RKIP localizes to the nucleus through a bipartite nuclear localization signal and interaction with importin α to regulate mitotic progression.

Raf kinase inhibitor protein (RKIP) is a multifunctional modulator of intracellular signal transduction. Although most of its functions have been considered cytosolic, we show here that the localization of RKIP is primarily nuclear in both growing and quiescent Madin-Darby canine kidney epithelial cells and in Cal-51 and BT-20 human breast cancer cells. We have identified a putative bipartite nuclear localization signal (NLS) in RKIP that maps to the surface of the protein surrounding a known regulatory region. Like classical NLS sequences, the putative NLS of RKIP is rich in arginine and lysine residues. Deletion of and point mutations in the putative NLS lead to decreased nuclear localization. Point mutation of all the basic residues in the putative NLS of RKIP particularly strongly reduces nuclear localization. We found consistent results in reexpression experiments with wildtype or mutant RKIP in RKIP-silenced cells. A fusion construct of the putative NLS of RKIP alone to a heterologous reporter protein leads to nuclear localization of the fusion protein, demonstrating that this sequence alone is sufficient for import into the nucleus. We found that RKIP interacts with the nuclear transport factor importin α in BT-20 and MDA-MB-231 human breast cancer cells, suggesting importin-mediated active nuclear translocation. Evaluating the biological function of nuclear localization of RKIP, we found that the presence of the putative NLS is important for the role of RKIP in mitotic checkpoint regulation in MCF-7 human breast cancer cells. Taken together, these findings suggest that a bipartite NLS in RKIP interacts with importin α for active transport of RKIP into the nucleus and that this process may be involved in the regulation of mitotic progression.

Outcomes of Partial Calcanectomy in an Academic Limb Salvage Center: A Multicenter Review.

Chronic nonhealing heel ulcerations have been established as an independent risk factor for major amputation, with poor rates of limb salvage success. Partial calcanectomy is a controversial limb salvage procedure reserved for patients with these heel ulcerations. We conducted a retrospective cohort study reviewing 39 limbs that underwent a partial calcanectomy from 2012 to 2018 to evaluate the proportion of patients healed, time to healing, ulcer recurrence, and postoperative functional level compared to the preoperative state. In addition, age, gender, body mass index, smoking status, coronary artery disease, diabetes mellitus, renal insufficiency, dialysis, peripheral arterial disease, method of closure, and percent of calcaneus resected were evaluated. Mean follow-up for our cohort was 2.3 years. We had a 1 year mortality rate of 11%, and a major amputation rate of 18%. Our results demonstrated a 77% healing rate with a median time to healing of 162 days. We found that patients who were closed primarily had a faster time to healing compared to patients who underwent closure by secondary intention. Our data showed that ulcer recurrence developed in 57% of healed limbs. We found that 76% of our patients were ambulatory postoperatively. These results suggest that partial calcanectomy is a viable limb salvage procedure with a predictable level of ambulation and function in a high-risk patient population.

Altered Brain Structure and Functional Connectivity Associated with Pubertal Hormones in Girls with Precocious Puberty.

Pubertal hormones play an important role in brain and psychosocial development. However, the role of abnormal HPG axis states in altering brain function and structure remains unclear. The present study is aimed at determining whether there were significant differences in gray matter volume (GMV) and resting state (RS) functional connectivity (FC) patterns in girls with idiopathic central precocious puberty (CPP) and peripheral precocious puberty (PPP). We further explored the correlation between these differences and serum pubertal hormone levels. To assess this, we recruited 29 idiopathic CPP girls and 38 age-matched PPP girls. A gonadotropin-releasing hormone (GnRH) stimulation test was performed, and pubertal hormone levels (including luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), prolactin, and cortisol) were assessed. All subjects underwent multimodal magnetic resonance imaging of brain structure and function. Voxel-based morphometry (VBM) analysis was paired with seed-to-voxel whole-brain RS-FC analysis to calculate the GMV and RS-FC in idiopathic CPP and PPP girls. Correlation analyses were used to assess the effects of pubertal hormones on brain regions with structural and functional differences between the groups. We found that girls with CPP exhibited decreased GMV in the left insula and left fusiform gyrus, while connectivity between the left and right insula and the right middle frontal gyrus (MFG), as well as the left fusiform gyrus and right amygdala, was reduced in girls with CPP. Furthermore, the GMV of the left insula and peak FSH levels were negatively correlated while higher basal and peak E2 levels were associated with increased bilateral insula RS-FC. These findings suggest that premature activation of the HPG axis and pubertal hormone fluctuations alter brain structure and function involved in the cognitive and emotional process in early childhood. These findings provide vital insights into the early pathophysiology of idiopathic CPP.

