Keratoconus (KC) is a progressive corneal thinning disease that manifests in puberty and worsens during pregnancy. KC onset and progression are attributed to diverse factors that include: environmental, genetics, and hormonal imbalances; however, the pathobiology remains elusive. This study aims to determine the role of corneal stroma sex hormone receptors in KC and their interplay with estrone (E1) and estriol (E3) using our established 3D in vitro model. Healthy cornea stromal cells (HCFs) and KC cornea stromal cells (HKCs), both male and female, were stimulated with various concentrations of E1 and E3. Significant changes were observed between cell types, as well as between males and females in the sex hormone receptors tested; androgen receptor (AR), progesterone receptor (PR), estrogen receptor alpha (ERα), and estrogen receptor beta (ERβ) using Western blot analysis. E1 and E3 stimulations in HCF females showed AR, PR, and ERβ were significantly upregulated compared to HCF males.
In contrast, ERα and ERβ had significantly higher expression in HKC’s females than HKC’s males. Our data suggest that the human cornea is a sex-dependent, hormone-responsive tissue that is significantly influenced by E1 and E3. Therefore, it is plausible that E1, E3, and sex hormone receptors are involved in the KC pathobiology, warranting further investigation.
Compatibility of Estradiol, Estriol, Estrone, Progesterone, and Testosterone Single Formulation in Fitalite, Versatile, or HRT Supreme Cream Base
In this work, we focus on three ready-to-use vehicles: Fitalite, Versatile, and HRT Supreme Cream Base. Fitalite is a natural, light, hydrophilic gel-cream that contains vitamin E and oil bodies from plant sources (phytosomes), providing antioxidant and skinmoisturizing properties. Versatile is a vanishing oil-inwater cream base which retains its consistency with a broad range and high concentrations of active pharmaceutical ingredients, dermaceutical ingredients, and solvents. Finally, HRT Supreme Cream Base is a paraben-free, dye-free, fragrance-free O/W emulsion base, formulated with a complex of botanical oils to soothe and provide moisture to dry and sensitive skin. In the current study, we evaluated the beyond-use date of formulations containing estradiol, estriol, estrone, progesterone, and testosterone in combination, compounded with these three vehicles.
Validated, stability-indicating high-performance liquid chromatography methods were used throughout a 180-day period. A beyond-use date of 180 days was observed for all vehicles stored both at refrigerated and at room temperature. The combination of five ingredients represents a worst-case scenario since there are more possibilities of cross reactions. Therefore, we expect the same or greater stability as individual ingredients are removed from the tested formulation. The extended beyond-use dates provide convenience for both the compounding pharmacist and the patient.
A technical and clinical evaluation of the new ThermoFisher BRAHMS unconjugated estriol and inhibin-A assays and their use in second trimester Down syndrome screening
To evaluate second-trimester Down syndrome screening performance of the new ThermoFisher BRAHMS GOLD unconjugated estriol (uE3) and inhibin-A assays. Serum samples were analyzed for levels of uE3 and inhibin-A using the ThermoFisher BRAHMS GOLD immunoanalyzer and compared to other platforms. Levels were transformed to multiples of the median (MoM) in unaffected pregnancies. Log10 MoM distributions in unaffected and Down syndrome pregnancies were assessed for central tendency (mean) and dispersion (SD). Empirical and estimated screening performances were determined. Correlation between BRAHMS and AutoDELFIA® uE3 and inhibin-A were 0.63 and 0.97, respectively, the respective mean difference was 31.3% [95%CI 50.2% to -112.8%] and -23.3% [95%CI -41.9% to -4.7%]. Passing-Bablok indicated significant systematic (-2.78 [95%CI -3.57 to -2.04]) and proportional bias (1.30 [95%CI 1.15 to -1.47]) between uE3 assays and significant proportional bias (0.71[95%CI 0.65-0.78]) between inhibin-A assays.
The uE3 and inhibin-A log10 MoM distribution mean [SD] in unaffected and Down syndrome pregnancies were 0.0024 [SD = 0.2341] and -0.0001 [SD = 0.2078], and -0.2028 [SD = 0.2495] and 0.3645 [SD = 0.2576], respectively. The new BRAHMS uE3 and inhibin-A assays had an 81-83% detection rate for Trisomy21 for a 5% false-positive rate. The new BRAHMS assays achieved the expected screening performance provided the risk estimation model is adjusted to account for the higher BRAHMS uE3 MoM measurement distribution variance.
Estriol dissolving microneedle patches for protection against ionizing radiation-induced injury
Estriol can be used to treat radiation-induced leukopenia by increasing peripheral blood leukocytes and therefore it plays an important role in radiation protection. However, only high-dose injectable suspensions are available when estriol is used to combat against ionizing radiation-induced injury. Intramuscular (i.m.) administration of estriol is very painful and inconvenient, and the lack of timely self-administered formulation greatly limits the wide application of estriol. This will facilitate quick response under emergent conditions in complementary with the available estriol formulations. Herein, we prepared estriol microneedle (MNs) patches for the convenient and efficient treatment of radiation-induced injury.
