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Bi3+ ions doped borate barium fluoride glasses with nominal compositions of (59-x)B2O3 + 20BaF2 + 20Li2O + 1Yb2O3 + xBi2O3 with x = 0 (BBLYBi-0), 1 (BBLYBi-1), 3 (BBLYBi-3), 5 (BBLYBi-5), and 7 (BBLYBi-7) mol% have been prepared by using well-known melt quenching method. Borate barium fluoride glasses doped with Bi2O3 have been investigated by means of X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and UV–Vis measurements. In addition to the MCNP5 simulation code and Phy-X software have been applied in order to evaluate the mass-(MACs) absorption coefficients as γ-ray absorption parameter of the BBLYBi-glasses. The XRD pattern for borate barium fluoride glasses doped with Bi2O3 verified the amorphous nature of the samples. The FTIR spectra exhibited many absorption bands with a combination of broad and sharp peaks. The inclusion of Bi3+ ions in the suggested glass networks leads to increase the density (ρ) from 3.2443 to 3.6824 g/cm3 and the molar volume (Vm) increases from 26.5254 cm3/mol to 30.9039 cm3/mol. The optical band gap reduced as increasing Bi3+ content; the direct optical gap ( E g Direct ) ranged from 3.98 ± 0.01 to 3.59 ± 0.01 eV, while the indirect optical ( E g Indirect ) ranged from 3.47 ± 0.01 to 2.90 ± 0.01 eV. The studied samples exhibited a decreasing in Urbach's energy, suggesting a decrease in the structural disorder inside the glass samples. The maximum MACs values were 49.491cm2/g, 44.043cm2/g, 37.713 cm2/g, 30.268 cm2/g, and 26.031 cm2/g for BBLYBi-0, BBLYBi-1, BBLYBi-3, BBLYBi-5, and BBLYBi-7 samples, respectively. Results of MCNP5 simulation code and Phy-X software confirm that the MACs of the studied glasses were enhanced as Bi3+ increasing in the glass networks. Therefore, the suggested glasses are appropriate for usage in optical and radiation shielding regions. [ABSTRACT FROM AUTHOR]

Context Patients with nonfunctioning adrenal adenomas (NFA) and mild autonomous cortisol secretion (MACS) demonstrate an increased risk of chronic kidney disease (CKD); however, factors associated with CKD are unknown. Objective We aimed to identify the factors associated with CKD and assess the effect of adrenalectomy on kidney function in patients with NFA or MACS. Methods A single-center cohort study of patients with NFA and MACS, 1999 to 2020, was conducted. MACS was diagnosed based on post dexamethasone suppression test (DST) cortisol greater than or equal to 1.8 mcg/dL. Age, sex, dysglycemia, hypertension, therapy with statin, angiotensin-converting enzyme inhibitor, or angiotensin II receptor blocker were included in the multivariable analysis. Outcomes included estimated glomerular filtration rate (eGFR) at the time of diagnosis with MACS or NFA and postadrenalectomy delta eGFR. Results Of 972 patients, 429 (44%) had MACS and 543 (56%) had NFA. At the time of diagnosis, patients with MACS had lower eGFR (median 79.6 vs 83.8 mL/min/1.73 m2; P <.001) than patients with NFA. In a multivariable analysis, factors associated with lower eGFR were older age, hypertension, and higher DST. In 204 patients (MACS: 155, 76% and NFA: 49, 24%) treated with adrenalectomy, postadrenalectomy eGFR improved in both groups starting at 18 months up to 3.5 years of follow-up. Factors associated with increased eGFR were younger age, lower preadrenalectomy eGFR, and longer follow-up period. Conclusion DST cortisol is an independent risk factor for lower eGFR in patients with adrenal adenomas. Patients with both MACS and NFA demonstrate an increase in eGFR post adrenalectomy, especially younger patients with lower eGFR pre adrenalectomy. [ABSTRACT FROM AUTHOR]

Background Pre–diabetes mellitus (DM) is associated with proteinuria, a risk factor for chronic kidney disease. While people with human immunodeficiency virus (HIV; PWH) have a higher risk of proteinuria than people without HIV (PWOH), it is unknown whether incident proteinuria differs by HIV serostatus among prediabetic persons. Methods The urine protein-to-creatinine ratio was measured at semiannual visits among men in the Multicenter AIDS Cohort Study since April 2006. Men with pre-DM on or after April 2006 and no prevalent proteinuria or use of antidiabetic medications were included. Pre-DM was defined as a fasting glucose level of 100–125 mg/dL confirmed within a year by a repeated fasting glucose or hemoglobin A1c measurement of 5.7%–6.4%. Incident proteinuria was defined as a urine protein-to-creatinine ratio (UPCR) >200 mg/g, confirmed within a year. We used Poisson regression models to determine whether incident proteinuria in participants with pre-DM differed by HIV serostatus and, among PWH, whether HIV-specific factors were related to incident proteinuria. Results Between 2006 and 2019, among 1276 men with pre-DM, proteinuria developed in 128 of 613 PWH (21%) and 50 of 663 PWOH (8%) over a median 10-year follow-up. After multivariable adjustment, the incidence of proteinuria in PWH with pre-DM was 3.3 times (95% confidence interval, 2.3–4.8 times) greater than in PWOH (P <.01). Among PWH, current CD4 cell count <50/µL (P <.01) and current use of protease inhibitors (P =.03) were associated with incident proteinuria, while lamivudine and integrase inhibitor use were associated with a lower risk. Conclusions Among men with pre-DM, the risk of incident proteinuria was 3 times higher in PWH. Strategies to preserve renal function are needed in this population. [ABSTRACT FROM AUTHOR]

Purpose: To identify the sperm preparation procedure that selects the best sperm population for medically assisted reproduction. Methods: Prospective observational study comparing the effect of four different sperm selection procedures on various semen parameters. Unused raw semen after routine diagnostic analysis was split in four fractions and processed by four different methods: (1) density gradient centrifugation (DGC), (2) sperm wash (SW), (3) DGC followed by magnetic activated cell sorting (MACS), and (4) using a sperm separation device (SSD). Each fraction was analyzed for progressive motility, morphology, acrosome index (AI), and DNA fragmentation index (DFI). Results: With DGC as standard of care in intraclass correlation coefficient analysis, only SSD was in strong disagreement regarding progressive motility and DFI [0.26, 95%CI (− 0.2, 0.58), and 0.17, 95%CI (− 0.19, 0.45), respectively]. When controlling for abstinence duration, DFI was significantly lower after both MACS and SSD compared to DGC [− 0.27%, 95%CI (− 0.47, − 0.06), p = 0.01, and − 0.6%, 95%CI (− 0.80, − 0.41), p < 0.001, respectively]. Further comparisons between SSD and MACS indicate significantly less apoptotic cells [Median (IQR) 4 (5), 95%CI (4.1, − 6.8) vs Median (IQR) 5 (8), 95%CI (4.9, − 9.2), p < 0.001, respectively] and dead cells [Median (IQR) 9.5 (23.3), 95%CI (13.2, − 22.4) vs Median (IQR) 22 (28), 95%CI (23.1, − 36.8), p < 0.001, respectively] in the SSD group. Conclusion: The selection of a population of highly motile spermatozoa with less damaged DNA from unprocessed semen is ideally performed with SSD. Question remains whether this method improves the embryological outcomes in the IVF laboratory. [ABSTRACT FROM AUTHOR]

