Possible retinotoxicity of long-term vardenafil treatment.

Phosphodiesterase (PDE) inhibitors - such as vardenafil - are used primarily for treating erectile dysfunction via increasing cyclic guanosine monophosphate (cGMP) levels. Recent studies have also demonstrated their significant cardioprotective effects in several diseases, including diabetes, upon long-term, continuous application. However, PDE inhibitors are not specific for PDE5 and also inhibit the retinal isoform. A sustained rise in cGMP in photoreceptors is known to be toxic; therefore, we hypothesized that long-term vardenafil treatment might result in retinotoxicity. The hypothesis was tested in a clinically relevant animal model of type 2 diabetes mellitus. Histological experiments were performed on lean and diabetic Zucker Diabetic Fatty rats. Half of the animals were treated with vardenafil for six months, and the retinal effects were evaluated. Vardenafil treatment alleviated rod outer segment degeneration but decreased rod numbers in some positions and induced changes in the interphotoreceptor matrix, even in control animals. Vardenafil treatment decreased total retinal thickness in the control and diabetic groups and reduced the number of nuclei in the outer nuclear layer. Müller cell activation was detectable even in the vardenafil-treated control animals, and vardenafil did not improve gliosis in the diabetic group. Vardenafil-treated animals showed complex retinal alterations with improvements in some parameters while deterioration in others. Our results point towards the retinotoxicity of vardenafil, even without diabetes, which raises doubts about the retinal safety of long-term continuous vardenafil administration. This effect needs to be considered when approving PDE inhibitors for alternative indications.

Reference data on estrogen metabolome in healthy pregnancy.

INTRODUCTION: Estrogen hormones and their metabolites are implicated in the maintenance of healthy pregnancy and adequate fetal development. Abnormal levels were related to increased risk of pregnancy complications, particularly preeclampsia. Our aims were (1) to develop a methodological platform for the comprehensive assessment of estrogen metabolome in pregnancy; (2) to collect healthy reference data for relevant elements of estrogen metabolome in each trimester; (3) to assess unconjugated fractions of the estrogen metabolome, (4) to assess the dominant metabolic pathways of estrogen compounds. METHODS: We enrolled healthy pregnant mothers between gestational week 5-15 (on the confirmation of pregnancy; 79 samples), gestational weeks 19-27 (70 samples), and gestational week 34-39 (54 samples). A method employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed to assess estrone, 17-beta-estradiol, estriol levels, and their metabolites as conjugated and unconjugated forms. Descriptive statistics were used to characterize the level of compounds in each trimester. RESULTS: Estrone, 17-beta-estradiol and estriol levels are dramatically increasing with the advancement of pregnancy. Measured levels were in a very wide range. 17-beta-estradiol is neither glucuronated nor sulphated. To the contrary, estriol and estrone are significantly conjugated; unconjugated fraction is <15% of total hormone levels in any trimester. Regarding metabolism, 4-methoxy-estradiol and 17-epiestriol were not detected. CONCLUSION: We concluded that (1) the levels of estrogen compounds and metabolites increase with advancing gestational age; (2) the wide ranges of levels challenge the establishment of a healthy reference range for clinical purposes; (3) 17-beta-estradiol is not conjugated significantly; (4) 4-methylation and 17-epimerization pathways of estrogens are negligible with our LC-MS/MS method.

Long-term recovery dynamics determined by the degree of the disturbance - Ten years tracking of aquatic macroinvertebrate recolonisation after an industrial disaster (Red Sludge Disaster, Hungary).