A biocompatible polymer, polyvinylpyrrolidone K90, was dissolved in an estriol solution of methanol and cast into a mold to obtain conical-shaped MNs. N-vinyl pyrrolidone was poured on the base of the MNs and photocured to enhance the mechanical strength of estriol MNs (EMNs). EMNs were easily pierced 200 μm into the mouse skin. More importantly, the EMNs tips were dissolved very quickly within 5 min so that the drugs could permeate across skin. Mouse models of ionizing radiation-induced injury were established with 6.5 Gy radiation of 60Co γ ray. Moreover, EMNs increased peripheral blood leukocytes in irradiated mice, protected the bone marrow hematopoietic system, and improved the survival rate of the irradiated mice to 80 %. EMNs are a promising transdermal drug delivery system that allows for easy, rapid administration and protects the body from damage caused by ionizing radiation.
Cortisol and estriol responses to awakening in the first pregnancy trimester: Associations with maternal stress and resilience factors
Background: Little is known about the maternal cortisol awakening response (CAR) in the first pregnancy trimester. Similarly unknown is how the CAR in early gestation relates to other steroid hormones, such as estriol. Maternal estriol in blood and urine is used to monitor fetal well-being since it is produced by the fetoplacental unit from fetal precursors. Low levels have been associated with maternal-fetal complications. We were recently able to show that estriol is measurable in maternal saliva from 6 weeks’ gestation onwards. However, its pattern following morning awakening and potential links with salivary cortisol in early gestation is relatively unknown. In this prospective study, we explored the cortisol and estriol responses to morning awakening in first-trimester pregnant women, the potential association of these endocrine variables with maternal stress and resilience factors, and their predictive value for the further pregnancy course.
Methods: Fifty-one women with an uncomplicated, singleton pregnancy responded to questionnaires measuring chronic and pregnancy-specific stress, emotional support, and daily uplifts at 6 weeks’ gestation. At 8 and 10 weeks, the women collected saliva samples immediately, 30, and 60 min after morning awakening. After 12 weeks, 40 women reported on the further pregnancy course, of whom 6 had developed complications.
Results: In response to morning awakening, cortisol levels increased significantly at 10 weeks (p = .04), while estriol levels decreased significantly at both 8 and 10 weeks (p < .001). A stronger cortisol increase was linked to a stronger estriol decrease at 8 (p = .03), but not at 10 weeks. Then, perceived emotional support at 6 weeks was negatively associated with cortisol baseline at 8 (p = .01) and positively with estriol baseline at 10 weeks (p = .03). Moreover, higher pregnancy-specific stress was related to a lower estriol baseline at 8 weeks (p = .047). Furthermore, compared to healthy women, those with complications at follow-up had already reported less emotional support (p = .03) and fewer daily uplifts (p = .03) at 6 weeks. These women also seemed to lack a significant estriol response to morning awakening at 8 weeks (p > .10).
Discussion: These findings advance our knowledge of cortisol and estriol secretion following morning awakening and encourage the investigation of E3 in addition to cortisol when researching prenatal stress and its consequences for maternal and fetal health.
Identification and evaluation of long-term markers is crucial in prolonging the detection window for anabolic steroid abuse in sport. Recently, sulfoconjugated epiandrosterone was identified as a potential long-term marker for the abuse of certain endogenous anabolic agents, including testosterone, which continues to be widely used as a performance enhancing agent in sport. To evaluate the applicability of epiandrosterone sulfate as a marker for testosterone use, administration studies were conducted with multiple modes of testosterone administration – transdermal, intramuscular, and subcutaneous.
A modified sample preparation method was used to collect both glucuronidated and sulfoconjugated analytes of interest. Carbon isotope ratio measurements from the administration studies are presented here. Epiandrosterone was less effective than the conventionally used target compounds for detection of the low dose application (transdermal gel). With intramuscular administration, epiandrosterone was more diagnostic than with transdermal administration, but it did not prolong the detection window more than the conventional target compounds.
With subcutaneous administration, the doses administered to the subjects were varied and the effect in the epiandrosterone values was dependent on the magnitude of the dose administered. Epiandrosterone does not appear to be a useful marker in the detection of low dose testosterone administration. It is responsive to higher dose administration, but it does not provide extension of the detection window relative to conventional target compounds.
Development and validation of a fast gas chromatography combustion isotope ratio mass spectrometry method for the detection of epiandrosterone sulfate in urine.
