As part of our quest for digital biomarkers of neuromuscular aging, and encouraged by recent findings in healthy volunteers, this study investigated if the instantaneous median frequency (IMDF) derived from back muscle surface electromyographic (SEMG) data monitored during cyclic back extensions could reliably differentiate between younger and older individuals with cLBP. A total of 243 persons with cLBP participated in three experimental sessions: at baseline, one to two days after the first session, and then again approximately six weeks later. During each session, the study participants performed a series of three isometric maximal voluntary contractions (MVC) of back extensors using a dynamometer. These were followed by an isometric back extension at 80% MVC, and—after a break—25 slow cyclic back extensions at 50% MVC. SEMG data were recorded bilaterally at L5 (multifidus), L2 (longissimus dorsi), and L1 . Linear mixed-effects models found the IMDF-SEMG time-course changes more rapidly in younger than in older individuals, and more prominently in male participants. The absolute and relative reliabilities of the SEMG time–frequency representations were well compared between older and younger participants. The results indicated an overall good relative reliability, but variable absolute reliability levels. IMDF-SEMG estimates derived from cyclic back extensions proved to be successful in reliably detecting differences in back muscle function in younger vs. older persons with cLBP. These findings encourage further research, with a focus on assessing whether an IMDF-SEMG-based index could be utilized as a tool to achieve the preclinical detection of back muscle aging, and possibly predict the development of back muscle sarcopenia. The decline in back muscle strength and power  with age may translate into disablement and adverse health outcomes . Aging mWe recently suggested that an index derived from surface electromyographic (SEMG) data collected during a fatigue test could be used as a digital biomarker of the elevated risk of an early onset of neuromuscular aging processes ,19. ThisTest protocols relying on the collection of data during isometric 80% MVC back extensions sustained for approximately 30 s have traditionally been used to derive the MDF-SEMG time-course ,27. HoweThis research sought to investigate, for the first time, if the IMDF-SEMG derived from recordings collected during a cyclic back extensor exercise protocol in exercise-naive persons would enable the detection of differences in back muscle function in younger vs. older male as well as female individuals with cLBP. In addition, the retest reliability of this method was investigated.The Ethics Committee of the City of Vienna—Number: “EK 11-064-VK-NZ” approved the study protocol. All participants provided written informed consent, in accordance with the Declaration of Helsinki directives.Study volunteers were recruited via word of mouth and by presenting the study to cLBP patients undergoing outpatient rehabilitation. All prospective participants received information that they would have access to six months of training at the Karl-Landsteiner Institute for Outpatient Rehabilitation Research after completion of the study.A total of 294 persons affected by non-specific cLBP , between 18 and 90 years old, volunteered to participate in the study. They all completed a short screening questionnaire to assess the location, duration, and intensity of pain, as well as functional limitations and co-morbidities affecting study participants. Thereafter, eligible persons with cLBP completed a medical examination performed by a Physical and Rehabilitation Medicine specialist. Persons with cLBP were included in the study if they were otherwise healthy and suffered from LBP with a minimum of 30 mm on a visual–analogue scale (0–100 mm), as recorded during a 12 week period prior to screening. Patients were not eligible to participate in the study if the following conditions applied to them: (1) the receipt of health care advice for headache episodes and more than five episodes within the past year (one or more lasting more than two days), (2) neck pain equal to or exceeding 30 mm on a visual–analogue scale (0–100 mm), (3) headache within the last six weeks, (4) peripheral neurological deficit, (5) spinal fracture, (6) infection, (7) cancer, (8) previous surgery involving the back region, (9) previous experience with trunk muscle strength testing, (10) the performance of exercise more than two times per week or at a competitive level, (11) an inability to follow German verbal instructions, and (12) a BMI exceeding 35 kg/m². Participants were instructed not to take analgesic drugs, muscle relaxants, or psycho-pharmaceuticals within two days of testing. A total of 243 patients were included in the study.The schedule of assessments and tasks were similar to those described in a previous study conducted in healthy volunteers by our research group . ParticiThe complete set of measurements was repeated in a second experimental session one to two days after the first, and further repeated in a third session approximately six weeks later.® health solutions, Helsinki, Finland) was used to assess maximum isometric back extension torque. The dynamometer is described elsewhere in detail [®, Cesena, Italy) exercise device was used to assess static and dynamic muscle fatigue. The TechnoGym® device is built similarly to the F110 DAVID® extension device, but it is equipped with a dorsal sacral pad instead of a back pad, and it therefore allows for unrestricted SEMG recording from the lower back.The F110 back-extension device  were used to collect SEMG data. After standard preparation of the skin, double-sided adhesive interfaces were used to attach the sensors. Accounting for the muscle fiber direction and according to the SENIAM guidelines and previous work [® software  was used for data collection.Active sensors with parallel-bar electrodes and triaxial accelerometers  attached to the dynamometer arm was used to monitor the trunk movements. Biomechanical variables  were calculated using the accelerometer data. The triaxial accelerometer gathered data with a dynamic range of ±2.0 g, a maximum resolution of 0.016 g/bit, and a bandwidth of dc-50 Hz. The accelerometer data were sampled at 160 Hz with a resolution of 8 bits using the EMG Works® software.A triaxial accelerometer  in an unsupported upright seated position. The order was alternated between the right and left hand. A rest interval of 20 s followed each test. Tests were considered invalid and thus repeated if they differed more than 10%.Questionnaires: During each session, participants rated their perceived back muscle fatigue level on an 11 point Borg visual–analogue scale with anchors 0  and 10 (most severe fatigue ever experienced), and completed the International Physical Activity Questionnaire (IPAQ) , as wellIn order to familiarize the participants with the equipment and the test procedures, they performed a warm-up exercise at a low load. Then, they completed two consecutive isometric MVC tests under supervision by an experimenter who provided instructions and encouragement in a standardized manner. Further trials were carried out until consistent data were collected. If the second MVC test varied by more than 10%, or if the peak effort was achieved later than 3 s after the onset of the contraction, the MVC test was repeated. The best out of two consistent trial scores was stored.Participants performed a static back extension at 80% MVC for 30 s with their trunks 30° ante-flexed and in a seated position. After resting for 10 min and performing a few trials without external load in order to familiarize themselves with the cyclic back exercise, participants were asked to perform 25 cyclic contractions with the device arm exerting a load equal to 50% MVC. Therefore, they repetitively flexed and extended their trunks between the upright sitting position and a 40° forward flexed position following the pace of a metronome and the verbal instructions provided by the experimenter. The movement velocity of the flexion–extension exercise bout was paced with a metronome set at 2 s intervals (2 s extension and 2 s flexion). If the participant was unable to perform 25 repetitions or the task-specific range of motion (ROM), or if the movement velocity could not be sustained, the test was stopped. If more than 15 consecutive repetitions were successfully performed, the data were considered suitable for analysis. Otherwise, the data were discarded. Immediately after the dynamic test, volunteers rated their perceived back muscle fatigue level using the BORG rating scale.MATLAB scripts  were used to analyze the SEMG data. In order to avoid transitory behaviors, the first three seconds of the static tests and the first eight seconds (first two cycles) of the dynamic tests were discarded. The SEMG data were filtered using a 20 Hz high-pass and a 450 Hz low-pass Butterworth filter. From the second to the fifth second of the sustained 80% MVC contraction, the root-mean-square (RMS) values of the SEMG data were estimated for one-second intervals, as described in previous work .For the concentric phases of the cyclic exercise, SEMG data segments were identified based on the accelerometer data monitored using the probe located on the arm of the Total Trunk device . The RMSElectromyographic representations of localized muscle fatigue were calculated from each series of 25 bursts of activity, by computing the least squares regression line of the IMDF estimates derived from each of the 25 EMG bursts of activity. The IMDF values of each exercise bout were averaged to obtain one IMDF value per cycle and phase. The slope of the regression line  served as the measure of localized muscle fatigue. The RMS values were normalized by the RMS value obtained during 4 s  of the 80% MVC test. A linear regression was derived from the RMS values, as described above. The regression line slope allowed us to estimate the rate of change in the RMS value of the SEMG data.To account for inter-individual differences that would affect the spectral SEMG representations, such as subcutaneous tissue thickness, and following previous recommendations , both thThe trunk range of motion was monitored by utilizing the sagittal angular displacement data derived from a geometrical procedure, using gravity as reference, as described in previous work ,50. To cThe following dependent variables were used in the analysis: MVC (trunk extensor strength), mean IMDF-SEMG onset values calculated from the linear regressions, and mean IMDF-slopes normalized using the IMDF-SEMG initial values. The independent variables were subjects’ age (older or younger than 50 years of age), sex, and test day (3 days). The R software environment for statistical computing was used for all statistical analyses .The sample size for the mixed ANOVA  was estiFor the main analyses that sought to identify age- and gender-specific differences, artefact-free SEMG data from the full set of electrodes derived from biomechanically comparable (velocity and acceleration) cyclic exercise tasks were available for 85 older (48 women) and 92 younger (39 women) participants. These data were selected to control for any confounders that could occur from group imbalances in the biomechanics of task performance or due to missing EMG variables. Our primary analysis was performed with the fitted data of the missing EMG variables of individual electrodes. The fitting of data could be achieved if the SEMG recordings of one of the two (paired) electrodes per segment were artifact free. Then, the artifact-free SEMG data were duplicated, and both the onset and the slope values were corrected using the left–right imbalance ratios of the respective lumbar segment obtained from the 177 LBP individuals with full datasets. Hence, data from a total of 108 older (60 women) and 114 younger (52 women) individuals with LBP were selected for analysis.p-values were Bonferroni corrected and considered as significant if p ≤ 0.01. If significant effects for “age”, “sex”, or their respective interactions were detected, post hoc analyses were conducted using estimated marginal means (EMMs) [Descriptive statistics was used to summarize the participants’ characteristics. A mixed effects model with fixed effects for “age”, “sex”, and “test number”, and random effects for “persons” were computed for each outcome variable, in order to evaluate age-specific and sex-specific between-group effects. As in , p-values (EMMs) . ResiduaLinear mixed models were used to test whether the variables test day, age, sex, pain intensity, physical activity behavior scores (IPAQ), AEQ-derived movement behavior, and disability score  affected the normalized IMDF fatigue slopes of each of the different electrode-recording sites.The Generalizability Theory (G-Theory) ,56,57 waA total of 177 out of the 243 LBP study participants showed artefact-free SEMG data for all six electrode-recording sites. The EMG recordings of 21 participants were not considered in the analysis, either because the data displayed biomechanical inconsistences when performing the exercise, or because the participants were unable to complete the task. The data of an additional 45 participants had artefacts affecting the SEMG data because of intermittent electrode detachment during the performance of the cyclic exercise; filtering significantly attenuated these artifacts. Overall, the recordings from 222 individuals were considered for the analyses herein presented. The reasons for the exclusion of individual SEMG signal recordings are summarized in For the study participants whose data were included in the analyses, all the values of the demographic variables , the functional scores (“maximum back extension strength” and “hand grip strength”), and the fatigue ratings at the end of the exercise task did not differ from those whose data had to be excluded from further analysis (as described above).All the other variables  revealed age- and/or sex-specific between-group differences .The biomechanical performance quality variables “trunk range of motion”, “peak movement velocity”, and “mean movement velocity” for the concentric portion of the cyclic back extension exercise were comparable between the age and sex subgroups. It is worth noticing that these biomechanical variables were approximately constant throughout the dynamic exercise in each of the two age and sex subgroups .p < 0.001), and in younger males vs. females (p < 0.001).The IMDF-SEMG slope values  demonstrated a significant shift toward the lower frequencies of the SEMG data collected during the exercise test, in all electrode sites. These slopes were generally greater in younger than older participants, and in males than females. A significant age- and sex-specific interaction, accompanied by significant age- and sex-specific effects, was observed for the L2 electrodes. A Post hoc analyses of this electrode site revealed significantly steeper slopes in younger vs. older male participants (p ≤ 0.01). The comparison of the results for younger and older women showed significantly steeper slopes for the L5 recording site. The imbalance scores were comparable in younger and older participants, as well as in male and female CLBP participants. The factor repetition of test days revealed a significant effect on the data collected from L1 between test sessions one and two, and in the data collected from L2, and for all electrodes between test sessions two and three. The results showing age- and sex-specific differences in the IMDF-SEMG data are summarized in Significant age-specific as well as sex-specific effects were observed for all electrodes . The L5 electrode-recording site and the L2 electrode-recording site displayed the most negative slope values. Post hoc analyses revealed a larger decrease during the exercise in IMDF-SEMG values, in younger vs. older male participants, and in younger male vs. female ones for all electrodes (The absolute standard error of measurement (SEM) values derived from the IMDF-SEMG onset value estimates were found to be the highest for L5 and L2, and the lowest for the recording site associated with the most negative IMDF-SEMG slope. The absolute SEM values of the normalized IMDF-SEMG slopes were the largest for the recording site associated with the most negative IMDF-SEMG slope, and the smallest for L1. No major differences were observed between the two age subgroups. D-values for the IMDF-SEMG onset value estimates ranged between 0.72 and 0.84 for all the different recording sites, indicating good to excellent reliability. The corresponding D-values of the normalized IMDF-SEMG slope values ranged between 0.4 (L1) and 0.71. These D-values were comparable in younger and older cLBP individuals. Normalized IMDF-SEMG fatigue slope values were significantly smaller in older than younger individuals with cLBP, with differences that were more prominent in males than in females;Normalized IMDF-SEMG fatigue values were greater in younger male participants with cLBP vs. female participants with cLBP;Neither pain intensity nor health-related disablement had a major effect on the IMDF-SEMG slope;The relative reliability was overall moderate-to-good for the IMDF-SEMG slope values ; the absolute reliability indicated a small test–retest variability for both the IMDF-SEMG slope values and the IMDF-SEMG onset values.In our quest for a predictive, digital biomarker that could detect early signs of neuromuscular aging, we investigated whether parameters derived from the time–frequency representation of SEMG data could identify age-specific differences in individuals with cLBP who engaged in a moderately fatiguing cyclic back extension exercise. The main findings from the comparison of the data collected from a group of younger individuals and from older adults with cLBP were:A reliable interpretation of the SEMG variables depends on carefully controlling the level of generated force output , as well as controlling biomechanical performance variables ,58. MaxiThe biomechanical variables “range of motion” and “mean/peak movement velocity” were comparable between groups, and were approximately constant throughout the exercise. This is evidence that the overall biomechanical quality of the exercise was high, and that confounders potentially affecting the SEMG recordings due to the variability in biomechanical performance of the cyclic exercise unlikely affected the between-group comparisons of the SEMG-based parameters.At the end of the cyclic exercise, older participants reported significantly less back muscle fatigue than younger participants. This was observed even though the corresponding rating scores still indicated the fatigue to be moderate, and the work performed during the exercise was comparable between the two age groups. Such observations are consistent with previous research that found that muscle endurance during sustained and slow dynamic contractions is better preserved in older than younger healthy individuals ,60,61. IThe significant decrease in the IMDF-SEMG values for all the electrode sites was associated with RMS-SEMG onset values that exceeded those of the 80% MVC test by 20% to 30% for most of the electrode sites  or the deep mono- to oligo-segmental back muscles , which appeared to have almost no spindles . Given tThis study further sought to investigate the day-to-day retest reproducibility of SEMG fatigue-related variables. Depending on the recording site, indicators suggest a moderate-to-good relative reliability for the spectral SEMG variables. However, absolute reliability was found to be highly variable for both the normalized slopes and the SEMG onset values. This was observed for both age-specific subgroups. If one compares the results of the current study with those of earlier research in which healthy individuals performed cyclic free lifting exercises , or wherOne may argue that other factors than those related to the slowing in muscle fiber conduction velocity affected the age- and sex-specific differences in SEMG spectral compression observed during the cyclic exercise. In fact, changes in motor unit firing statistics with fatigue, and their correlation to each other, as well as changes in the spatial distribution of activated motor units, could affect the spectral-related fatigue changes over time. Regarding the firing statistics, the RMS-SEMG increase observed in this study was similar in older and younger participants, as well as in males and females. Thus, an increase in firing rate and/or the recruitment of additional motor units during the exercise would not explain the differences in the SEMG spectral compression. Considering that muscle fatigue may influence common drive, as demonstrated recently in intermittent, fatiguing 20% isometric knee extensions , an incrOther confounders of the SEMG spectral variables, such as non-travelling potentials and scattered innervation zones, most likely equally affect the data collected from the two groups ,76, and We attempted to record a full set of SEMG data during exercise testing from all 243 participants. We were successful in obtaining artefact-free recordings from all electrode sites in 177 individuals. One of the main reasons leading to SEMG artefacts was the intermittent loss of electrode-skin contact, which occurred most frequently for recording sites at L5 and L2. As such a loss of electrode recordings may have reduced the statistical power for detecting age-specific differences, we decided to analyze the fitted SEMG data. Findings from the analyses of the fitted data and those of the 177 participants with artefact-free EMG recordings were similar. Future research will have to address the electrode detachment problem when a cyclic exercise is to be performed.Monitoring the spectral compression of the SEMG data recorded from back muscles during a cyclic exercise at moderate efforts would allow one to detect the early signs of an accelerated neuromuscular aging process, particularly in males, and to a lower extent, in females with LBP. Notably, the dynamic test used in this study does not allow one to identify the specific underlying causes of the altered IMDF-SEMG fatigue-related behavior in older patients with cLBP. This could result from either the long-term-recruitment failure of innervated high threshold type II glycolytic muscle fibers , or fromAs in healthy, pain-free individuals, the observation of trends in IMDF-SEMG values during a dynamic test proved to be a successful approach for reliably distinguishing between back muscle function in younger vs. older men, and to a smaller extent, women with cLBP. These findings provide motivation for further developing this approach into a screening test that would be readily available in the clinic, and intended to early detect aging back muscles prior to more obvious age-related muscle function changes typically observed in the target population. The early detection of aging back muscles would be relevant to the design of therapeutic interventions that are intended to prevent an early aging process and disability in older individuals.


