bluuebunny/crossref_metadata_2025 huggingface dataset:
Exploring therapeutic potential of Woodfordia fruticosa (L.) Kurz leaf and bark focusing on antioxidant, antithrombotic, antimicrobial, anti‐inflammatory, analgesic, and antidiarrheal properties
Authors: Rahman Md. Mahfuzur , Soma Mahfuza Afroz , Sultana Nahid , Hossain Md. Jamal , Sufian Md. Abu , Rahman M. Oliur , Rashid Mohammad A.
Published: October 2023

Background and Aims

The study aimed to evaluate the pharmacological properties of methanolic extracts of leaves and barks of Woodfordia fruticosa (L.) Kurz (family: Lythraceae) focusing on antioxidant, thrombolytic, anti‐inflammatory, antibacterial, analgesic, and antidiarrheal effects.

Methods

1,1‐Diphenyl‐2‐picrylhydrazyl (DPPH) free radical scavenging assay, clot lysis, disc diffusion, and membrane stabilizing methods were employed to assess in vitro antioxidant, thrombolytic, antibacterial, and anti‐inflammatory properties of the leaf and bark methanolic extracts (ME) of W. fruticosa and different organic solvents, that is, petroleum ether (PE), dichloromethane (DCM), chloroform (CL), and aqueous (AQ) fractions. In addition, in vivo central and peripheral analgesic and antidiarrheal activities of both crude extracts were evaluated at two doses (200 and 400 mg/kg of body weight [bw]).

Results

All the extracts and fractions showed promising antioxidant properties by scavenging DDPH free radicals with IC50 of 6.11–20.79 μg/mL. AQ fraction (41.24%) of leaves and ME (44.90%) of bark exerted notable in vitro thrombolytic activity. The CL fraction of leaves and AQ fraction of the bark showed 43.16% and 45.37% inhibition of RBC hemolysis, respectively, compared to the inhibition of RBC hemolysis by aspirin in a hypotonic‐induced membrane stabilizing assay. Besides, both extracts were observed to provide significant (p < 0.001) central and peripheral analgesic responses at both doses of 200 and 400 mg/kg bw. Furthermore, both doses of bark extract (p < 0.001) and the 400 mg/kg bw of leaf extract (p < 0.05) were observed to possess statistically significant antidiarrheal activity. Additionally, in an in vivo acute toxicity investigation, both extracts had a median lethal dose (LD50) greater than 5000 mg/kg bw, indicating their safety level.

Conclusion

The current study proves the ethnomedicinal uses of W. fruticosa; however, further studies are required for phytochemical screening to isolate the responsible bioactive compounds and discover the lead molecules from the plant species.
Does organizational support affect the governance performance of the farm irrigation system? Evidence from the Yellow River basin in China
Authors: Fu Yangqi , Zhu Yuchun
Published: October 2023

The engagement of farmers in the governance of farm irrigation systems is crucial for meeting irrigation demands and enhancing farm productivity. Theoretically, grassroots organizations' concerns and support for farmers will encourage them to participate in the activities of the organization, thus improving the management performance of the irrigation system. This paper examines the influence of organizational aid on the governance of agricultural irrigation systems using field survey data in the Yellow River basin, China. The seemingly unrelated regression model is used to explore the group disparities under distinct organizational circumstances and farm household characteristics. The study demonstrates that the software and hardware support provided by organizations can promote improved management performance of irrigation systems. In addition, considering the heterogeneity of organizational environments, such support is more conducive to enhancing the governance of systems in organizations with more cooperative atmospheres and higher degrees of fairness. From the perspective of the heterogeneity of system users, providing organizational support to growers with a low degree of nonagricultural transformation and labour shortage is more conducive to improving governance performance. Consequently, establishing an excellent organizational atmosphere and targeting organizational support to farmers is a viable means to optimize the governance of irrigation systems.
Overview of the Fluoroquinolone Antibiotics
Authors: Just Paul M.
Published: October 2023

The fluoroquinolones represent an important advance in antimicrobial therapy. Commercially available products in the United States now include norfloxacin, ciprofloxacin, ofloxacin, enoxacin, and lomefloxacin. Although they share a common mechanism of action, they differ significantly in their antimicrobial spectrum of activity, their pharmacokinetic characteristics, and, to a lesser degree, their safety profiles. These compounds are generally highly effective against aerobic gram‐negative and many gram‐positive isolates; their activity is more limited against anaerobic bacteria. Quinolone‐resistant bacteria have been isolated, but most do not appear to pose a clinically significant problem at this time. The agents are effective in the treatment of a wide range of infections. Although some, such as ciprofloxacin and enoxacin, have been associated with clinically significant interactions with theophylline derivatives, others such as ofloxacin and lomefloxacin appear to have a limited propensity for such interactions.
Urinary Tract Infections in the Elderly
Authors: McCue Jack D.
Published: October 2023

Urinary tract infection (UTI) is the most common infectious disease of the elderly and is especially prevalent in debilitated, institutionalized older individuals. Unlike UTI in younger women, which tends to be related to frequency of sexual intercourse and is uncomplicated, in the elderly it is more difficult to treat and its pathogenesis is related to abnormal bladder function, bladder outlet obstruction, vaginal and urethral atrophy, use of long‐term indwelling catheters, and puddling related to bed rest. The spectrum of organisms causing infection relates to the ecology of the patients' environments; those residing in nursing homes and especially with permanent indwelling catheters tend to have a greater variety of pathogenic organisms, many of which may be relatively antibiotic resistant. Short‐course antibiotic therapy is less effective in older patients, and relapse or recurrence is more common regardless of the duration of treatment. Asymptomatic bacteriuria is common in older patients with abnormal bladder function. The clinical significance of asymptomatic bacteriuria generally is minor, and treatment is not beneficial.
Pharmaceutical Care and Specialty Practice
Authors: Hepler Charles D.
Published: October 2023