A ten-year-long examination of macroinvertebrate community recovery was conducted following a catastrophic spill of highly alkaline red sludge (pH >13) into lowland streams. Our primary objective was to compare recovery patterns after coarse- and fine-grain disturbances, focusing on two aspects: i) trend analysis to reveal long-term changes of six community parameters, and ii) variation analyses to assess parameter changes over time. We conducted statistical analysis on long-term data series of macroinvertebrates obtained from quantitative samples collected at four sections with varying degrees of disturbance along the impacted stream sections. We developed a comprehensive theoretical framework comprising a series of sequential phases: Ramp-up, Overshoot, and Oscillation Phases. i) A trend analysis revealed that disturbances show a gradual recovery pattern, while variance analyses showed an asymptotic convergence to an equilibrium. ii) Evaluating these trends across phases unveiled that the initial recovery phase exhibited a steep trajectory, lasting 4-9 months, irrespective of disturbance severity. Coarse-grain disturbances induced a remarkable Overshoot phenomenon across all community metrics. The more severe the disturbance, the greater the height and duration of the Overshoot. Our results suggest that the presence or absence of Overshoot can serve as an indicator for coarse-grain disturbances in the context of large and infrequent disturbances (LID). The entire recovery process lasts for 2.5-3 years irrespective of the severity of the LID. In conclusion, a minimum survey duration of two and half years is deemed imperative to capture the phases of recovery, and changes associated with LID are not expected to extend beyond the three-year threshold. The theoretical framework, including Overshoot parameters, may assist future studies in comparing recovery patterns of different LID types. Furthermore, our theoretical framework is likely to be applicable to other groups of organisms given a sufficiently long monitoring of recovery, influenced also by the length of reproductive cycles.

Arthroscopic capsular release is more effective in pain relief than conservative treatment in patients with frozen shoulder.

BACKGROUND: Frozen shoulder is a common medical condition, but the ideal therapeutic method is yet to be determined. Our aim was to analyze the pain-relieving effect of different treatment options used for the management of this disease. METHODS: Medical records of 59 patients (22 male, 37 female, average age: 55.5 years ±9.9) with early stage primary frozen shoulder were evaluated, their demographic data, physical examination, concomitant diseases and treatment specific data were registered. Life quality and the level of pain were assessed using the Oxford Shoulder Score (OSS) and Numeric Rating Scale (NRS). Different treatment modalities and their effect on pain relief were recorded. Any existing correlation between life quality, pain and demographic data, concomitant diseases or the therapeutic method used was investigated. RESULTS: The level of pain measured on NRS improved from 7.9 ± 1.6 to 1.9 ± 2.2. The most effective therapeutic method in terms of pain relief was surgery, followed by physiotherapy and intraarticular steroid injection (NRS score after treatment: 2 - p < 0.0001; 3.3 - p < 0.0001; 4.9 - p < 0.0001, respectively). Non-steroidal anti-inflammatory drugs (NSAIDs) did not reduce pain significantly. OSS improved from 24 to 43.6 and was not affected by the investigated variables, time to recovery was not influenced by the demographic data, the type of treatment or concomitant diseases. CONCLUSIONS: Arthroscopic capsular release, physiotherapy and intraarticular steroid injection outperformed physical therapy and NSAID treatment in terms of pain relief. Despite of slight but persistent post-therapeutic pain found in half of the cases, treatment was considered satisfactory by the patients. Nor patient specific neither therapy specific data had a significant effect on the course of the disease.

Sarcomas Harboring EWSR1::PATZ1 Fusions: A Clinicopathologic Study of 17 Cases.

