bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of Clusterin/Sulfated Glycoprotein‐2 Under Conditions of Heat Stress in Rat Sertoli Cells and a Mouse Sertoli Cell Line
Authors: CLARK ANN M. , GRISWOLD MICHAEL D.
Published: October 2023

ABSTRACT: Clusterin is the major protein produced by rat Sertoli cells and is deposited onto sperm membranes; however, its function is unknown. In order to gain insight into the regulation of clusterin in Sertoli cells, the objective of the present study was to develop a model where the expression of clusterin could be affected in Sertoli cells in vitro. Rat Sertoli cells and mouse Sertoli cells (MSC1) were cultured under heat stress conditions (41°C) for up to 48 hours. The mRNA for clusterin in Sertoli cells was compared to that in human epitheliod cancer cells (A431) to determine if clusterin expression was regulated in a testis‐specific manner. The mRNA for heat shock protein 70 (HSP70) was also examined as it is a known stress‐regulated gene. Expression of HSP70 mRNA was increased in all three cell types by 4 hours after the start of heat stress. Clusterin mRNA was increased over that of controls by 4 hours in heat‐stressed A431 cells but did not significantly increase in MSC1 or Sertoli cells until 12 hours (P < 0.05). The induction of clusterin mRNA in MSC1 cells continued for at least 48 hours and required the sustained exposure of cells to the 41°C temperature. The increase in the amount of clusterin mRNA was not due to an increase in transcript half‐life, as determined by the addition of actinomycin D to the media of control vs. heat‐stressed MSC1 cells. From the development of this in vitro model, we have seen that the timing of induction of clusterin by heat stress is Sertoli cell specific and is different than that of HSP70. This response in surviving cells during heat stress may be protective in that clusterin would bind to toxic compounds or solubilize cellular debris released by degenerating cells.
Evidence of Proacrosin Molecule Abnormality as a Possible Cause of Low Acrosin Activity and Unexplained Failure of Fertilization In Vitro
Authors: SHIMIZU YASUSHI , KODAMA HTOEYA , FUKUDA JUN , TANAKA TOSHINOBU
Published: October 2023

ABSTRACT: The aim of this study was to investigate whether the molecular abnormality of proacrosin is associated with an unexplained failure of in vitro fertilization (IVF) and low acrosin activity in human infertility. Seventeen infertile men, whose spermatozoa appeared to be normal but did not fertilize using a conventional IVF technique, were included in this study. Proacrosin molecules from each patient were screened with electrophoresis and Western blotting using either anti‐human proacrosin or anti‐boar acrosin polyclonal antibodies. The bands of proacrosin appeared equally on the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) at an identical site and reacted equally with human proacrosin antibodies in all patients studied. The anti‐boar acrosin antibodies cross‐reacted with the proacrosin bands of the patients in all but two cases. The two patients also demonstrated lower levels of total acrosin activity and different peptide maps of the isolated proacrosin. In conclusion, the molecular abnormality of proacrosin, which is reflected by a lack of cross‐reactivity to the anti‐boar acrosin antibodies, is associated with low acrosin activity and may explain some cases of unexplained failure of human IVF treatment.
Seminal Plasma Lactoferrin Concentrations in Normal and Abnormal Semen Samples
Authors: BUCKETT WILLIAM M. , LUCKAS MURRAY J. M. , GAZVANI MEHMET R. , AIRD IAN A. , LEWIS‐JONES D. IWAN
Published: October 2023

ABSTRACT: Although the iron‐chelating protein lactoferrin is secreted by the seminal vesicles, the precise role of lactoferrin in semen is unclear. This study aimed to determine whether there is any association between seminal lactoferrin concentrations and normal and abnormal semen samples with and without leucocytospermia. Lactoferrin concentrations were measured by radial immunodiffusion of semen samples from 368 men attending a regional andrology referral center. Routine seminal analysis, including the presence of leucocytospermia, was also performed. Results showed increased seminal lactoferrin in samples showing oligospermia (13.3 mg/100 ml) and oligoasthenospermia (13.4 mg/100 ml) compared to normospermic samples (11.2 mg/100 ml). There were no significant differences in seminal lactoferrin between normospermic samples and azoospermic samples or asthenospermic samples with normal sperm density. Although there was a trend toward increased lactoferrin concentration with leucocytospermia, this was not significant. Possible causes for raised lactoferrin in association with oligospermia are discussed.
Comparison Between Testosterone Enanthate‐Induced Azoospermia and Oligozoospermia in a Male Contraceptive Study. V. Localization of Higher 5α‐Reductase Activity to the Reproductive Tract in Oligozoospermie Men Administered Supraphysiological Doses of Testosterone
Authors: ANDERSON R. A. , KELLY R. W. , WU F. C. W.
Published: October 2023

