Unexplained Etiology Research Articles

A validated heart-specific model for splice-disrupting variants in childhood heart disease

BackgroundCongenital heart disease (CHD) is the most common congenital anomaly. Almost 90% of isolated cases have an unexplained genetic etiology after clinical testing. Non-canonical splice variants that disrupt mRNA splicing through the loss or creation of exon boundaries are not routinely captured and/or evaluated by standard clinical genetic tests. Recent computational algorithms such as SpliceAI have shown an ability to predict such variants, but are not specific to cardiac-expressed genes and transcriptional isoforms.MethodsWe used genome sequencing (GS) (n = 1101 CHD probands) and myocardial RNA-Sequencing (RNA-Seq) (n = 154 CHD and n = 43 cardiomyopathy probands) to identify and validate splice disrupting variants, and to develop a heart-specific model for canonical and non-canonical splice variants that can be applied to patients with CHD and cardiomyopathy. Two thousand five hundred seventy GS samples from the Medical Genome Reference Bank were analyzed as healthy controls.ResultsOf 8583 rare DNA splice-disrupting variants initially identified using SpliceAI, 100 were associated with altered splice junctions in the corresponding patient myocardium affecting 95 genes. Using strength of myocardial gene expression and genome-wide DNA variant features that were confirmed to affect splicing in myocardial RNA, we trained a machine learning model for predicting cardiac-specific splice-disrupting variants (AUC 0.86 on internal validation). In a validation set of 48 CHD probands, the cardiac-specific model outperformed a SpliceAI model alone (AUC 0.94 vs 0.67 respectively). Application of this model to an additional 947 CHD probands with only GS data identified 1% patients with canonical and 11% patients with non-canonical splice-disrupting variants in CHD genes. Forty-nine percent of predicted splice-disrupting variants were intronic and > 10 bp from existing splice junctions. The burden of high-confidence splice-disrupting variants in CHD genes was 1.28-fold higher in CHD cases compared with healthy controls.ConclusionsA new cardiac-specific in silico model was developed using complementary GS and RNA-Seq data that improved genetic yield by identifying a significant burden of non-canonical splice variants associated with CHD that would not be detectable through panel or exome sequencing.

Women’s fertility and vitamin D: Could hypovitaminosis D biomarkers correlate with the disease, and explain the unexplained female factor infertility?

Objective: Vitamin D (Vit D) deficiency correlates women reproductive pathophysiology. We analyzed Vit D deficiency biomarkers in infertile women. Patients & method: This case-control study enrolled 80 infertile women polycystic ovary syndrome (PCOS), and other etiologies; anovulation and unexplained and 25 controls. Serum calcidiol and calcitriol were determined by ELISA and their direct ratio was calculated. Results: 72% of controls and 92.5% of patients had Vit D deficiency/insufficiency (calcidiol mean ± SDM of 33.50±22.10 vs 20.26±5.226 ng/mL and an AUC of 0.808±0.048). Calcitriol had an AUC of 0.909±0.031 that more effectively distinguished patients and etiologies (53.49±23.30 pg/mL) from controls (114.0±43.20 pg/mL; P<0.001). 4 other etiology cases and 17 controls had calcitriol levels ≥100 pg/mL. 64% of controls (4.090±0.020) and 16.25% of patients [2.634±0.855, P<0.04; 5 PCOS (3 primary/2 secondary), 3 secondary unexplained, and 5 others (one primary tubal, one primary/one secondary peritoneal, one primary/one cervical and one primary tubal had a normal ratio ≥3.333 at an AUC of 0.740±0.065. All biomarkers revealed patient levels ~50% lower than controls; lowest in PCOS and unexplained etiologies. Conclusion: Vit D levels are significantly reduced in infertile women; lowest in PCOS and unexplained etiology, for all biomarkers, where calcitriol was the optimal predictor of both infertility and etiology. Keywords: Female infertility, Polycystic ovary syndrome; unexplained infertility; Vitamin D; 25-Hydroxy-cholecalceferol, 1; 25-Dihydroxyl-cholecalceferol.

P-419 Relevance of intravenous immunoglobulins in patients with unexplained recurrent pregnancy loss that occurred at the same gestational weeks every time