The practice of pharmaceutical care is a possible goal for all pharmacists' professional maturation. Six issues exist in which pharmaceutical care may represent a radical departure from some contemporary clinical practices. Because no profession is capable of providing pharmaceutical care singlehandedly at an acceptable level of quality, it is necessary to construct systems that organize the necessary variety of professionals and patients for managing drug therapy. In such a system, pharmacists can organize specialty practice around a strong general practice of pharmacy that is able to coordinate drug therapy and manage for outcomes, organize around the practice of medicine as if pharmacy were a medical specialty, or merge into the organization of a hospital or other healthcare organization.
Changing Trends in Antihypertensive Therapy in Two New England Communities During the 1980s
Authors: Hume Anne Lamont , Barbour Marilyn McFarland , Willey Cynthia J. , Assaf Annlouise R. , Lapane Kate L. , Carleton Richard A.
Published: October 2023

The prevalence rates and correlates of antihypertensive drug use among individuals with hypertension were determined using data derived from five biennial population‐based surveys conducted between 1981 and 1990 in two New England communities. Point prevalence estimates were determined for nine categories of antihypertensive agents at five time points, and were analyzed by age and sex using multiple logistic regression. In the first cross‐sectional survey, the prevalence of use per 1000 individuals with hypertension was 235.4 for diuretics, 57.1 for β‐blockers, 65.5 for combination products, 29.2 for central α‐agonists, 2.8 for peripheral α‐antagonists, and 8.4 each for adrenergic blockers and direct vasodilators. The prevalence rates for calcium channel blockers and angiotensin‐converting enzyme inhibitors increased sharply between the third and fourth survey cycles. Significant age‐ and sex‐related differences in antihypertensive use were detected.
Understanding and Treating Dyslipidemia Associated With Noninsulin‐Dependent Diabetes Mellitus and Hypertension
Authors: McKenney James M.
Published: October 2023

Hypertension and diabetes appear to increase coronary heart disease risk in part by causing an abnormality in lipid metabolism. Most affected are patients with familial dyslipidemic hypertension (FDH) and noninsulin‐dependent diabetes mellitus (NIDDM). The lipid disorders most often encountered in these patients are increased levels of triglycerides, very low‐density lipoprotein (VLDL) cholesterol, and small, dense low‐density lipoprotein (LDL) cholesterol, and low levels of high‐density lipoprotein (HDL) cholesterol. These abnormalities appear to result from increased hepatic secretion of VLDL particles due to increased concentrations of free fatty acids and glucose, reduced VLDL clearance due to reduced activity of lipoprotein lipase, and reduced LDL clearance due to glycosylation of ligand proteins. Treatment of the dyslipidemia associated with FDH should follow the guidelines from the National Cholesterol Education Program. Treatment in men and women with NIDDM should be considered when LDL cholesterol levels are 130 mg/dl or above, triglyceride levels are 200 mg/dl or above, or non‐HDL cholesterol levels are 160 mg/dl or greater. Aggressive lifestyle changes should be initiated first, including weight loss in obese patients, control of glucose levels in those with NIDDM, avoidance of antihypertensive drugs that may worsen lipid levels in patients with FDH, and eating a diet restricting saturated fat and cholesterol. Addition of lipid‐altering drugs should be considered if such changes do not achieve effective lipid control. The agent should be tailored to the patient's lipid profile, in general by using bile acid resins, niacin, or reductase inhibitors to lower LDL cholesterol and gemfibrozil or niacin to lower triglycerides. Niacin should be avoided in patients with NIDDM.
Differences in Serum Concentrations of and Responses to Generic Verapamil in the Elderly
Authors: Carter Barry L. , Noyes Margaret A. , Demmler Richard W.
Published: October 2023

Study Objective. To determine if there are greater differences in bioavailability of generic verapamil at steady state in elderly patients than in healthy young subjects.

Design. Double‐blind, randomized, three‐way crossover study.

Setting. Clinical research center at a general teaching hospital.

Subjects. Eight young, healthy subjects between ages 18 and 45 years, and eight elderly, hypertensive patients over 65 years of age.

Interventions. Each participant was randomly assigned to receive brand‐name verapamil or one of two generic preparations. Young subjects received each of the three preparations 80 mg twice/day for 7 days separated by a 1‐week washout. Elderly patients took the three preparations for 21 days. Serum drug levels and blood pressure were measured and electrocardiograms performed 13 times after the last dose of drug.

Main Results. In young subjects, the pharmacokinetics and pharmacodynamics of the three preparations were nearly identical. There was, however, a great deal of variability in the bioavailability of the generic products. In the elderly patients, one generic product had a maximum serum concentration and area under the curve that were 77% and 43% greater, respectively, than the brand‐name product (p<0.02). Assessments of bioequivalency (based on confidence intervals) demonstrated that this generic drug's bioavailability was from 4–118% and peak serum concentration from 10–139% greater than the brand‐name drug. This was associated with greater prolongation of the PR intervals.