Treatment for a large symptomatic syrinx associated with a Chiari 1 is predominately surgical, via a foramen magnum decompression (FMD), with the aim to normalise cerebrospinal fluid (CSF) movement. Whilst theories of underlying hyperdynamic states in Chiari 1 and Syringomyelia exist, to date there is no effective medical treatment to reverse Syringomyelia.A 17-year-old female was referred with a seven-month history of gradually progressive impaired temperature sensation in her left upper limb. She had also been concomitantly diagnosed with thyrotoxicosis. Magnetic resonance imaging (MRI) confirmed a Chiari 1 with a large syrinx. The patient preferred to avoid surgery in the first instance. She underwent treatment for her thyrotoxicosis. The eight-month, 20- and 36-month follow-up MRI scans demonstrated a gradual resolution of the Chiari 1 malformation and the syrinx.Whilst there have been reports of Chiari 1 malformation association with hyperthyroidism, this is the first report describing syrinx resolution following treatment of thyrotoxicosis. Hyperdynamic circulation can result in syrinx formation through various mechanisms. We hypothesise that the treatment of thyrotoxicosis resulted in normalisation of CSF pulse amplitude and subsequent syrinx resolution. Hyperthyroidism evaluation may be explored in studies of CM1 and Syrinx or other CSF disorders. Natural history of SyringomyeliaSyringomyelia is a cystic cavitation of the spinal cord that is commonly associated with Chiari I malformation CM1) [ [1]. TheManagement of SyringomyeliaIn light of the unpredictable natural history, the management questions that Klekamp et al. raised in 2001 remain relevant today, “Should every patient with Chiari I malformation (and a Syrinx) undergo surgery? Should surgery be postponed until symptoms start to develop?” . An onliSpontaneously resolving syrinxesFurther complicating decision-making is the ever-increasing literature of large syrinxes spontaneously resolving . Herein,HypothesisWe hypothesise that this patient had a hyperdynamic CSF pulsatility and that the treatment of this patient’s thyrotoxicosis resulted in normalisation of CSF dynamics, an ultimately syrinx resolution.Clinical presentationWe present a case of a 17-year-old female with a large syrinx and CM1. She was referred with a seven-month history of gradually progressive impaired temperature sensation in her left upper limb. She had also been concomitantly diagnosed with thyrotoxicosis. She had a family history, with her father also having a CM1 treated with FMD and ventriculoperitoneal (VP) shunt.Examination findingsClinical findings relieved normal power throughout, reduced sensation in left C4-T2 dermatomes in the pinprick and temperature modalities; and absent left biceps and supinator reflexes and reduced left triceps reflex.ImagingMagnetic resonance imaging (MRI) Figure  confirmeManagementAt the first clinic appointment, all treatment options were discussed, including surgery, given the size of the syrinx and her symptomatology. However, a conservative surveillance approach was preferred by the patient. Concomitantly, treatment for her thyrotoxicosis was commenced by her endocrinology team. This treatment initially started with 40mg Carbimazole twice daily, the dose of which was titrated down in line with her blood results. Her presenting blood test results were T3 4.6 nmol/L , T4 225 nmol/L  and thyroid-stimulating hormone (TSH) 0.1 mU/L , consistent with her disease. Her bloods improved to near normal following treatment at 20 months showed T3 1.2 nmol/L, T4 110nml/L and TSH 5.6mU/L. OutcomeThe patients eight-month follow-up MRI Figure  demonstrWe present a case of a large spontaneously resolving syrinx, where resolution coincided with the treatment of thyrotoxicosis. Whilst spontaneous resolution of syringomyelia without surgery is a well-reported phenomenon in the literature, particularly since the widespread use of MRI, its pathophysiology remains unknown. This discussion describes various potential mechanisms through which hyperthyroidism may result in syrinx formation.Aetiology of syrinx formation and resolutionAetiologies for syrinx formation include pathologies with increased intracranial pressure (ICP) , CM1, trauma, multiple sclerosis, spinal tumours and idiopathic aetiology .The first report of an MRI-verified spontaneous resolution of syringomyelia was in 1990 by Sudo et al. ,3,8.Different hypotheses have been proposed for the spontaneous resolution of syringomyelia. In children, the restoration of normal CSF dynamics and thus correction of the syrinx has been attributed to enlargement of the posterior fossa space with age . In adulCardiac cycle and syrinx formationHyperdynamic CSF flow at the craniocervical junction may be a causative factor in the development of syrinx in CM1. Pulsatile CSF movement during a cardiac cycle can contribute to syrinx progression . FurtherOldfield et al. explored the relationship between cardiac systole and syringomyelia with dynamic MRI . The dowWe postulate that this patient had a large CSF pulse amplitude secondary to the hyperdynamic state of thyrotoxicosis, thus developing a syrinx as per Oldfield’s theory. The treatment of the hyperthyroid state thus resulted in the normalisation of CSF dynamics, an ultimate syrinx resolution.Could hyperthyroidism precipitate a syrinx?There are few reports in the literature of CSF dynamic disturbance due to hyperthyroidism. One case reported by Herwig et al. describes a 32-year-old woman suffering from Grave’s and new hydrocephalus that then resolved after propranolol and carbimazole . SimilarHyperthyroidism has been linked to an increase in cerebral blood flow and therefore cerebral blood volume (CBV) and ICP, as well as to increased CSF pulsations ,20. MoreIn this case study, the increased metabolic turnover could have resulted in (i) the increased production of CSF by the choroid plexus, (ii) increased CBV and ICP, (iii) possible abnormal PA due to hyperdynamic state and ultimately and a CM1 and Syrinx formation. The key element in our case report, however, is the fact the syrinx disappeared with medical treatment of the thyrotoxicosis.Strengths and limitationsThis is a case report, and we acknowledge that association is not equivocal to causality. Nonetheless, we consider this case to be valuable, given the correlation between syrinx resolution and concomitant timely treatment. Furthermore, there is evidence in the literature to support the theory that a hyperdynamic state may have been driving abnormal CSF dynamics and this syrinx formation.Hyperdynamic circulation and raised pulse amplitude is a known factors in the development of syrinxes with CM1. We hypothesise that the treatment of this patient’s thyrotoxicosis resulted in the normalisation of CSF flow dynamics. Similar cases of spontaneous syrinx resolution may reveal alternative treatment options and elucidate the pathophysiology of this complex condition.


We report a case of bilateral neonatal testicular torsion, with an extravaginal and a contralateral intravaginal testicular torsion.A 5‐day‐old boy with bilateral scrotal swelling and palpable induration was diagnosed with bilateral neonatal testicular torsion by color Doppler ultrasonography. The right testis was black with 360‐degree extravaginal torsion of the spermatic cord, and the left testis was brown with 90‐degree intravaginal torsion. We repaired the torsion and incised the tunica albuginea to reduce intratesticular pressure. The left testis became pink in color, but the right testis remained unchanged. Based on the pathological findings of the intraoperative biopsy of tissue specimens from both testes, we performed a right orchiectomy and preserved the left testis.Our experience suggests that testicular color improvement after fasciotomy and pathological findings of intraoperative testicular biopsy may indicate testicular preservation. Rare case of bilateral neonatal testicular torsion, with an extravaginal and a contralateral intravaginal testicular torsion.Testicular torsion in neonates differs from that in non‐neonates in terms of pathogenesis and clinical manifestations and has a very low testicular salvage rate.The patient was born naturally at 39–5/7 weeks' gestation to a mother who received regular prenatal care. The pregnancy was uncomplicated, and the mother had no past medical history. The patient's Apgar scores were 9 at 1 min and 10 at 5 min after birth. The initial newborn physical examination showed no abnormalities. The genitalia were noted on the medical record as being normal. However, on Day 2 after birth, bilateral scrotal swelling and palpable induration of the testis were observed, and he was referred to our hospital on Day 5. The entire scrotum was swollen, the right testis was enlarged and palpable with induration, and the left testis was mildly enlarged and partially palpable with induration. Laboratory examinations were unremarkable. Color Doppler ultrasonography showed fluid components and septal structures around the right testis with no blood flow inside the testis Fig. . The lefNeonatal testicular torsion is a very rare disease.In the present case, the types of testicular torsion suggested that the right testis had a failure of fixation to the scrotal subcutaneous tissue, and the left testis had an abnormal attachment to the tunica vaginalis.The rate of testicular sparing surgery for neonatal testicular torsion is very low, at 3–9%, compared to those for such torsion in infants or older children.A coincidental occurrence of bilateral testicular torsion with different pathogenesis is very rare. In the present case, the spread of hemorrhagic necrosis suggested that the right testicular torsion occurred prenatally and the left testicular torsion occurred during parturition. In the case of a bilateral testicular torsion, it is difficult to decide whether to remove or preserve the testes. Our experience suggests that testicular color improvement after fasciotomy and intraoperative pathological findings of the remaining seminiferous tubules could be indicators of testicular preservation.Yusuke Kirihana: Conceptualization; writing – original draft. Yuichi Sato: Writing – review and editing. Junya Hata: Writing – review and editing. Hitomi Imai: Writing – review and editing. Yuki Yoshida: Writing – review and editing. Kanako Matsuoka: Writing – review and editing. Seiji Hoshi: Writing – review and editing. Tomoyuki Koguchi: Writing – review and editing. Soichiro Ogawa: Writing – review and editing. Yoshiyuki Kojima: Writing – review and editing.The authors declare no conflict of interest.Not applicable.Not applicable.Not applicable.