The purpose of the present study was in vitro determination of the combined effects of doxorubucin and 5-fluorouracil by 2D and 3D culture conditions on breast cancer using MCF-7 cell line and CSCs isolated from these cells. In the first stage of this study, CSC isolation and their characterization were performed. In the next experimental period, the antiproliferative effects of 5-Fu and Dox on the MCF-7 and CSCs were demonstrated on 2D. To evaluate the synergistic/antagonistic effects of these chemotherapeutics, the CI was calculated. Additionally, 3D tumor spheroids were used as another model. In the last step, qRT-PCR analysis was performed to examine apoptosis-related gene expressions. In this study, it was clearly seen that CSCs obtained from the breast cancer cell line express stemness factors. In addition, the antiproliferative effects of 5-Fu and Dox on breast cancer and associated CSCs were very clear. Their synergistic effects were determined by CI values. Moreover, it was seen that combined theraphy changed the expression levels of genes related to apoptosis. Additionally, it was molecularly demonstrated that 3D tumoroids were more resistant than the others. In conclusion, the polychemotherapeutic approach was much more effective than the monotherapy. The fact that this effect was seen not only in breast cancer cells, but also in breast cancer stem cells. In addition, it was very promising that the results obtained were similar in both two-dimensional and three-dimensional tumoroids. [ABSTRACT FROM AUTHOR]

ObjectivesGlucocorticoid cosecretion is more common in primary aldosteronism (PA) than previously thought. Chronic subtle cortisol excess in patients with mild autonomous cortisol secretion (MACS) negatively affects bone health. This study aimed to evaluate the impact of MACS on bone density and turnover markers in PA patients.MethodsPatients with PA and MACS (n = 50), confirmed by a 1-mg dexamethasone suppression test (DST) with a cortisol cutoff of ≥1.8 µg/dL without symptoms of overt Cushing, were compared to age- and sex-matched patients with PA without MACS (non-MACS, n = 50). Lumbar volumetric bone mineral density (vBMD) was extracted by a novel convolutional neural network (CNN)-based framework (SpineQ software v1.0) applied to routine CT data, incorporated into the diagnostic protocol for PA. Additionally, bone turnover markers—including osteocalcin, bone-specific alkaline phosphatase, N-terminal propeptide of type I collagen, and carboxy-terminal crosslinked telopeptide of type I collagen were evaluated between the groups.ResultsMedian cortisol after DST was 1.1 µg/dL (30.3 nmol/L) [IQR: 0.5 µg/dL (13.8 nmol/L)] in the non-MACS group and 2.5 µg/dL (69.0 nmol/L) [IQR: 1.4 µg/dL (38.5 nmol/L)] in the MACS group (p < 0.001). Patients with MACS had significantly lower vBMD values compared to the non-MACS group (106.4 mg/cm³ vs. 116.6 mg/cm³, p = 0.038). Cortisol after DST negatively correlated with vBMD (Spearman’s r=−0.33, p=0.00042). No significant differences in bone turnover markers were found, and classifications based on visible lesions on CT or PA-lateralization via adrenal venous sampling did not reveal any significant differences in these markers (p > 0.05 for all comparisons).ConclusionDespite non-significant differences in bone turnover markers between patients with PA with or without MACS, CT scans revealed significantly reduced vBMD in PA and MACS patients, indicating compromised bone health and vBMD significantly negatively correlated with cortisol post DST. Thus, opportunistic evaluation of vBMD in routine CT screenings could aid in the early detection of bone alterations in MACS and help mitigate potential long-term adverse effects on bone health.

The thermonuclear reaction rates and Maxwellian-averaged cross-sections (MACS) for the 84 Kr(n, γ ) 85 Kr and 86 Kr(n, γ ) 87 Kr processes were examined using the statistical model code Talys v1.96. The effects of nuclear level densities (NLDs) on Maxwellian-averaged cross-sections and neutron capture rates are examined both quantitatively and qualitatively. The present Talys-based MACS and radiative capture rates for 84 Kr(n, γ ) 85 Kr and 86 Kr(n, γ ) 87 Kr processes agree well with the earlier reported findings. The statistical model’s nuclear properties (level density and gamma ray strength) were fine-tuned to reproduce the existing experimental nuclear data. [ABSTRACT FROM AUTHOR]

Erythroid progenitor cells (EPCs) can differentiate into erythroid precursors and eventually in to mature red blood cells. Cluster of differentiation (CD)34 is a known surface marker for identification of primitive human erythroid progenitor cells including EPCs. CD34+ cells can be isolated from peripheral blood samples using magnetic-assisted cell sorting (MACS) or by flowcytometry-mediated cell sorting. There are no reported studies from South Asia in which CD34+-derived EPCs have been isolated and purified from the peripheral blood samples obtained from healthy adult humans. The aim of the present study was to isolate CD34+ cells, differentiate CD34+ cells in to EPCs, purify and characterize CD34+-derived EPCs from peripheral blood samples from healthy adult human subjects. Blood samples were obtained from apparently healthy Pakistani adult volunteer human subjects. Human peripheral blood mononuclear cells (PBMNCs) were separated from the blood sample by density gradient centrifugation. CD34+ cells were isolated from PBMNCs by MACS using CD34+ selection kit. CD34+ cells were differentiated in to EPCs by using erythroid expansion supplement. Purity of the CD34+-derived EPCs was assessed through immune fluorescence microscopy using monoclonal antibodies against erythroid specific surface antigens CD34, CD235a and CD71. The purity of CD34+-derived EPCs was determined to be 95%-98%. Cord blood samples contain immature cells, which can result in contamination of PBMNCs layer. Thus, obtaining EPCs from CD34+ peripheral blood cells of healthy adult humans is superior as the risk of contamination is minimal. The method of isolation and purification of CD34+-derived EPCs used in this study is simpler, user friendly and is more efficient as compared to other techniques. Human EPCs differentiated by this protocol can then be used to study normal and abnormal erythropoiesis. [ABSTRACT FROM AUTHOR]