Normal behavioral discrimination of envelope statistics in budgerigars with kainate-induced cochlear synaptopathy.

Cochlear synaptopathy is a common pathology in humans associated with aging and potentially sound overexposure. Synaptopathy is widely expected to cause "hidden hearing loss," including difficulty perceiving speech in noise, but support for this hypothesis is controversial. Here in budgerigars (Melopsittacus undulatus), we evaluated the impact of long-term cochlear synaptopathy on behavioral discrimination of Gaussian noise (GN) and low-noise noise (LNN) signals processed to have a flatter envelope. Stimuli had center frequencies of 1-3kHz, 100-Hz bandwidth, and were presented at sensation levels (SLs) from 10 to 30dB. We reasoned that narrowband, low-SL stimuli of this type should minimize spread of excitation across auditory-nerve fibers, and hence might reveal synaptopathy-related defects if they exist. Cochlear synaptopathy was induced without hair-cell injury using kainic acid (KA). Behavioral threshold tracking experiments characterized the minimum stimulus duration above which animals could reliably discriminate between LNN and GN. Budgerigar thresholds for LNN-GN discrimination ranged from 40 to 60ms at 30dB SL, were similar across frequencies, and increased for lower SLs. Notably, animals with long-term 39-77% estimated synaptopathy performed similarly to controls, requiring on average a ∼7.5% shorter stimulus duration (-0.7±1.0dB; mean difference ±SE) for LNN-GN discrimination. Decision-variable correlation analyses of detailed behavioral response patterns showed that individual animals relied on envelope cues to discriminate LNN and GN, with lesser roles of FM and energy cues; no difference was found between KA-exposed and control groups. These results suggest that long-term cochlear synaptopathy does not impair discrimination of low-level signals with different envelope statistics.

Assessing the efficacy of a graded pulmonary rehabilitation protocol in mechanically ventilated patients following brainstem hemorrhage.

BACKGROUND: The objective of this study is to assess the impact of an early-graded pulmonary rehabilitation training program on patients undergoing mechanical ventilation due to brainstem hemorrhage. METHODS: Eighty patients receiving mechanical ventilation due to brainstem hemorrhage at our hospital's neurosurgery department between August 2022 and October 2023 were enrolled as participants. A sampling table was generated based on the order of admission, and 80 random sequences were generated using SPSS software. These sequences were then sorted in ascending order, with the first half designated as the control group and the second half as the intervention group, each comprising 40 cases. The control group received standard nursing care for mechanical ventilation in brainstem hemorrhage cases, while the intervention group underwent early-graded pulmonary rehabilitation training in addition to standard care. This intervention was conducted in collaboration with a multidisciplinary respiratory critical care rehabilitation team. The study compared respiratory function indices, ventilator weaning success rates, ventilator-associated pneumonia incidence, mechanical ventilation duration, and patient discharge duration between the 2 groups. RESULTS: The comparison between patients in the observation group and the control group regarding peak expiratory flow and maximum inspiratory pressure on days 1, 3, 5, and 7 revealed statistically significant differences (P < .05). Additionally, there was a statistically significant interaction between the main effect of intervention and the main effect of time (P < .05). The success rate of ventilator withdrawal was notably higher in the observation group (62.5%) compared to the control group (32.5%), with a statistically significant difference (P < .05). Moreover, the incidence rate of ventilator-associated pneumonia was significantly lower in the observation group (2.5%) compared to the control group (17.5%) (P < .05). Furthermore, both the duration of mechanical ventilation and hospitalization were significantly shorter in the observation group compared to the control group (P < .05). CONCLUSION: Early-graded pulmonary rehabilitation training demonstrates effectiveness in enhancing respiratory function, augmenting the ventilator withdrawal success rate, and reducing both the duration of mechanical ventilation and hospitalization in mechanically ventilated patients with brainstem hemorrhage. These findings suggest the potential value of promoting the application of this intervention in clinical practice.