Firmicutes/Bacteroidetes (F/B) ratio and controlling the number of certain gut bacteria. TT also alleviated HFD‐induced abnormalities of the gut microbiota. The Muribaculaceae, Lachnospiraceae NK4A136_group, Alistipes, and Odoribacter families were identified as the major beneficial gut microorganisms using Spearman's correlation analysis. Fecal microbiota transplantation (FMT) demonstrated that TT's anti‐obesity and gut microbiota‐modulating benefits might be transmitted to mice on an HFD, demonstrating that one of TT's targets for preventing obesity is the gut microbiota. TT also increased the amount of short‐chain fatty acids (SCFAs) in the feces, including acetic, propionic, and butyric acids. These results indicate the possible development of TT as a prebiotic to combat obesity and associated disorders. These results suggest that TT may act as a prebiotic against obesity and its associated diseases.It has been shown that Tibetan tea (TT) inhibits obesity and controls lipid metabolism. The fundamental processes by which TT prevents obesity are yet entirely unknown. Consequently, this research aimed to ascertain if TT may prevent obesity by modifying the gut flora. Our research demonstrated that TT prevented mice from gaining weight and accumulating fat due to the high‐fat diet (HFD), decreased levels of blood total cholesterol (TC), triglycerides (TG), and low‐density lipoprotein cholesterol (LDL‐C), and raised levels of high‐density lipoprotein cholesterol (HDL‐C). Adipogenesis‐related genes such as acetyl‐Coenzyme A carboxylase 1 , fatty acid synthase , sterol regulatory element‐binding protein‐1c , CCAAT/enhancer‐binding protein α , stearoyl‐CoA desaturase 1 , and peroxisome proliferator‐activated receptor γ  had their expression downregulated by lowering the  We were able to demonstrate that Tibetan tea (TT) dramatically inhibited high‐fat diet‐induced weight gain, fat accumulation, hyperglycemia, and hyperlipidemia in mice, as well as control the expression of genes involved in lipid metabolism. The way that TT controls the gut microbiota produces short‐chain fatty acids and controls the expression of genes involved in lipid synthesis may be responsible for its positive effects on obesity. Our research gave TT's potential as a functional beverage for the prevention and treatment of obesity a new direction. Weight Bacteroidota, Firmicutes, Proteobacteria, and Actinobacteria   is a type of dark tea, mainly produced in Ya'an, Sichuan Province, China, and has a history of about 1000 years , catechin, epigallocatechin gallate (EGCG), (−)‐epigallocatechin (EGC), and epicatechin (EC) were purchased from Yuanye Biotechnology Co., Ltd. Methanol, anhydrous ethanol, acetonitrile, and phenol reagents  were purchased from Sigma‐Aldrich. Other reagents such as acetic acid, propionic acid, butyric acid, and isobutyric acid are of analytical grade .2.2TT was extracted as described by Gong et al.  with minThe quantification of tea polyphenols, total flavonoids, and soluble sugar in TT water extracts was carried out by the Folin–Ciocalteu colorimetric method  NCD_TT_H, which received standard chow and was treated with 400 mg/kg of TT extract; (3) HFD, which received high‐fat chow and was treated with 0.9% saline; (4) HFD_TT_L, which received high‐fat feed and was treated with 100 mg/kg of TT extract; (5) HFD_TT_M group, which received high‐fat feed and was treated with 200 mg/kg of TT extract; (6) HFD_TT_H group, which received high‐fat feed and was treated with 400 mg/kg of TT extract. The weight of the mice was noted weekly throughout the 9‐week trial. After a 12‐h fast, mice were slaughtered at the end of the ninth week, and blood and fat samples were taken. The other tissue was snap‐frozen in liquid nitrogen, and the epididymal fat was preserved in 4% formaldehyde for further study.At eight weeks old and 20 ± 2 g each, 60 male C57BL/6J mice were purchased from SiPeiFu Biotechnology Co., Ltd. in Beijing, China. They were given access to food, water, and a 12‐h light/dark cycle while being housed in a controlled environment with a temperature of 22 ± 2°C and a humidity level of 55% ± 5%. The Sichuan Industrial Institute of Antibiotics' Ethical Committee at Chengdu University gave its approval for the animal trials . After a week of acclimatization, the mice were randomly divided into six groups, each with ten mice: (1) 2.4NCD, NCD_TT_H, HFD, and HFD_TT_H groups were fed for 2 months and then used as donor mice. The feces of each of the four groups of mice were collected in sterile cages; 200 mg of feces were resuspended in 2 mL of sterile saline, vortexed for 10 s, and then centrifuged at 800 g for 3 min. Eight‐week‐old male recipient mice were fed HFD and gavaged with fresh graft samples (100 μL per mouse) daily for two months.Fecal microbiota transplantation (FMT) was performed according to the method described by Chang et al. . Briefly2.5According to the protocol of the kit , the serum concentrations of high‐density lipoprotein cholesterol (HDL‐C), low‐density lipoprotein cholesterol (LDL‐C), total cholesterol (TC), and triglyceride (TG) were measured.2.6The mice fasted for 12 h at week 8. 1g of glucose per kilogram of body weight was gavaged, and blood glucose levels were checked at 0, 15, 30, 60, 90, and 120 min later.2.7Through the use of the Trizol reagent procedure, total RNA was extracted from the tissue. Using Beijing Labgic Technology Co., Ltd.'s Reverse Transcription kit (which includes a dsDNase), cDNA was produced from an equal volume of total RNA. A StepOnePlusTM Real‐Time PCR Detection System from Applied Biosystems was used to evaluate the generated cDNA.CtΔΔ technique.The Table 2.8A 4% formaldehyde solution was used to fix freshly removed mouse epididymal fat overnight. Later, it underwent dehydration, embedding, sectioning, and morphological analysis using hematoxylin and eosin staining.2.9g at 4°C for 10 min and performing a static extraction at 4°C for 30 min, the supernatant was recovered. 0.04 M HCl  and the supernatant were combined before standing overnight. Before analysis, the supernatant was put into a clean micro gas‐phase vial after being filtered via a 0.22 μm membrane. Gas chromatography and mass spectrometry (GC–MS) equipment from Perkin Elmer was used to analyze the samples (Perkin Elmer Technologies).Short‐chain fatty acids were detected in feces using the technique previously described . Thermo Fisher Scientific, microspectrophotometer was used to quantify the DNA content and integrity, and agarose gel electrophoresis was used to corroborate the results further. Using two universal primer pairs , the V3‐V4 hypervariable portions of the bacterial 16S rRNA gene were amplified. Over a 25‐L response. AMPure XP beads (Beckman Coulter Co.) were used to purify the PCR products, and a Qubit dsDNA assay kit was used to quantify them. The concentrations were then modified for sequencing. OE Biotechnology Ltd. carried out sequencing and analysis of the 16S rRNA gene amplicon in Shanghai, China.2.11p < .05 was used to represent the data as mean ± SD (standard deviation). The studies utilized GraphPad Prism 9.0 (San Diego).The difference was shown to be significant using one‐way ANOVA and the Tukey test. A statistically significant value of 33.1The chemical composition and content of TT water extracts are shown in Table 3.2p < .05).The HFD mice were noticeably heavier than the other five groups after 9 weeks, as shown in Figure 3.3HFD group, the mice's blood glucose levels were lower after TT intervention. The impact of TT on the serum biochemical parameters in mice is shown in Figure p < .05).Blood lipids and glucose levels are likely to change as obesity progresses. We performed a glucose tolerance test and determined the AUC values in the trial's eighth week Figure . Accordi3.4HFD group had the lowest chao1 values, observed‐species values, and goods coverage values compared to the other five groups, according to the alpha diversity analysis  Figure , the mics Figure , and theBacteroidota, Firmicutes, Deferribacterota, Desulfobacterota, and Proteobacteria were among the prominent species identified in histograms showing the relative abundance of gut microbiomes at the phylum level . After TT intervention, animals had substantially increased amounts of butyric acid, propionic acid, and acetic acid in their feces compared to the HFD group. Similar to this, TT significantly altered branched‐chain fatty acids (BCFAs), mostly by increasing the quantity of isobutyric acid; however, isovaleric acid did not change.The impact of TT on the level of SCFAs in mouse feces is shown in Table 3.6NCD group, the NCD_TT_H group's expression of ACC1, C/EBPα, Fas, PPARγ, SCD1, and SREBP‐1c was considerably lower (p < .05). The expression of six genes was also significantly lower in the three TT intervention groups than in the HFD group (p < .05). Interestingly, in comparison to the HFD group, there was a dose‐dependent reduction in the expression of ACC1, C/EBPα, Fas, and SREBP‐1c  into HFD‐fed animals. Then, we examined obesity‐related characteristics and metabolic alterations in glucose and lipids levels. The findings demonstrated that fecal transplantation from the NCD group, NCD_TT_H group, and HFD_TT_H group decreased the body weight  were used in the investigation of the diversity. According to PCoA, there were significant differences between the intestinal microbiotas of the four mouse groups Figure . For PC1d Figure , and thed Figure . The alps Figure . After tHFD_HFD group changed the makeup of the gut microbiota at the phylum level by raising the F/B ratio, which fell following the transplantation of mouse feces from TT‐fed animals ; data curation (supporting); formal analysis (lead); funding acquisition (lead); investigation (supporting); methodology (lead); project administration (lead); resources (lead); supervision (lead); validation ; visualization ; writing – original draft ; writing – review and editing (lead). Tangcong Chen: Conceptualization (lead); data curation (lead); formal analysis (lead); investigation (lead); methodology (lead); supervision (lead); validation (lead); visualization (lead); writing – original draft (lead); writing – review and editing (lead). Lifen Huang: Conceptualization ; data curation ; formal analysis ; investigation ; supervision ; validation ; visualization ; writing – original draft (supporting); writing – review and editing (supporting). Yiyuan Zhang: Conceptualization (supporting); data curation ; formal analysis (supporting); investigation (supporting); methodology (supporting); validation (supporting); visualization (supporting). Yanjiao Feng: Data curation (supporting); formal analysis (supporting); investigation (supporting); methodology (supporting); validation (supporting); visualization (supporting); writing – original draft (supporting). Qijun Liu: Formal analysis (supporting); investigation (lead); methodology (supporting); visualization (supporting); writing – original draft (supporting). Xiaojing Yin: Data curation (supporting); formal analysis (supporting); investigation (supporting); validation (supporting); visualization (supporting); writing – original draft (supporting). Shaokui Qu: Conceptualization (supporting); data curation (supporting); formal analysis (supporting); investigation (supporting); validation (supporting); visualization (supporting); writing – original draft (supporting). Chen Yang: Conceptualization (supporting); data curation (supporting); formal analysis (supporting); investigation (supporting); methodology (supporting); resources (supporting). Jianghong Wan: Formal analysis ; investigation ; methodology . Li liang: Conceptualization (supporting); funding acquisition ; project administration (supporting); supervision (supporting). Jun Yan: Conceptualization ; formal analysis ; funding acquisition (lead); methodology ; project administration (lead); resources ; supervision . Wei Liu: Conceptualization ; formal analysis ; funding acquisition (lead); project administration (lead); resources ; supervision .The authors declare no conflicts of interest.The Sichuan Industrial Institute of Antibiotics at Chengdu University's Protection of Laboratory Animals committee examined and approved the animal research .Figure S1.Click here for additional data file.Table S1.Click here for additional data file.