Background: A recent cross-sectional study showed that both comorbidities and mortality in patients with adrenal incidentaloma (AI) are tied to sex. However, few longitudinal studies evaluated the development of arterial hypertension, hyperglycemia, dyslipidemia and bone impairment in patients with AI. The aim of this study is to analyze the impact of sex in the development of these comorbidities during long-term follow-up. Methods: We retrospectively evaluated 189 patients (120 females, 69 males) with AI, from four referral centers in Italy and Croatia. Clinical characteristics, comorbidities and cortisol after 1-mg dexamethasone suppression test (1-mg DST) were assessed at baseline and at last follow-up visit (LFUV). Median followup was 52 (Interquartile Range 25-86) months. Results: The rates of arterial hypertension and hyperglycemia increased over time both in females (65.8% at baseline versus 77.8% at LFUV, p=0.002; 23.7% at baseline versus 39.6% at LFUV, p<0.001; respectively) and males (58.0% at baseline versus 69.1% at LFUV, p=0.035; 33.8% at baseline versus 54.0% at LFUV, p<0.001; respectively). Patients were stratified in two groups using 1.8 µg/dl as cut-off of cortisol following 1-mg DST: non-functional adrenal tumors (NFAT) and tumors with mild autonomous cortisol secretion (MACS). In the NFAT group (99 patients, females 62.6%), at baseline, we did not observe any difference in clinical characteristics and comorbidities between males and females. At LFUV, males showed a higher frequency of hyperglycemia than females (57.6% versus 33.9%, p=0.03). In the MACS group (89 patients, females 64.0%), at baseline, the prevalence of hypertension, hyperglycemia and dyslipidemia was similar between sexes, despite females were younger (60, IQR 55-69 versus 67.5, IQR 61-73, years; p=0.01). Moreover, females presented higher rates of bone impairment (89.3% versus 54.5%, p=0.02) than males. At LFUV, a similar sexrelated pattern was observed. Conclusion: Patients with AI frequently develop arterial hypertension and hyperglycemia and should be periodically checked for these comorbidities, regardless of sex. In patients with MACS, the lack of difference between sexes in the frequency of cardiometabolic comorbidities despite that females are younger, and the higher frequency of bone impairment in females, suggest a sex-specific effect of cortisol. [ABSTRACT FROM AUTHOR]

The neutron capture rates and Maxwellian-averaged cross-sections (MACS) for 90Zr(n,γ)91Zr and 92Zr(n,γ)93Zr processes have been computed within the framework of Talys v1.96. The effects of phenomenological nuclear level density (NLD) parameters and the gamma strength functions (GSFs) on Maxwellian-averaged cross-sections and neutron capture rates are examined both quantitatively and qualitatively. The present model-based computed data for MACS and reaction rates gives a good comparison with the existing literature. The fine-tuning of the statistical model's nuclear properties (level density and gamma-ray strength) to reproduce experimental data will allow the detailed investigation of the s-process network. [ABSTRACT FROM AUTHOR]

ObjectivesThis study of people with HIV (PWH) and those without HIV conducted during the COVID-19 pandemic in the United States in 2020 examines the impact of posttraumatic stress disorder (PTSD) on COVID-19 burden, defined as pandemic-related disruptions.MethodsData consisted of survey responses on PTSD among participants (N = 2434) enrolled in the Multicenter AIDS Cohort Study (MACS) and the Women's Interagency HIV (WIHS) cohorts. Unadjusted and adjusted regression models were used to examine the association of PTSD with COVID-19 burden (overall and domain-specific burdens). Quasi-Poisson regression models were used to assess associations with the COVID-19 burden score and 2 domain-specific burdens: (1) changes in resources and (2) interruptions in health care. Analyses was adjusted for age, race/ethnicity, HIV serostatus, current smoking status, number of comorbidities, education, and study regions.ResultsStudy participants were a median age of 58 (interquartile range, 52-65) years. In both bivariate and multivariable models, PTSD severity was associated with greater overall COVID-19 burden. PTSD severity was associated with the number of resource changes and number of interruptions in medical care. These findings were also consistent across cohorts (MACS/WIHS) and across HIV serostatus, suggesting a greater risk for COVID-19 burden with greater PTSD severity, which remained significant after controlling for covariates.ConclusionsThis study builds on emerging literature demonstrating the impact of mental health on the burden and disruption associated with the COVID-19 pandemic, providing context specific to PWH. The ongoing pandemic requires structural and social interventions to decrease disruption to resources and health resource needs among these vulnerable populations.

Background: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees of differentiation of cell subpopulations poses a considerable challenge in preclinical studies. The cells at a pluripotent-like stage represent a stem cell population of interest for many researchers worldwide, which is worthy of identification, isolation, and functional characterization. In the current study, we asked whether Wharton's jelly-derived MSCs (WJ-MSCs) which express stage-specific embryonic antigen-4 (SSEA-4) can be considered as a pluripotent-like stem cell population. Methods: SSEA-4 expression in different culture conditions was compared and the efficiency of two cell separation methods were assessed: Magnetic Activated Cell Sorting (MACS) and Fluorescence Activated Cell Sorting (FACS). After isolation, SSEA-4+ cells were analyzed for the following parameters: the maintenance of the SSEA-4 antigen expression after cell sorting, stem cell-related gene expression, proliferation potential, clonogenicity, secretome profiling, and the ability to form spheres under 3D culture conditions. Results: FACS allowed for the enrichment of SSEA-4+ cell content in the population that lasted for six passages after sorting. Despite the elevated expression of stemness-related genes, SSEA-4+ cells neither differed in their proliferation and clonogenicity potential from initial and negative populations nor exhibited pluripotent differentiation repertoire. SSEA-4+ cells were observed to form smaller spheroids and exhibited increased survival under 3D conditions. Conclusion: Despite the transient expression of stemness-related genes, our findings could not fully confirm the undifferentiated pluripotent-like nature of the SSEA-4+ WJ-MSC population cultured in vitro. [ABSTRACT FROM AUTHOR]

Simple Summary: Couples attending infertility clinics may suffer failed attempts and need consecutive treatments to achieve a healthy newborn, proving that there is room for improvement among the techniques we currently use. Selection of the right sperm—the most physiologically competent one—to be injected inside the egg among all available within the ejaculate is crucial to create good-quality embryos and increase cycle success rates. It is probably the step with the highest number of possibilities to choose from influencing the final outcomes. Apoptotic sperm, those undergoing programmed cell death, do not differ morphologically from healthy ones, their presence has been described as elevated in the ejaculates of infertile men, and they can fertilize the egg in both natural and artificial conception. In the latter, they can be unconsciously chosen by the embryologist and result in a poor-quality embryo that will stop developing or fail to implant. MACS enables the removal of apoptotic sperm from an ejaculate, thus leaving the non-apoptotic available to be microinjected. The existing literature on the topic provides conflicting evidence of variable quality that needs to be scrutinized and interpreted in order to define what is the benefit, if any, of using this technology and if this fits all infertile patients. We aim to provide clinicians and patients with a more accurate interpretation on how, when, and by how much, the use of MACS may impact their reproductive chances under an evidence-based medicine approach. Sperm selection of the most competent sperm is a promising way to enhance reproductive outcomes. Apoptosis is the programmed cell death process to maintain tissue homeostasis, and MACS sperm selection of non-apoptotic cells enables the removal of apoptotic sperm from an ejaculate, thus leaving the non-apoptotic available to be microinjected, but given the associated costs of adding these sperm selection steps to the routine practice, there is a need for a careful examination of the literature available to answer questions such as who can benefit from this MACS, how significant this improvement is, and how robust the evidence and data available supporting this choice are. Thus, the aim of this narrative review was to objectively evaluate the available evidence regarding the potential benefits of the use of MACS. From the literature, there are controversial results since its implementation as an in vitro fertilization add-on, and this may be explained in part by the low quality of the evidence available, wrong designs, or even inadequate statistical analyses. We concluded that the benefits of adding MACS are unclear, and further methodologically sound research on specific populations is much needed before offering it clinically. [ABSTRACT FROM AUTHOR]

