A pilot study exploring time- and dose-dependent DNA damage and chromosomal instability caused by benzo[a]pyrene in two urothelial cell types.

Environmental and occupational exposure to polycyclic aromatic hydrocarbons (PAHs) is associated with adverse health effects in humans. Uncertainty exists regarding the causation of urinary bladder cancer by benzo[a]pyrene (B[a]P) due to a lack of sufficient data. In this work, we focused on in-vitro DNA damage and the formation of micronuclei and chromosomal aberrations as predictors of cancer risk, applying a wide range of dosages and time periods to quantify the onset, intensity, and duration of the response. We chose two urothelial cell types to compare susceptibility and the ability to increase the malignity of a pre-existing bladder cancer: a cancer cell line (T24) and a pooled sample of primary urinary bladder epithelia cells (PUBEC) from pigs. The highest level of DNA damage assessed by comet assay was observed following 24-h treatment in both cell types, whereas PUBEC cells were clearly more susceptible. Even 4-h treatment induced DNA damage in PUBEC cells with benchmark doses of 0.0027 µM B[a]P and 0.00023 µM after 4-h and 24-h exposure, respectively. Nearly no effect was observed for periods of 48 h. The frequency of micronucleus formation increased more markedly in T24 cells, particularly with 24-h treatment. In PUBEC cells, 48-h exposure notably induced the formation of nucleoplasmic bridges and nuclear buds. Even though only one biological replicate was studied due to the sophisticated study design, our results give a strong indication of the potential of B[a]P to induce and increase malignity in human-relevant cell types.

Crystallizing galactocele: A rare case report and review of literature.

ABSTRACT: Crystallizing galactocele is an uncommon condition that produces a viscous, chalky substance on fine needle aspiration cytology. (FNAC). Both the diagnosis and the management of this illness include the use of FNAC. Here, we discuss the case of a 25-year-old nursing woman who experienced left breast edema lump for two years. The upper outer quadrant of the leftt breast was involved by the hard, small, non-tender, and movable enlargement. The lesion's FNAC produced a thick, milky, and chalky substance. Numerous semi-transparent crystals of various sizes and shapes with angulated edges could be seen in cytological smears against a background of granular and amorphous proteinaceous material. A diagnosis of crystallizing galactocele was made on the basis of the patient's clinical history of lactation and characteristic cytological findings. Due to the rarity of this condition-to the best of our knowledge, less than ten cases of crystallizing galactocele have been documented in medical literature.

Genome sequence of bubaline herpesvirus-1 associated with pustular vulvovaginitis in Indian water buffalo.

We report here the genome sequence of a bubaline herpesvirus 1 isolated from Indian water buffalo. The bubaline herpesvirus 1 strain S102_1 was isolated in 2021 from a Murrah buffalo heifer with clinical presentation of pustular vulvovaginitis.

Stimuli-responsive and self-healing supramolecular Zn(II)-guanosine metal-organic gel for Schottky barrier diode application.

Spontaneous formation of a supramolecular metal-organic hydrogel using unsubstituted guanosine as a ligand and Zn2+ ions is reported. Guanosine, in the presence of NaOH, self-assembled into a stable G-quadruplex structure, which underwent crosslinking through Zn2+ ions to afford a stable hydrogel. The gel has been characterized using several spectroscopic as well as microscopic studies. The hydrogel demonstrated excellent stimuli responsiveness towards various chemicals and pH. Furthermore, the gel exhibited intrinsic thixotropic behavior and showed self-healing and injectable properties. The optical properties of the Zn-guanosine metallo-hydrogel suggested a semiconducting nature of the gel, which has been exploited for fabricating a thin film device based on a Schottky diode interface between metal and a semiconductor. The fabricated device shows excellent charge transport characteristics and linear rectifying behavior. The findings are likely to pave the way for newer research in the area of soft electronic devices fabricated using materials synthesized by employing simple biomolecules.

Genetic analysis for semen quality traits in buffalo bulls.