Abstract Study question Are intravenous immunoglobulins (IVIG) effective for the selected patients who experienced unexplained recurrent pregnancy loss (RPL) that occurred at the same gestational weeks every time? Summary answer IVIG before and after the gestational week-limit (GWL), where miscarriage occurred at the same gestational weeks every time, may improve pregnancy outcomes in RPL patients. What is known already There is no established treatment for patients with unexplained RPL. Some investigators have indicated that IVIG may exhibit therapeutic efficacy in women with unexplained RPL. Since it has been reported that live birth rate significantly increased in women with four or more RPL of unexplained etiology, who received a high dose IVIG at 4-5 weeks of gestation in 2022, ESHRE Guideline Group on RPL (2022) has changed the recommendation to the following: the use of repeated and high doses of IVIG very early in pregnancy may improve live birth rate in women with four or more unexplained RPL. Study design, size, duration We performed a retrospective study between September 2013 to March 2023 in two fertility centers. We included 164 patients in the study who experienced 2 or more miscarriages between 5 and 21 weeks of gestation that occurred at the same gestational weeks every time, who had all negative results for our routine RPL work up and still unsuccessful result after treating these conditions, and whose products of conceptus (POC) revealed at least one normal karyotype. Participants/materials, setting, methods IVIG (Venoglobulin IH 5%, Japan Blood Products Organization, 0.4g/kg) injection was performed twice before and after 1 week of GWL. We divided 164 patients into IVIG group and non-IVIG group by their own choice. The primary outcome was live birth rate, and the secondary outcome was modified live birth rate excluding pregnancies with abnormal fetal karyotype, maternal and neonatal complications. All participants provided written informed consent, and Institutional Review Board approval was obtained. Main results and the role of chance IVIG group (n = 116) and non-IVIG group (n = 48) showed no significant differences in the background of patients on BMI (20.3±3.7 and 21.1±2.8), age (37.4±4.2 and 36.7±3.2) and Childbirth history (20.7% and 23.0%), respectively (average±SD), except for the history of past miscarriages. Average numbers of previous miscarriages on IVIG group and non-IVIG group were 3.2±1.3 and 2.5±0.8 (p < 0.001), respectively. After multivariate analysis for BMI, age, Childbirth history and previous miscarriages, live birth rate of IVIG group (70.7%, 82/116) was significantly higher than that of non-IVIG group (60.4%, 29/48; Odds ratio 3.40, 95% CI: 1.49-7.74, p = 0.003). Similarly, modified live birth rate of IVIG group (73.9%, 82/111) was significantly higher than that of non-IVIG group (63.0%, 29/46; Odds ratio 3.40, 95% CI: 1.49-7.74, p = 0.003). Treatment outcomes when IVIG was used for each number of previous miscarriages, the modified birth rate was 88.3% for two times miscarriages, 77.0% for three times, and 57.1% for four or more miscarriages. Maternal complications were observed in 2 cases in IVIG group (chest tightness, obstetric critical bleeding). There were no significant differences in the birth weight, gestational weeks, preterm birth, or fetal growth restriction of newborns between two groups. Limitations, reasons for caution Since retrospective nature and we did not perform POC tests for all patients whose pregnancies had ended in miscarriage, we could not assert the effectiveness of IVIG. We need to conduct randomized controlled trial using placebo group to demonstrate the IVIG efficacy before and after the GWL using euploid blastocysts. Wider implications of the findings IVIG before and after the GWL adjusted for each patient may improve pregnancy outcomes in RPL patients. Although IVIG is expensive, the current method requires only two IVIG injections, that may be acceptable for much more patients who want to avoid miscarriages and who didn’t use IVIG for economic reasons. Trial registration number NA

ADGRL1 variants: From developmental and epileptic encephalopathy to genetic epilepsy with febrile seizures plus.

To expand the phenotypic spectrum of ADGRL1 and explore the correlation between epilepsy and the ADGRL1 genotype. We performed whole-exome sequencing in a cohort of 115 families (including 195 males and 150 females) with familial febrile seizure or epilepsy with unexplained aetiology. The damaging effects of variants was predicted using protein modelling and multiple in silico tools. All reported patients with ADGRL1 pathogenic variants were analysed. One new ADGRL1 variant (p.Pro753Leu) was identified in one family with genetic epilepsy with febrile seizures. Further analysis of 12 ADGRL1 variants in 16 patients revealed that six patients had epilepsy. Epilepsy types ranged from early-onset epileptic encephalopathy to genetic epilepsy with febrile seizures plus (GEFS+). All four variants associated with epilepsy were located in the non-helix or sheet region of ADGRL1. Three of the four epilepsy-associated variants were missense variants. Thus, all three variants located in the G-protein-coupled receptor autoproteolytic-inducing domain exhibited epilepsy. We found one new missense variant of ADGRL1 in one family with GEFS+. ADGRL1 may be a potential candidate or susceptibility gene for epilepsy. ADGRL1-associated epilepsy ranged from benign GEFS+ to severe epileptic encephalopathy; the genotypes and variant locations may help explain the phenotypic heterogeneity of patients with the ADGRL1 variant.

Bridging the Diagnostic Gap between Histopathologic and Hysteroscopic Chronic Endometritis with Deep Learning Models.