In doping control, to confirm the exogenous origin of exogenously administered anabolic androgenic steroids (AAS), a gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) analysis is performed. Recently published work suggests epiandrosterone sulfate (EpiAS) as a promising IRMS target compound for the detection of AAS, capable of prolonging the detection window. However, EpiAS is only excreted in urine in its sulfoconjugated form while all other IRMS target compounds are excreted glucuronidated, meaning that EpiAS cannot be incorporated in the existing IRMS methods.
A separate extensive sample preparation needs to be performed on this compound with a different hydrolysis and extraction procedure and a different liquid chromatography (LC) clean-up. The current work presents a new, fast and easy to implement EpiAS IRMS method. The approach was based on the direct GC analysis of non-hydrolysed EpiAS, making the solid phase extraction, hydrolysis and acetylation step redundant.
Sample preparation consisted of a simple liquid liquid extraction, followed by LC fraction collection. A population study was performed to check the compliance with the criteria drafted by the World Anti-Doping Agency (WADA). To verify the applicability of the developed approach, the method was applied to the samples of four administration studies (i.e., dehydroepiandrosterone (DHEA), testosterone gel (T gel), androstenedione (ADION) and intramuscular testosterone undecanoate. In contrast to previously published data, the strength of EpiAS as target compound and the prolongation of the detection window in comparison with the conventional IRMS target compounds was less pronounced.
Synthesis and bioevaluation of novel steroidal isatin conjugates derived from epiandrosterone/androsterone.
Steroids are classes of natural products widely distributed in nature, which have been demonstrated to exhibit broad biological functions, and have also attracted increasing interest from bioorganic and pharmaceutical researches. In order to develop novel chemical entities as potential cytotoxic agents, a series of steroidal isatin conjugations derived from epiandrosterone and androsterone were efficiently prepared and characterized, and all these obtained compounds were screened for their potential cytotoxic activities.
The preliminary bioassay indicated that most of the newly synthesized compounds exhibited good cytotoxic activities against human gastric cancer (SGC-7901), melanoma (A875), and hepatocellular liver carcinoma (HepG2) cell lines compared with 5-fluorouracil (5-FU), which might be considered as promising scaffold for further development of potential anticancer agents.
The quantitation of 7beta-hydroxy-epiandrosterone in the plasma and seminal plasma of men with different degrees of fertility.
7beta-hydroxy-epiandrosterone (7beta-OH-EpiA) is an endogenous androgen metabolite that has been shown to exert neuroprotective, anti-inflammatory and anti-estrogenic effects. However, to the best of our knowledge no information is available about this androgen steroid in relation to sperm quality. We analyzed 7beta-OH-EpiA in plasma and seminal plasma using a newly developed isotope dilution ultra-high performance liquid chromatography – mass spectrometry method. Validation met the requirements of FDA guidelines. Levels of 7beta-OH-EpiA were measured in 191 men with different degrees of infertility. One-way analysis of variance followed by multiple comparison and correlation analysis adjusted for age, BMI and abstinence time were performed to evaluate the relationships between this steroid and sperm quality.
Concentrations of 7beta-OH-EpiA in seminal plasma were significantly higher in severely infertile men in comparison with healthy men and slightly infertile men. The same trend was found when blood plasma was evaluated. Furthermore, plasma 7beta-OH-EpiA negatively correlated with sperm concentration (-0.215; p<0.01) and total count (-0.15; p<0.05). Seminal 7beta-OH-EpiA was negatively associated with motility (-0.26; p<0.01), progressively motile spermatozoa (-0.233; p<0.01) and nonprogressively motile spermatozoa (-0.188; p<0.05). 7beta-OH-EpiA is associated with lower sperm quality and deserves more research in that respect.
Epiandrosterone sulfate prolongs the detectability of testosterone, 4-androstenedione, and dihydrotestosterone misuse by means of carbon isotope ratio mass spectrometry.
In the course of investigations into the metabolism of testosterone (T) by means of deuterated T and hydrogen isotope ratio mass spectrometry, a pronounced influence of the oral administration of T on sulfoconjugated steroid metabolites was observed. Especially in case of epiandrosterone sulfate (EPIA_S), the contribution of exogenous T to the urinary metabolite was traceable up to 8 days after a single oral dose of 40 mg of T. These findings initiated follow-up studies on the capability of EPIA_S to extend the detection of T and T analogue misuse by carbon isotope ratio (CIR) mass spectrometry in sports drug testing. Excretion study urine samples obtained after transdermal application of T and after oral administration of 4-androstenedione, dihydrotestosterone, and EPIA were investigated regarding urinary concentrations and CIR.
With each administered steroid, EPIA_S was significantly depleted and prolonged the detectability when compared to routinely used steroidal target compounds by a factor of 2 to 5. In order to simplify the sample preparation procedure for sulfoconjugated compounds, enzymatic cleavage by Pseudomonas aeruginosa arylsulfatase was tested and implemented into CIR measurements for the first time. Further simplification was achieved by employing multidimensional gas chromatography to ensure the required peak purity for CIR determinations, instead of sample purification strategies using liquid chromatographic fractionation. Taking into account these results that demonstrate the unique and broad applicability of EPIA_S for the detection of illicit administrations of T or T-related steroids, careful consideration of how this steroid can be implemented into routine doping control analysis appears warranted.