Conclusions. A generic verapamil product that is bioequivalent in young subjects may not be bioequivalent in elderly subjects. Testing of drugs with complex pharmacokinetics in young subjects may not reliably predict the clinical outcomes in elderly hypertensive patients.
Institution‐Specific Patterns of Infection and Gram's Stain as Guides for Empiric Treatment of Patients Hospitalized With Typical Community‐Acquired Pneumonia
Authors: Belliveau Paul , Hickingbotham Nancy , Maderazo Eufronio G. , Mazens‐Sullivan Mary , Robinson Ann
Published: October 2023

Study Objective. To determine the appropriateness of cefazolin as empiric treatment of typical, as opposed to atypical, bacterial community‐acquired pneumonia at our institution.

Design. Combination of retrospective chart review and prospective determination of microbial susceptibilities and cefazolin‐associated cost savings.

Setting. General acute‐care referral hospital.

Patients. We evaluated the charts of patients discharged with a diagnosis of community‐acquired pneumonia over a 10‐year period. Gram's stains and culture results of sputum samples processed over 2 months were analyzed to determine the ability of the stains to predict positive Haemophilus influenzae cultures. The susceptibility and β‐lactamase status of clinical isolates of H. influenzae were determined. Cost savings of cefazolin as empiric treatment for community‐acquired pneumonia were evaluated.

Measurements and Main Results. The frequency of H. influenzae pneumonia at our institution was 15% of the three major bacterial community‐acquired pneumonias. Gram's stain was highly accurate in predicting the presence or absence of Haemophilus sp in sputum. Five patients had positive outcomes with cefazolin treatment despite being diagnosed with H. influenzae pneumonia. The organism isolates demonstrated intermediate sensitivity to cefazolin and 85% were β‐lactamase negative. Our program that encourages empiric use of cefazolin over cefuroxime for typical bacterial community‐acquired pneumonia has allowed a modest projected annual cost savings of $24,000.

Conclusions. We concluded that when Gram's stain of sputum does not show Haemophilus sp in patients with typical bacterial community‐acquired pneumonia, empiric treatment with cefazolin is appropriate and results in cost savings.
Faculty Have Few Incentives for Becoming Board Certified in Pharmacotherapy
Authors: Wagner Mary L. , Wagner Bertil K.J. , Smith Curtis L.
Published: October 2023

A telephone survey of all colleges of pharmacy in the United States (including Puerto Rico) and Canada was conducted to assess the incentives offered for pharmacy practice faculty to become board‐certified pharmacotherapy specialists (BCPS). Board certification is not a requirement for employment at any college of pharmacy; however, it plays a part in promotion, tenure, and merit salary increases at several schools. Fewer than half of the schools provide at least partial reimbursement of examination‐related expenses. Sixteen have initiated BCPS study groups. We perceive that faculty must have more incentives to take the examination. It may be difficult to change institutional policies regarding merit increases or promotion and tenure, but colleges of pharmacy can do more to decrease the cost burden and to promote study groups within the department.
Subcapsular Hematoma After Laparoscopic Cholecystectomy, Associated With Ketorolac Administration
Authors: Erstad Brian L. , Rappaport William D.
Published: October 2023

Ketorolac is the first injectable nonsteroidal antiinflammatory drug used as an analgesic in the perioperative period. Its adverse effect profile is different from that of the opioid analgesics; in particular, in its lack of respiratory depressive actions. However, ketorolac has risks associated with its perioperative administration, including episodes of substantial gastrointestinal bleeding. A patient undergoing elective laparoscopic cholecystectomy developed a subcapsular hepatic hematoma shortly after receiving a dose of injectable ketorolac. No evidence of parenchymal injury was found on laparoscopy which argues against iatrogenic trauma. Clinicians should be aware that ketorolac may cause or aggravate bleeding, and it should be used with caution in perioperative patients.
Thrombolytic Therapy for Pulmonary Embolism: Reversal of Shock in the Early Postoperative Period
Authors: Nasraway Stanley A. , Kabani Noormahal , Lawrence Kenneth R.
Published: October 2023

Life‐threatening acute pulmonary embolism (PE) associated with circulatory shock requires effective therapy directed at removing the obstruction to flow in the pulmonary vasculature and improving hemodynamics. Options for treatment are pulmonary embolectomy and thrombolytic therapy. Although safe and effective, thrombolytic therapy is relatively contraindicated within 10 days of major surgery due to the risk of bleeding. Intravenous streptokinase was administered to a man on the third postoperative day for treatment of a massive PE associated with circulatory shock. Within 2 hours of initiating therapy the patient experienced marked improvement in hemodynamics and tissue perfusion. No bleeding complications were noted. This case demonstrates that intravenous thrombolytics may be administered safely to patients who have recently undergone surgery. It also underscores that the decision to apply the therapy when relatively contraindicated must be made on an individual patient basis and thus ensure that potentially life‐saving therapy is not withheld from those who require it most.
Hospital and Pharmacy Characteristics Associated With Mortality Rates in United States Hospitals
Authors: Pitterle Michael E. , Bond C.A. , Raehl Cynthia L. , Franke Todd
Published: October 2023