This study was attempted to estimate the genetic parameters of semen quality traits in buffalo bulls. The study data consisted of 10975 ejaculates from 45 Murrah buffalo bulls (aged 24-72 months) used for breeding program during year 2010 to 2020. Semen quality traits (ejaculate volume, concentration of sperm, mass activity, initial and post-thaw motility, number of sperms per ejaculate, motile sperm number and discard rates) were studied. It was observed that average ejaculate volume was 2.82 ± 1.45 mL with mean concentration of 1040.12 ± 523.26 million/mL. Higher heritability was observed for number of sperms per ejaculate, number of motile sperm and sperm concentration. Significant phenotypic correlation was obtained between volume and number of sperms per ejaculate as well as volume and number of motile sperms. Likewise, significant phenotypic correlation was evident between sperm concentration with sperm number per ejaculate. Highest phenotypic correlation was obtained between sperm count per ejaculate and motile sperm count. Estimated genetic trends showed significant change in volume and motile sperm number. In conclusion, this study ascertains that genetic parameters of semen traits can be considered during the selection of buffalo bulls in breeding program.

Self-Managed Abortion in the United States.

Purpose of Review: This review aims to provide up-to-date information about self-managed abortion in the USA. Recent Findings: Evidence indicates that there is growing demand for self-managed abortion in the USA as obstacles to facility-based care increase, especially since the Supreme Court overturned Roe v. Wade. Self-managed abortion with medications is safe and effective. Summary: Based on a nationally representative survey, the lifetime prevalence of self-managed abortion in the USA in 2017 was estimated to be 7%. People experiencing barriers to abortion care, including people of color, people with lower incomes, people in states that have restrictive abortion laws, and those living farther from facilities providing abortion care are more likely to attempt self-managed abortion. While people may use a range of methods to self-manage abortion, there is growing use of safe and effective medications, including mifepristone used together with misoprostol and misoprostol used alone; use of traumatic and dangerous methods is rare. While many people attempt to self-manage their abortion because of barriers to facility-based care, others have a preference for self-care because it is convenient, accessible, and private. While the medical risks of self-managed abortion may be few, the legal risks may be significant. Sixty-one people have been criminally investigated or arrested between 2000 and 2020 for allegedly self-managing their abortion or helping someone else do so. Clinicians play an important role in providing evidence-based information and care to patients considering or attempting self-managed abortion, as well as minimizing legal risks.

Obstacles to Care Mount 1 Year After Dobbs Decision.

This Viewpoint discusses the ramifications of the Dobbs v Jackson Women's Health Organization decision by the US Supreme Court and the state of reproductive health care in the US.

Epsilon poly-lysine in buffalo semen extender: A step towards reducing the development of antibiotic resistance.

The use of antibiotics in semen extenders can contribute to the development of antibiotic resistance. The objective of the study was to evaluate epsilon-polylysine (Ɛ-PL) as a substitute for antibiotics in the buffalo semen extender. For this, 20 semen ejaculates were collected from four Murrah buffalo bulls. Each ejaculate was divided into three equal aliquots and extended into an egg yolk-based semen extender containing either antibiotics (strepto-penicillin) or different concentrations of Ɛ-PL (0.64 and 1.28 g/L) to make the final concentration 80 million sperm/mL and cryopreserved as per the standard procedure. The antibiogram sensitivity test confirmed that Ɛ-PL is an effective antimicrobial against microbes present in buffalo semen ejaculates. Furthermore, the addition of Ɛ-PL in the semen extender significantly reduces the colony forming unit (CFU)/mL in cryopreserved semen equivalent to strepto-penicillin. The sperm motility and kinematic parameters assessed by a computer-assisted sperm analyser showed that Ɛ-PL did not inhibit either sperm motility not kinematic parameters of cryopreserved sperm. The flow-cytometric evaluation of frozen-thawed sperm revealed interesting results. The extender supplemented with Ɛ-PL protected sperm acrosome and mitochondrial membrane potential greater than the extender supplemented with strepto-penicillin. Further, Ɛ-PL reduced significantly the production of superoxide anions from mitochondria during the cryopreservation process. In this way, Ɛ-PL may be a suitable alternative to antibiotics in semen extenders. In conclusion, Ɛ-PL at a concentration of 0.64 g/L acts as an effective antimicrobial as well as antioxidant in semen extender for cryopreservation of buffalo sperm.