Chronic endometritis (CE) is an inflammatory pathologic condition of the uterine mucosa characterized by unusual infiltration of CD138(+) endometrial stromal plasmacytes (ESPCs). CE is often identified in infertile women with unexplained etiology, tubal factors, endometriosis, repeated implantation failure, and recurrent pregnancy loss. Diagnosis of CE has traditionally relied on endometrial biopsy and histopathologic/immunohistochemistrical detection of ESPCs. Endometrial biopsy, however, is a somewhat painful procedure for the subjects and does not allow us to grasp the whole picture of this mucosal tissue. Meanwhile, fluid hysteroscopy has been recently adopted as a less-invasive diagnostic modality for CE. We launched the ARCHIPELAGO (ARChival Hysteroscopic Image-based Prediction for histopathologic chronic Endometritis in infertile women using deep LeArninG mOdel) study to construct the hysteroscopic CE finding-based prediction tools for histopathologic CE. The development of these deep learning-based novel models and computer-aided detection/diagnosis systems potentially benefits infertile women suffering from this elusive disease.

Endoscopically diagnosed hookworm infestation in an adult with chronic iron deficiency anaemia

Despite advances in hookworm control and prevention campaigns, the prevalence remains high in sub-Saharan Africa. Iron deficiency anaemia is a major complication in those with prolonged infection, a high hookworm burden, and undernutrition, though most infected people are asymptomatic. While infected patients are commonly diagnosed by the presence of ova or cysts in the stool, gastrointestinal (GI) endoscopy is required for those with negative stool tests and chronic iron deficiency anaemia. A 48-year-old female with symptoms of anaemia for nearly two years presented to Mbarara Regional Referral Hospital with worsening palpitations, easy fatigability, and dizziness over a period of one month. She was given blood transfusions on two occasions. She lives near the lake where she also gets water for drinking and domestic use. We advise GI endoscopy for all patients with chronic iron deficiency anaemia of unexplained aetiology on standard non-invasive testing. Hookworm’s mass empirical treatment policy in endemic areas should be further emphasised.

Expanding the clinical phenotype and genetic spectrum of GEMIN5 disorders: Early-infantile developmental and epileptic encephalopathies.

Several biallelic truncating and missense variants of the gem nuclear organelle-associated protein 5 (GEMIN5) gene have been reported to cause neurodevelopmental disorders characterized by cerebellar atrophy, intellectual disability, and motor dysfunction. However, the association between biallelic GEMIN5 variants and early-infantile developmental and epileptic encephalopathies (EIDEEs) has not been reported. This study aimed to expand the phenotypic spectrum of GEMIN5 and explore the correlations between epilepsy and molecular sub-regional locations. We performed whole-exome sequencing in two patients with EIDEE with unexplained etiologies. The damaging effects of variants were predicted using multiple in silico tools and modeling. All reported patients with GEMIN5 pathogenic variants and detailed neurological phenotypes were analyzed to evaluate the genotype-phenotype relationship. Novel biallelic GEMIN5 variants were identified in two unrelated female patients with EIDEE, including a frameshift variant (Hg19, chr5:154284147-154284148delCT: NM_015465: c.2551_c.2552delCT: p.(Leu851fs*30)), a nonsense mutation (Hg19, chr5:154299603-154299603delTinsAGA: NM_015465: c.1523delTinsAGA: p.(Leu508*)), and two missense variants (Hg19, chr5:154282663T>A: NM_015465: c.2705T>A: p.(Leu902Gln) and Hg19, chr5:154281002C>G: NM_015465: c.2911C>G: p.(Gln971Glu)), which were inherited from asymptomatic parents and predicted to be damaging or probably damaging using in silico tools. Except p.Leu508*, all these mutations are located in tetratricopeptide repeat (TPR) domain. Our two female patients presented with seizures less than 1 month after birth, followed by clusters of spasms. Brain magnetic resonance imaging suggests dysgenesis of the corpus callosum and cerebellar hypoplasia. Video electroencephalogram showed suppression-bursts. Through a literature review, we found 5 published papers reporting 48 patients with biallelic variants in GEMIN5. Eight of 48 patients have epilepsy, and 5 patients started before 1 year old, which reminds us of the relevance between GEMIN5 variants and EIDEE. Further analysis of the 49 GEMIN5 variants in those 50 patients demonstrated that variants in TPR-like domain or RBS domain were more likely to be associated with epilepsy. We found novel biallelic variants of GEMIN5 in two individuals with EIDEE and expanded the clinical phenotypes of GEMIN5 variants. It is suggested that the GEMIN5 gene should be added to the EIDEE gene panel to aid in the clinical diagnosis of EIDEE and to help determine patient prognosis.