An omics investigation into chronic widespread musculoskeletal pain reveals epiandrosterone sulfate as a potential biomarker.
Chronic widespread musculoskeletal pain (CWP) is common, having a population prevalence of 10%. This study aimed to define the biological basis of the CWP/body mass association by using a systems biology approach. Adult female twins (n = 2444) from the TwinsUK registry who had extensive clinical, anthropometric, and “omic” data were included. Nontargeted metabolomics screening including 324 metabolites was carried out for CWP and body composition using dual-energy X-ray absorptiometry. The biological basis of these associations was explored through a genome-wide association study and replicated in an independent population sample (Cooperative Health Research in the Region of Augsburg [KORA] study, n = 2483). A causal role for the genetic variants identified was sought in CWP using a Mendelian randomisation study design.
Fat mass/height2 was the body composition variable most strongly associated with CWP (TwinsUK: P = 2.4 × 10(-15) and KORA: P = 1.59 × 10(-10)). Of 324 metabolites examined, epiandrosterone sulfate (EAS) was highly associated with both CWP (P = 1.05 × 10(-09) in TwinsUK and P = 3.70 × 10(-06) in KORA) and fat mass/height2. Genome-wide association study of EAS identified imputed single nucleotide polymorphism rs1581492 at 7q22.1 to be strikingly associated with EAS levels (P ≤ 2.49 × 10(-78)), and this result was replicated in KORA (P = 2.12 × 10(-9)). Mendelian randomization by rs1581492 genotype showed that EAS is unlikely to be causally related to CWP. Using an agnostic omics approach to focus on the association of CWP with body mass index, we have confirmed a steroid hormone association and identified a genetic variant upstream of the CYP genes, which likely controls this response. This study suggests that steroid hormone abnormalities result from pain rather than causing it, and EAS may provide a biomarker that identifies subgroups at risk of CWP.
An omics investigation into chronic widespread musculoskeletal pain reveals epiandrosterone sulfate as a potential biomarker.
Chronic widespread musculoskeletal pain (CWP) is common, having a population prevalence of 10%. This study aimed to define the biological basis of the CWP/body mass association by using a systems biology approach. Adult female twins (n = 2444) from the TwinsUK registry who had extensive clinical, anthropometric, and “omic” data were included. Nontargeted metabolomics screening including 324 metabolites was carried out for CWP and body composition using dual-energy X-ray absorptiometry. The biological basis of these associations was explored through a genome-wide association study and replicated in an independent population sample (Cooperative Health Research in the Region of Augsburg [KORA] study, n = 2483). A causal role for the genetic variants identified was sought in CWP using a Mendelian randomisation study design.
Fat mass/height2 was the body composition variable most strongly associated with CWP (TwinsUK: P = 2.4 × 10(-15) and KORA: P = 1.59 × 10(-10)). Of 324 metabolites examined, epiandrosterone sulfate (EAS) was highly associated with both CWP (P = 1.05 × 10(-09) in TwinsUK and P = 3.70 × 10(-06) in KORA) and fat mass/height2. Genome-wide association study of EAS identified imputed single nucleotide polymorphism rs1581492 at 7q22.1 to be strikingly associated with EAS levels (P ≤ 2.49 × 10(-78)), and this result was replicated in KORA (P = 2.12 × 10(-9)). Mendelian randomization by rs1581492 genotype showed that EAS is unlikely to be causally related to CWP. Using an agnostic omics approach to focus on the association of CWP with body mass index, we have confirmed a steroid hormone association and identified a genetic variant upstream of the CYP genes, which likely controls this response. This study suggests that steroid hormone abnormalities result from pain rather than causing it, and EAS may provide a biomarker that identifies subgroups at risk of CWP.
Currently, the only definitive method for diagnosing ovarian cancer involves histological examination of tissue obtained at time of surgery or by invasive biopsy. Blood has traditionally been the biofluid of choice in ovarian cancer biomarker discovery; however, there has been a growing interest in exploring urinary biomarkers, particularly as it is non-invasive. In this systematic review, we present the diagnostic accuracy of urinary biomarker candidates for the detection of ovarian cancer. A comprehensive literature search was performed using the MEDLINE/PubMed and EMBASE, up to 1 st April 2021. All included studies reported the diagnostic accuracy using sensitivity and/or specificity and/or receiver operating characteristics (ROC) curve. Risk of bias and applicability of included studies were assessed using the QUADAS-2 tool.