Epithelioid haemangioendothelioma is a rare type of cancer with an unpredictable disease course and very few treatment options. To better understand how this type of cancer develops and to uncover possible lines of treatment, it is critical to have experimental approaches to study this cancer. Here, we describe and compare the model systems that are currently available to study this disease. The research undertaken and the discoveries made using each of these experimental models is presented, and the advantages and disadvantages of each model are discussed.Epithelioid haemangioendothelioma (EHE) is a rare sarcoma of the vascular endothelium with an unpredictable disease course. EHE tumours can remain indolent for long period of time but may suddenly evolve into an aggressive disease with widespread metastases and a poor prognosis. Two mutually exclusive chromosomal translocations define EHE tumours, each involving one of the transcription co-factors TAZ and YAP. The TAZ-CAMTA1 fusion protein results from a t translocation and is present in 90% of EHE tumours. The remaining 10% of EHE cases harbour a t translocation, resulting in the YAP1-TFE3 (YT) fusion protein. Until recently, the lack of representative EHE models made it challenging to study the mechanisms by which these fusion proteins promote tumorigenesis. Here, we describe and compare the recently developed experimental approaches that are currently available for studying this cancer. After summarising the key findings obtained with each experimental approach, we discuss the advantages and limitations of these different model systems. Our survey of the current literature shows how each experimental approach can be utilised in different ways to improve our understanding of EHE initiation and progression. Ultimately, this should lead to better treatment options for patients. WWTR1 gene) and yes-associated protein 1 (YAP1) are the transcriptional co-activators downstream of the Hippo pathway which facilitate the onset of these characteristics [Dysregulation of the Hippo pathway is observed in a vast array of cancers, and is attributed to the development of multiple cancer cell characteristics ,2. Theseeristics ,4,5. Thieristics ,6,7,8. Teristics . The reseristics ,10,11. Teristics ,13,14,15Until recently, the lack of representative EHE models made it challenging to study the mechanisms by which TC and YT promote tumorigenesis. This is a significant issue with rare diseases, such as EHE, due to the scarcity of patient samples. Here, we discuss currently available EHE model systems and how they have furthered our understanding of EHE biology. We also speculate on how these models could be used and further developed to uncover novel aspects of EHE biology.EHE is a rare sarcoma which arises from vascular endothelial cells ,16. EHE A recurrent tp36;q25) chromosomal translocation was first identified in EHE patient samples in 2001 . This wa chromosoCDKN2A, which encodes key positive regulators of cell growth arrest and senescence [The known TAZ/YAP fusion proteins present in EHE are mutually exclusive of each other. In addition to the disease-defining TAZ/YAP fusion proteins, analysis of EHE patient samples have revealed that at least 50% harbour a secondary mutation ,30. The nescence . Other cnescence ,30. ThesDrosophila  and Warts  resulted in the overgrowth of imaginal discs and the related adult organs [Given that all EHE tumours are defined by expression of a TAZ/YAP fusion protein, it is evident that they are central to EHE oncogenesis ,14,15,29t organs ,42. The t organs ,43,44. Tt organs ,46. LATSt organs ,48.(CTGF) and cysteine rich angiogenic inducer 61 (CYR61) [TAZ/YAP must interact with other transcription factors as they do not contain a DNA binding domain themselves ,5. This  (CYR61) . TAZ/YAP (CYR61) ,50. Here (CYR61) . Example (CYR61) ,51,52,53 (CYR61) ,57,58,59In recent years, a range of model systems for investigating EHE have been reported, and together have greatly improved our understanding of the disease ,61,62,63Seminal studies on the regulation and function of TC have made use of immortalised cell lines ,38,60 F. Cell liThe first functional study into TC introduced its ectopic expression into human embryonic kidney (HEK293) and murine fibroblast (NIH3T3) cell lines, prior to analysis of cell proliferation and transcriptional landscape . This waCTGF, a known transcriptional target of wild-type TAZ/YAP [Another key finding of this study determined that TC initiates a predominantly ‘TAZ-like’ transcriptional programme, through its interaction with TEAD4 . Further TAZ/YAP . CTGF pr TAZ/YAP ,38. This TAZ/YAP . This is TAZ/YAP ,61,62,63Further investigation revealed that TC and YT occupy both TEAD and non-TEAD transcription factor binding motifs, including early growth response gene 2 (EGR2) motifs for TC, and microphthalmia-associated transcription factor motif (MiTF) motifs for YT . MoreoveAnother key advantage of cell line-based models is the ability to introduce xenografts into mouse models. This permits in vivo tumorigenesis and metastasis studies, while being less time and resource intensive than developing a GEMM. Merritt et al. took advantage of this, creating mouse xenograft models with the same TC or YT expressing NIH3T3 or SW872 cells used in in vitro experiments . These mOne drawback of the cell line-based models discussed above is the lack of contextual relevance to EHE, as they are not of endothelial origin. This is of particular importance, given the cell-context-specific functions of wild-type TAZ/YAP ,69,70. AThis EHE model utilises a doxycycline-inducible system for inducing TC expression. This prevents TC from interfering with the differentiation process; however also permits the study of the immediate consequences of TC expression, which is not possible with other EHE models. Investigations using this model provided evidence that TC expression causes hypertranscription, leading to DNA damage in endothelial cells . As homoCdkn2a deletion, the most common secondary mutation in EHE tumours, permitted senescence bypass and increased cell growth [Another benefit of this model was that secondary mutations could be introduced into the endothelial cells relatively easily, using CRISPR/Cas9 methods . We uncol growth ,30,63. Tl growth ,30. MoreCdh5 promoter, resulting in a mouse model that expressed TC specifically in the endothelium. Furthermore, TC expression could be repressed upon doxycycline administration [WWTR1 promoter, as is the case in EHE [In addition to the in vitro models described above, genetically engineered mouse models (GEMMs) of EHE have been developed . The avastration . These mstration ,13,39. Me in EHE ,39.Cdh5 or the Rosa26 promoter to allow endothelial specific or ubiquitous expression, respectively [A second GEMM overcame this limitation by using a flip-excision (FLEx) system and Cre recombinase . The FLEectively . This moectively ,38,39,61Cdkn2a knockdown to EHE tumourigenesis [Cdkn2a gene were crossed with those harbouring the TC transgene. This resulted in mice with endothelial-specific loss of functional p16INK and p19ARF protein expression, with TC expression induced by tamoxifen administration [Cdkn2a caused reduced latency to tumour development and increased Ki67+ fraction, while maintaining the EHE specific transcriptome [One GEMM was further developed to determine the contribution of igenesis ,62. Herestration ,62. Thiscriptome .Cdkn2a-null mice [Only one GEMM investigating YT-driven tumourigenesis has been described to date . This waull mice . Transcrull mice ,11,38. Aull mice ,11,24.Cdkn2a deletion, and one which was heterozygous [Cdkn2a was not possible.The development of an EHE cell line is important, as it would encompass the benefits of the cell line-based models while being in the correct cellular context. To date, attempts to derive an EHE cell line by expanding cells from tumour samples have proven unsuccessful. One group has been able to generate three EHE cell lines of murine origin, by ex vivo expansion of tumours generated by the GEMM described above ,62 Figu. This alThese novel EHE cell lines were validated to confirm their relevance to the human disease and findings from previously described model systems . RNA-seqRecent advances mean that there are now a range of model systems available for investigating EHE . These uCDKN2A to EHE development, as it is conceivable that these would affect tumour growth to varying degrees. One way to achieve this is to use the CRISPR/Cas9-mediated approach adopted to study CDKN2A deletion in the stem cell-based model [Most current studies have focussed on characterising TC function and regulation in EHE development. While this is understandable, as TC is the most common driver fusion protein, future investigation should expand this to include YT-driven EHE, and the rarer TAZ fusion proteins present in cardiac EHEs. The currently available models could utilise the same methods to express the alternative fusion proteins, which would allow comparative studies. This would build on previous studies comparing the transcriptome and chromatin landscape of TC and YT expressing cells, and YT to other YAP fusion proteins ,10,11,24ed model . This woOne previously unexplored area of EHE biology is the contribution of the tumour microenvironment, which would be possible using one of the EHE GEMMs. This would likely uncover novel functions of TC, particularly as wild-type TAZ/YAP has been implicated in crosstalk with the microenvironment ,82,83,84


Cerebral ventricular shunt infections caused by methicillin-resistant Staphylococcus aureus (MRSA), especially strains with elevated minimum inhibitory concentration (MIC) values, have a poor prognosis. Monitoring serum vancomycin (VCM) levels with therapeutic drug monitoring and maintaining high VCM concentrations in the cerebrospinal fluid (CSF) are critical to treatment success. However, there have been a few reports about the CSF penetration and the pharmacokinetics of VCM in children.Here, we report the case of a pediatric patient with cysto-peritoneal shunt-related meningitis caused by MRSA with an MIC of 2 μg/mL. The adequate VCM concentration was maintained by monitoring the VCM concentration in the CSF via the external ventricular drain, and frequent blood taking was avoided. VCM showed a good CSF penetration in our patient, and she was discharged without complications.Therapeutic drug monitoring of VCM concentration in the CSF may result in successful treatment even if MRSA shows a higher MIC. Therapeutic drug monitoring of VCM concentration in the CSF may also reduce the side effects. Post-neurosurgical central nervous system (CNS) infections can occur in 1%–8.6% of cases and can be treated, but antimicrobial-resistant pathogens are a growing concern and limit treatment options . TherapeVancomycin (VCM) has a time-dependent activity with limited penetration in the cerebrospinal fluid (CSF) because of its hydrophilicity and large molecular size . TherefoStaphylococcus aureus (MRSA). However, these recommendations are primarily based on data from adult populations. They also propose a second-line drug if the strain’s minimum inhibitory concentration (MIC) value is 2 μg/mL .–15.12–15Several studies have reported that intraventricular use of VCM may improve treatment outcomes without severe side effects in adult patients –23. HoweIn our patient, although the strain isolated from the CSF showed MIC of VCM of 2 μg/mL, successful treatment with intravenous VCM was achieved by monitoring the concentration in both the serum and CSF. The penetration rate was sufficiently high and the CSF trough levels were above the MIC during treatment.High serum VCM concentration can cause complications such as nephrotoxicity, ototoxicity, and vasculitis . We coulIn conclusion, monitoring the VCM concentration in the CSF and its serum concentration as indicators may help make decisions about the optimal dosage, changing second-line drugs, and reducing the frequency of side effects.