We attempted to determine hospital and pharmacy characteristics associated with mortality rates in 4864 United States hospitals. Data were obtained from the Health Care Financing Administration, the American Hospital Association, and the National Clinical Pharmacy Services survey. Univariate and multivariate regression models were used to determine which hospital characteristics were associated with mortality. A similar regression analysis was performed on 718 hospitals for which detailed pharmacy information was available. In a multivariate regression model, some characteristics of 4864 hospitals associated with reduced mortality rates were high‐technology index (R2=0.09, p<0.001), severity of illness (R2=0.048, p<0.001), number of hospital beds (R2=0.016, p<0.001), and medical personnel (R2=0.012, p<0.001). This analysis accounted for 41% of the mortality rate variance. For the 718 hospitals and pharmacies, some of these characteristics were high‐technology index (R2=0.157, p<0.001), severity of illness (R2=0.07, p<0.001), number of pharmacists/average daily census (R2=0.021, p<0.001), and combined hospitalwide clinical pharmacy services (R2=0.016, p<0.01). The results of this analysis were similar to those in the only other large study in this area, but that excluded pharmacy characteristics. This is the first study to show a statistically significant association between pharmacist and pharmacy variables and reduced hospital mortality rates.
Dyslipidemias in Patients With Diabetes Mellitus: Classification and Risks and Benefits of Therapy
Authors: Oki Julie C.
Published: October 2023

To characterize the lipid and lipoprotein abnormalities in patients with diabetes mellitus and evaluate the risks and benefits of marketed pharmacologic therapies, a MEDLINE search of the National Library of Medicine data base was performed of studies published from January 1966 to March 1994. Clinical trials assessing effects on lipids and lipoproteins, and adverse effects of marketed lipid‐lowering agents were extracted. Reviews and other relevant articles were included if they provided information regarding lipid and lipoprotein metabolism or guidelines on the treatment of dyslipidemias in patients with diabetes mellitus. An extensive review of clofibrate was not included. The most common dyslipidemia in patients with poorly controlled insulin‐dependent diabetes mellitus (IDDM) is combined elevated triglyceride and cholesterol levels, with reduced high‐density lipoprotein (HDL) cholesterol (mixed hyperlipidemia). Hypertriglyceridemia combined with a reduced HDL cholesterol is the most common dyslipidemia in patients with noninsulin‐dependent diabetes mellitus, but essentially any pattern of dyslipidemia may be present. Small and dense low‐density lipoprotein (LDL), glycosylation of lipoproteins, and increased oxidized lipoproteins may be present in patients with diabetes mellitus; all contribute to accelerated atherosclerotic cardiovascular disease. Insulin therapy generally corrects quantitative lipid abnormalities in patients with IDDM, so drug treatment is seldom indicated. Diet, exercise, and insulin or oral sulfonylureas will improve hypertriglyceridemia and low HDL concentrations, but do not always return them to normal. Drug therapy is indicated when nonpharmacologic measures are inadequate. It is administered based on the effects of each agent on lipids and lipoproteins, patient age, adverse effect profile, patient tolerability, and drug‐disease and drug‐drug interactions. A fibric acid derivative is the drug of choice for marked hypertriglyceridemia in patients with diabetes mellitus. Niacin can worsen glycemic control, but it may be required in severe hypertriglyceridemia, hypercholesterolemia, or mixed hyperlipidemia. Bile‐acid binding resins may accentuate hypertriglyceridemia but may be useful in selected patients with marked hypercholesterolemia and normal triglycerides. Hydroxymethylglutaryl coenzyme A reductase inhibitors are preferred in patients with elevated LDL cholesterol and mild hypertriglyceridemia. Patients with marked lipid abnormalities or mixed hyperlipidemias may require carefully dosed combinations of lipid‐lowering drugs.
Influence of Bile Acid Replacement on Cyclosporine Absorption in a Patient With Jejunoileal Bypass
Authors: Kino Kathryn J. , Wittkowsky Ann K.
Published: October 2023

Jejunoileal bypass reportedly inhibits the absorption of cyclosporine. In patients with malabsorption syndromes, bile acid replacement has been given to enhance the absorption of cyclosporine. A candidate for heart transplantation with New York Heart Association class IV heart failure and a jejunoileal bypass received cyclosporine alone and with concomitant ursodiol administration to determine whether therapeutic cyclosporine plasma concentrations would be attainable after heart transplantation. There were no differences in peak concentration, time to peak, area under the serum concentration versus time curve, or bioavailability of oral cyclosporine when administered alone or with ursodiol. Plasma cyclosporine concentrations were consistent with those in the general population.
The Acrosome Reaction‐Inducing Effect of Human Follicular and Oviductal Fluid
Authors: de JONGE CHRISTOPHER J. , BARRATT CHRISTOPHER L. R. , RADWANSKA EWA , COOKE IAN D.
Published: October 2023

The human sperm acrosome reaction (AR) can be induced in vitro by a variety of naturally occurring and synthetic compounds. The present investigation determined the effect of human natural cycle periovulatory follicular (hFF) and oviductal fluids (hOF) on the human sperm AR in dose—response fashion using the synchronous AR assay. When hFF (30% v/v) or hOF (40% v/v) was added to non‐capacitated spermatozoa, no significant (P > 0.05) increase in the % AR was detected in comparison to the non‐treatment control. When either of these compounds was added (10%, 20%, and 30% v/v hFF; 20%, 30%, and 40% v/v hOF) to capacitated spermatozoa (3 hour incubation), a significant (P < 0.05) stimulation of the AR was detected. Bovine oviductal fluid (bOF) was tested (20%, 30%, and 40% v/v) to determine if it might have an effect similar to hOF. In contrast to hOF, the highest concentration of bOF (40% v/v) tested failed to stimulate a significant (P > 0.05) increase in the % AR of capacitated spermatozoa in comparison to control. Inhibitors of protein kinases A (KT5720) and C (calphostin C) were tested to determine their effects on the hFF‐induced AR. In comparison to hFF treatment alone, the kinase A inhibitor KT5720 (50 nM, 100 nM) prevented hFF (20%) stimulation of the AR when added at the end of the capacitation period and 5 minutes prior to the addition of inducer. Similarly, the kinase C inhibitor calphostin C (50 nM, 100 nM) prevented hFF (20%) stimulation of the AR. The present data demonstrate that periovulatory hFF and hOF stimulate the human sperm AR. hFF appears to stimulate the AR by activating multiple signal transduction pathways.
Germ Cell‐Conditioned Medium Contains Multiple Factors that Modulate the Secretion of Testins, Clusterin, and Transferrin by Sertoli Cells
Authors: PINEAU CHARLES , SYED VIQAR , BARDIN C. WAYNE , JÉGOU BERNARD , CHENG C. YAN
Published: October 2023