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BackgroundA recent cross-sectional study showed that both comorbidities and mortality in patients with adrenal incidentaloma (AI) are tied to sex. However, few longitudinal studies evaluated the development of arterial hypertension, hyperglycemia, dyslipidemia and bone impairment in patients with AI. The aim of this study is to analyze the impact of sex in the development of these comorbidities during long-term follow-up.MethodsWe retrospectively evaluated 189 patients (120 females, 69 males) with AI, from four referral centers in Italy and Croatia. Clinical characteristics, comorbidities and cortisol after 1-mg dexamethasone suppression test (1-mg DST) were assessed at baseline and at last follow-up visit (LFUV). Median follow-up was 52 (Interquartile Range 25-86) months.ResultsThe rates of arterial hypertension and hyperglycemia increased over time both in females (65.8% at baseline versus 77.8% at LFUV, p=0.002; 23.7% at baseline versus 39.6% at LFUV, p

Background: High heterogeneity of mesenchymal stem/stromal cells (MSCs) due to different degrees of differentiation of cell subpopulations poses a considerable challenge in preclinical studies. The cells at a pluripotent-like stage represent a stem cell population of interest for many researchers worldwide, which is worthy of identification, isolation, and functional characterization. In the current study, we asked whether Wharton’s jelly-derived MSCs (WJ-MSCs) which express stage-specific embryonic antigen-4 (SSEA-4) can be considered as a pluripotent-like stem cell population.Methods: SSEA-4 expression in different culture conditions was compared and the efficiency of two cell separation methods were assessed: Magnetic Activated Cell Sorting (MACS) and Fluorescence Activated Cell Sorting (FACS). After isolation, SSEA-4+ cells were analyzed for the following parameters: the maintenance of the SSEA-4 antigen expression after cell sorting, stem cell-related gene expression, proliferation potential, clonogenicity, secretome profiling, and the ability to form spheres under 3D culture conditions.Results: FACS allowed for the enrichment of SSEA-4+ cell content in the population that lasted for six passages after sorting. Despite the elevated expression of stemness-related genes, SSEA-4+ cells neither differed in their proliferation and clonogenicity potential from initial and negative populations nor exhibited pluripotent differentiation repertoire. SSEA-4+ cells were observed to form smaller spheroids and exhibited increased survival under 3D conditions.Conclusion: Despite the transient expression of stemness-related genes, our findings could not fully confirm the undifferentiated pluripotent-like nature of the SSEA-4+ WJ-MSC population cultured in vitro.

A formula for Maxwellian-Averaged Cross-Sections (MACS) at 30 keV is derived. Based on the presented formula, a linear relationship was found between the log ( MACS A 1. 4 ) and (N − Z) 2 A 2 of the target nucleus. From this relationship, a good agreement between the calculated MACS and the experimental data can be observed. It seems therefore possible to use this systematic law in order to analyze the experimental data. It would be useful in estimating the MACS of neighboring isotopes for which no experimental data are available. [ABSTRACT FROM AUTHOR]

BackgroundStudies have examined patterns of substance use among Men who have Sex with Men (MSM), but few have examined factors predicting transitioning from one substance use pattern to another. We investigated transitioning from one substance use pattern to another over a 12-year period (2004-2016) among the Multicenter AIDS Cohort Study participants.MethodAlcohol, marijuana, heroin, cocaine, poppers, uppers (e.g., methamphetamines) and erectile dysfunction(ED) medications use in the last 6 months from 3568 US MSM was dichotomized (no/yes) to classify participants into substance use classes at each follow up visit. We fit latent transition models to calculate transition probabilities of moving from one substance use class to another over a 3, 4 and 6-year time period. Then fit regression models to identify factors associated with the probability of each participant staying in or moving from the same substance use class.ResultsOverall, cocaine and ED medication use declined but marijuana and heroin use increased over 2004-2016. We observed most participants (84.6 %-100 %) stayed in the same class. Increased age was associated with transition from the Minimal-use class to the Alcohol-only class (aOR = 1.06,95 %CI:1.01-1.13;p < 0.01) and non-White MSM reported lower odds of moving from the Alcohol-only class to the Alcohol-Popper class (aOR = 0.50,95 %CI:0.30-0.82;p

Medications are known to affect the thyroid physiology and are a known cause of hypothyroidism. There is an ever-growing list of medications that affect the thyroid by 1 or more mechanisms. Mifepristone is presently used for the treatment of mild autonomous cortisol secretion (MACS). Hypothyroidism is not a known side effect of this medication. We present a 71-year-old woman with newly diagnosed impaired fasting glucose, dyslipidemia, and osteopenia presenting with a 3-year history of unintentional 15-pound weight gain (despite exercise and a good diet) and increased anxiety. Her physical examination was pertinent for mild lower extremity edema, easy bruising, and skin thinning. Workup revealed adrenocorticotropic hormone (ACTH)-independent MACS from bilateral micronodular hyperplasia of the adrenals. Since she was not a surgical candidate, medical management with mifepristone was chosen. While on mifepristone, she complained of excessive fatigue, a workup done revealed new-onset hypothyroidism. Given her symptoms and bloodwork, she was started on levothyroxine. After stopping mifepristone, she was biochemically and clinically euthyroid and was eventually off levothyroxine. The mechanism by which mifepristone induces hypothyroidism is unknown. Except for a multicenter case series suggesting that mifepristone increases thyroid hormone requirements in patients with central hypothyroidism, to the best of our knowledge, the literature on euthyroid patients developing hypothyroidism secondary to mifepristone is scarce. In conclusion, while the hypothyroidism seems reversible our case highlights the importance of getting baseline thyroid function tests (TFTs) and repeating them while on the medication. Treatment of hypothyroidism is based on symptoms and bloodwork. [ABSTRACT FROM AUTHOR]

PURPOSE: This study aimed to identify factors, including degree of disability, that contribute to the caregiver burden of raising children with cerebral palsy in Sri Lanka. METHODS: Participants were caregivers of children with cerebral palsy attending the pediatric neurology clinic of the only tertiary care center in southern Sri Lanka. The locally validated Caregiver Difficulties Scale (CDS) was administered, and demographic information was obtained in a structured interview. Disability data was accessed through the medical record. RESULTS: Of 163 caregivers who participated in this study, 133 (81.2%) demonstrated a moderate to high level of burden, and 91 (55.8%) were at high risk for psychological burden. In the bivariate analysis, caregiver burden significantly correlated with degree of physical disability based on the Gross Motor Function Classification System (GMFCS) and the Manual Ability Classification System (MACS), the presence of medical co-morbidities, and having two or more children. However, only the GMFCS level and number of children remained significant predictors of caregiver burden after controlling for confounding effects. CONCLUSION: Raising a child with cerebral palsy in Sri Lanka is likely to cause caregiver burden, particularly if they have a high level of disability or one or more siblings. Monitoring caregiver burden as part of routine cerebral palsy management is important, which allows targeting psychosocial support to families most in need. [ABSTRACT FROM AUTHOR]