Periodontitis, Dental Procedures, and Young-Onset Cryptogenic Stroke

Periodontitis is associated with an increased risk of ischemic stroke, and the risk may be particularly high among young people with unexplained stroke etiology. Thus, we investigated in a case-control study whether periodontitis or recent invasive dental treatments are associated with young-onset cryptogenic ischemic stroke (CIS). We enrolled participants from a multicenter case-control SECRETO study including adults aged 18 to 49 y presenting with an imaging-positive first-ever CIS and stroke-free age- and sex-matched controls. Thorough clinical and radiographic oral examination was performed. Furthermore, we measured serum lipopolysaccharide (LPS) and lipotechoic acid (LTA) levels. Multivariate conditional regression models were adjusted for stroke risk factors, regular dentist visits, and patent foramen ovale (PFO) status. We enrolled 146 case-control pairs (median age 41.9 y; 58.2% males). Periodontitis was diagnosed in 27.5% of CIS patients and 20.1% of controls (P < 0.001). In the fully adjusted models, CIS was associated with high periodontal inflammation burden (odds ratio [OR], 95% confidence interval) with an OR of 10.48 (3.18–34.5) and severe periodontitis with an OR of 7.48 (1.24–44.9). Stroke severity increased with the severity of periodontitis, having an OR of 6.43 (1.87–23.0) in stage III to IV, grade C. Invasive dental treatments performed within 3 mo prestroke were associated with CIS, with an OR of 2.54 (1.01–6.39). Association between CIS and invasive dental treatments was especially strong among those with PFO showing an OR of 6.26 (1.72–40.2). LPS/LTA did not differ between CIS patients and controls but displayed an increasing trend with periodontitis severity. Periodontitis and recent invasive dental procedures were associated with CIS after controlling for multiple confounders. However, the role of bacteremia as a mediator of this risk was not confirmed.

Anatomical and topographic substantiation of biopsy of intraarticular structures in infectious polyarthritis of cats

Currently, infectious polyarthritis in cats is a very urgent problem in veterinary medicine. Worldwide, one of the most common causes of cat euthanasia is polyarthritis of unexplained etiology, which is not amenable to empirical and symptomatic treatment. The difficulty in diagnosing polyarthritis in cats lies in the large number of infectious agents that can penetrate into joint tissues with blood and lymph flow, while not all regions of the world have a full-fledged diagnosis of these diseases. The second reason complicating the diagnosis of polyarthritis in cats is the absence of infectious agents in the synovial fluid during its examination by various methods such as PCR, bacteriological culture, and ELISA diagnostics. The aim of the study was to consider an open method of biopsy of intraarticular structures in cats with infectious polyarthritis and to identify the safest places of material selection for patients with anatomical and topographic justification. The study was conducted in the Network of veterinary centers "Kotonai" and at the Department of Animal Anatomy of the St. Petersburg State University of Veterinary Medicine. The object of the study was cats in the number of six individuals. The average age of the patients ranged from 3 months to 9 years. There were no severe postoperative complications requiring repeated treatment in the studied patients. The clinical result in these cases can be called positive, despite the presence of minor complications in the early postoperative period. The diagnosis in most patients was established on the basis of examination of biopsies of the articular capsule and articular cartilage. For the collection of material in cats with polyarthritis and altered anatomy in the joint area, the safest localization is craniolateral access to the wrist joint, caudolateral access to the hock joint and cranial access to the hock joint.

Intrauterine fetal death: A hospital based cross sectional study

Introduction: Intrauterine fetal death (IUFD) is not only a medical concern but also a deeply emotional and societal one which affects the intrinsic human desire for healthy and thriving offspring. Therefore, it is important to meticulously identify the potential causal factors contributing to fetal deaths, which will help to ascertain the likelihood of its recurrence, and subsequently, to strategize for prevention or necessary interventions.&#x0D; Objectives: The objective of the study is to describe the socio-epidemiological and etiological characteristics ofIUFD.Methodology: This descriptive cross-sectional study was conducted in the Department of Obstetrics and Gynaecology, Birat Medical College and Teaching Hospital, Biratnagar for a duration of 16 months from May 11st, 2022 to August 31st, 2023. All pregnant women who had IUFD with gestational age of 22 weeks and above were included in the study.&#x0D; Results: During the study period there were total of 5,404 deliveries and among them 73 deliveries were IUFD (13.5 per 1000 live birth). The majority of IUFD, specifically 37 cases (50.4%), were attributed to unexplained etiology. The most common coexisting condition was hypertensive disorder of pregnancy contributing 20.5% followed by Intra-Uterine Growth Retardation (IUGR) in 11(15.1%) cases. &#x0D; Conclusion: There is a significant burden of stillbirths, which will have effect in physical and psychological well-being along with socio-economic impact for grieving parents. Though hypertensive disorder, IUGR and diabetes were the major identified co-morbidities, more than half of the stillbirths were unexplained. This warrants the need of consistent antenatal care (ANC) visits and timely hospital admission for its aversion.

Bone and Tooth: Substrates for Determining Drowning: A New Diagnostic Procedure in Forensic Medicine Practice?