Twenty seven studies were included in the narrative synthesis. Protein/peptide biomarkers were most commonly described (n=18), with seven studies reporting composite scores of multiple protein-based targets. The most frequently described urinary protein biomarker was HE4 (n=5), with three studies reporting a sensitivity and specificity >80%. Epigenetic (n=1) and metabolomic/organic compound biomarkers (n=8) were less commonly described. Overall, six studies achieved a sensitivity and specificity of >90% and/or an AUC >0.9. Evaluation of urinary biomarkers for the detection of ovarian cancer is a dynamic and growing field. Currently, the most promising biomarkers are those that interrogate metabolomic pathways and organic compounds, or quantify multiple proteins. Such biomarkers require external validation in large, prospective observational studies before they can be implemented into clinical practice.
Rapid electrostatic DNA enrichment for sensitive detection of Trichomonas vaginalis in clinical urinary samples
Estimated to be the most common non-viral sexually transmitted infection globally, Trichomonas vaginalis (TV) can lead to pelvic inflammatory disease, pregnancy complications, and increased risk of acquiring and transmitting HIV. Once diagnosed, TV infection can be treated with oral antibiotics; however, infected individuals are often asymptomatic and do not seek treatment. The WHO and others have identified a need for point-of-care tests to expand access to TV testing and screening; ideal test characteristics include high sensitivity and specificity and the ability to use urine as a sample type, rather than invasively collected swab samples. Here, we report on a proof-of-concept prototype for rapid, electrostatic enrichment of DNA from urine samples and demonstrate the use of large volumes of urine to increase sensitivity of downstream nucleic acid amplification testing.
We developed an internally controlled thermophilic helicase-dependent amplification (tHDA) assay with lateral flow immunoassay readout and demonstrate that this tHDA assay can be performed directly on our DNA capture filter. We validated our method using clinical urine samples with qPCR-quantified TV loads. Using 62 clinical urine samples and a simple sample processing device, our tHDA assay displayed 96.6% sensitivity and 100% specificity. Our analytical limit of detection was found to be approximately 7 genomic equivalents of TV DNA per mL of sample when 1 mL of sample was tested, comparable to existing isothermal tests for TV. Using large-volume simulated samples (40 mL of buffered urine with spiked-in TV DNA), we also demonstrated that sensitivity could be improved 28-fold to 0.25 genomic equivalents of TV DNA per mL, with a sample processing time of only 2 minutes.
Nanomaterials-Based Urinary Extracellular Vesicles Isolation and Detection for Non-invasive Auxiliary Diagnosis of Prostate Cancer
Extracellular vesicles (EVs) are natural nanoparticles secreted by cells in the body and released into the extracellular environment. They are associated with various physiological or pathological processes, and considered as carriers in intercellular information transmission, so that EVs can be used as an important marker of liquid biopsy for disease diagnosis and prognosis. EVs are widely present in various body fluids, among which, urine is easy to obtain in large amount through non-invasive methods and has a small dynamic range of proteins, so it is a good object for studying EVs. However, most of the current isolation and detection of EVs still use traditional methods, which are of low purity, time consuming, and poor efficiency; therefore, more efficient and highly selective techniques are urgently needed. Recently, inspired by the nanoscale of EVs, platforms based on nanomaterials have been innovatively explored for isolation and detection of EVs from body fluids.
These newly developed nanotechnologies, with higher selectivity and sensitivity, greatly improve the precision of isolation target EVs from urine. This review focuses on the nanomaterials used in isolation and detection of urinary EVs, discusses the advantages and disadvantages between traditional methods and nanomaterials-based platforms, and presents urinary EV-derived biomarkers for prostate cancer (PCa) diagnosis. We aim to provide a reference for researchers who want to carry out studies about nanomaterial-based platforms to identify urinary EVs, and we hope to summarize the biomarkers in downstream analysis of urinary EVs for auxiliary diagnosis of PCa disease in detail.
Rational Design of Phe-BODIPY Amino Acids as Fluorogenic Building Blocks for Peptide-based Detection of Urinary Tract Candida Infections
Fungal infections caused by Candida species are among the most prevalent in hospitalized patients. However, current methods for the detection of Candida fungal cells in clinical samples rely on time-consuming assays that hamper rapid and reliable diagnosis. Herein, we describe the rational development of new Phe-BODIPY amino acids as small fluorogenic building blocks and their application to generate fluorescent antimicrobial peptides for rapid labelling of Candida cells in urine.
We have used computational methods to analyse the fluorogenic behaviour of BODIPY-substituted aromatic amino acids and performed bioactivity and confocal microscopy experiments in different strains to confirm the utility and versatility of peptides incorporating Phe-BODIPYs. Finally, we have designed a simple and sensitive fluorescence-based assay for the detection of Candida albicans in human urine samples.