Drosophila abnormal spindle (asp) gene was discovered about 40 years ago and shown to be required for both mitotic and meiotic cell division. Subsequent studies showed that asp is highly conserved and that mutations in its human ortholog ASPM  are the most common cause of autosomal recessive primary microcephaly. This finding greatly stimulated research on ASPM and its fly and mouse (Aspm) orthologs. The three Asp orthologous proteins bind the microtubules (MTs) minus ends during cell division and also function in interphase nuclei. Investigations on different cell types showed that Asp/Aspm/ASPM depletion disrupts one or more of the following mitotic processes: aster formation, spindle pole focusing, centrosome-spindle coupling, spindle orientation, metaphase-to-anaphase progression, chromosome segregation, and cytokinesis. In addition, ASPM physically interacts with components of the DNA repair and replication machineries and is required for the maintenance of chromosomal DNA stability. We propose the working hypothesis that the asp/Aspm/ASPM genes play the same conserved functions in Drosophila, mouse, and human cells. Human microcephaly is a genetically heterogeneous disorder caused by mutations in 30 different genes that play a variety of functions required for cell division and chromosomal DNA integrity. Our hypothesis postulates that ASPM recapitulates the functions of most human microcephaly genes and provides a justification for why ASPM is the most frequently mutated gene in autosomal recessive primary microcephaly.The  Most of these genes are involved in different aspects of mitotic division, including centriole biogenesis, centrosome-driven MT nucleation, kinetochore assembly and function, and spindle formation. However, a fraction of them are required for chromatin condensation and remodeling, DNA repair, and chromosome stability [ASPM are responsible for more than 40% of MCPH cases [ meiosis . Moleculle poles . The inty (MCPH) ,4,5. MCPy (MCPH) ,7,8). ToASPM gene is its role in tumor development. ASPM expression is upregulated in several cancers, including prostate cancer, glioblastoma, and hepatocellular carcinoma, and increased ASPM expression is associated with tumor progression and poor clinical prognosis. It has also been shown that siRNA-mediated ASPM depletion strongly inhibits tumor cell proliferation .Drosophila, mouse, and human cell types. These studies reveal that Asp, Aspm, and ASPM tend to have cell type-specific requirements. However, despite this fact and some apparently conflicting results, we propose a model that suggests a common mitotic function for the three orthologous proteins. We believe that this model explains most of the phenotypes associated with asp and ASPM mutations, providing a basis for future studies on the conserved asp/Aspm/ASPM gene.Although the Asp protein and its human (ASPM) and mouse (Aspm) orthologs have been studied for many years, a precise functional comparison between the three proteins has never been made. Here, we review the studies on the mitotic roles of these proteins in different https://www.ncbi.nlm.nih.gov; accessed on 20 December 2022) contains more than 450 orthologs of these proteins. Here we will focus on Drosophila Asp, mouse Aspm, and human ASPM, which are functionally best characterized. When we refer to one of the three orthologs, we will use its specific gene/protein name. To refer to the three orthologs together, we will indicate them as Asp/ASPM proteins. The three proteins have different lengths, although they contain the same structural domains. The difference in size  is mainly due to the number of isoleucine-glutamine (IQ) motifs that varies from 24 in Drosophila to 60 and 81 in mice and humans, respectively    domain with a putative MT-binding function . It has ASPM transgene in Aspm mutant mice can rescue all mutant phenotypes, including the reduction in brain size and the massive loss of germ cells in both males and females [asp and Aspm can rescue the mutant phenotypes elicited by ASPM mutation has never been investigated.Although the structural features of the Asp/ASPM proteins suggest that they might be interchangeable, we have very little data addressing this issue. We know that the expression of human  females . HoweverDrosophila embryos [asterless , cnn or dd4 (encoding a γ-tubulin ring component) [asp mutant metaphases are of similar intensity to those of controls [Early studies using antibodies against an Asp N terminal fragment of 512 amino acids showed that Asp localizes to the polar regions of the spindles and to the telophase central spindle in syncytial  embryos ,24, epit embryos , S2 tiss embryos ,21,26,27 embryos ,29. Specmponent) , (ii) thIn addition to the spindle poles, Asp accumulates at the central spindle. The central spindle, or intercellular bridge, is a prominent MT bundle that forms during late telophase. It consists of antiparallel MTs with the plus ends interdigitating at the center of the bundle, where they associate with many different proteins generating a discrete structure currently called the midbody. These proteins form a dense cluster (midbody ring) that impedes the access of anti-tubulin antibodies, resulting in a dark zone after tubulin immunostaining . Asp assDrosophila pupal notum [Besides its accumulation at the spindle poles and central spindle extremities, antibody staining revealed a weak Asp signal along the spindle MTs of different cell types, including larval neuroblasts, spermatocytes, and S2 cells ,26,28. Ial notum .Drosophila cells. Aspm localizes to the spindle poles throughout mitosis, accumulating in the immediate vicinity of the γ-tubulin signal of centrosomes. Aspm staining does not overlap the γ-tubulin immunoreactivity, and Aspm is absent from centrosomes during interphase [Drosophila ortholog, mouse Aspm appears to localize to spindle regions enriched in MT minus ends, consistent with the observation that Aspm preferentially associates with the MT minus ends in vitro [Aspm localization in neuroepithelial (NE) mouse cells is very similar to the Asp localization in terphase . In additerphase . Thus, lin vitro .Early studies on U2OS tissue culture cells suggested that human ASPM co-localizes with the centrosomes at the spindle poles and is enriched at the centrosomes in interphase nuclei . SimilarDrosophila and mouse counterparts, but there are some conflicting results about its precise localization to this structure. Using a commercial anti-human ASPM antibody and an antibody raised against amino acids 1-418 of rat ASPM, Paramasivam et al. [ASPM localizes to the central spindle like its m et al.  showed tm et al.  showed tm et al. . ASPM exon. In HeLa cells, GFP-ASPM accumulated to the spindle poles throughout mitosis; it also accumulated at central spindle extremities but was absent from the midbody. Importantly, Jiang et al. [More recently, Jiang et al.  analyzedg et al.  also shoCollectively the extant results indicate that the orthologous Asp/ASPM proteins exhibit very similar, if not identical, localization patterns. They do not appear to be integral centrosome components but accumulate to the spindle poles, where they are likely to bind and crosslink the minus ends of the MTs that detach from the centrosomes  with frequent metaphase-like figures and almost no anaphases. In addition, they showed frequent hyperploid and polyploid cells [asp mutant brains exhibit a MI three- to six-fold higher than wild-type brains, in asp double mutants with either rod or mad2, which encode SAC components, the MI drops to the wild-type level [asp mutant cells. It has been proposed that SAC signaling might be caused by centrosome misplacement that would influence the dynamic interaction between MTs and kinetochores . Althouypes see  below.Drosophila somatic cells. However, an analysis of cell division in live NE cells  revealed that they neither exhibit a metaphase delay nor a cytokinesis defect but show errors in chromosome segregation [Drosophila cell types.Besides brain cells, an increased MI and a metaphase delay phenotype following Asp depletion have been observed in S2 cells ,48 but nregation . Thus, iAspm mutants lacking either exons 8–28 or 26–28 of the gene (wild-type Aspm contains 28 exons), the NE cells of the embryonic dorsal telencephalon showed relative frequencies of prometaphases/metaphases and anaphases/telophases comparable to those of wild-type cells, indicating that loss of the Aspm function does not result in a detectable defect in mitotic progression [ASPM was removed using the CRISPR/Cas9 technology did not reveal any abnormality in mitotic progression [Defects in mitotic progression have not been detected in most Aspm- and ASPM-depleted cells. In mouse gression . Howevergression . There agression . Furthergression . asp in the control of chromosome segregation was first detected by genetic analysis of sex chromosome segregation in asp mutant males [asp mutations do not disrupt chromosome pairing at meiosis I but alter chromosome migration towards the poles [asp mutant males revealed that they have different sizes, indicative of different chromosomal contents [The role of nt males . These mhe poles . Consistcontents ,24,28,42asp is also required for mitotic chromosome segregation. The hyperploid cells observed in larval brains of asp mutants [asp mutations generated by heterozygous mothers showed a substantial increase in cuticular clones derived by the loss of a marked Y chromosome compared to their heterozygous sibs [asp mutant mothers  [asp mutant larvae showed that 23% of the anaphases exhibit errors in segregation [Several results indicate that  mutants  suggest ous sibs . Abnormal stage) . Moreoveregation .To the best of our knowledge, no clear defects in chromosome segregation have been described in ASPM- or Aspm-deficient cells. Drosophila embryos from asp mutant mothers, the centrosomes are frequently dissociated from the spindle poles [asp mutants, spindle poles are often unfocused, and centrosomes are frequently detached from the poles [asp mutants recruit normal amounts of γ-tubulin and Cnn, and asp asterless (asl) double mutants that lack functional centrosomes assemble bipolar spindles with unfocused poles [asp mutants further showed that the centrosomes detached from the poles can move randomly within the dividing cell so that, in some cases, one of the daughter cells inherits both centrosomes [asp mutant brains exhibit hemi-spindle-like structures associated with a single centrosomal signal [Drosophila asp mutants [Drosophila asp mutants, which also showed asters smaller than in the wild type [Consistent with their localization during cell division, all Asp orthologues control the organization of the spindle poles. In le poles . Similarhe poles . Moreoveed poles . An analtrosomes . This obl signal . Furtherl signal ,26,48,53 mutants , as well mutants . Finallyild type . ASPM gene [ASPM gene to RNAi against CDK5RAP2/CEP215, a cnn ortholog responsible for human microcephaly [Although Aspm accumulates at the spindle poles in different types of mouse cells, no defects have been reported in spindle pole focusing or aster formation upon Aspm loss ,32,49. ISPM gene . Howeverocephaly . More thocephaly . Interesasp/ASPM deficient