Soft tissue sarcomas harboring EWSR1::PATZ1 are a recently recognized entity with variable morphology and a heterogeneous immunohistochemical profile. We studied 17 such tumors. The tumors occurred in 12 men and 5 women (median age, 50 years; range, 15-71 years), involved the thoracoabdominal soft tissues (14 cases; 82%), lower extremities (2 cases; 12%), and tongue (1 case; 6%), and ranged from 0.7 to 11.3 cm (median, 4.7 cm). All but 1 patient received complete surgical resection; 7 were also treated with neoadjuvant chemo/radiotherapy. All cases showed typical features of EWSR1::PATZ1 sarcoma, including uniform round to spindled cells, fibromyxoid matrix, fibrous bands, hyalinized vessels, and pseudoalveolar/microcystic spaces. Unusual features, seen in a subset of cases, included degenerative-appearing nuclear atypia, epithelioid cytomorphology, mature fat, abundant rhabdomyoblasts, high mitotic activity, and foci with increased cellularity and nuclear atypia. Positive immunohistochemical results were desmin (16/17, 94%), MyoD1 (13/14, 93%), myogenin (6/14, 43%), GFAP (10/10, 100%), S100 protein (15/17, 88%), SOX10 (7/13, 54%), keratin (10/17, 59%), CD99 (4/11, 36%), H3K27me3 (retained expression 9/9, 100%), p16 (absent expression 1/4, 25%), and p53 (wild type 3/3, 100%). Fusion events included EWSR1 exon 8::PATZ1 exon 1 (14/17, 82%), EWSR1 exon 9::PATZ1 exon 1 (2/17, 12%), and EWSR1 exon 7::PATZ1 exon 1 (1/17, 6%). No evaluated tumor had alterations of CDKN2A/B and/or TP53, or MDM2 amplification. Clinical follow-up (16 patients: median, 13.5 months; range, 1-77 months) showed distant metastases in 3 patients (1/3 at time of presentation) and no local recurrences. At the time of last follow-up, 14 patients were disease free, 1 was alive with disease, 1 was dead of disease (at 13 months), and 1 had an indeterminant pulmonary nodule. We conclude that the morphologic spectrum of EWSR1::PATZ1 is broader than has been previously appreciated. Although more long-term follow-up is needed, the prognosis of these very rare sarcomas may be more favorable than previously reported.

Anisotropy, Anatomical Region, and Additional Variables Influence Young's Modulus of Bone: A Systematic Review and Meta-Analysis.

The importance of finite element analysis (FEA) is growing in orthopedic research, especially in implant design. However, Young's modulus (E) values, one of the most fundamental parameters, can range across a wide scale. Therefore, our study aimed to identify factors influencing E values in human bone specimens. We report our systematic review and meta-analysis based on the recommendation of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guideline. We conducted the analysis on November 21, 2021. We included studies investigating healthy human bone specimens and reported on E values regarding demographic data, specimen characteristics, and measurement specifics. In addition, we included study types reporting individual specimen measurements. From the acquired data, we created a cohort in which we performed an exploratory data analysis that included the explanatory variables selected by random forest and regression trees methods, and the comparison of groups using independent samples Welch's t test. A total of 756 entries were included from 48 articles. Eleven different bones of the human body were included in these articles. The range of E values is between 0.008 and 33.7 GPa. The E values were most heavily influenced by the cortical or cancellous type of bone tested. Measuring method (compression, tension, bending, and nanoindentation), the anatomical region within a bone, the position of the bone within the skeleton, and the bone specimen size had a decreasing impact on the E values. Bone anisotropy, specimen condition, patient age, and sex were selected as important variables considering the value of E. On the basis of our results, E values of a bone change with bone characteristics, measurement techniques, and demographic variables. Therefore, the evaluation of FEA should be performed after the standardization of in vitro measurement protocol. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Role of CB1 Cannabinoid Receptors in Vascular Responses and Vascular Remodeling of the Aorta in Female Mice.

Both the endocannabinoid system (ECS) and estrogens have significant roles in cardiovascular control processes. Cannabinoid type 1 receptors (CB1Rs) mediate acute vasodilator and hypotensive effects, although their role in cardiovascular pathological conditions is still controversial. Estrogens exert cardiovascular protection in females. We aimed to study the impact of ECS on vascular functions. Experiments were performed on CB1R knockout (CB1R KO) and wild-type (WT) female mice. Plasma estrogen metabolite levels were determined. Abdominal aortas were isolated for myography and histology. Vascular effects of phenylephrine (Phe), angiotensin II, acetylcholine (Ach) and estradiol (E2) were obtained and repeated with inhibitors of nitric oxide synthase (NOS, Nω-nitro-L-arginine) and of cyclooxygenase (COX, indomethacin). Histological stainings (hematoxylin-eosin, resorcin-fuchsin) and immunostainings for endothelial NOS (eNOS), COX-2, estrogen receptors (ER-α, ER-ß) were performed. Conjugated E2 levels were higher in CB1R KO compared to WT mice. Vasorelaxation responses to Ach and E2 were increased in CB1R KO mice, attenuated by NOS-inhibition. COX-inhibition decreased Phe-contractions, while it increased Ach-relaxation in the WT group but not in the CB1R KO. Effects of indomethacin on E2-relaxation in CB1R KO became opposite to that observed in WT. Histology revealed lower intima/media thickness and COX-2 density, higher eNOS and lower ER-ß density in CB1R KO than in WT mice. CB1R KO female mice are characterized by increased vasorelaxation associated with increased utilization of endothelial NO and a decreased impact of constrictor prostanoids. Our results indicate that the absence or inhibition of CB1Rs may have beneficial vascular effects.