Objective: Prospective data on determinants of muscle strength impairment and quality of life in patients with various subtypes and severity of endogenous hypercortisolism are lacking. Design: Single-center cross-sectional study, 2019 to 2022. Methods: Patients with Cushing syndrome (CS) and mild autonomous cortisol secretion (MACS) were assessed with clinical and biochemical severity scores, muscle function (nondominant hand grip strength and sit-to-stand test), and quality of life (Short Form-36 [SF36] and CushingQoL). Referent subjects were recruited from the local population undergoing abdominal imaging for reasons other than suspected adrenal disorder. Results: Of 164 patients, 81 (49%) had MACS, 14 (9%) had adrenal CS, 60 (37%) had pituitary CS, and 9 (5%) had ectopic CS. Median age was 53 years (interquartile range: 42-63 years), and 126 (77%) were women. The SF36 mental component score was similarly low in patients with MACS vs CS, but physical component score was lower in CS when compared to MACS (mean of 34.0 vs 40.5, P = .001). Compared to MACS, patients with CS had lower scores on the standardized CushingQoL (mean of 47.1 vs 34.2, P < .001). Compared to referent subjects, patients with MACS demonstrated reduced muscle strength, similar to patients with CS (mean sit to stand Z-score of -0.47 vs -0.54, P = .822). Clinical severity (r = -0.22, P = .004) but not biochemical severity was associated with sit-to-stand test performance. Conclusions: Both patients with overt CS and MACS demonstrate reduced muscle strength and low quality of life. The clinical severity score utilized is associated with both physical and psychosocial components of CushingQoL and with the physical component of SF36. [ABSTRACT FROM AUTHOR]

Genome editing technologies hold great promise for numerous applications including the understanding of cellular and disease mechanisms and the development of gene and cellular therapies. Achieving high editing frequencies is critical to these research areas and to achieve the overall goal of being able to manipulate any target with any desired genetic outcome. However, gene editing technologies sometimes suffer from low editing efficiencies due to several challenges. This is often the case for emerging gene editing technologies, which require assistance for translation into broader applications. Enrichment strategies can support this goal by selecting gene edited cells from non-edited cells. In this review, we elucidate the different enrichment strategies, their many applications in non-clinical and clinical settings, and the remaining need for novel strategies to further improve genome research and gene and cellular therapy studies. [ABSTRACT FROM AUTHOR]

Lung cancer is one of the most common cancers in the world and a leading cause of death. Cancer stem cells (CSCs) are the cells responsible for tumor initiation. CSCs features include self-renewal capacity, differentiation, high invasion-migration, and resistance to chemotherapy. CSCs are characterized by specific surface markers. CD133 is widely used as a marker of CSCs in many cancers, including lung cancer. CD133-positive cells are an important marker of lung cancer stem cells because they show characteristics of lung cancer CSCs. CSCs are less sensitive and/or resistant to conventional treatments, remaining viable and regenerating tumors after treatment. Treatments that can target CSCs will be able to destroy CSCs more effectively, leading to the disappearance of tumors. Currently, attempts are being made to develop therapeutics that can effectively target CSCs. These studies concern the isolation and characterization of CSCs. In this study, CD133-positive CSCs and CD133-negative cells were isolated from H460 cells, a large cell lung cancer model, by MACS method. CSCs properties were investigated by tumorSphere formation assay (2% agar plate), hanging drop assay and qPCR method. CD133 positivity of H460 cells was determined as 0.3%. It has been observed that CD133-positive cells show CSCs-specific self-renewal properties. It was determined that the CD133 gene expression difference between CD133-positive CSCs and CD133-negative cells was approximately 25-fold. It has been observed that CD133-positive CSCs express more pluripotency genes (such as OCT-4, SOX-2 and KLF-4) compared to CD133-negative cells. [ABSTRACT FROM AUTHOR]

Magnetic cell sorting technology stands out because of its speed, simplicity, and ability to process large cell numbers. However, it also suffers from a number of drawbacks, in particular low discrimination power, which results in all-or-none selection outcomes limited to a bulk separation of cell populations into positive and negative fractions, as well as the modest purity of the selected cells and the inability to select subpopulations of cells with high expression of a surface marker. In the present study, we developed a simple solution to this problem and confirmed the effectiveness of this approach by multiple experiments with the magnetic selection of transduced cell populations. Murine NIH 3T3 cells were transduced with the bicistronic retroviral vector constructs co-expressing fluorescent reporter proteins EGFP (enhanced green fluorescent protein) or DsRed-Express 2 and LNGFR (low-affinity nerve growth factor receptor) as surface selection markers. The effects of the magnetic selection of transduced cells with anti-LNGFR Micro Bead (MB) doses ranging from 0.5 to 80 µL have been assessed. Low doses of MBs favored the depletion of weakly positive cells from the population, resulting in the higher expression levels of EGFP or DsRed-Express2 reporters in the selected cell fractions. Low MB doses also contributed to the increased purity of the selected population, even for samples with a low initial percentage of positive cells. At the same time, high MB doses resulted in the increased yield and a more faithful representation of the original expression profiles following selection. We further demonstrate that for populations with fairly narrow distribution of expression levels, it is possible to achieve separation into high- and low-expressing subsets using the two-stage selection scheme based on the sequential use of low and high doses of MBs. For populations with broad expression distribution, a one-stage selection with low or high doses of MBs is sufficient for a clear separation of low- and high-expressing subsets in the column-retained and flow-through fractions, respectively. This study substantially extends the potential of magnetic cell sorting, and may open new possibilities in a number of biomedical applications. [ABSTRACT FROM AUTHOR]

The maintenance of redox homeostasis is associated with a healthy status while the disruption of this mechanism leads to the development of various pathological conditions. Bioactive molecules such as carbohydrates accessible to the microbiota (MACs), polyphenols, and polyunsaturated fatty acids (PUFAs) are food components best characterized for their beneficial effect on human health. In particular, increasing evidence suggests that their antioxidant ability is involved in the prevention of several human diseases. Some experimental data indicate that the activation of the nuclear factor 2-related erythroid 2 (Nrf2) pathway—the key mechanism in the maintenance of redox homeostasis—is involved in the beneficial effects exerted by the intake of PUFAs and polyphenols. However, it is known that the latter must be metabolized before becoming active and that the intestinal microbiota play a key role in the biotransformation of some ingested food components. In addition, recent studies, indicating the efficacy of the MACs, polyphenols, and PUFAs in increasing the microbial population with the ability to yield biologically active metabolites (e.g., polyphenol metabolites, short-chain fatty acids (SCFAs)), support the hypothesis that these factors are responsible for the antioxidant action on the physiology of the host. The underlying mechanisms through which MACs, polyphenols, and PUFAs might influence the redox status have not been fully elucidated, but based on the efficacy of SCFAs as Nrf2 activators, their contribution to the antioxidant efficacy of dietary bioactives cannot be excluded. In this review, we aimed to summarize the main mechanisms through which MACs, polyphenols, and PUFAs can modulate the host's redox homeostasis through their ability to directly or indirectly activate the Nrf2 pathway. We discuss their probiotic effects and the role played by the alteration of the metabolism/composition of the gut microbiota in the generation of potential Nrf2-ligands (e.g., SCFAs) in the host's redox homeostasis. [ABSTRACT FROM AUTHOR]