Age and duration of hypertension are associated with carotid artery tortuosity.

Objective: Tortuosity of the carotid artery is a common angiographic finding that may impact blood flow and neuronal function. However, information on its prevalence and risk factors remains limited. In this study, we determined to explore the factors affecting carotid artery tortuosity. Methods: The head and neck computed tomography angiography (CTA) imaging and cerebral angiography data performed at the Second Affiliated Hospital of Wenzhou Medical University between January 2019 and September 2021 were collected, and a total of 356 cases were enrolled in the study after screening. Carotid artery tortuosity refers to the angle between the two adjacent segments of the carotid artery, from the opening of the aortic arch on either side to the external orifice of the carotid canal, being less than 150°. A retrospective analysis was performed to compare the general information, laboratory indicators, personal history, and medical history between the two groups. The χ2 test, t-test, and Mann-Whitney U-test were performed to compare the parameters between the two groups. If there were significant differences between the groups, multivariate logistic regression was performed to analyze the factors affecting carotid artery tortuosity. Results: A total of 222 of the 356 cases were determined to have carotid artery tortuosity, accounting for 63.6%. There were statistically significant differences in age, body mass index (BMI), duration of diabetes and hypertension, levels of low-density lipoprotein cholesterol (LDL-C), diastolic blood pressure, history of ischemic and hemorrhagic stroke, and the usage of antihypertensive drugs between the two groups. Multivariate logistic regression analysis of the above factors showed that age (OR = 5.063, 95% CI 2.963-10.26, p < 0.001) and duration of hypertension (OR = 2.356, 95% CI 1.353-8.625, p = 0.021) were associated with a higher incidence of carotid artery tortuosity. Compared to patients who did not consume antihypertensive drugs, the incidence of carotid artery tortuosity was significantly less (OR = 0.094, 95% CI 0.002-0.713, p = 0.019) in those consuming antihypertensive drugs. Conclusion: Carotid artery tortuosity is a relatively common carotid artery disease. The incidence of carotid artery tortuosity may increase with age and the duration of hypertension. The consumption of antihypertensive drugs may reduce the incidence of carotid artery tortuosity.

Sound elicits stereotyped facial movements that provide a sensitive index of hearing abilities in mice.

Sound elicits rapid movements of muscles in the face, ears, and eyes that protect the body from injury and trigger brain-wide internal state changes. Here, we performed quantitative facial videography from mice resting atop a piezoelectric force plate and observed that broadband sounds elicited rapid and stereotyped facial twitches. Facial motion energy (FME) adjacent to the whisker array was 30 dB more sensitive than the acoustic startle reflex and offered greater inter-trial and inter-animal reliability than sound-evoked pupil dilations or movement of other facial and body regions. FME tracked the low-frequency envelope of broadband sounds, providing a means to study behavioral discrimination of complex auditory stimuli, such as speech phonemes in noise. Approximately 25% of layer 5-6 units in the auditory cortex (ACtx) exhibited firing rate changes during facial movements. However, FME facilitation during ACtx photoinhibition indicated that sound-evoked facial movements were mediated by a midbrain pathway and modulated by descending corticofugal input. FME and auditory brainstem response (ABR) thresholds were closely aligned after noise-induced sensorineural hearing loss, yet FME growth slopes were disproportionately steep at spared frequencies, reflecting a central plasticity that matched commensurate changes in ABR wave 4. Sound-evoked facial movements were also hypersensitive in Ptchd1 knockout mice, highlighting the use of FME for identifying sensory hyper-reactivity phenotypes after adult-onset hyperacusis and inherited deficiencies in autism risk genes. These findings present a sensitive and integrative measure of hearing while also highlighting that even low-intensity broadband sounds can elicit a complex mixture of auditory, motor, and reafferent somatosensory neural activity.