A Prospective Multicenter Trial to Evaluate Urinary Metabolomics for Non-invasive Detection of Renal Allograft Rejection (PARASOL): Study Protocol and Patient Recruitment
Background: In an earlier monocentric study, we have developed a novel non-invasive test system for the prediction of renal allograft rejection, based on the detection of a specific urine metabolite constellation. To further validate our results in a large real-world patient cohort, we designed a multicentric observational prospective study (PARASOL) including six independent European transplant centers. This article describes the study protocol and characteristics of recruited better patients as subjects.
Methods: Within the PARASOL study, urine samples were taken from renal transplant recipients when kidney biopsies were performed. According to the Banff classification, urine samples were assigned to a case group (renal allograft rejection), a control group (normal renal histology), or an additional group (kidney damage other than rejection).
Results: Between June 2017 and March 2020, 972 transplant recipients were included in the trial (1,230 urine samples and matched biopsies, respectively). Overall, 237 samples (19.3%) were assigned to the case group, 541 (44.0%) to the control group, and 452 (36.7%) samples to the additional group. About 65.9% were obtained from male patients, the mean age of transplant recipients participating in the study was 53.7 ± 13.8 years. The most frequently used immunosuppressive drugs were tacrolimus (92.8%), mycophenolate mofetil (88.0%), and steroids (79.3%). Antihypertensives and antidiabetics were used in 88.0 and 27.4% of the patients, respectively. Approximately 20.9% of patients showed the presence of circulating donor-specific anti-HLA IgG antibodies at time of biopsy. Most of the samples (51.1%) were collected within the first 6 months after transplantation, 48.0% were protocol biopsies, followed by event-driven (43.6%), and follow-up biopsies (8.5%). Over time the proportion of biopsies classified into the categories Banff 4 (T-cell-mediated rejection [TCMR]) and Banff 1 (normal tissue) decreased whereas Banff 2 (antibody-mediated rejection [ABMR]) and Banff 5I (mild interstitial fibrosis and tubular atrophy) increased to 84.2 and 74.5%, respectively, after 4 years post transplantation. Patients with rejection showed worse kidney function than patients without rejection.
Conclusion: The clinical characteristics of subjects recruited indicate a patient cohort typical for routine renal transplantation all over Europe. A typical shift from T-cellular early rejections episodes to later antibody mediated allograft damage over time after renal transplantation further strengthens the usefulness of our cohort for the evaluation of novel biomarkers for allograft damage.
Background: The sarcopenia index (SI), calculated as the ratio of serum creatinine to cystatin C levels, reflects skeletal muscle mass and strength. Patients with hip fracture (HF) and sarcopenia have poor functional outcomes, and many require long-term care after surgery. We hypothesized that the SI can predict preoperative and early postoperative functional outcomes.
Methods: Preoperative serum creatinine and cystatin C were measured to calculate the SI for patients with surgically treated HF (n = 130, mean age: 87.8 ± 6.9 years). Walking ability before and 2 weeks after surgery was assessed, and patients were dichotomized into independent and assistance groups. To assess the validity of the SI, we examined its correlation with the quality [computed tomography (CT) value] and quantity (cross-sectional area) of the muscles around the hip on the non-operated side, which were preoperatively measured using CT. Receiver operating characteristic (ROC) analysis was performed to evaluate the prognostic value of the SI.
Results: The SI of the preoperative independent (n = 77) and assistance groups (n = 53) significantly differed (70.2 ± 12.4 and 60.1 ± 9.8, respectively, P < 0.000001). At 2 weeks after surgery, the SI was significantly higher in the independent group (n = 31, 73.0 ± 14.9) than in the assistance group (n = 99, 64.0 ± 10.7, P = 0.0003). In the preoperative independent group, 28 could walk independently after surgery (SI: 74.8 ± 14.0) while 49 required assistance (SI: 67.7 ± 10.6, P = 0.01). For patients with femoral neck fracture (FNF), the SIs were significantly higher in the postoperative independent group (78.6 ± 15.7) than in the postoperative assistance group (63.2 ± 10.9, P = 0.002). Logistic regression analysis showed that the odds ratio (95% confidence interval) of the SI for postoperative walking ability was 0.95 (0.91-0.99, P = 0.03). The correlations of SIs with CT values and cross-sectional areas were as follows: iliopsoas at the apex of the femoral head, r = 0.40, P < 0.001 and r = 0.49, P < 0.001, respectively; rectus femoris at the level of the lessor trochanter, r = 0.26, P = 0.007 and r = 0.37, P < 0.001, respectively. ROC analysis for predicting postoperative walking ability in preoperative independent patients with HF and FNF revealed areas under the curve (95% confidence interval) of 0.63 (0.50-0.76) and 0.80 (0.65-0.96), respectively.
Conclusions: In patients with HF, the SI correlated with preoperative walking ability and could predict postoperative walking ability. Among patients who could walk independently before surgery, those with high SIs could walk independently early in the postoperative period. The SI is beneficial for estimating walking ability in patients with HF.