cells, the MTs would be normally nucleated by the centrosome but would not be held in its vicinity after their release from this organelle, leading to reduced aster size and centrosome detachment from the spindle poles  undergo a series of symmetric divisions , thereby expanding the NPC pool. After increasing their number, NPCs divide asymmetrically , generating another NPC and a cell that will differentiate into a neuron. Defects in spindle orientation and cell cycle/mitotic delays can result in premature differentiation leading to a reduction in the final neuron number and in brain size [Aspm knockdown, the spindle axes of NE cells were parallel to the surface of the neuroepithelium less frequently than in the wild type, indicating a precocious switch from proliferative to neurogenic divisions, accompanied by a reduction of the expansion of the NE progenitor pool [ASPM-dependent human microcephaly [In both ain size . Many stain size ,58,59. Ctor pool . This suocephaly . Howeverocephaly . To explocephaly . Consistocephaly . Aspm deocephaly .Drosophila model. Human tissue culture cells normally divide with the spindle axis parallel to the substratum . However, in U2OS osteosarcoma cells subjected to ASPM RNAi, there was a significant increase in spindles with axes perpendicular to the substrate compared to controls [Drosophila adults and pharate adults (adults still encased in the puparium) homozygous for asp mutations showed a striking reduction of the head size, while the abdomen and thorax were of normal size [asp mutant larvae, there was a premature switch of spindle positioning towards an oblique/perpendicular position, with 50% of the cells undergoing an asymmetrical division [asp mutations on myosin-II distribution resulting in defective tissue architecture [The roles of Asp/ASPM proteins in the control of spindle orientation have also been evaluated in cultured human cells and in the controls . Similarcontrols ,54. Drosmal size . To addrdivision . Noteworitecture .Drosophila asp mutant larvae, the cells of the central brain arrest in metaphase due to the SAC activity prevented the study of the role of Asp in telophase and cytokinesis. However, in male meiotic cells, the SAC is weak and causes only a slight delay of anaphase onset [asp mutant males, the central spindle is often disorganized, leading to mislocalization of midbody proteins, defective formation of the actin-based contractile ring and failures in cytokinesis [The fact that in se onset . Consistokinesis ,28. Drosophila pupal notum [asp mutations affect the cytokinesis of NE cells of the Drosophila optic lobe [Aspm mutant mice. No cytokinesis defects were described in a study on ASPM-depleted U2OS human cells [ASPM knockout human HCT116 cells [Drosophila and mammalian systems. The mechanisms underlying Asp-dependent cytokinesis failure have been investigated only in Drosophila meiotic cells. In asp mutant males, a substantial fraction of telophases displayed central spindles with irregularly arranged MTs and abnormally shaped contractile rings [Asp-dependent cytokinesis failures were not reported in several studies on S2 tissue culture cells ,21,26 anal notum . In additic lobe . Cytokintic lobe ,32 nor itic lobe  of Aspm an cells , and the16 cells . However16 cells . Thus, tle rings ,28. Thesle rings ,62 . Mud complex . In epit complex . Cherry-s of Mud . In epits of Mud . Howevers of Mud .mud mutants [The requirement of Asp for Mud localization is not conserved, as ASPM is not needed for NuMA localization in human cells . In addi mutants . Moreove mutants ,68,69. T mutants ,24,26,70 mutants , Asp doe mutants . Asp acc mutants ,53. It h mutants , a proce mutants . This su mutants .ASPM physically associates with the MCPH protein WDR62 (MCPH2) . In HeLaDrosophila ortholog of CITK is also required for proper spindle orientation in dividing larval neuroblasts [ASPM physically interacts with Citron kinase (CITK) in both coimmunoprecipitation experiments and in situ proximity ligation assay (PLA) ,54. The KATNB1) cause defects in spindle architecture and result in microcephaly [Drosophila Kat80 affects neuroblast division in the optic lobe, resulting in reduced brain size [An additional ASPM interacting partner is katanin, an MT-severing complex consisting of the p60 ATPase and the p80 regulatory subunit . Mutatioocephaly ; similarain size . ASPM anain size . In diviain size . ASPM anain size . These aain size .Drosophila Asp, in addition to its prominent accumulation at the spindle poles, also localizes along the spindle MTs. A relatively weak Asp signal in the nonpolar region of prometaphase and metaphase spindles has been observed in different Drosophila cell types, including larval neuroblasts, epithelial cells, and S2 cells [As mentioned above, S2 cells ,25,26,28S2 cells ,21,25. IS2 cells , this fiS2 cells ,25. ASPMS2 cells ,35,50.Drosophila studies, the localization of Asp in interphase cells was not addressed. The current results suggest that Asp is enriched in interphase nuclei of some cell types, such as S2 cells, epithelial cells, and possibly male meiotic cells. In contrast, Asp does not seem to localize to the embryo and larval neuroblast nuclei [In most t nuclei ,25,27. St nuclei ,35,76. It nuclei .Consistent with their nuclear localization, both human and mouse ASPM/Aspm have been implicated in DNA repair and replication. Early studies reported that RNAi-mediated depletion of ASPM results in a significant reduction of BRCA1, a protein that binds DNA and participates in the repair of double-strand breaks (DSBs) ,77. ConvASPM has also been implicated in DNA replication. Affinity purification experiments showed that FLAG-GFP-ASPM interacts with several proteins involved in DNA replication, including the five subunits of the replication factor complex (RFC1-5), the BRCA2 factor involved in both DSB repair and protection of the replication forks, the MCM5 core subunit of the MCM complex that promotes double-strand DNA unwinding at the replication origins, the TopBP1 topoisomerase, and the RAD9 and RAD17 factors that together with TopBP1 associate to the replication forks, facilitating activation of the ATR-CHK1 kinases . DespiteAspm mutant mice, the Cyclin E level was reduced compared to controls, causing a cell cycle delay, particularly at the G1-S transition, and an overall reduction of the cerebral cortex. Restoration of the normal Cyclin E level, or treatment with the proteasome inhibitor MG132, rescued the cell cycle delay in mutant Aspm mice but did not correct the abnormal orientation of neurogenic mitoses [Cyclin E, which is required for the G1-S transition, mostly resides in the nucleus and also mediates the nuclear localization of its binding partner Cdk2 . Coimmun mitoses . Thus, ADrosophila Asp functions in interphase nuclei, an issue that deserves future studies, as current results indicate that fly Asp is a good model to investigate the functions of human ASPM.In summary, it is clear that besides its role in spindle assembly, ASPM/Aspm has additional functions in interphase nuclei of both human and mouse cells, where it interacts with a number of proteins required for DNA replication and/or repair and cell cycle progression. ASPM/Aspm also controls genome stability, especially under stress conditions such as exposure to ionizing radiation or disturbances in DNA replication. It is currently unknown whether ASPM gene [ASPM functions that are phenotypically hidden in ASPM-deficient HCT116 cells become evident upon the downregulation of CDK5RAP2. These findings and the other current data prompt us to propose a working hypothesis on the mitotic roles of ASPM and its Drosophila and mouse orthologues. We suggest that these genes play the same conserved mitotic functions in different cell types of Drosophila, mouse, and humans and that these functions are phenotypically manifest or hidden depending on the cellular context. Mouse and human Aspm/ASPM also have clear roles in DNA replication and repair. Whether Drosophila Asp is involved in these processes has never been addressed. However, we speculate that Drosophila Asp might be involved in DNA metabolism just as its mouse and human counterparts. The current results indicate that Asp and its mammalian orthologous proteins exhibit virtually identical localization patterns during cell division. They localize to the spindle poles next to or around the centrosomes and to the outer sides of the central spindle, two spindle regions enriched in MT minus ends. In vitro studies have also shown that Asp/ASPM orthologous proteins preferentially bind the MT minus ends. While the results of the Asp/ASPM localization studies are in good agreement, except for a few conflicting findings discussed earlier , the resSPM gene . These cASPM, suggesting that mutations in this gene have many diverse inhibitory effects on neural progenitor growth and could be, therefore, particularly effective in hampering neurogenesis. This hypothesis also provides a justification for why ASPM is the most frequently mutated gene in human microcephaly.Our working hypothesis impacts the models for the etiology of ASPM-dependent microcephaly. As mentioned earlier, MCPH is a genetically heterogeneous disorder caused by mutations in at least 30 different genes (,7,8,82;


Astrocytes become activated with certain infections, and this might alter the brain to trigger or worsen depressed mood. Indeed, astrocytes are chronically activated in people with HIV infection (PWH), who are much more frequently depressed than people without HIV (PWoH). A particularly disabling component of depression in PWH is apathy, a loss of interest, motivation, emotion, and goal-directed behavior. We tested the hypothesis that depression and apathy in PWH would be associated with higher levels of a biomarker of astrocyte activation, glial fibrillary acidic protein (GFAP), in cerebrospinal fluid (CSF).We evaluated PWH in a prospective observational study using the Beck Depression Inventory-II (BDI-II) and additional standardized assessments, including lumbar puncture. We measured GFAP in CSF with a customized direct sandwich ELISA method. Data were analyzed using ANOVA and multivariable regression.Participants were 212 PWH, mean (SD) age 40.9±9.14 years, median (IQR) nadir and current CD4 199  and 411 , 65.1% on ART, 67.3% virally suppressed. Higher CSF GFAP correlated with worse total BDI-II total scores , and with worse apathy scores . The correlation between apathy/depression and GFAP was not in fluenced by other factors such as age or HIV suppression status.Astrocyte activation, reflected in higher levels of CSF GFAP, was associated with worse depression and apathy in PWH. Interventions to reduce astrocyte activation -- for example, using a peptide-1 receptor (GLP-1R) agonist -- might be studied to evaluate their impact on disabling depression in PWH. Despite viral suppression on combination antiretroviral therapy (ART), people with HIV (PWH) suffer from a higher prevalence of depression than the general population. Depression is the most common psychiatric comorbidity in HIV –3 and apThis cross-sectional study evaluated PWH at 6 US centers in CNS AntiRetroviral Effects Research (CHARTER), a prospective longitudinal cohort, between 