IntroductionDiatom tests are rarely used during autopsy to confirm drowning as the cause of death (COD) because of limitations of the current literature involving these techniques. Instead, experts rely on physical examination by the pathologist. Due to interpretive concerns regarding Diatom tests, they are often insufficient in establishing a diagnosis, but offer the potential to be an extremely useful diagnostic tool with further understanding. The aim of study is to optimize “Diatom Tests” for use in forensic medicine in Bosnia and Herzegovina. MethodsA randomized prospective experimental study was conducted, using albino Wistar rat models (Rattus norvegicus), at the Veterinary Facility, University of Sarajevo. Thirty-two adult albino rats, were used and distributed into groups as follows: Group A (6 deceased rats with COD other than drowning, but due to mechanical asphyxia, which were then submerged for 1 hour after death); Group B (6 deceased rats with COD other than drowning, but due to mechanical asphyxia, which were then submerged for 72 hours after death); Group C (6 rats that were immediately autopsied after drowning, with COD determined as drowning); Group D (6 rats that underwent a 48-hour postmortem period after drowning); Group E (COD: drowning, post-mortem 72hrs after death, remained submerged in water until PM).Live algological material was collected for the research of the systematics of algae from the Bosna river, Sarajevo, and transported to the University of Sarajevo (Department of Biology, Faculty of Science). Periphyllon, epiphyllon and epipelon were used to collect phytobenthos. The material was fixed with 4% formalin solution. Laboratory processing of diatoms was performed using the methods described by Hustedt (16). In the process of obtaining pure diatom valves, part of the material is digested with potassium permanganate (KMnO4), sulfuric acid (H2SO4), and oxalic acid (C2H2O4). In the next step, the cleaned diatom valves were mounted in Canadian balsam. A light microscope under 1000x magnification (Best Scope 2020) was used to evaluate and analyze the species. The identification of diatoms was performed using the reference of Cantonati et al (17). The nomenclature of diatom species was performed according to Guiry & Guiry's worldwide electronic internet database. ResultsNo diatoms were found in Groups A and B. However, Navicula sp. and Sellaphora sp. cf., were discovered during bone analysis of Group C where rats were immediately autopsied after drowning. Hantzschia amphioxus taxon was present in Group D, which underwent a 48-hour postmortem period after drowning and before samples were taken.In Groups C and D, where drowning was the COD, Diatoma vulgaris i Pinnularia major, Achnanthidium minutissimum i Melosira varians were present in the tooth samples. ConclusionOptimization of the “Diatom Test” method could potentially lead to its future use as a routine method within experimental settings. This experimental study is a starting point that guides forenscic medicine pracitioners towards the optimization of tests and sampling in cases of unexplained etiology, where preserved soft tissue structures is not available. In these cases, teeth and bones serve as accessible materials for diagnosing COD, alongside standardized nonspecific findings in the absence of organs for micro- and macroanalysis.

Armor Piercing Projectiles Based on Depleted Uranium and the Consequences of Their Use for the Environment and People

The intention of the collective West to supply the armed forces of Ukraine with armor-piercing shells with cores (penetrators) made of depleted uranium (DU), is changing the situation in the zone of special military operation (SVO). A new damaging factor is introduced into combat operations – uranium-238 (238U), one of the longest-lived natural radioactive isotopes of uranium. The purpose of the review is to identify the signs and consequences of the use of armor-piercing projectiles based on depleted uranium. Materials and research methods. The sources available through the PubMed, Google Scholar and Russian Electronic Library databases were analyzed. Research results. NATO uses DU in 20-, 25-, 30-, 105-, 120- and 140-mm caliber projectiles. The cores are made from recycled DU, which is a waste from the production of nuclear weapons. Due to man-made isotopes, it is more radioactive than DU from natural uranium. When such a projectile hits an armored object, a large amount of respirable radioactive and toxic dust of black uranium oxides, small fragments and fragments of the penetrator, remaining in the armored vehicles and around it, is formed. One 120 mm projectile produces approximately 950 g of black dust. Almost 99 % of the internal dose received by the military will come from alpha particles, the most dangerous to health. Projectiles that miss their targets sink deep into the soil, their penetrators corrode for decades, releasing soluble uranium compounds into underground water sources. In areas where DU shells were used, mass diseases of «unexplained etiology» are observed among military personnel and civilians, reducing their life expectancy and fertility. Discussion of results and conclusions. The first signs of the use of shells with DU, which can be installed on the battlefield: round holes in the armor of tanks and the presence of solid black dust around them and in the tank itself. In case of fires in the warehouses of such shells, due to other oxidation conditions, crumbling yellow dust is formed. When examining it, it is necessary to pay attention to the presence of elevated concentrations of 236U. The fact that a soldier was hit by DU can be confirmed by the presence of uranium in his urine. The use of DU shells on the territory of the Russian Federation, in terms of its consequences for people and nature, is the use of radiological weapons, a disguised form of nuclear warfare. And it must be treated accordingly.