Our Experiences with Asparaginase Activity Measurements in Children with Lymphoblastic Diseases.

BACKGROUND: Asparaginase is a key component of chemotherapy protocols for the treatment of lymphoblastic malignancies among children. Adequate asparagine depletion is an important factor to achieve optimal therapeutic outcomes. METHODS: Over a 3.5 year period, 106 patients were monitored for asparaginase activity (329 samples) in a single center of the Hungarian Pediatric Oncology-Hematology Group. In Hungary, three asparaginase products are available: native E. coli ASNase (Kidrolase), a pegylated form of this enzyme (Pegaspargase) and another native product from Erwinia chrysanthemi (Erwinase). A retrospective data analysis was performed. RESULTS: In 81% (268/329) of our patients, AEA levels were in the optimal therapeutic range of over 100 IU/L. Of 106 patients, 13 (12%) were diagnosed with 'silent inactivation'. CONCLUSIONS: Monitoring of AEA can help to identify patients with 'silent inactivation' and their asparaginase therapy can thus be optimized.

[Changes of violent suicide attempts during the first two years of COVID-19 pandemic in Dr. Manninger Jeno National Traumatology Center, Hungary.] / A violens öngyilkossági kísérletek számának változása a COVID­19-járvány elso két évében a Dr. Manninger Jeno Baleseti Központban.

INTRODUCTION: During the pandemic years in Hungary, the completed suicide rates have risen significantly. Violent suicide attempts represent the majority of completed suicides. OBJECTIVE: In our study, we analyzed the change of the number of inpatients treated in Dr. Manninger Jeno National Traumatology Center between 2016 and 2021 due to violent suicide attempts, focusing on the trend in the first two years of the pandemic outbreak. METHOD: We used an interrupted time-series analysis with Prais-Winsten regression, controlling autoagressive and seasonal effects, to estimate the effect of the pandemic on the violent suicide attempt rates in our sample. RESULTS: In the first two pandemic years, the number of inpatients treated in Dr. Manninger Jeno National Traumatology Center due to violent suicide attempts rose significantly, compared to the previous years. After the rapid rise observed in 2020, decreasing numbers were seen in 2021. DISCUSSION AND CONCLUSION: Analyzing the numbers of violent suicide attempts between 2016 and 2021, an increase in the number of attempts was observed during the first two pandemic years. Orv Hetil. 2023; 164(26): 1003-1011.

Subvastus Approach Supporting Fast-Track Total Knee Arthroplasty Over the Medial Parapatellar Approach: A Systematic Review and Network Meta-Analysis.

BACKGROUND: Numerous surgical approaches are being used to perform total knee arthroplasty (TKA). This systematic review and network meta-analysis aimed to compare surgical approaches used in TKA regarding postoperative outcomesat different time points. METHODS: We performed a literature search from medical database inception until October 2, 2021. We searched for randomized controlled trials (RCTs) investigating patients undergoing TKA and comparing at least 2 surgical approaches regarding early postsurgical clinical outcomes (range of motion [ROM], pain on a visual analog scale, and Knee Society Score [KSS]). We included 33 RCTs in our networks. Using paired and network meta-analysis, we calculated pooled mean differences (MDs) with 95% CIs by comparing surgical approaches to the medial parapatellar method. RESULTS: The subvastus (SV) method performed the best on days 1 (MD = 6.99; CI: 1.08; 12.89), 3 (MD = 8.00; CI: 2.08; 13.92), 4 (MD = 27.01; CI: 18.09; 35.92), and 6 (MD = 27.22; CI: 18.38; 36.07) for ROM improvement. Regarding the decrease in pain, the mini SV approach offered significantly lower pain values on days 1 (MD = -1.98; CI: -2.93; -1.03), 3 (MD = -0.85; CI: -1.49; -0.22), and 7 (MD = -1.90; CI: -2.23; -1.57). The differences decreased as time passed. Furthermore, the SV and mini-SV methods performed the best regarding total, knee and function KSS. CONCLUSION: Quadriceps-sparing approaches, especially the SV and mini-SV, are superior to the other approaches in the early postsurgical period, but the differences decrease as time passes.