A cadaveric study to know the anatomical and radiological correlation between the mastoid air cell system in relation to its volume and morphology. This is a rare and one of its kind of cadaveric study on temporal bone in which comparison of pre dissection and post cortical mastoidectomy dissection on x-ray mastoid in relation to their dimensions. To study the anatomical and radiological correlation of mastoid air cell system in relation to its morphology using pre and post dissection x-ray measurements and dissection method. 30 adult cadaveric, temporal bone cortical mastoidectomy dissections were performed and X-ray mastoid with a pre and post mastoid dissection measurements using vernier caliper was done. Further 3-D analysis of volume of mastoid cavity compared with a post dissection digital radiographic measurements was carried out. On statistical analysis, mean surface area of MACS, shortest length between sigmoidsinus and posterior wall of EAC, also shortest distance between dural plate and mastoid tip, in pre and post dissection x-ray mastoid and in direct mastoid cavity measurements were not found to be significant. Mastoidectomy being the treatment of choice in day to day practise in many cases this study hopes to add up to the present understanding for the MACS dynamics and assesses the possible anatomical variations that can exist. This study helps us to find the approximate time required for surgery to perform cortical mastoidectomy. [ABSTRACT FROM AUTHOR]

Sperm selection of the most competent sperm is a promising way to enhance reproductive outcomes. Apoptosis is the programmed cell death process to maintain tissue homeostasis, and MACS sperm selection of non-apoptotic cells enables the removal of apoptotic sperm from an ejaculate, thus leaving the non-apoptotic available to be microinjected, but given the associated costs of adding these sperm selection steps to the routine practice, there is a need for a careful examination of the literature available to answer questions such as who can benefit from this MACS, how significant this improvement is, and how robust the evidence and data available supporting this choice are. Thus, the aim of this narrative review was to objectively evaluate the available evidence regarding the potential benefits of the use of MACS. From the literature, there are controversial results since its implementation as an in vitro fertilization add-on, and this may be explained in part by the low quality of the evidence available, wrong designs, or even inadequate statistical analyses. We concluded that the benefits of adding MACS are unclear, and further methodologically sound research on specific populations is much needed before offering it clinically.

Human perivascular stem/stromal cells (PSC) are a multipotent mesenchymal progenitor cell population defined by their perivascular residence. PSC are increasingly studied for their application in skeletal regenerative medicine. PSC from subcutaneous white adipose tissue are most commonly isolated via fluorescence-activated cell sorting (FACS), and defined as a bipartite population of CD146+CD34-CD31-CD45- pericytes and CD34+CD146-CD31-CD45- adventitial cells. FACS poses several challenges for clinical translation, including requirements for facilities, equipment, and personnel. The purpose of this study is to identify if magnetic-activated cell sorting (MACS) is a feasible method to derive PSC, and to determine if MACS-derived PSC are comparable to our previous experience with FACS-derived PSC. In brief, CD146+ pericytes and CD34+ adventitial cells were enriched from human lipoaspirate using a multistep column approach. Next, cell identity and purity were analyzed by flow cytometry. In vitro multilineage differentiation studies were performed with MACS-defined PSC subsets. Finally, in vivo application was performed in nonhealing calvarial bone defects in Scid mice. Results showed that human CD146+ pericytes and CD34+ adventitial cells may be enriched by MACS, with defined purity, anticipated cell surface marker expression, and capacity for multilineage differentiation. In vivo, MACS-derived PSC induce ossification of bone defects. These data document the feasibility of a MACS approach for the enrichment and application of PSC in the field of tissue engineering and regenerative medicine. Impact Statement Our findings suggest that perivascular stem/stromal cells, and in particular adventitial cells, may be isolated by magnetic-activated cell sorting and applied as an uncultured autologous stem cell therapy in a same-day setting for bone defect repair.

Lung cancer is one of the most common cancers in the world and a leading cause of death. Cancer stem cells (CSCs) are the cells responsible for tumor initiation. CSCs features include self-renewal capacity, differentiation, high invasion-migration, and resistance to chemotherapy. CSCs are characterized by specific surface markers. CD133 is widely used as a marker of CSCs in many cancers, including lung cancer. CD133-positive cells are an important marker of lung cancer stem cells because they show characteristics of lung cancer CSCs. CSCs are less sensitive and/or resistant to conventional treatments, remaining viable and regenerating tumors after treatment. Treatments that can target CSCs will be able to destroy CSCs more effectively, leading to the disappearance of tumors. Currently, attempts are being made to develop therapeutics that can effectively target CSCs. These studies concern the isolation and characterization of CSCs. In this study, CD133-positive CSCs and CD133-negative cells were isolated from H460 cells, a large cell lung cancer model, by MACS method. CSCs properties were investigated by tumorSphere formation assay (2% agar plate), hanging drop assay and qPCR method. CD133 positivity of H460 cells was determined as 0.3%. It has been observed that CD133-positive cells show CSCs-specific self-renewal properties. It was determined that the CD133 gene expression difference between CD133-positive CSCs and CD133-negative cells was approximately 25-fold. It has been observed that CD133-positive CSCs express more pluripotency genes (such as OCT-4, SOX-2 and KLF-4) compared to CD133-negative cells.

Objective: Breast cancer is associated with a 5% genetic predisposition in women. Mitochondria play a role in important cellular events such as metabolism, cell death, and inflammation. Recent studies have highlighted precursor micro-RNA (pre-miRNA) to be located in mitochondria as well as mature miRNA. This study aims to reveal the occurrence of mitochondrial miRNA (mitomiR) in breast cancer cells. Materials and Methods: The study has prepared the mitochondrial fractions using the magnetic-activated cell sorting (MACS) method and performed small RNA (sRNA) sequencing. Results: The study has identified known and novel mitomiR sequences aligned to the mitochondrial genome. Conclusion: Identifying new mitomiRs can provide significant contributions and illuminate the molecular mechanism underlying mitomiR biogenesis. [ABSTRACT FROM AUTHOR]