Relationship Between Serum Uric Acid-to-Creatinine Ratio and the Risk of Metabolic-Associated Fatty Liver Disease in Patients with Type 2 Diabetes Mellitus
Purpose: To investigate the association between serum uric acid-to-creatinine ratio (SUA/Cr) and the risk of developing metabolic-associated fatty liver disease (MAFLD) in patients with type 2 diabetes mellitus (T2DM).
Patients and methods: Overall, 1434 patients with T2DM who were admitted to Hebei General Hospital from January 2019 to December 2019 were selected as the study subjects. According to abdominal ultrasound findings, patients were divided into two groups: MAFLD group and non-MAFLD group. A total of 734 patients were diagnosed with MAFLD. Participants were divided into three study groups according to the SUA/Cr ratio. Chi-square test and one-way analysis of variance were used to perform a comparison between groups. The relationship between SUA/Cr ratio and MAFLD risk was analyzed using correlation analysis and regression analysis. Furthermore, subgroup analyses were performed to verify the robustness of the results.
Results: The detection rate of MAFLD in patients with T2DM was 51.2%, and the detection rate of progressive liver fibrosis in T2DM patients with MAFLD was 36.6%. A significantly higher SUA/Cr ratio was seen in the MAFLD group than in the non-MAFLD group. After adjusting for confounding factors, multivariate logistic regression analysis revealed that the SUA/Cr ratio was an independent risk factor for MAFLD development. Stronger correlations were found in participants with a body mass index ranging between 23 and 28 kg/m2, HbA1C >7%, or female sex.
Conclusion: An elevated SUA/Cr index is independently correlated with an increased risk of MAFLD in Chinese adults with T2DM.
Early prediction of COVID-19-associated Acute Kidney Injury: Are serum NGAL and serum Cystatin C levels better than serumcreatinine?
Background: Coronavirus disease-2019 (COVID-19) is associated with a high risk of acute kidney injury (AKI), often requiring renal replacement therapy (RRT). Serum Cystatin C (sCysC) and serum Neutrophil Gelatinase-Associated Lipocalin (sNGAL) are emerging biomarkers for kidney injury, and were suggested to be superior to serum creatinine (sCr) in several clinical settings. Moreover, elevated sCysC is associated with disease severity and mortality in COVID-19. We aimed to assess the utility of sCysC and sNGAL for predicting COVID-19-associated AKI, need for RRT, and need for intensive care unit (ICU) admission, when measured at presentation to the emergency department (ED).
Methods: Patients presenting to the ED with laboratory-confirmed COVID-19 were included. The primary outcome was development of COVID-19-associated AKI, while the secondary outcomes were need for RRT and ICU admission.
Results: Among 52 COVID-19 patients, 22 (42.3%) developed AKI with 8/22 (36.4%) requiring RRT. Both sCr and sCysC demonstrated excellent performance for predicting AKI (AUC, 0.86 and 0.87, respectively) and need for RRT (AUC, 0.94 and 0.95, respectively). sNGAL displayed acceptable performance for predicting AKI (AUC, 0.81) and need for RRT (AUC, 0.87).
Conclusions: SCr and sCysC measured at ED presentation are both highly accurate predictors of AKI and need for RRT, whereas sNGAL demonstrated adequate diagnostic performance. While sCyC was previously shown to be superior to sCr as a diagnostic biomarker of kidney injury in certain etiologies, our findings demonstrate that sCr is comparable to sCyC in the context of predicting COVID-19-associated AKI. Given the high sensitivity of these biomarkers for predicting the need for RRT, and as sCysC is associated with mortality in COVID-19 patients, we recommend their measurement for enabling risk stratification and early intervention.
The effect of gender-affirming hormone treatment on serumcreatinine in transgender and gender-diverse youth: implications for estimating GFR
Background: Equations for estimated glomerular filtration rate (eGFR) based on serum creatinine include terms for sex/gender. For transgender and gender-diverse (TGD) youth, gender-affirming hormone (GAH) treatment may affect serum creatinine and in turn eGFR.
Methods: TGD youth were recruited for this prospective, longitudinal, observational study prior to starting GAH treatment. Data collected as part of routine clinical care were abstracted from the medical record.
Results: For participants designated male at birth (DMAB, N = 92), serum creatinine decreased within 6 months of estradiol treatment (mean ± SD 0.83 ± 0.12 mg/dL to 0.76 ± 0.12 mg/dL, p < 0.001); for participants designated female at birth (DFAB, n = 194), serum creatinine increased within 6 months of testosterone treatment (0.68 ± 0.10 mg/dL to 0.79 ± 0.11 mg/dL, p < 0.001). Participants DFAB treated with testosterone had serum creatinine similar to that of participants DMAB at baseline, whereas even after estradiol treatment, serum creatinine in participants DMAB remained higher than that of participants DFAB at baseline. Compared to reference groups drawn from the National Health and Nutritional Examination Survey, serum creatinine after 12 months of GAH was more similar when compared by gender identity than by designated sex.