ABSTRACT: In view of the evidence showing that germ cells regulate Sertoli cell (SC) function, the aim of this study was to examine if germ cell (GC)‐conditioned media contained multiple biological factors that affect SC secretory functions. Total GC were isolated from adult rat testes. Pachytene spermatocytes (SPC) and early spermatids (SPT) were enriched to about 90% pure by centrifugal elutriation. GC, SPC, and SPT were cultured in serum‐free medium for 20 hours with a viability greater than 95%. Conditioned media derived from these cells were fractionated by anion‐exchange high‐performance liquid chromatography (HPLC). An aliquot from each of these fractions was resolved by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and proteins were visualized by silver staining. The patterns of protein staining using media from GC, SPC, and SPT were similar. Bioassays of these column fractions on SC showed that the transferrin stimulatory, the testins inhibitory, and the clusterin inhibitory activities were eluted from the anion‐exchange HPLC column in overlapping fractions. To determine whether these activities were confined to one or several molecules, further fractionations were performed. Eleven liters of GC‐conditioned medium were fractionated by sequential HPLC using anion‐exchange, gel permeation, and reversed‐phase columns. Throughout the entire fractionation scheme, the HPLC fractions were bioassayed using primary SC‐enriched cultures prepared from 20‐day‐old rats by incubating SC with aliquots of these fractions for 24 hours to monitor their effects on SC secretory function. The concentrations of transferrin, clusterin, and testins were quantified by specific radioimmunoassays. These studies showed that the transferrin stimulatory activity can be fractionated into four peaks (I, IIa, IIb, and IIc); clusterin inhibitory activity into three peaks (A, B, and C); and testins inhibitory activity into two peaks (1 and 2). Some of these bioactivities were eluted in overlapping fractions such as I and B, IIb and 1, and IIc and 2, whereas A, C, and IIa were not associated with any other assayed activities. In summary, additional GC modulators of SC function were identified for the first time, these include clusterin and testins inhibitors. The previously identified transferrin stimulatory activity was also resolved into multiple molecular forms.
Analysis of the Responses of Human Spermatozoa to A23187 Employing a Novel Technique for Assessing the Acrosome Reaction
Authors: AITKEN R. JOHN , BUCKINGHAM DONNA W. , FANG HUANG GUI
Published: October 2023

ABSTRACT: Protocols for the use of A23187 in assessing the ability of human spermatozoa to acrosome‐react and exhibit sperm—oocyte fusion have been developed and the results compared in two independent cohorts of infertile patients. Both bioassays were found to depend upon such factors as the dose and formulation of A23187, the duration of exposure, and the amount and type of protein used to supplement the medium. An optimal protocol for the hamster oocyte penetration test comprised a 3‐hour exposure to 1.23–2.5 μM ionophore and gave penetration rates of 93.2 ± 3.2% (11.26 ± 1.27 sperm/egg) for a group of 33 fertile donors compared with 63.0 ± 5.4% (4.73 ± 0.81 sperm/egg) for a cohort of 56 patients consulting for infertility (P < 0.001). Higher doses (5.0–10.0 μM) of A23187 caused an inhibition of sperm‐oocyte fusion in association with a loss of motility, although the integrity of sperm plasma membrane did not appear to be compromised and high rates (∼80%) of acrosome reaction were observed. A protocol for assessing the ability of viable human spermatozoa to acrosome‐react in response to A23187 was developed, employing a fluorescein‐conjugated lectin in concert with the hypoosmotic swelling test, which gave values of 20.1 ± 2.6% and 13.6 ± 1.6% for groups of fertile donors (n = 29) and infertile patients (n = 32) respectively (P < 0.05). Although only acrosome‐reacted spermatozoa were capable of fusing with zona‐free hamster oocytes, there was no significant correlation between the proportion of acrosome‐reacted cells and the levels of sperm‐oocyte fusion observed in two independent groups of patients, indicating that these bioassays are measuring different aspects of human sperm function. These results have implications for the way in which the responses of human spermatozoa to ionophore treatment are quantified and interpreted.
Synthesis of Docosahexaenoyl Coenzyme A in Human Spermatozoa
Authors: JONES ROBERT E. , PLYMATE STEPHEN R.
Published: October 2023