Exposure to per- and polyfluoroalkyl substances in early pregnancy and risk of cerebral palsy in children

BackgroundMost cerebral palsy (CP) cases have an unexplained etiology, but a role for environmental exposures has been suggested. One purported environmental risk factor is exposure to endocrine-disrupting pollutants specifically per- and polyfluoroalkyl substances (PFAS). ObjectivesWe investigated the association between prenatal PFAS exposures and CP in Swedish children. MethodsIn this case-control study, 322 CP cases, 343 population controls, and 258 preterm controls were identified from a birth registry in combination with a CP follow-up program from 1995 to 2014 and linked to a biobank which contains serum samples from week 10–14 of pregnancy. Maternal serum concentrations of four PFAS compounds: perfluorohexane sulfonate (PFHxS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluorooctane sulfonate (PFOS) were analyzed using liquid chromatography-tandem-mass-spectrometry. We estimated odds ratios (ORs) and 95 % confidence intervals (CIs) for CP and each PFAS in quartiles and as continuous variables controlling for various sociodemographic and lifestyle factors. ResultsIn crude and adjusted analyses, we did not find consistent evidence of associations between serum PFHxS, PFOA, PFNA, PFOS and concentrations in early pregnancy and CP, except in preterm infants. The ORs comparing the highest PFAS quartiles to the lowest were 1.05 (95 % CI: 0.63–1.76), 0.96 (95 % CI: 0.55–1.68), 0.71 (95 % CI: 0.41–1.25), and 1.17 (95 % CI: 0.61–2.26), for PFHxS, PFOA, PFNA, and PFOS, respectively. Some positive associations were observed for preterm infants, but the results were imprecise. Similar patterns were observed in analyses treating PFAS as continuous variables. ConclusionsIn this study, we found little evidence that early pregnancy prenatal exposure to PFHxS, PFOA, PFNA, or PFOS increases the risk of CP. However, some positive associations were observed for preterm cases and warrant further investigation.

High doses of intravenous immunoglobulin stimulate regulatory T cell and suppress natural killer cell in women with recurrent pregnancy loss

The aim was to evaluate whether natural killer (NK) cells and regulatory T (Treg) cells were involved in mechanisms underlying beneficial effects of a high dose of intravenous immunoglobulin (IVIG) on recurrent pregnancy losses (RPL) of unexplained etiology. In a double-blind, randomized, placebo-controlled trial of IVIG (400 mg/kg, for 5 days in 4–6 weeks of gestation) in women with RPL, blood samples were collected pre-infusion, one week after infusion (1 w), and eight weeks of gestation/when miscarried (8 w). Levels of NK and Treg cells in peripheral blood were compared between women with IVIG (n = 50) and placebo (n = 49), and between women with IVIG who gave live birth (n = 29) and those who had miscarriage with normal chromosome (n = 12). Effector Treg cell percentages in IVIG group at 1 w (mean 1.43 % vs. 1.03 %) and at 8 w (1.91 % vs. 1.18 %) were higher than those in placebo group (p < 0.01). Total Treg cell percentages in IVIG group at 1 w (4.75 % vs. 4.08 %) and at 8 w (5.55 % vs. 4.47 %) were higher than those in placebo group (p < 0.05). In women with live birth, total Treg cell percentages increased at 8 w (5.52 %, p < 0.001) compared with pre-infusion (4.54 %) and 1 w (4.47 %), while NK cell activity decreased at 1 w (20.18 %, p < 0.001) compared with pre-infusion (26.59 %). IVIG increased Treg cell percentages and suppressed NK cell activity very early in pregnancy, and these were associated with subsequent live birth. Stimulation of Treg cells and suppression of NK cell activity very early in pregnancy may be a mechanism of pharmacological effects of high dose IVIG.

Unearths IFNB1 immune infiltrates in SOP-related ossification of ligamentum flavum pathogenesis

