primer concentration assays, and reaction amplification conditions such as volume and reaction time. We also evaluated the qPCR protocol by comparing the results of 140 cassava accession evaluations using three diagnostic methodologies (DAS-ELISA, end-point PCR, and qPCR) for CsCMV. Our protocol established that qPCR technique analysis is ten-times more sensitive in detecting CsCMV compared to end-point PCR, showing a maximum detection level of 77.97 copies/µL of plasmid, with 76 min of reaction time. The comparison allowed us to verify the level of CsCMV detection through the techniques evaluated, concluding that qPCR was more sensitive and allowed the quantification of viral concentration. The optimized qPCR protocol will be used to accelerate diagnostic screening of cassava germplasm for the presence or absence of CsCMV to ensure safe movement and distribution of disease-free germplasm.