ABSTRACT The synthesis of docosahexaenoyl coenzyme A (22: 6‐CoA) was studied in a long‐chain fatty acid: CoASH ligase (AMP)‐enriched fraction from human spermatozoa and was compared to palmitoyl CoA (16:0‐CoA) synthesis. The pH optimum for 22:6 activation was 8.4, which was identical to the value obtained with 16: 0. The Km for ATP was 0.5 mM when 22:6 was the acyl substrate; however, when 16:0 was incubated with the ligase preparation, the Km for ATP was 2.9 mM. When CoASH was varied and 22:6 was the fatty acyl acceptor, a pattern of negative cooperativity was observed. This was confirmed by a downwardly concave double‐reciprocal plot, a Hill coefficient of 0.63, and an Rs in excess of 150. The Hill coefficient with 16:0 and CoASH was 0.94. Palmitic acid was demonstrated to be a competitive inhibitor of 22:6‐CoA synthesis. Based upon these data, we conclude that the kinetics of spermatozoan ligase are complex, and, in addition, these data support the hypothesis that 22:6 may regulate ligase activity, and therefore free fatty acid utilization, in sperm.
Effects of Epidermal Growth Factor on Human Sperm Cell Function
Authors: NAZ RAJESH K. , KAPLAN PAUL
Published: October 2023

ABSTRACT: The effects of human recombinant epidermal growth factor (EGF) on the fertilizing capacity of human sperm were investigated. At lower concentrations (0.1–10 nM) EGF did not significantly (P > 0.05) affect human sperm penetration of zona‐free hamster oocytes (SPA). At higher concentrations (25–100 nM), EGF significantly (P= 0.01 to < 0.001) decreased the human sperm penetration rates in SPA. At higher concentrations (≥25 nM), EGF also significantly inhibited the spontaneous as well as calcium ionophore‐induced acrosome reaction and release of acrosin from human sperm. There was no effect of EGF on percent sperm motility, but at higher concentrations (≥25 nM) it significantly affected various sperm motility characteristics especially velocity and amplitude of lateral head displacement. EGF was detected by radioimmunoassay as well as radioreceptor assay in seminal plasma of fertile men. However, there were no statistical differences between the levels of EGF or EGF/transforming growth factor (TGF‐α) in seminal plasma of fertile, infertile, and immunoinfertile men. Also, there was no significant correlation of the EGF or EGF/TGF‐α levels with total sperm number, sperm motility characteristics, and penetration rates in SPA in these patients. These results indicate that EGF has either no effect or an inhibitory/negative effect on the human sperm cell capacitation and/or acrosome reaction, especially at higher concentrations (>25 nM).
Protection by Gonadal Steroid Hormones Against Procarbazine‐Induced Damage to Spermatogenic Function in LBNF1 Hybrid Rats
Authors: PARCHURI N. , WILSON G. , MEISTRICH M. L.
Published: October 2023

ABSTRACT: Some anti‐cancer drugs, such as procarbazine (PCZ), are associated with irreversible damage to spermatogenic function and cause sterility in men. In the present study, protection of spermatogenesis by gonadal steroid hormones during PCZ treatment was investigated in male rats. LBNF1 hybrid rats were chosen for these studies because the response of the testis to PCZ was more uniform than in a stock of Sprague‐Dawley outbred rats. Mature male LBNF1 rats were subcutaneously implanted with cholesterol (C)‐, testosterone (T)‐, and/or estradiol‐17β (E)‐ containing capsules, and 5–10 weeks later given one or four weekly intraperitoneal injections of PCZ at a dose of 200–250 mg/kg of body weight. Hormone capsules were removed 24 hours after the last PCZ injection and the animals were killed 10–12 weeks later. Testicular weights, sonication‐resistant sperm head counts, and quantitative testicular histology revealed protection of the spermatogenic epithelium from PCZ toxicity by gonadal steroid hormones when compared with the C controls. Protection was observed against both single and multiple injections of PCZ. A combination of T and E capsules provided better protection than T alone. This model system is suitable for studies of the mechanism of protection of spermatogenesis from chemotherapy‐induced damage.
Antifertility Effects of Neem (Azadirachta indica) Oil in Male Rats by Single Intra‐Vas Administration: An Alternate Approach to Vasectomy
Authors: UPADHYAY SHAKTI N. , DHAWAN SUMAN , TALWAR G. P.
Published: October 2023

ABSTRACT: An alternate approach to vasectomy for long‐term male contraception following a single intra‐vas application of a traditional plant (Azadirachta indica) product having immunomodulatory properties is described. Male Wistar rats of proven fertility were given a single dose (50 μl) of neem oil in the lumen of the vas deferens on each side; control animals received the same volume of peanut oil. Animals were put on continuous mating 4 weeks after the treatment, with females of proven fertility. While the control animals impregnated the female partners, all males treated with neem oil remained infertile throughout the 8 months of observation period. Epididymal and vas histology were normal without any inflammatory changes or obstruction. The intra‐vas administration of neem oil resulted in a block of spermatogenesis without affecting testosterone production; the seminiferous tubules, although reduced in diameter, appeared normal and contained mostly early spermatogenic cells. No anti‐sperm antibody could be detected in the serum. Unilateral administration of neem oil in the vas resulted in a significant reduction of testicular size and spermatogenic block only on the side of application; the draining lymph node cells of the treated side also showed enhanced proliferative response to in vitro mitogen challenge. These results indicate that the testicular effects following intra‐vas application of neem oil may possibly be mediated by a local immune mechanism.
Validation of an Acrosomal Stain for Equine Sperm that Differentiates between Living and Dead Sperm
Authors: CASEY PATRICK J. , HILLMAN ROBERT B. , ROBERTSON KATHRYN R. , YUDIN ASHLEY I. , LIU IRWIN K. M. , DROBNIS ERMA Z.
Published: October 2023