Perspectives on an early abortion ban in a restrictive US state: A qualitative exploration.

OBJECTIVES: To gain a deeper understanding of perspectives on abortion and early abortion bans in a restrictive US state. STUDY DESIGN: We conducted a qualitative study using semistructured Zoom interviews with residents of the US state Georgia's 6th Congressional District. Potential participants first completed a screening tool to recruit people who held "middle-of-the-spectrum" views on abortion based on two abortion questions on a 5-point Likert scale. The interviews focused on participants' thoughts and feelings on abortion and Georgia's early abortion ban. We transcribed, coded, and analyzed the interviews, and present a subset of themes. RESULTS: We interviewed 28 people from March to May 2020. Participants often described holding complex views on abortion shaped by a range of lived experiences, values, and identities. They lamented the "black-and-white" nature of the national abortion discussion, which they felt oversimplified the issue and did not represent their views. Participants discussed the importance of experiences that allowed them to empathize with people who choose abortion, even when they personally felt they would make a different decision in a similar situation. Based on these experiences, many participants emphasized the importance of separating their own views on abortion from what needed to be regulated for others. However, participants often demonstrated a lack of understanding about the extent to which HB481 makes abortion inaccessible in Georgia. CONCLUSIONS: Our results indicate that, even in states traditionally labeled as restrictive or hostile towards abortion, many people express an openness to understand others' experiences and hold complex and multifaceted views. IMPLICATIONS: Our laws and policies at the state level profoundly affect the practice of medicine and access to care. Our study furthers our understanding of how a sample of people in a restrictive US state think and feel about abortion and early abortion bans. These results can be used to support educational efforts, policies, and communication practices that better reflect the complex views of the public.

Self-managed Abortion in the US.

This Viewpoint details how clinicians can treat patients who have self-managed an abortion without putting the patient at risk for criminalization.

Ionic Liquid-Mediated Interfacial Polymerization for Fabrication of Reverse Osmosis Membranes.

This study revealed the effects of incorporating ionic liquid (IL) molecules: 1-ethyl, 1-butyl, and 1-octyl-3-methyl-imidazolium chlorides with different alkyl chain lengths, in interfacial polymerization (IP) on the structure and property (i.e., permeate-flux and salt rejection ratio) relationships of resulting RO membranes. The IL additive was added in the aqueous meta-phenylene diamine (MPD; 0.1% w/v) phase, which was subsequently reacted with trimesoyl chloride (TMC; 0.004% w/v) in the hexane phase to produce polyamide (PA) barrier layer. The structure of resulting free-standing PA thin films was characterized by grazing incidence wide-angle X-rays scattering (GIWAXS), which results were correlated with the performance of thin-film composite RO membranes having PA barrier layers prepared under the same IP conditions. Additionally, the membrane surface properties were characterized by zeta potential and water contact angle measurements. It was found that the membrane prepared by the longer chain IL molecule generally showed lower salt rejection ratio and higher permeation flux, possibly due to the inclusion of IL molecules in the PA scaffold. This hypothesis was supported by the GIWAXS results, where a self-assembled surfactant-like structure formed by IL with the longest aliphatic chain length was detected.

Detrimental NFKB1 missense variants affecting the Rel-homology domain of p105/p50.