Associations between vertebral bone marrow fat and sagittal spine alignment as assessed by chemical shift-encoding-based water-fat MRI.

BACKGROUND: The relationship between sagittal spine alignment and vertebral bone marrow fat is unknown. We aimed to assess the relationship between vertebral bone marrow fat and sagittal spine alignment using chemical shift-encoding-based water-fat magnetic resonance imaging (MRI). METHODS: A total of 181 asymptomatic volunteers were recruited for whole spine X-ray and lumbar MRI. Spine typing was performed according to the Roussouly classification and measurement of vertebral fat fraction based on the chemical shift-encoding-based water-fat MRI. One-way analysis of variance (ANOVA) was used to analyze the differences in vertebral fat fraction between spine types. The post hoc least significant difference (LSD) test was utilized for subgroup comparison after ANOVA. RESULTS: Overall, the vertebral fat fraction increased from L1 to L5 and was the same for each spine type. The vertebral fat fraction was the highest in type 1 and lowest in type 4 at all levels. ANOVA revealed statistically significant differences in fat fraction among different spine types at L4 and L5 (P < .05). The post hoc LSD test showed that the fat fraction of L4 was significantly different (P < .05) between type 1 and type 4 as well as between type 2 and type 4. The fat fraction of L5 was significantly different between type 1 and type 3, between type 1 and type 4, and between type 2 and type 4 (P < .05). CONCLUSION: Our study found that vertebral bone marrow fat is associated with sagittal spine alignment, which may serve as a new additional explanation for the association of sagittal alignment with spinal degeneration.

Changes in Expression of Tumor Suppressor Gene RKIP Impact How Cancers Interact with Their Complex Environment.

Tumor microenvironment (TME) is the immediate environment where cancer cells reside in a tumor. It is composed of multiple cell types and extracellular matrix. Microenvironments can be restrictive or conducive to the progression of cancer cells. Initially, microenvironments are suppressive in nature. Stepwise accumulation of mutations in oncogenes and tumor suppressor genes enables cancer cells to acquire the ability to reshape the microenvironment to advance their growth and metastasis. Among the many genetic events, the loss-of-function mutations in tumor suppressor genes play a pivotal role. In this review, we will discuss the changes in TME and the ramifications on metastasis upon altered expression of tumor metastasis suppressor gene RKIP in breast cancer cells.

Human RAMP1 overexpressing mice are resistant to migraine therapies for motion sensitivity: a mouse model of vestibular migraine.

Both enhanced motion-induced nausea and increased static imbalance are \observed symptoms in migraine and especially vestibular migraine (VM). Motion-induced nausea and static imbalance were investigated in a mouse model, nestin/hRAMP1, expressing elevated levels of human RAMP1 in the CNS, which enhances CGRP signaling in the nervous system. Behavioral surrogates such as the motion-induced thermoregulation and postural sway center of pressure (CoP) assays were used to assess motion sensitivity. Tail vasodilation analysis revealed that this model exhibits an increased sensitivity to CGRP's effects at lower doses compared to control mice. In addition, the nestin/hRAMP1 mice exhibit a higher dynamic range in postural sway than their wildtype counterparts, along with increased sway observed in nestin/hRAMP1 male mice that was not present in male littermate controls. Results from migraine blocker experiments were challenging to interpret, but the data suggests that olcegepant is incapable of reversing CGRP-induced alterations in the nestin/hRAMP1 mice, while rizatriptan was ineffective in both the nestin/hRAMP1 and control mice. The results indicate that overexpression of hRAMP1 leads to heightened endogenous CGRP signaling. Results also suggest that both olcegepant and rizatriptan are ineffective in reducing CGRP-triggered nausea and sway in this hypersensitive CGRP mouse model. This study suggests that hypersensitivity to CGRP may be a mouse model for difficult to treat cases of vestibular migraine.