Conclusion: GAH treatment leads to changes in serum creatinine within 6 months of treatment. Clinicians should consider a patient’s hormonal exposure when estimating kidney function via eGFR and use other methods to estimate GFR if eGFR based on serum creatinine is concerning.
Recent proof signifies that genomic individuality of neurons, characterised by DNA-content material variation, is a widespread if not common phenomenon in the human mind that happens naturally however may also present aberrancies which have been linked to the pathomechanism of Alzheimer’s illness and associated neurodegenerative issues.
Etiologically, this genomic mosaic has been prompt to come up from defects of cell cycle regulation that will happen both throughout mind growth or in the mature mind after terminal differentiation of neurons. Here, we purpose to attract consideration in the direction of one other mechanism that can provide rise to genomic individuality of neurons, with far-reaching penalties.
This mechanism has its origin in the transcriptome moderately than in replication defects of the genome, i.e., somatic gene recombination of RNA. We proceed to develop the idea that somatic gene recombination of RNA supplies a physiological course of that, by integration of intronless mRNA/ncRNA into the genome, permits a specific practical state on the degree of the person neuron to be listed.
By insertion of outlined RNAs in a somatic recombination course of, the presence of particular mRNA transcripts inside a particular temporal context could be “frozen” and can function an index that may be recalled at any later level in time. This permits data associated to a particular neuronal state of differentiation and/or exercise related to a reminiscence hint to be fastened. We recommend that this course of is used all through the lifetime of every neuron and might need each advantageous and deleterious penalties.
Genomic Indexing by Somatic Gene Recombination of mRNA/ncRNA – Does It Play a Role in Genomic Mosaicism, Memory Formation, and Alzheimer’s Disease?
Culturomics-based genomics sheds mild on the ecology of the brand new haloarchaeal genus Halosegnis.
The growth of tradition-unbiased strategies has revolutioned our understanding of microbial ecology, particularly by the illustration of the huge hole between the environmentally considerable microbial variety and that accessible by cultivation.
However, tradition-primarily based approaches usually are not solely essential for understanding the evolutionary, metabolic and ecological milieu of microbial variety, but additionally for the event of novel biotechnological functions.
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Mouse anti-PD-1 antibody (RMP1-14) is a recombinant antibody where the original Rat IgG2a backbone of clone RMP1-14 has been changed to Mouse IgG2a. This antibody recognizes an epitope on Mouse PD-1
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Description: Mouse anti-mouse PD-L1 antibody (10F.9G2) is a recombinant antibody where the original rat IgG2b backbone of 10F.9G2 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse CD274
Description: Mouse anti-mouse PD-L1 antibody (10F.9G2) is a recombinant antibody where the original rat IgG2b backbone of 10F.9G2 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse CD274
Description: Mouse anti-mouse PD-L1 antibody (10F.9G2) is a recombinant antibody where the original rat IgG2b backbone of 10F.9G2 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse CD274
Description: Mouse anti-mouse PD-L1 antibody (10F.9G2) is a recombinant antibody where the original rat IgG2b backbone of 10F.9G2 has been changed to Mouse IgG1. This antibody recognizes an epitope on Mouse CD274
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Mouse Anti-Mouse PD-1 In Vivo Antibody (29F.1A12) LALAPG
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Mouse Anti-Mouse PD-1 In Vivo Antibody (29F.1A12) LALAPG
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
Mouse Anti-Mouse PD-1 In Vivo Antibody (29F.1A12) LALAPG
Description: Anti-PD-1 In Vivo Antibody - Low Endotoxin (29F.1A12) recognizes an epitope on Mouse PD-1. Despite its predicted molecular weight, PD-1 often migrates at higher molecular weight in SDS-PAGE.
In this research, we used a culturomics-primarily based strategy in order to isolate novel microbial taxa from hypersaline environments (i.e. Isla Cristina and Isla Bacuta salterns in Huelva, Spain). We managed to acquire axenic cultures of 4 haloarchaeal strains that belong to a new haloarchaeal genus, and to acquire their genomic sequences.
The phylogenomic and phylogenetic analyses (along with AAI, ANI and digital DDH indices) confirmed that the isolates represent two new species, for which we suggest the names Halosegnis longus sp. nov. and Halosegnis rubeus sp. nov.
The genomic-primarily based metabolic reconstructions indicated that members of this new haloarchaeal genus have photoheterotrophic cardio way of life with a typical salt-in signature. 16S rRNA gene sequence reads abundance profiles and genomic recruitment analyses revealed that the Halosegnis genus has a worldwide geographical distribution, reaching excessive abundance (as much as 8%) in habitats with intermediate salinities.