Most of the currently known heterozygous pathogenic NFKB1 (Nuclear factor kappa B subunit 1) variants comprise deleterious defects such as severe truncations, internal deletions, and frameshift variants. Collectively, these represent the most frequent monogenic cause of common variable immunodeficiency (CVID) identified so far. NFKB1 encodes the transcription factor precursor p105 which undergoes limited proteasomal processing of its C-terminal half to generate the mature NF-κB subunit p50. Whereas p105/p50 haploinsufficiency due to devastating genetic damages and protein loss is a well-known disease mechanism, the pathogenic significance of numerous NFKB1 missense variants still remains uncertain and/or unexplored, due to the unavailability of accurate test procedures to confirm causality. In this study we functionally characterized 47 distinct missense variants residing within the N-terminal domains, thus affecting both proteins, the p105 precursor and the processed p50. Following transient overexpression of EGFP-fused mutant p105 and p50 in HEK293T cells, we used fluorescence microscopy, Western blotting, electrophoretic mobility shift assays (EMSA), and reporter assays to analyze their effects on subcellular localization, protein stability and precursor processing, DNA binding, and on the RelA-dependent target promoter activation, respectively. We found nine missense variants to cause harmful damage with intensified protein decay, while two variants left protein stability unaffected but caused a loss of the DNA-binding activity. Seven of the analyzed single amino acid changes caused ambiguous protein defects and four variants were associated with only minor adverse effects. For 25 variants, test results were indistinguishable from those of the wildtype controls, hence, their pathogenic impact remained elusive. In summary, we show that pathogenic missense variants affecting the Rel-homology domain may cause protein-decaying defects, thus resembling the disease-mechanisms of p105/p50 haploinsufficiency or may cause DNA-binding deficiency. However, rare variants (with a population frequency of less than 0.01%) with minor abnormalities or with neutral tests should still be considered as potentially pathogenic, until suitable tests have approved them being benign.

Comparison of various delignification/desilication pre-treatments and indigenous fungal cellulase for improved hydrolysis of paddy straw.

The efficient removal of lignin and silica from paddy straw is essential for its volarization into biofuels and other value-added products. In this work, different chemical pre-treatments viz. acid/alkali, organosolv and deep eutectic solvents were carried out to assess the extent of delignification and desilication of paddy straw. Maximum lignin and silica removal of 96.08 and 95.51% was observed with two step acid (0.5% sulphuric acid) followed by alkali (4% sodium hydroxide) pre-treatment with significantly low total lignin (2.30%) and silica content (0.80%) of the treated straw residue. The treated straw residue contained significantly high holocellulose (91.65%), cellulose (75.01%) and hemicellulose content (16.64%). Among the four indigenous fungal isolates, Penicillium mallochii (JS17) cellulase showed better accessibility for the treated straw residue with maximum release of 504.18 mg g-1 of reducing sugars and saccharification efficiency of 56.90%. The two-step acid/alkali pre-treatment of paddy straw was highly effective for removing lignin and silica from paddy straw, thereby, resulting in enhanced enzymatic accessibility of the substrate and more efficient hydrolysis of cellulose into fermentable sugars.

Increased Seroprevalence and Improved Antibody Responses Following Third Primary SARS-CoV-2 Immunisation: An Update From the COV-AD Study.

Background: Patients with primary and secondary antibody deficiency are vulnerable to COVID-19 and demonstrate diminished responses following two-dose SARS-CoV-2 vaccine schedules. Third primary vaccinations have been deployed to enhance their humoral and cellular immunity. Objectives: To determine the immunogenicity of the third primary SARS-CoV-2 immunisation in a heterogeneous cohort of patients with antibody deficiency. Methods: Participants enrolled in the COV-AD study were sampled before and after their third vaccine dose. Serological and cellular responses were determined using ELISA, live-virus neutralisation and ELISPOT assays. Results: Following a two-dose schedule, 100% of healthy controls mounted a serological response to SARS-CoV-2 vaccination, however, 38.6% of individuals with antibody deficiency remained seronegative. A third primary SARS-CoV-2 vaccine significantly increased anti-spike glycoprotein antibody seroprevalence from 61.4% to 76.0%, the magnitude of the antibody response, its neutralising capacity and induced seroconversion in individuals who were seronegative after two vaccine doses. Vaccine-induced serological responses were broadly cross-reactive against the SARS-CoV-2 B.1.1.529 variant of concern, however, seroprevalence and antibody levels remained significantly lower than healthy controls. No differences in serological responses were observed between individuals who received AstraZeneca ChAdOx1 nCoV-19 and Pfizer BioNTech 162b2 during their initial two-dose vaccine schedule. SARS-CoV-2 infection-naive participants who had received a heterologous vaccine as a third dose were significantly more likely to have a detectable T cell response following their third vaccine dose (61.5% vs 11.1%). Conclusion: These data support the widespread use of third primary immunisations to enhance humoral immunity against SARS-CoV-2 in individuals with antibody deficiency.