Trajectories of General Health Status and Depressive Symptoms Among Persons With Cognitive Impairment in the United States.

ObjectivesTo identify and examine heterogeneous trajectories of general health status (GHS) and depressive symptoms (DS) among persons with cognitive impairment (PCIs). Methods: We use group-based trajectory models to study 2361 PCIs for GHS and 1927 PCIs for DS from the National Health and Aging Trends Survey 2011-2018, and apply multinomial logistic regressions to predict identified latent trajectory group memberships using individual characteristics. Results: For both GHS and DS, there were six groups of PCIs with distinct trajectories over a 7-year period. More than 40% PCIs experienced sharp declines in GHS, and 35.5% experienced persistently poor GHS. There was greater heterogeneity in DS trajectories with 55% PCIs experiencing improvement, 16.4% experiencing persistently high DS, and 30.5% experiencing deterioration. Discussion: The GHS trajectories illustrate the heavy burden of poor and declining health among PCIs. Further research is needed to understand the factors underlying stable or improving DS despite declining GHS.

De novo brain arteriovenous malformation formation and development: A case report.

BACKGROUND: Brain arteriovenous malformation (AVM), an aberrant vascular development during the intrauterine period, is traditionally considered a congenital disease. Sporadic reports of cases of de novo AVM formation in children and adults have challenged the traditional view of its congenital origin. CASE SUMMARY: In this report, we have presented the case of a child with a de novo brain AVM. Magnetic resonance imaging and magnetic resonance angiography of the brain showed no AVM at the age of 5 years and 2 mo. Brain AVM was first detected in this child at the age of 7 years and 4 mo. The brain AVM was significantly advanced, and hemorrhage was seen for the first time at the age of 12 years and 8 mo. There was further progression in the AVM, and hemorrhage occurred again at the age of 13 years and 5 mo. Genetic analysis of this patient revealed a mutation in the NOTCH2 (p.Asp473Val) gene. CONCLUSION: In short, our case has once again confirmed the view that brain AVM is an acquired disease and is the result of the interaction of genes, environment, and molecules.

Combined foliar and soil selenium fertilizer improves selenium transport and the diversity of rhizosphere bacterial community in oats.

Agronomic selenium (Se) biofortification of grain crops is considered the best method for increasing human Se intake, which may help people alleviate Se-deficiency. To investigate the efficiency of agronomic Se biofortification of oat, four Se fertilizer application treatments were tested: topsoil (T), foliar (S), the combination of T and S (TS), and control without Se application (CK). Compared with CK, TS significantly increased the 1000-grain weight, grain yield, Se contents in all parts of oats, contents of soil available N, K, and organic matter by 18%, 8.70%, 19.7-60.2%, 6.00%, 8.02%, and 17.95%, respectively. Leaves, roots, and ears had the highest conversion rate of exogenous Se in S (644.63%), T (416.00%), and TS (273.20%), respectively. TS also increased the activities of soil urease, alkaline phosphatase, and sucrose and the diversity of soil bacterial communities. TS and T increased the relative abundance of bacteria involved in the decomposition of organic matter, such as Actinobacteria, Gemmatimonadetes, Chloroflexi, and Bacteroidetes positively correlated with soil nutrients and enzyme activities, and reduced Proteobacteria and Firmicutes negatively correlated with them, Granulicella, Bacillus, Raoultella, Lactococcus, Klebsiella, and Pseudomonas. Furthermore, TS significantly increased the relative abundance of Planctomycetes, Chlorobi, Nitrospinae, Nitrospirae, Aciditeromonas, Gemmatimonas, Geobacter, and Thiobacter. T significantly increased the abundance of Lysobacter, Holophaga, Candidatus-Koribacter, Povalibacter, and Pyrinomonas. S did not significantly change the bacterial communities. Thus, a combined foliar and soil Se fertilizer proved conducive for achieving higher yield, grain Se content, and improving Se transport, the diversity of rhizosphere bacterial community, and bacterial functions in oats.