BackgroundOssification of ligamentum flavum (OLF) is a hidden, indolent disease condition with variable unexplained etiology and pathology. Growing evidences show a correlation between senile osteoporosis (SOP) and OLF, but the fundamental relationship between SOP and OLF remains unclear. Therefore, the purpose of this work is to investigate unique SOP-related genes and their potential functions in OLF. MethodsGene Expression Omnibus (GEO) database was utilized to gather the mRNA expression data (GSE106253) and then analyzed by R software. A variety of methods, including ssGSEA, machine learning (LASSO and SVM-RFE), GO and KEGG enrichment, PPI network, transcription factor enrichment analysis (TFEA), GSEA and xCells were employed to verified the critical genes and signaling pathways. Furthermore, ligamentum flavum cells were cultured and used in vitro to identify the expression of the core genes. ResultsThe preliminary identification of 236 SODEGs revealed their involvement in BP pathways associated with ossification, inflammation, and immune response, including the TNF signaling pathway, PI3K/AKT signaling pathway and osteoclast differentiation. Four down-regulated genes (SERPINE1, SOCS3, AKT1, CCL2) and one up-regulated gene (IFNB1) were among the five hub SODEGs that were validated. Additionally, they were performed by ssGSEA and xCell to show the relationship of immune cells infiltrating in OLF. The most fundamental gene, IFNB1, which was only found in the classical ossification- and inflammation-related pathways, suggested that it may affect OLF via regulating the inflammatory response. In vitro experiment, we found that IFNB1 expression was dramatically higher in cells cocultured with osteogenic induction than in controls. ConclusionAs far as we are concerned, this is the first observation using transcriptome data mining to reveal distinct SOP-related gene profiles between OLF and normal controls. Five hub SODEGs were ultimately found using bioinformatics algorithms and experimental verification. These genes may mediate intricate inflammatory/immune responses or signaling pathways in the pathogenesis of OLF, according to the thorough functional annotations. Since IFNB1 was discovered to be a key gene and was connected to numerous immune infiltrates in OLF, it is possible that IFNB1 expression has a substantial impact on the pathogenesis of OLF. Our research will give rise to new possibilities for potential therapeutics that target SOP reverent genes and immune-associated pathways in OLF.

Inhibition of TMUB1 blocks apoptosis and NF-κB pathway-mediated inflammation in recurrent spontaneous abortion.

Approximately 50% of cases with recurrent spontaneous abortion (RSA) have unexplained etiology. Aberrant expression of transmembrane and ubiquitin-like domain containing 1 (TMUB1) is closely related to a series of diseases, including RSA. However, the function and underlying mechanism of TMUB1 in the occurrence of RSA has not been described. TMUB1 expression was detected in the placental villous tissues of 30 women with normal miscarriages and 12 women with RSA. The pregnant mice were injected intraperitoneally with lipopolysaccharide (LPS) to induce abortion. Human chorionic trophoblast cells were treated with LPS. Pathological analysis of placental tissues was performed by hematoxylin and eosin staining. TMUB1 was highly expressed in the placental villous tissues of RSA patients compared to the patients who underwent induced abortions. After LPS administration, the mice exhibited high embryo absorption and pathological alterations, as well as presented an increase in inflammation and apoptosis (the etiology of RSA induction) in placental tissues. Moreover, the upregulated expression of TMUB1 was also found in placental tissues of LPS-induced mice, and further investigation showed that TMUB1 deficiency blocked embryo loss as well as inhibited apoptotic rate and inflammation after LPS activation. Furthermore, we found that the loss of TMUB1 suppressed the phosphorylation of IkappaB kinase (IKK) α/β and attenuated cytoplasmic-nuclear translocation of nuclear factor-κB (NF-κB) p65 in LPS-induced cells. Our results indicate that TMUB1 may involve in the modulation of apoptosis and NF-κB pathway-mediated inflammation in RSA. Therefore, TMUB1 may develop as a potential biomarker for RSA treatment.

A Rare Cause of Small Bowel Obstruction: Congenital Peritoneal Encapsulation

Congenital peritoneal encapsulation (CPE) is a rare condition in which part or all of the small intestine is surrounded by an accessory peritoneal layer congenitally. Although it rarely causes small bowel obstruction, it is usually asymptomatic and the diagnosis is mostly made incidentally during surgery or autopsy. A 41-year-old male patient presented to the emergency department with diffuse and cramping pain lasting for approximately 8 hours. He had nausea and vomiting. No gas or faeces output for 72 hours. Abdominal computed tomography (CT) showed dilated abdominal small intestines and findings consistent with obstruction. The patient was hospitalized with the diagnosis of ileus. Decompression was performed with a nasogastric tube. It was decided to perform diagnostic laparoscopic surgery for the patient who did not respond to 24-hour observation and medical treatment. Laparoscopic examination revealed a thin membrane covering the small intestine from the terminal ileum to the middle of the jejunal segment on the right side of the abdomen. All adhesions were separated, the small intestines were released from the pressure of the accessory peritoneum and placed in the abdomen. The patient was discharged without complications on the 6th postoperative day. CPE should be considered in small bowel obstructions of unexplained etiology. Laparoscopic evaluation is effective in diagnosing CPE, but in cases where the long small bowel segment is affected, as in our case, we think that it would be appropriate to switch to open surgery to prevent morbidity, as well as the necessity of separating all bands

Successful Pregnancy and Delivery at Term Following Intravenous Immunoglobulin Therapy with Heparin for Unexplained Recurrent Pregnancy Loss Suspected of Immunological Abnormalities: A Case Report and Brief Literature Review