CXCL13 expressed on inflamed cerebral blood vessels recruit IL-21 producing TFH cells to damage neurons following stroke.

BACKGROUND: Ischemic stroke is a leading cause of mortality worldwide, largely due to the inflammatory response to brain ischemia during post-stroke reperfusion. Despite ongoing intensive research, there have not been any clinically approved drugs targeting the inflammatory component to stroke. Preclinical studies have identified T cells as pro-inflammatory mediators of ischemic brain damage, yet mechanisms that regulate the infiltration and phenotype of these cells are lacking. Further understanding of how T cells migrate to the ischemic brain and facilitate neuronal death during brain ischemia can reveal novel targets for post-stroke intervention. METHODS: To identify the population of T cells that produce IL-21 and contribute to stroke, we performed transient middle cerebral artery occlusion (tMCAO) in mice and performed flow cytometry on brain tissue. We also utilized immunohistochemistry in both mouse and human brain sections to identify cell types and inflammatory mediators related to stroke-induced IL-21 signaling. To mechanistically demonstrate our findings, we employed pharmacological inhibitor anti-CXCL13 and performed histological analyses to evaluate its effects on brain infarct damage. Finally, to evaluate cellular mechanisms of stroke, we exposed mouse primary neurons to oxygen glucose deprivation (OGD) conditions with or without IL-21 and measured cell viability, caspase activity and JAK/STAT signaling. RESULTS: Flow cytometry on brains from mice following tMCAO identified a novel population of cells IL-21 producing CXCR5+ CD4+ ICOS-1+ T follicular helper cells (TFH) in the ischemic brain early after injury. We observed augmented expression of CXCL13 on inflamed brain vascular cells and demonstrated that inhibition of CXCL13 protects mice from tMCAO by restricting the migration and influence of IL-21 producing TFH cells in the ischemic brain. We also illustrate that neurons express IL-21R in the peri-infarct regions of both mice and human stroke tissue in vivo. Lastly, we found that IL-21 acts on mouse primary ischemic neurons to activate the JAK/STAT pathway and induce caspase 3/7-mediated apoptosis in vitro. CONCLUSION: These findings identify a novel mechanism for how pro-inflammatory T cells are recruited to the ischemic brain to propagate stroke damage and provide a potential new therapeutic target for stroke.

Clinical and diagnostic relevance of asymmetric and symmetric dimethyl arginine (ADMA/SDMA). / Az aszimmetrikus és a szimmetrikus dimetilált arginin (ADMA/SDMA) klinikai és diagnosztikai jelentosége.