Effect of mitochondria-targeted antioxidant on the regulation of the mitochondrial function of sperm during cryopreservation.

Sperm mitochondrion is one of the major susceptible organelles that get damaged during cryopreservation. The study aimed to minimize mitochondrial dysfunction and oxidative stress during sperm cryopreservation using mitochondria-specific antioxidants. For this, semen was collected from five buffalo bulls (3 ejaculates/bull). The ejaculates were diluted in an low-density lipoprotein-based extender and divided into four equal aliquots. Mitochondria-targeted antioxidant (MitoQ) was added at a final concentration of 0 (control), 0.02, 0.2 and 2 µM separately in each aliquotes and cryopreserved. The addition of MitoQ at a concentration of 0.02 µM improved post-thaw sperm motility, plasma membrane integrity and able to sustain sperm motility for a longer time. To investigate MitoQ's effects on mitochondrial function, we measured mitochondrial membrane potential (MMP) using JC-1 dye, superoxide production using Mitosox assay, and lipid peroxidation by TBARS assay. The supplementation of 0.02 µM MitoQ in the extender prevented the significant reduction of MMP and reduced superoxide production resulting in lower lipid peroxidation of sperm plasma membrane after cryopreservation. Further, we found that a higher concentration of MitoQ decreases MMP and increases mitochondrial superoxide production. In conclusion, MitoQ @ 0.02 µM can alleviate oxidative stress by regulating mitochondrial functionality in spermatozoa during cryopreservation.

SARS-CoV-2 Vaccine Responses in Individuals with Antibody Deficiency: Findings from the COV-AD Study.

BACKGROUND: Vaccination prevents severe morbidity and mortality from COVID-19 in the general population. The immunogenicity and efficacy of SARS-CoV-2 vaccines in patients with antibody deficiency is poorly understood. OBJECTIVES: COVID-19 in patients with antibody deficiency (COV-AD) is a multi-site UK study that aims to determine the immune response to SARS-CoV-2 infection and vaccination in patients with primary or secondary antibody deficiency, a population that suffers from severe and recurrent infection and does not respond well to vaccination. METHODS: Individuals on immunoglobulin replacement therapy or with an IgG less than 4 g/L receiving antibiotic prophylaxis were recruited from April 2021. Serological and cellular responses were determined using ELISA, live-virus neutralisation and interferon gamma release assays. SARS-CoV-2 infection and clearance were determined by PCR from serial nasopharyngeal swabs. RESULTS: A total of 5.6% (n = 320) of the cohort reported prior SARS-CoV-2 infection, but only 0.3% remained PCR positive on study entry. Seropositivity, following two doses of SARS-CoV-2 vaccination, was 54.8% (n = 168) compared with 100% of healthy controls (n = 205). The magnitude of the antibody response and its neutralising capacity were both significantly reduced compared to controls. Participants vaccinated with the Pfizer/BioNTech vaccine were more likely to be seropositive (65.7% vs. 48.0%, p = 0.03) and have higher antibody levels compared with the AstraZeneca vaccine (IgGAM ratio 3.73 vs. 2.39, p = 0.0003). T cell responses post vaccination was demonstrable in 46.2% of participants and were associated with better antibody responses but there was no difference between the two vaccines. Eleven vaccine-breakthrough infections have occurred to date, 10 of them in recipients of the AstraZeneca vaccine. CONCLUSION: SARS-CoV-2 vaccines demonstrate reduced immunogenicity in patients with antibody deficiency with evidence of vaccine breakthrough infection.

Ameliorative Effect of Acetyl L-carnitine in Alzheimer's Disease via Downregulating of Homocysteine Levels in Hyperhomocysteinemia Induced Cognitive Deficit in Mouse Model.