ABSTRACT: An acrosomal staining technique that can differentiate between living and dead sperm was developed for equine sperm. The fluoresceinated lectin Pisum sativum agglutinin (FITC‐PSA) was used to identify the presence or absence of acrosomal contents, while the supravital nuclear dye Hoechst 33258 (H258) was used to assess viability. The accuracy of the FITC‐PSA acrosomal stain was tested by comparing the percentage of sperm that had lost their acrosomal contents, detected by the staining method, with that detected by transmission electron microscopy (TEM). Following capacitation in vitro, the acrosomal status of sperm induced to acrosome react with A23187 and of control sperm were very similar with the staining technique and TEM, confirming the accuracy of the FITC‐PSA acrosomal stain. We investigated the relationship between viability as measured by exclusion of H258 and motility as measured by three methods: one subjective and two objective. Although there was a good correlation between viability and motility as measured by all three methods (r = 0.88, 0.85, 0.75), there was always a proportion of viable sperm that were nonmotile. The physiology of the viable, nonmotile sperm was further investigated by comparing for individual sperm the viability as measured by exclusion of H258 with the mitochondrial function as measured by rhodamine 123. A good correlation (r = 0.99) was found to exist between viability and mitochondrial function, indicating that viable, nonmotile sperm possess functional mitochondria and confirming the ability of supravital staining to distinguish between living and dead sperm. We determined that 29–81% of the sperm in semen that had lost their acrosomal contents were in fact dead. Thus, this acrosomal staining technique can provide more relevant endpoints for future investigations of capacitation, the acrosome reaction, and sperm handling techniques in the horse.
Histone acetylation regulates ORMDL3 expression‐mediated NLRP3 inflammasome overexpression during RSV‐allergic exacerbation mice
Authors: Cheng Qi , He Fanghan , Zhao Wenqi , Xu Xianhong , Shang Yunxiao , Huang Wanjie
Published: October 2023

Whether respiratory syncytial virus (RSV) infection in early life may induce orosomucoid 1‐like protein 3 (ORMDL3) and lead to NOD‐like receptor protein 3 (NLRP3) inflammasome overexpression in asthma, which could be alleviated by the inhibition of HAT p300. First, we explored the relationship between RSV, ORMDL3, and recurrent wheezing in the future through clinical data of infants with RSV‐induced bronchiolitis. Then, we used bronchial epithelium transformed with Ad12‐SV40 2B (BEAS‐2B) and an asthmatic mouse model of repeated RSV infection and OVA sensitization and challenge (rRSV + OVA) in early life to assess the effects of ORMDL3 on NLRP3 inflammasome and that of histone acetylation on ORMDL3 regulation. ORMDL3 overexpression is the independent risk factor of recurrent wheezing in RSV‐bronchiolitis follow‐up. In BEAS‐2B, ORMDL3‐induced NLRP3 inflammasome expression. BEAS‐2B infected by RSV resulted in overexpression of ORMDL3 and NLRP3 inflammasome and histone hyperacetylation, while ORMDL3‐small interfering RNA and C646 interfered could decrease NLRP3 inflammasome. ORMDL3 overexpression in mouse lung increased NLRP3 inflammasome. The expression of ORMDL3 and NLRP3 inflammasome significantly increased, with histone hyperacetylation in the lung in rRSV + OVA mice. p300 and acetylH3 bound to ORMDL3 promoter. In C646 + rRSV + OVA mice, C646 alleviated lung inflammation and overexpression of ORMDL3 and NLRP3 inflammasome. RSV activated ORMDL3 overexpression through histone hyperacetylation and induced NLRP3 inflammasome expression.
Salmonella engages CDC42 effector protein 1 for intracellular invasion
Authors: Bandyopadhyay Sheila , Zhang Xiao , Ascura Andrea , Edelblum Karen L. , Bonder Edward M. , Gao Nan
Published: October 2023

Human enterocytes are primary targets of infection by invasive bacterium Salmonella Typhimurium, and studies using nonintestinal epithelial cells established that S. Typhimurium activates Rho family GTPases, primarily CDC42, to modulate the actin cytoskeletal network for invasion. The host intracellular protein network that engages CDC42 and influences the pathogen's invasive capacity are relatively unclear. Here, proteomic analyses of canonical and variant CDC42 interactomes identified a poorly characterized CDC42 interacting protein, CDC42EP1, whose intracellular localization is rapidly redistributed and aggregated around the invading bacteria. CDC42EP1 associates with SEPTIN‐7 and Villin, and its relocalization and bacterial engagement depend on host CDC42 and S. Typhimurium's capability of activating CDC42. Unlike CDC42, CDC42EP1 is not required for S. Typhimurium's initial cellular entry but is found to associate with Salmonella‐containing vacuoles after long‐term infections, indicating a contribution to the pathogen's intracellular growth and replication. These results uncover a new host regulator of enteric Salmonella infections, which may be targeted to restrict bacterial load at the primary site of infection to prevent systemic spread.
Appendicular lean mass index changes in patients with Duchenne muscular dystrophy and Becker muscular dystrophy
Authors: Wong Brenda L. , Summer Suzanne , Horn Paul S. , Rutter Meilan M. , Rybalsky Irina , Tian Cuixia , Shellenbarger Karen C. , Kalkwarf Heidi J.
Published: October 2023

Introduction

Mutations in the 79 exons of the dystrophin gene result in muscle wasting and weakness of varying clinical severity, ranging from severe/typical Duchenne muscular dystrophy (DMD) to intermediate DMD and mild Becker muscular dystrophy (BMD), depending on the frameshift of the mutation. We previously reported that males with DMD have progressively declining appendicular lean mass (ALM) and ALM index (ALMI) with age and worsening functional motor ability compared with healthy controls. These indices have not been studied in patients with intermediate DMD and BMD phenotypes and across DMD genotypes. In this study, we compared age‐related trajectories of ALM and ALMI of patients who had (1) BMD without functional mobility deficits with patients who had DMD at different stages of disease and healthy controls; (2) a DMD intermediate phenotype with patients who had a typical DMD phenotype; and (3) DMD categorized by genotype.