Introduction: Graft-versus-host disease (GvHD) is a life-threatening syndrome commonly associated with hematopoietic stem cell transplantation (HSCT). Preventing the incidence of GVHD after HSCT along with minimizing long-term immunosuppression is currently under investigation with regulatory T cells (Tregs). As Tregs are a low-frequency population and the yield of all memory Tregs is not sufficient for clinical application, an initial Treg expansion is essential. Methods: Thirty milliliters of peripheral blood from the β-thalassemia major (beta-TM) patients and healthy controls were obtained and Tregs were isolated using MACS. Isolated cells were cultured in the presence of rapamycin and rIL-2 followed by activated with anti-CD3/CD28-coated beads. To evaluate Treg plasticity, expanded Tregs were cultured in a medium containing IL1β, IL6, TGFβ, and IL2, with or without 500 nM rapamycin for 72 h. To assess the functional properties of Tregs, CFSE dilution assays were performed to evaluate the ability of in vivo expanded Tregs from beta-TM patients. Statistical analysis was performed using paired t-test and independent t-test, with the aid of SPSS version 12.0. p-value ≤0.05 was considered significant. Results: The percentage of Tregs isolated from the control group was significantly higher than the Tregs isolated from patients (p-value = 0.01), which is probably due to the iron overload in beta-TM patients as a result of continuous blood transfusion. Also, the percentage of Tregs after 5 days of expansion had a significant increase in both groups compared to before expansion (p-value = 0.03). Our results also showed that the expansion of Tregs after 72 h in the presence of inflammatory cytokines and in the absence of rapamycin led to the increase in the intracellular expression of IL-17 (p-value = 0.01), while intracellular expression of IL-17 remained low following the addition of 100 nM rapamycin to the culture medium (pvalue = 0.073). The results of the functional evaluation of expanded Tregs showed relatively differences in both patient and control groups. Thus, expanded Tregs inhibited the proliferation of responder T cells in a dose-dependent manner in the control group (p-value = 0.028), while in the patient group this inhibitory effect was not significant (p-value = 0.055). Conclusion: Tregs isolated from beta-TM patients have poorer inhibitory performance than Tregs isolated from healthy individuals. Also, we concluded that rapamycin stabilizes the Treg population by inhibiting the production of IL-17, all necessitating the administration of appropriate immunosuppressive drugs in patients receiving Treg therapy. [ABSTRACT FROM AUTHOR]

In aging, the brain is more vulnerable to injury and neurodegenerative disease, but the mechanisms responsible are largely unknown. Evidence now suggests that neuroinflammation, mediated by resident brain astrocyte and microglia populations, are key players in the generation of inflammatory responses and may influence both age related processes and the initiation/progression of neurodegeneration. Consequently, targeting these cell types individually and collectively may aid in the development of novel disease-modifying therapies. We have optimized and characterized a protocol for the effective sequential isolation of both microglia and astrocytes from the adult mouse brain in young and aged mice. We demonstrate a technique for the sequential isolation of these immune cells by using magnetic beads technology, optimized to increase yield and limit potential artifacts in downstream transcriptomic applications, including RNA-sequencing pipelines. This technique is versatile, cost-effective, and reliable for the study of responses within the same biological context, simultaneously being advantageous in reducing mice numbers required to assess cellular responses in normal and age-related pathological conditions. [ABSTRACT FROM AUTHOR]

Objective: Sperm parameters and DNA integrity are crucial factors in ART outcomes. This study compared four sperm preparation methods (microfluidics, MACS, zeta potential, and swim-Up) for sorting spermatozoa with normal parameters and chromatin integrity., Methods: This study evaluated semen samples from 25 couples with male factor infertility. The semen samples were divided into four portions: one prepared by MACS, one by zeta potential, the other by microfluidics, and the last by swim-up. After preparation, sperm viability, motility, and morphology were assessed based on the WHO guidelines. DNA intergrity was assessed by SDF assay, and the CMA3 staining test was used to evaluate sperm chromatin packaging., Results: Compared to other preparation techniques, microfluidic preparation significantly improved sperm parameters, including motility, viability, morphology, and DNA integrity as well as chromatin packaging (p-value <0.05). The results also demonstrated that sperm motility, viability, and sperm DNA integrity as well as chromatin packaging, were not significantly different after preparation with MACS and Zeta potential methods. However, the MACS and Zeta methods produced improved sperm parameters and better DNA integrity than the swim-up method., Conclusions: Our results indicate that microfluidics can improve sperm quality compared to other methods of sperm preparation. When the microfluidic chip is not available, considering the similar results of sperm preparation by MACS and Zeta potential methods, it is preferred to use the Zeta method for the ART cycle due to its simplicity and cost-effectiveness.

Alexandra Kasim,1,&ast; Gabby Elbaz-Greener,2,3,&ast; Amjad Shalabi,4– 6 Erez Kachel,4– 6 Liza Grosman-Rimon,4,5 Michael Jerdev,4 offer amir,2– 5 Shemy Carasso2– 5 1Department of Radiology, B Padeh Medical Center, Poriya, Israel; 2Department of Cardiology, Hadassah Medical Center, Jerusalem, Israel; 3Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel; 4Cardiovascular Institute, B Padeh Medical Center, Poriya, Israel; 5The Azrieli Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel; 6Department of Cardiac Surgery, B Padeh Medical Center, Poriya, Israel&ast;These authors contributed equally to this workCorrespondence: Gabby Elbaz-GreenerDepartment of Cardiology, Hadassah Medical Center, Room Number 114, Jerusalem, IsraelTel +97226776564Fax +97226411028Email Gabby@hadassah.org.ilBackground: Severe mitral annulus calcification (MAC) is believed to bear high operative and post-operative risk during mitral valve replacement (MVR) surgery, including longer surgery time, post-surgical valvular leaks and increased rate of embolic phenomena. We hypothesized that quantification of mitral calcium in pre-operative chest computerized tomography (CCT), performed to assess aortic root before cross-clamping may help in risk assessment of adverse intraoperative and postoperative outcomes in patients undergoing MVR.Methods: We included patients who underwent MVR between the years 2015 and 2018 at Poriya medical center. Preoperative CCT was performed using Philips iCT 256 and Agatston mitral annulus calcium score (MACS) was retrospectively calculated using Philips Intellispace portal version 8.0. Patients were divided into MACS quintiles; 1– 3 quintiles were grouped (Low MACS) and compared to the 4– 5 quintiles (High MACS) group for demographic, clinical operative and post-operative parameters.Results: A total of 66 patients had MVR, out of which 61% were males, with mean age of 64± 9. Concomitant coronary or valvular procedures were done in 60% of patients. The median MACS was 43. High MACS (≥ 854) was not associated with longer bypass or cross clamp times. No differences in the MVR results were found between the groups. There were 6 post-operative embolic events; 1 mesenteric and 5 cerebral, which were not associated with MACS.Conclusion: MACS did not seem to be related to adverse outcomes in MVR. Due to a low event rate and probable pre-selection of patients without extreme mitral annulus calcifications our results should be confirmed in larger prospective study.Keywords: mitral annulus calcification, MACS, mitral valve replacement, MVR

Background: The primary function of testicular Leydig cells (LCs) is to produce testosterone (T). In vitro culture of the cells represents a very important approach to study androgen production and its regulations. Various methods have been developed for the enrichment of the cells from the testes. However, getting cells in large numbers with high purity and viability is still challenging. Here, we describe a new way to isolate LCs from rat testes in large quantity with high purity and viability. Methods: Enzymatic digested testicular cells from adult rats were labelled with prolactin receptor (PRLR) antibody. The positive cells were isolated by magnetic‐activated cell sorting (MACS) protocol. Purified LCs were tested in vitro for their steroidogenic (T production) and non‐steroidogenic (25‐OH‐vitamin D production and Insl3 and Cyp2r1expressions) functions in the presence of Luteinizing Hormone (LH) for up to 24 h. Results: Reanalysis of scRNA‐seq data indicates that Prlr expression is highly specific in LCs of adult rat testis. MACS procedure based on PRLR expression was able to isolate LCs with very high yield (about 106 cells/testis), high purity (about 95%) and viability (> 93%). Purified LCs retained high steroidogenic and non‐steroidogenic functions in responding to maximal LH stimulations, with more than 10‐fold increases in T production in 3 h and 42% and 103% increases in Insl3 and Cyp2r1 expressions in 24 h. Discussion and conclusion: We have established an excellent way to purify high quality LCs from adult rat testis that can serve as a useful tool to study the physiology, pharmacology and toxicology of the cells in vitro. [ABSTRACT FROM AUTHOR]