About 60% of cases of recurrent pregnancy loss have unexplained etiology. Immunotherapy for unexplained recurrent pregnancy loss is still unestablished. A 36-year-old woman, not obese, had a stillbirth at 22 gestational weeks and a spontaneous abortion at 8 weeks. She had been examined for recurrent pregnancy loss at previous clinics with no significant findings. When she visited our clinic, a hematologic test showed a Th1/Th2 ratio imbalance. Ultrasonography, hysteroscopy, and semen analysis showed no abnormalities. She successfully conceived by embryo transfer in hormone replacement therapy cycle. However, she had a miscarriage at 19 weeks. The baby had no deformities, but a chromosomal test was not performed, according to the parents' will. The placenta pathologically suggested hemoperfusion problems. Her and her husband's chromosomal tests showed normal karyotypes. Other examinations revealed a repeated Th1/Th2 ratio imbalance and a high resistance index of uterine radial artery blood flow. She was administered low-dose aspirin, intravenous immunoglobulin, and unfractionated heparin after the second embryo was transferred. Her baby was healthily born by cesarean section at 40 weeks. Intravenous immunoglobulin therapy can be a choice for recurrent miscarriage without risk factors because it has clinically beneficial influences on the patient's immunological aberration.

The persistent viral infections in the development and severity of myalgic encephalomyelitis/chronic fatigue syndrome

BackgroundMyalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a multifactorial disease with an unexplained aetiology in which viral infections are possible trigger factors.The aim of this study was to determine the involvement of human herpesvirus (HHV)-6A/B, HHV-7, and parvovirus B19 (B19V) in the etiopathogenesis of ME/CFS.Methods200 patients with clinically diagnosed ME/CFS and 150 apparently healthy individuals were enrolled in this study. Single-round, nested, and quantitative real-time polymerase chain reactions (PCR) were used to detect the presence and load of HHV-6A/B, HHV-7, and B19V. HHV-6A and HHV-6B were distinguished by PCR and restriction analysis. Immunoenzymatic assays were applied to estimate the presence of virus-specific antibodies and the level of cytokines.ResultsHHV-6A/B, HHV-7, and B19V specific antibodies were detected among patients and healthy individuals in 92.1% and 76.7%, 84.6% and 93.8%, and 78% and 67.4% of cases. HHV-6B had 99% of HHV-6 positive patients.Latent HHV-6A/B, HHV-7, and B19V infection/co-infection was observed in 51.5% of the patients and 76.7% of the healthy individuals, whereas active–45% of the ME/CFS patients and 8.7% of healthy individuals. HHV-6A/B load in patients with a persistent infection/co-infection in a latent and active phase was 262 and 653.2 copies/106 cells, whereas HHV-7 load was 166.5 and 248.5 copies/106 cells, and B19V-96.8 and 250.8 copies/106 cells, respectively.ME/CFS patients with persistent infection in an active phase had a higher level of pro-inflammatory cytokines (interleukin(IL)-6, tumor necrosis factor-alpha(TNF-α) and IL-12) and anti-inflammatory (IL-10) than with a persistent infection in a latent phase. A significant difference was revealed in the levels of TNF-α, IL-12, and IL-10 among the patient groups without infection, with latent infection/co-infection, active single, double and triple co-infection. The levels of TNF-α, IL-12, and IL-10 are significantly higher in patients with severe compared with a moderate course of ME/CFS.ConclusionsSignificantly more persistent HHV-6A/B, HHV-7, and B19V infection/co-infection in an active phase with a higher viral load and elevated levels of pro- and anti-inflammatory cytokines among patients with ME/CFS than healthy individuals indicate the importance of these infections/co-infections in ME/CFS development. The presence of these infections/co-infections influences the ME/CFS clinical course severity.

Ulcerative colitis – recent and potential methods of treatment - review

Introduction and purpose: Ulcerative colitis (UC, colitis ulcerosa) belongs to a group of inflammatory bowel diseases of unexplained etiology. It is a diffuse nonspecific inflammation of the mucosa of the rectum or rectum and colon, leading to ulceration in more severe cases. Despite a decline in the incidence of ulcerative colitis in developed countries, the prevalence continues to rise due to increasingly predominant benign course of the disease with a low mortality rate. The purpose of this review is to summarize latest directions and discoveries in research of ulcerative colitis. Materials and methods: This review is based on articles published in Pubmed between 2017 and 2022 by searching keywords ulcerative colitis treatment, ulcerative colitis guidelines.State of knowledge: Currently, many independent scientific societies represent a consensus position on the standard management of patients with UC, while recent therapeutic options are constantly emerging due to intensive research. These options require the involvement of multiple medical centers to determine their position in future guidelines.Conclusion: Although ulcerative colitis is a disease that is diagnosed and detected relatively easily, due to its chronic nature, relatively high prevalence in the population and lack of causal treatment options, it is an important direction for further research.