2003–2008. Inclusion criteria were HIV seropositivity and ability to complete the protocol. Participants who had severe neuropsychiatric comorbidities  were excluded. All study procedures were approved by local Institutional Review Boards and all participants provided written informed consent for the study procedures, including future use of data and biospecimens.All participants were comprehensively evaluated with standardized assessments including lumbar puncture, phlebotomy, neuromedical history and examination, and laboratory testing. A trained clinical examiner interviewed and examined participants to collect information such as antiretroviral treatments, nadir CD4+ T cell counts, and history of diabetes mellitus.Depressive symptoms were assessed using the Beck Depression Inventory (BDI-II), a validated survey of 21 questions that assess depressive symptoms and their severity . Higher HIV infection was diagnosed using an enzyme-linked immunosorbent assay with Western blot confirmation. HIV RNA in plasma was measured using commercial assays and deemed undetectable at a lower limit of quantification (LLQ) of 50 copies/mL. CD4+ T lymphocytes were measured by flow cytometry and nadir CD4+ T lymphocyte count was assessed by self-report.CSF GFAP in picograms per milliliter (pg/mL)was measured by a customized direct sandwich ELISA method, with a mouse monoclonal antibody cocktail against GFAP  as the capturing antibody and a rabbit polyclonal anti-GFAP antibody  as the detection antibody. GFAP protein standards  were used to standardize concentration curves.10 transformation was used to normalize CSF GFAP values. The Pearson correlation coefficient was used to measure the relationship of GFAPlevels to indices of depressed mood and apathy. We applied ANOVA when the distribution of the outcome variable was not significantly different from normal. When distributions significantly deviated from normal, non-parametric analyses were conducted. Follow-up analyses used recursive partitioning to identify informative GFAP cut-offs. We adjusted for testing multiple related outcomes using the Benjamini Hochberg procedure. When potential statistically confounding variables such as age and demographic and disease variables were significantly related to both the predictor (CSF GFAP level) and outcomes of interest , we evaluated these further in multivariable regression analyses. Relevant covariates considered included demographics, HIV disease and treatment parameters, and antidepressant treatments. Analyses were conducted using JMP Pro version 15.0.0 .Demographic and clinical characteristics were summarized using means and standard deviations, medians, and interquartile ranges or percentages, as appropriate. Log10 CSF GFAP level was 3.47±0.0781pg/mL, and the mean BDI-II score was 12.2±9.84, with 39.2% having a BDI-II>13, reflecting at least mild depression.The sample included 212 participants with a mean±SD age 40.9±9.14 years, female17.9%, black40.6%, non-Hispanic white 47.6%,Hispanic 8.96%, other race/ethnicity 2.83%, non-Hispanic white 47.6%, median (IQR) duration of HIV infection 7.3  years, current CD4 411 , nadir CD4199 , plasma HIV RNA suppressed (<50 copies/mL) in32.7%, CSF HIV RNA suppressed in62.3%. The mean±SD log10CSF GFAP . In a multivariable regression predicting apathy scores, the interaction between age and GFAP was not significant(p=0.888), while both main effects of both GFAP (p=0.0442) and age (p=0.0197) were significant. Apathy scores were not related to sex or ethnicity. Older PWH also had worse overall depressed mood . In a multivariable regression predicting BDI-II total score, the main effect of age (p=0.164) and the interaction of age with GFAP (p=0.846) were not significant.Several variables were significantly related to depression parameters and GFAP levels. Older individuals had both worse apathy  and higher logSex and ethnicity were not significantly related toBDI-II total score. However, both sex and ethnicity were related to CSF GFAP. Males had higher GFAP levels than females  and whites had higher levels than the other ethnicities . In a multivariable regression predicting BDI-II total score, GFAP was statistically significant (p=0.0165), while sex and the interaction term were not(ps>0.25).In a similar regression for ethnicity, GFAP was significant (0.0239), while ethnicity(p=0.237) and the interaction (p=0.891) were not.The proportion of participants taking antidepressant medications was 32%. The odds of taking antidepressants for those with a BDI-II>13 was 2.50 . Antidepressant use was associated with worse apathy scores  and higher GFAP levels .PWH both on and off antidepressant medications contributed to the relationship between GFAP and apathy: for those on at least one antidepressant: r=0.123, p=0.295, N=67; for those not on any antidepressants: r=0.139, p=0.0988, N=142;interaction p=0.895.PWH with detectable plasma viral loads had worse total BDI-II scores as well as worse scores on the cognitive , but not apathy, somatic or affective items . Both virally suppressed and unsuppressed PWH contributed to the relationship between GFAP and apathy: for suppressed PWH r=0.126, p=0.302, N=69; for not suppressed r=0.262, p=0.0016, N=142; interaction p=0.388. Detectable viral load was not associated with CSF GFAP. In a multivariable regression predicting BDI-II cognitive scores, the main effects of both GFAP (p=0.00139) and detectable viral load were significant (p=0.00316), but their interaction was not (p=0.438). Findings were similar for the BDI-II total score (data not shown). Those with suppressed CSF HIV RNA had better apathy scores . Higher GFAP correlated with higher CSF total protein , but CSF protein did not relate to apathy scores . GFAP was not influenced by CSF leukocyte count . Apathy scores did not correlate with current  or nadir CD4+ T cells , or plasma viral suppression .In a multivariable regression predicting BDI-II, GFAP was significant while being on an antidepressant and its interaction with GFAP were not (ps=0.153 and 0.359). In a stepwise multivariable regression  predicting BDI-II total score from CSF GFAP, age, sex, ethnicity, nadir, and current CD4+ T-cell count and viral suppression, the model selected CSF GFAP (p=0.00915) and lack of viral suppression(p=0.00998) as the best correlates .PWH with worse depression (higher BDI-II scores) had worse HIV-MOS physical health summary scores , and worse mental health summary scores . Similarly, higher CSF GFAP correlated with worse physical and mental  health scores. The proportion of participants reporting dependence in instrumental activities of daily living (IADLs) was 15.6%; participants with a BDI-II>13 had 11.9-fold higher odds of being dependent . There was a 3% increase in the odds of having detectable plasma viral load per one-unit increase in BDI-II scores increased . Similarly, the odds of having detectable CSF viral load increased as BDI-II scores increased .This is the first study to show that PWH with worse apathy and other attributes of depressed mood had higher levels of GFAP in CSF. Since in the central nervous system GFAP is found only in astrocytes, and since its expression is upregulated in activated astrocytes , higher We suggest that the impact of astrocyte activation on depression is via neurotoxicity . AstrocyStrengths of this work include the careful characterization of depressed mood and the consideration of a range of potential confounding factors, to which the primary findings were robust. The cohort was multicenter and racially diverse, enhancing generalizability. Limitations of this study include its cross-sectional design, limiting causal inference. Based on existing knowledge, a causal link between astrocyte activation, as indexed by CSF GFAP, and depressed mood is plausible; however, it is conceivable that changes in activity, diet and other lifestyle factors associated with depression might lead to astrocyte activation (reverse causation). Statistical confounds were not detected in this study; however, an unmeasured variable might account for the association between GFAP and depressed mood. The effect sizes demonstrated here were small, albeit statistically significant. Females were underrepresented. The rate of viral suppression was lower than in many modern cohorts; however, after adjustment for viral suppression, elevated CSF GFAP levels were still significantly associated with depressed mood. Antidepressant medications could have been taken for reasons other than depressed mood, such as for neuropathic pain.These findings raise the possibility of interventions, potentially influencing pathways that might affect depressed mood –51. For


Any If For any regular We study  X we denote by S(X) the group of permutations of X. The group maximal cofinitary subgroups. Here, a permutation cofinitary if it has only finitely many fixed points. A group cofinitary if all of its non-identity elements are cofinitary. The group G is moreover said to be maximal if no proper supergroup is cofinitary.Given a set mentclass2pt{minim example , 12). Ofmaximal cofinitary groups. Throughout the paper, we fix a regular, uncountable cardinal maximal it there are no proper An important area of set theory, which has been of increased interest in the past decades, is the study of analogues of combinatorial sets of reals in the higher Baire spaces, mentclass2pt{minimlies see , 6.If is a specifically centered partial order,and is a family of dense subsets of whereand then there is a filter such that for allA main tool in our analysis of the structure of G is a maximal cofinitary group , we estG so that the orbits of G are exactly  There iWe continue by recalling some facts about the cardinal entclass1pt{minimaeader to  for moreentclass1pt{minimastrong intersection property if for any subfamily pseudointersection number forLet entclass1pt{minimaWe will not need this definition of canonical lower bounds if there is a function Let The significance of this definition is that the forcing axiom for the class of [Theorem 1.8 of ] If \docThe point for us here is that we will show the forcing notion introduced in  is a \doA be an index set and denote by good words i.e. those that are either the power of a singleton or start and end with a different letter. Given a mapping A to induces acofinitary representation if the image of a in A. If each if if if We also define Let A and B be disjoint sets and let ) from which the maximality of G follows. We claim that By interweaving the proofs of the above result and Theorem G which is universal for groups of size Assume G can be universal for groups of size at most Kastermans’ original theorem on the If Enumerate At stage 0 first use Now suppose we have constructed a This completes the construction. Let G has any particular set of Again this theorem can be proved with the added assumption that the universal We finish by recording some open questions on the structure of Is it consistent that there is a partition of This seems to be unknown even in the The next has to do with the analogue of another theorem of Kastermans. Kastermans also showed in [Is it consistent that there is a locally finite Finally we repeat the observation noted after the proof of Theorem Fix