Background: The cases with unexplained infertility may have an abnormality in their sperm chromatin structure. Sperm selection methods can be used to separate sperm with low DNA fragmentation. The purpose of this study was to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) with magneticactivated cell sorting (MACS) in assisted reproductive techniques in cases with unexplained infertility. Methods: The semen samples were collected from couples with unexplained infertility. After semen analysis and sperm DNA fragmentation (SDF) evaluations, samples were prepared with swim-up method. The rates of SDF in different fractions including raw semen (n=20), swim-up (n=20), only motile sperm after swim-up (swim-up selection) (n=20), MACS sperm selection (n=20), only motile sperm after MACS (MACS selection) (n=20), and PICSI sperm selection (n=16) were evaluated. Also, the main sperm characteristics and fine morphology of sperm suspension after MACS were assessed. Statistical analysis was performed using GraphPad Prism. The p<0.05 was considered statistically significant. Results: DNA fragmentation index (DFI) values in PICSI and MACS groups were significantly reduced as compared to the swim-up group. The rate of this reduction was more pronounced in MACS (58.20±13.02) than PICSI (36.57±15.52) group. Also, our results showed that MACS resulted in decreased sperm motility, with no alteration in their fine morphology. Conclusion: MACS was found to be more efficient in reduction of SDF rates than PICSI. However, none of the sperm selection techniques can not totally eliminated the spermatozoa with DNA fragmentation in the final sperm sample. [ABSTRACT FROM AUTHOR]

• Liquid chromatography has been employed in plasma and urine as an adjunct in clinical diagnosis. • Administration of contraceptive medications may alter cortisol levels. • Variations in cortisol levels have been observed among different patient cohorts. • New cutoff values for diagnostic classification are needed when applying HPLC-MS. Cortisol is a glucocorticoid hormone, which is involved in cardiovascular, metabolic, inflammatory and behavioral processes in the human body. Immunoassays, used for routine analysis of analytes, usually lead to erroneous quantification due to the low specificity of these methods. In this study, we developed LC-MS/MS methods with surrogating matrices for evaluating cortisol in both urine and plasma samples and validated them according to Brazilian Health Regulatory Agency guidelines. Urine samples were prepared with an enzymatic hydrolysis stage, followed by a solid-phase extraction (C18 cartridges) using dichloromethane:methanol 9:1 as elution solvent. Plasma samples were prepared by protein precipitation with acetonitrile, using 50 μL of sample. HPLC was performed using a C8 column under 300 mL min−1 flow gradient conditions with water and methanol, both containing 5 mM ammonium formate and 0.1 % formic acid. Mass spectrometer with electrospray ionization in positive mode and selected reaction monitoring as detection technique were employed. Calibration curves were linear over a concentration range of 1–200 ng mL-1 for urine (r2 = 0.9950) and 0.5–300 ng mL-1 for plasma (r2 = 0.9970). The methods were selective, showed suitable precision, accuracy, and sensibility (limit of quantification = 0.85 ng mL−1 for urine and 0.15 ng mL−1 for plasma). Validated methods were successful applied to 22 real samples and a cohort of patients [n = 63 urines and n = 79 plasmas (from the Clementino Fraga Filho University Hospital, Rio de Janeiro, Brazil)]. [ABSTRACT FROM AUTHOR]

The maintenance of redox homeostasis is associated with a healthy status while the disruption of this mechanism leads to the development of various pathological conditions. Bioactive molecules such as carbohydrates accessible to the microbiota (MACs), polyphenols, and polyunsaturated fatty acids (PUFAs) are food components best characterized for their beneficial effect on human health. In particular, increasing evidence suggests that their antioxidant ability is involved in the prevention of several human diseases. Some experimental data indicate that the activation of the nuclear factor 2-related erythroid 2 (Nrf2) pathway—the key mechanism in the maintenance of redox homeostasis—is involved in the beneficial effects exerted by the intake of PUFAs and polyphenols. However, it is known that the latter must be metabolized before becoming active and that the intestinal microbiota play a key role in the biotransformation of some ingested food components. In addition, recent studies, indicating the efficacy of the MACs, polyphenols, and PUFAs in increasing the microbial population with the ability to yield biologically active metabolites (e.g., polyphenol metabolites, short-chain fatty acids (SCFAs)), support the hypothesis that these factors are responsible for the antioxidant action on the physiology of the host. The underlying mechanisms through which MACs, polyphenols, and PUFAs might influence the redox status have not been fully elucidated, but based on the efficacy of SCFAs as Nrf2 activators, their contribution to the antioxidant efficacy of dietary bioactives cannot be excluded. In this review, we aimed to summarize the main mechanisms through which MACs, polyphenols, and PUFAs can modulate the host’s redox homeostasis through their ability to directly or indirectly activate the Nrf2 pathway. We discuss their probiotic effects and the role played by the alteration of the metabolism/composition of the gut microbiota in the generation of potential Nrf2-ligands (e.g., SCFAs) in the host’s redox homeostasis.

Magnetic cell sorting technology stands out because of its speed, simplicity, and ability to process large cell numbers. However, it also suffers from a number of drawbacks, in particular low discrimination power, which results in all-or-none selection outcomes limited to a bulk separation of cell populations into positive and negative fractions, as well as the modest purity of the selected cells and the inability to select subpopulations of cells with high expression of a surface marker. In the present study, we developed a simple solution to this problem and confirmed the effectiveness of this approach by multiple experiments with the magnetic selection of transduced cell populations. Murine NIH 3T3 cells were transduced with the bicistronic retroviral vector constructs co-expressing fluorescent reporter proteins EGFP (enhanced green fluorescent protein) or DsRed-Express 2 and LNGFR (low-affinity nerve growth factor receptor) as surface selection markers. The effects of the magnetic selection of transduced cells with anti-LNGFR Micro Bead (MB) doses ranging from 0.5 to 80 µL have been assessed. Low doses of MBs favored the depletion of weakly positive cells from the population, resulting in the higher expression levels of EGFP or DsRed-Express2 reporters in the selected cell fractions. Low MB doses also contributed to the increased purity of the selected population, even for samples with a low initial percentage of positive cells. At the same time, high MB doses resulted in the increased yield and a more faithful representation of the original expression profiles following selection. We further demonstrate that for populations with fairly narrow distribution of expression levels, it is possible to achieve separation into high- and low-expressing subsets using the two-stage selection scheme based on the sequential use of low and high doses of MBs. For populations with broad expression distribution, a one-stage selection with low or high doses of MBs is sufficient for a clear separation of low- and high-expressing subsets in the column-retained and flow-through fractions, respectively. This study substantially extends the potential of magnetic cell sorting, and may open new possibilities in a number of biomedical applications.