Összefoglaló. Régóta folynak kutatások olyan újabb biomarkerek azonosítására, amelyek segítik a krónikusan progrediáló, úgynevezett civilizációs betegségek - például cardiovascularis kórképek, vesefunkció-beszukülés - korai felismerését. Az aszimmetrikus és a szimmetrikus dimetil-arginin (ADMA és SDMA) ketto azon paraméterek közül, amelyek biológiai hatásai évtizedek óta ismertek ugyan, ám biomarkerként egyelore nem terjedtek el a humán orvosi-diagnosztikai gyakorlatban. A fehérjearginin-metiltranszferázok katalizálta folyamatban L-argininbol keletkezo vegyületek a nitrogén-monoxid-szintáz aktivitásának gátlói. Mivel a nitrogén-monoxid számos biológiai folyamat kulcsszereploje - gátolja az érpálya simaizomsejtjeinek relaxációját, csökkenti a thrombocytaaggregációt, és gyulladáscsökkento hatást fejt ki -, termelodésének zavarai megnövelik a magas vérnyomás és cardiovascularis betegségek kialakulásának kockázatát. Áttekinto közleményünkben az ADMA és az SDMA mint lehetséges új diagnosztikai markerek, valamint a társadalmi és orvosszakmai szempontból is kihívást jelento betegségek kapcsolatának bemutatását tuztük ki célul. Orv Hetil. 2022; 163(13): 500-505. Summary. Research has long been underway to identify additional biomarkers that will help in the early detection of chronic diseases of civilization, such as cardiovascular disease and renal impairment. Asymmetric and symmetric dimethyl arginine (ADMA and SDMA), two of the parameters whose biological effects have been known for decades, have not yet been widely used as biomarkers in human medical-diagnostic practice. In a process catalyzed by protein arginine methyltransferases, compounds derived from L-arginine are inhibitors of nitric oxide synthase activity. Because nitric oxide is a key player in many biological processes - for instance, inhibiting the relaxation of vascular smooth muscle cells, reducing platelet aggregation, and having anti-inflammatory effect -, disturbances in its production increase the risk of developing high blood pressure and cardiovascular disease. Therefore, in our review paper, we aimed to present the relationship between ADMA and SDMA as possible new diagnostic markers and socially and physically challenging diseases. Orv Hetil. 2022; 163(13): 500-505.

Adrenal, Gonadal and Peripherally Steroid Changes in Response to Extreme Physical Stress for Characterizing Load Capacity in Athletes.

Athletes are often exposed to extreme physical stress during training or competitions. The consequent activation of the hypothalamus-hypophysis-adrenal (HPA) axis results in intensified steroid hormone production in the adrenal cortex. We determined the impact of an acute extreme physical stress on adrenal and gonadal steroidogenesis in healthy male professional athletes (n = 40). The subjects underwent an extreme physical load test until total voluntary fatigue between 14:00 and 18:00 when the hormone levels are relatively stable. Blood was taken before the start (baseline), at the peak load (peak), and 30 min following completion of the exercise (recovery). The vital parameters, lactate levels, and blood levels of the 14 steroid hormones were recorded. The multivariate statistical analysis of the results revealed that all monitored hormone levels increased upon stress. Significant changes in steroid concentrations were detected at peak versus baseline, peak versus recovery, and at baseline versus recovery. The mineralocorticoid (including aldosterone and corticosterone), glucocorticoid (11-deoxycortisol and cortisol), and androgen (androstenedione, dehydroepiandrosterone, and dehydroepiandrosterone sulfate) pathways, as well as gonadal testosterone synthesis are activated simultaneously under extreme physical load. The profiling of adrenal and gonadal steroid biosynthesis in athletes may help the characterization of their loading capacity.

Sex-related differences of early cardiac functional and proteomic alterations in a rat model of myocardial ischemia.

BACKGROUND: Reduced cardiovascular risk in premenopausal women has been the focus of research in recent decades. Previous hypothesis-driven experiments have highlighted the role of sex hormones on distinct inflammatory responses, mitochondrial proteins, extracellular remodeling and estrogen-mediated cardioprotective signaling pathways related to post-ischemic recovery, which were associated with better cardiac functional outcomes in females. We aimed to investigate the early, sex-specific functional and proteomic changes following myocardial ischemia in an unbiased approach. METHODS: Ischemia was induced in male (M-Isch) and female (F-Isch) rats with sc. injection of isoproterenol (85 mg/kg) daily for 2 days, while controls (M-Co, F-Co) received sc. saline solution. At 48 h after the first injection pressure-volume analysis was carried out to assess left ventricular function. FFPE tissue slides were scanned and analyzed digitally, while myocardial proteins were quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using isobaric labeling. Concentrations of circulating steroid hormones were measured with LC-MS/MS. Feature selection (PLS and PLS-DA) was used to examine associations among functional, proteomic and hormonal datasets. RESULTS: Induction of ischemia resulted in 38% vs 17% mortality in M-Isch and F-Isch respectively. The extent of ischemic damage to surviving rats was comparable between the sexes. Systolic dysfunction was more pronounced in males, while females developed a more severe impairment of diastolic function. 2224 proteins were quantified, with 520 showing sex-specific differential regulation. Our analysis identified transcriptional, cytoskeletal, contractile, and mitochondrial proteins, molecular chaperones and the extracellular matrix as sources of disparity between the sexes. Bioinformatics highlighted possible associations of estrogens and their metabolites with early functional and proteomic alterations. CONCLUSIONS: Our study has highlighted sex-specific alterations in systolic and diastolic function shortly after ischemia, and provided a comprehensive look at the underlying proteomic changes and the influence of estrogens and their metabolites. According to our bioinformatic analysis, inflammatory, mitochondrial, chaperone, cytoskeletal, extracellular and matricellular proteins are major sources of intersex disparity, and may be promising targets for early sex-specific pharmacologic interventions.