Different spinal subtypes with varying characteristics of lumbar disc degeneration at specific level with age: a study based on an asymptomatic population.

BACKGROUND: The relationship between spinal sagittal subtypes and lumbar disc degeneration is unclear. Thus, we aimed to investigate the relationship between lumbar intervertebral disc degeneration and age in asymptomatic healthy individuals with different sagittal alignments. METHODS: In this cross-sectional observational study, we examined 209 asymptomatic young and middle-aged volunteers (123 women and 86 men) who were divided into the following three groups according to age: groups A (20-30 years), B (31-40 years), and C (41-50 years). The volunteers underwent full-spine standing lateral radiography and magnetic resonance imaging (MRI, 3.0 T) of the lumbar spine. Based on panoramic radiography, two observers measured the spinopelvic parameters and classified the spine into Roussouly subtypes. The degree of disc degeneration was assessed based on T2-weighted images according to the Pfirrmann classification. RESULTS: There was a statistically significant difference in the degree of degeneration of type I spine between groups B and C at L4-L5 (P < 0.03) and L5-S1 (P < 0.01) and between groups A and C at L1-L2 (P < 0.04) and L4-L5 (P < 0.01). The degeneration degree of type II spine at all levels were significantly different between groups A and C. No statistically significant difference was found between groups A and B in all subtypes except for type II spine at L1-L2 (P < 0.04). A significant difference was found at four levels between groups B and C in type III spine (P < 0.05) and between groups A and C. For type IV spine, there was a significant difference in the degree of degeneration at L4-L5 (P < 0.02) between groups A and C. Moreover, almost all single parameters were not strongly correlated with the degree of disc degeneration. CONCLUSION: The different spinal subtypes have characteristics of lumbar disc degeneration at specific levels with age. We considered that spinal classification could be used as a predictor of lumbar disc degeneration. Our data may be helpful to increase awareness of the relationship between spinal subtypes and lumbar disc degeneration. LEVEL OF EVIDENCE: 3.

Evaluation and validation of the diagnostic value of the apparent diffusion coefficient for differentiating early-stage endometrial carcinomas from benign mimickers at 3T MRI.

Previous researchers obtained various apparent diffusion coefficient (ADC) cutoff values to differentiate endometrial carcinoma from benign mimickers with 1.5T magnetic resonance imaging (MRI). Few studies have used 3T MRI or validated the effectiveness of these cutoff ADC values prospectively. This study was designed in two stages to obtain a cutoff ADC value at 3T MRI and to validate prospectively the role of the ADC value. First, we conducted a retrospective study of 60 patients to evaluate the diagnostic value of ADC by obtain a theoretical cutoff ADC value for differentiating between benign and malignant endometrial lesions. Student's t test revealed that ADC values for stage I endometrial carcinomas were significantly lower than those for benign lesions. The area under the curve value of the receiver operating characteristic curve was 0.993, and the cutoff ADC value was 0.98 × 10-3 mm2/s. The sensitivity, specificity, and overall accuracy of diagnosing stage I endometrial carcinoma were 100%, 97.1%, and 98.3%, respectively. Second, we conducted a prospective study of 26 patients to validate the use of the cutoff ADC value obtained in the study's first stage. The sensitivity, specificity, and overall accuracy for differentiating malignant from benign endometrial lesions based on the cutoff ADC value obtained earlier were as follows: radiologist 1 attained 86.67%, 100.0%, and 92.31%, respectively; radiologist 2 attained 86.67%, 91.0%, and 88.5%, respectively. Our results suggest that ADC values could be a potential biomarker for use as a quantitative and qualitative tool for differentiating between early-stage endometrial carcinomas and benign mimickers.