Methods

We conducted a retrospective review of ALM and ALMI data from 499 patients (ages 5–23 years) with DMD (466 typical and 33 intermediate) and 46 patients (ages 5–21 years) with BMD (without functional mobility deficits and functional mobility score of 1). Patients were grouped according to age reflecting disease stage (ages 5 to <7, 7 to <10, 10 to <14, and 14 to <20 years) and genotype (mutations in exons 1–30, 31–44, 45–62, and 63–79).

Results

ALM and ALMI trajectories of patients with BMD paralleled those of healthy controls until adolescence, in contrast to patients with DMD. ALMI Z‐scores of patients with BMD remained within ±2 SD without decline while those of patients with DMD fell below −2 SD around age 12 years. Patients with BMD had increasing ALM and ALMI with age, with peak accrual between ages 10 to <14 years. ALMI declined after age 14 years for those with intermediate DMD compared with 10 years for patients with typical DMD. Patients with mutations in exons 63–79 had a greater decline in ALMI as compared with those with other genotypes after age 10 years.

Conclusions

Age‐related changes in ALMI in patients with BMD and intermediate DMD differ from those with typical DMD, reflecting their clinical phenotypes. ALM and ALMI should be further studied in patients with BMD and DMD subtypes for their potential value as surrogate markers to characterize the severity of BMD and DMD and inform clinical care decisions and clinical trial designs.
Suppressors of cGAS‐STING are downregulated during fin‐limb regeneration and aging in aquatic vertebrates
Authors: Mathavarajah Sabateeshan , Thompson Andrew W. , Stoyek Matthew R. , Quinn T. Alexander , Roy Stéphane , Braasch Ingo , Dellaire Graham
Published: October 2023

During the early stages of limb and fin regeneration in aquatic vertebrates (i.e., fishes and amphibians), blastema undergo transcriptional rewiring of innate immune signaling pathways to promote immune cell recruitment. In mammals, a fundamental component of innate immune signaling is the cytosolic DNA sensing pathway, cGAS‐STING. However, to what extent the cGAS‐STING pathway influences regeneration in aquatic anamniotes is unknown. In jawed vertebrates, negative regulation of cGAS‐STING activity is accomplished by suppressors of cytosolic DNA such as Trex1, Pml, and PML‐like exon 9 (Plex9) exonucleases. Here, we examine the expression of these suppressors of cGAS‐STING, as well as inflammatory genes and cGAS activity during caudal fin and limb regeneration using the spotted gar (Lepisosteus oculatus) and axolotl (Ambystoma mexicanum) model species, and during age‐related senescence in zebrafish (Danio rerio). In the regenerative blastema of wounded gar and axolotl, we observe increased inflammatory gene expression, including interferon genes and interleukins 6 and 8. We also observed a decrease in axolotl Trex1 and gar pml expression during the early phases of wound healing which correlates with a dramatic increase in cGAS activity. In contrast, the plex9.1 gene does not change in expression during wound healing in gar. However, we observed decreased expression of plex9.1 in the senescing cardiac tissue of aged zebrafish, where 2′3′‐cGAMP levels are elevated. Finally, we demonstrate a similar pattern of Trex1, pml, and plex9.1 gene regulation across species in response to exogenous 2′3′‐cGAMP. Thus, during the early stages of limb‐fin regeneration, Pml, Trex1, and Plex9.1 exonucleases are downregulated, presumably to allow an evolutionarily ancient cGAS‐STING activity to promote inflammation and the recruitment of immune cells.
Disability education and implications for genetic counselor training
Authors: Douglas Lauren , Czerwinski Jennifer , Vukovic Rose , Ashfaq Myla , Lunstroth Rebecca , Wagner Chelsea
Published: October 2023

The genetic counseling (GC) community has faced criticism about the duality of promoting patient autonomy while also advocating for individuals with disabilities. This study assessed the attitudes of the disability community and GCs to identify content that should be included in GC disability education and evaluate the landscape of GC disability education. Members of the disability community and GCs completed an electronic survey distributed through electronic listservs and partnering organizations. A total of 672 responses were analyzed from both the disability community (n = 596) and the GC community (n = 76). Members of the disability community noted differences in GC comfort level discussing different aspects of disability with GCs being perceived as being very knowledgeable about medical aspects 71% of the time versus 49% of the time when discussing social/lifestyle aspects of disability. This discordance was reflected in GCs reported comfort level in discussing medical aspects (89%) and social aspects of disability (65%) during a session. Most GC respondents (71%) felt they received adequate knowledge during their disability education and variation was reported in the execution of disability education by training programs. Disability education content recommendations from the disability community and GCs included emphasizing four key aspects of disability: medical, social/lifestyle, lived experience, and the disability rights movement. Respondents of both cohorts stressed the inclusion of and exposure to persons with disabilities in disability education to understand the lived experience of persons with disabilities. The disability community identified additional disability education content to be included such as empathy training, family hardships, and mental health. The results of this study have practice implications and provide a foundation for training expectations to ensure future GCs possess the necessary skills to improve the quality of services provided to families and persons with disabilities.