AIMS: The study was aimed at exploring the role of Acetyl L-Carnitine supplementation attenuating dementia and degradation of cognitive abilities in Hyperhomocysteinemia induced AD manifestations in the mouse model. BACKGROUND: Alzheimer's disease (AD) is a neurological disorder that is marked by dementia, and degradation of cognitive abilities. There is great popularity gained by natural supplements as the treatment for AD, due to the higher toxicities of synthetic drugs. Hyperhomocysteinemia causes excitotoxicity to the cortical neurons, which brought us to the point that amino acids possibly have a role in causing cholinergic deformities, which are an important etiological parameter in AD. Acetyl L-Carnitine a methyl donor with the presence of three chemically reactive methyl groups linked to a nitrogen atom was found to possess neuroprotective activity against experimental models of AD. OBJECTIVE: The objective of the present investigation was to investigate and evaluate the pharmacological effect of Acetyl L-Carnitine against hyperhomocysteinemia induced Alzheimer's disease (AD) in the mouse model. MATERIALS AND METHODS: The animals were divided into normal control (vehicle-treated), HHcy (dl-Homocysteine thiolactone treated) negative control, test group i.e., low dose (50mg/kg, p.o) of acetyl L-carnitine (L-ALC), high dose (100mg/kg,p.o) of acetyl L-carnitine (H-ALC), L-ALC+ SOV (Sodium orthovanadate) and H-ALC+SOV. HHcy was induced by administration of dl-Homocysteine thiolactone (dl-HCT; 1 g/kg, p.o.) on day-1 to day-15 of experimental schedule to all animals except normal control. The changes in the behaviour pattern of the animals due to neuroinflammation, and cholinergic dysfunction were examined in rotarod, novel objective recognition, passive avoidance, elevated plus maze, and morris water maze analysis. Biochemical investigation includes the estimation of total homocysteine (tHcy), Creatinine Kinase (CK), Acetylcholinesterase (AChE), thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and IL-6 and TNF-α. RESULTS: Supplementation of ALC in mouse considerably lowered the HHcy-induced AD manifestations in the experimental animals. It was found that ALC and SOV successfully diminished the behaviour abnormalities and lessened the Hcy-induced alteration in systemic Hcy levels, CK activity, and cholinergic dysfunction with improved bioenergetics in the Prefrontal cortex of the mice. CONCLUSION: ALC was found to improve the HHcy-induced cognitive disabilities which was found to be associated with the decreased systemic levels of Hcy, CK, and cholinergic abnormalities. It also combats the oxidative stress-induced neuroinflammation with diminished pro-inflammatory markers in the pre frontal cortex. The outcomes collectively indicate ALC's potential to be used as a supplementation in the pharmacotherapy of AD.

Hormonal Contraception: Systemic Estrogen and Progestin Preparations.

Combined hormonal contraception (CHC) are short-acting, reversible methods containing both estrogen and progestin. Available CHC methods include combined oral contraceptives, transdermal patches, and vaginal rings. The combined oral contraceptive remains the most commonly used contraceptive method in the United States. The general principles of CHC will be reviewed, including mechanism of action and effectiveness. Unless otherwise stated, these principles apply to all CHCs. When discussing clinical studies and specific considerations related only to pills, patches, or rings, the method(s) will be specified. Words that specify sex are used when discussing studies in which sex was specified.

IGF-1 supplementation in semen affects mitochondrial functional and calcium status of buffalo sperm following cryopreservation.

This study was designed to examine the effects of seminal insulin-like growth factor-1 (IGF-1) supplementation on structural and functional properties of buffalo sperm post cryopreservation. Semen ejaculates from buffalo bulls (n = 6) were proportioned into four aliquots and diluted with egg yolk-based extender. Prior to equilibration, IGF-1 was added to extender as four treatments: group IGF0 (no supplementation), IGF150 (150 ng/mL), IGF250 (250 ng/mL) and IGF350 (350 ng/mL). The extended semen was transferred into 0.25 mL mini-straws, equilibrated (4 °C at 4 h), and cryopreserved. Total sperm motility was greater (P < 0.05) when there was the IGF150 treatment compared with values for other groups. Furthermore, with the IGF150 treatment there was the least and greatest (P < 0.05) mitochondrial superoxide status and membrane potential, respectively. Similarly, with the IGF150 treatment there was a greater (P < 0.05) sperm membrane integrity with a lesser (P < 0.05) calcium status compared to values for the other groups. In conclusion, seminal IGF-1 supplementation affects the structural and functional properties of buffalo sperm following cryopreservation.