Identification of disease- and headache-specific mediators and pathways in migraine using blood transcriptomic and metabolomic analysis.

BACKGROUND: Recent data suggest that gene expression profiles of peripheral white blood cells can reflect changes in the brain. We aimed to analyze the transcriptome of peripheral blood mononuclear cells (PBMC) and changes of plasma metabolite levels of migraineurs in a self-controlled manner during and between attacks. METHODS: Twenty-four patients with migraine were recruited and blood samples were collected in a headache-free (interictal) period and during headache (ictal) to investigate disease- and headache-specific alterations. Control samples were collected from 13 age- and sex-matched healthy volunteers. RNA was isolated from PBMCs and single-end 75 bp RNA sequencing was performed using Illumina NextSeq 550 instrument followed by gene-level differential expression analysis. Functional analysis was carried out on information related to the role of genes, such as signaling pathways and biological processes. Plasma metabolomic measurement was performed with the Biocrates MxP Quant 500 Kit. RESULTS: We identified 144 differentially-expressed genes in PBMCs between headache and headache-free samples and 163 between symptom-free patients and controls. Network analysis revealed that enriched pathways included inflammation, cytokine activity and mitochondrial dysfunction in both headache and headache-free samples compared to controls. Plasma lactate, succinate and methionine sulfoxide levels were higher in migraineurs while spermine, spermidine and aconitate were decreased during attacks. CONCLUSIONS: It is concluded that enhanced inflammatory and immune cell activity, and oxidative stress can play a role in migraine susceptibility and headache generation.

Relevance of a Novel Circuit-Level Model of Episodic Memories to Alzheimer's Disease.

The medial temporal lobe memory system has long been identified as the brain region showing the first histopathological changes in early Alzheimer's disease (AD), and the functional decline observed in patients also points to a loss of function in this brain area. Nonetheless, the exact identity of the neurons and networks that undergo deterioration has not been determined so far. A recent study has identified the entorhinal and hippocampal neural circuits responsible for encoding new episodic memories. Using this novel model we describe the elements of the episodic memory network that are especially vulnerable in early AD. We provide a hypothesis of how reduced reelin signaling within such a network can promote AD-related changes. Establishing novel associations and creating a temporal structure for new episodic memories are both affected in AD. Here, we furnish a reasonable explanation for both of these previous observations.

Episodic Memories: How do the Hippocampus and the Entorhinal Ring Attractors Cooperate to Create Them?

The brain is capable of registering a constellation of events, encountered only once, as an episodic memory that can last for a lifetime. As evidenced by the clinical case of the patient HM, memories preserving their episodic nature still depend on the hippocampal formation, several years after being created, while semantic memories are thought to reside in neocortical areas. The neurobiological substrate of one-time learning and life-long storing in the brain, that must exist at the cellular and circuit level, is still undiscovered. The breakthrough is delayed by the fact that studies jointly investigating the rodent hippocampus and entorhinal cortex are mostly targeted at understanding the spatial aspect of learning. Here, we present the concept of an entorhinal cortical module, termed EPISODE module, that could explain how the representations of different elements constituting episodic memories can be linked together at the stage of encoding. The new model that we propose here reconciles the structural and functional observations made in the entorhinal cortex and explains how the downstream hippocampal processing organizes the representations into meaningful sequences.

Development of a methodology to make individual estimates of the precision of liquid chromatography-tandem mass spectrometry drug assay results for use in population pharmacokinetic modeling and the optimization of dosage regimens.

BACKGROUND: The clinical value of therapeutic drug monitoring can be increased most significantly by integrating assay results into clinical pharmacokinetic models for optimal dosing. The correct weighting in the modeling process is 1/variance, therefore, knowledge of the standard deviations (SD) of each measured concentration is important. Because bioanalytical methods are heteroscedastic, the concentration-SD relationship must be modeled using assay error equations (AEE). We describe a methodology of establishing AEE's for liquid chromatography-tandem mass spectrometry (LC-MS/MS) drug assays using carbamazepine, fluconazole, lamotrigine and levetiracetam as model analytes. METHODS: Following method validation, three independent experiments were conducted to develop AEE's using various least squares linear or nonlinear, and median-based linear regression techniques. SD's were determined from zero concentration to the high end of the assayed range. In each experiment, precision profiles of 6 ("small" sample sets) or 20 ("large" sample sets) out of 24 independent, spiked specimens were evaluated. Combinatorial calculations were performed to attain the most suitable regression approach. The final AEE's were developed by combining the SD's of the assay results, established in 24 specimens/spiking level and using all spiking levels, into a single precision profile. The effects of gross hyperbilirubinemia, hemolysis and lipemia as laboratory interferences were investigated. RESULTS: Precision profiles were best characterized by linear regression when 20 spiking levels, each having 24 specimens and obtained by performing 3 independent experiments, were combined. Theil's regression with the Siegel estimator was the most consistent and robust in providing acceptable agreement between measured and predicted SD's, including SD's below the lower limit of quantification. CONCLUSIONS: In the framework of precision pharmacotherapy, establishing the AEE of assayed drugs is the responsibility of the therapeutic drug monitoring service. This permits optimal dosages by providing the correct weighting factor of assay results in the development of population and individual pharmacokinetic models.

Metabolic and catecholamine response to sympathetic stimulation in early-treated adult male patients with phenylketonuria.

PURPOSE: Defective function of phenylalanine hydroxylase in phenylketonuria (PKU) results in the accumulation of phenylalanine (Phe) and the reduction of tyrosine (Tyr) in the blood, interfering in the normal development and function of organs and tissues in the body. Tyr is the precursor of catecholamines, secreted in response to stress by the adrenal medulla and paraganglia. The aim of this study was to evaluate plasma catecholamine and amino acid response to an escalating series of sympathetic stress tests in PKU patients. METHODS: Twelve males with classical PKU (aged 18-41 years) and ten healthy male controls were included in this study. The subjects were exposed to three different sympathetic stress stimulations: cold pressor, isometric handgrip, and peak treadmill tests to exhaustion. Physiological, metabolic, and hormonal changes were determined. RESULTS: Aerobic capacity (VO2max) was significantly lower in the PKU group (p = 0.018); however, relative VO2max was similar in the two groups during the spiroergometric test. No significant differences in norepinephrine or in epinephrine response were found between the two groups during the different stimulation tests. Blood Phe increased significantly in the PKU group compared with controls (p = 0.027) during the spiroergometric test, while Tyr levels remained stable in both groups. CONCLUSION: PKU itself might not influence stress-induced catecholamine changes. Only strenuous exercise increased blood Phe levels in PKU subjects.

[Estrogen metabolism during pregnancy]. / Az ösztrogénmetabolizmus várandósság alatt.

The extensive metabolism of estrogen hormones, where oxidized forms, structural isomers and conjugated products appear in many tissues locally as well as in systemic circulation, is believed to be associated with a number of diseases. Targeted estrogen metabolomic studies have been largely associated with postmenopausal, malignant advert immune conditions. Although the role of estriol in maintaining pregnancy and the biological activity of estrogen metabolites is known, a relatively small number of publications have addressed the formation and transformation of these compounds during pregnancy. The aim of this study is to present in detail the formation and progression of estrogen metabolites during pregnancy and to summarize the knowledge of their role in undesirable processes occurring during gestation. Orv Hetil. 2019; 160(26): 1007-1014.