ABSTRACT: Sex steroid contraceptive regimens result in incomplete suppression of spermatogenesis in 30–45% of Caucasian men. The basis for this is unclear, but differences in the activity of 5α‐reductase (5αR) have been demonstrated. Two isoforms of 5αR have been described: 5αR1 is found in skin, whereas the predominant form in reproductive tissues is 5αR2. To investigate possible contributions of these isoenzymes, we have investigated androgen‐dependent changes in seminal plasma androgens (5αR2) and sebum production (5αR1) during administration of a supraphysiological dose (200 mg IM weekly) of testosterone enanthate (TE) to 33 normal men.

Eighteen men rapidly (<20 weeks treatment) became azoospermia, the remainder having a mean sperm density of 2.0 ± 0.6 × 106 at that time. The concentrations of testosterone and 3α, 17β‐andros‐tanediol glucuronide (AdiolG) were lower in seminal plasma than in blood but rose by a similar degree (100%) after 16 weeks TE treatment in both groups. There were no differences in seminal‐plasma concentration of testosterone or AdiolG between azoospermic and Oligozoospermie responders, either pretreatment or after 16 weeks TE treatment. Although the concentrations of dihydrotestosterone (DHT) were similar in seminal plasma and blood pre‐ and posttreatment, there was a selective increase in seminal plasma DHT concentration in the Oligozoospermie responders from 2.12 ± 0.29 to 2.94 ± 0.33 nmol/L (P < 0.05), while there was no significant change in the azoospermic responders (2.18 ± 0.31–2.54 ± 0.27 nmol/L) after 16 weeks of TE treatment. Dihydrotestosterone in seminal plasma is primarily derived from 5αR activity in the epididymis. The concentration of prostaglandin E2 (PGE2) in seminal plasma was unchanged during TE treatment. Sebum excretion was increased during TE treatment, but there were no differences between azoospermic and Oligozoospermie responders pretreatment or after 16 weeks TE treatment. These results are consistent with the hypothesis that incomplete suppression of spermatogenesis during TE treatment is associated with a relatively higher 5αR activity in the reproductive tract (epididymis and/or testis) during TE treatment. As the predominant form of 5αR in the reproductive tract is 5αR2 (type 2), we conclude that the increase in activity derives from this form of the enzyme, rather than the type 1 form (5αR1) predominantly found in nongenital skin.
Development and Characterization of a Prepubertal Rat Sertoli Cell Line, 93RS2
Authors: JIANG CHENGYU , HALL SUSAN J. , BOEKELHEIDE KIM
Published: October 2023

ABSTRACT: Sertoli cells in the seminiferous epithelium provide both structural and nutritional support to germ cells during spermatogenesis. Primary Sertoli cells in culture are an effective tool for the in vitro study of Sertoli cell function; however, primary cultures are inherently variable, time consuming to prepare, expensive, and wasteful of animals. We therefore developed a Sertoli cell line, called 93RS2, by immortalizing primary Sertoli cells derived from prepubertal rats with SV40 tsA255. This cell line proliferates at the permissive temperature (32°C) and has enhanced expression of a differentiated Sertoli cell phenotype at the nonpermissive temperature (40–41 °C). Cytogenetic analysis demonstrated that 93RS2 has 42 chromosomes per cell, the same as a normal rat. mRNA analysis showed that this cell line, when cultured at a nonpermissive temperature, exhibited increased expression of transferrin in the presence of testosterone and enhanced expression of sulfated glycoprotein‐2. A tumorigenicity assay showed that 93RS2 cells were temperature‐dependent for growth in soft agar and were capable of forming tumors in nude mice. In conclusion, this rat 93RS2 cell line should be useful for the study of Sertoli cell function.
Low Density Lipoprotein Receptor‐Related Protein‐1 Expression in the Testis: Regulated Expression in Sertoli Cells
Authors: IGDOURA SULEIMAN A. , ARGRAVES W. SCOTT , MORALES CARLOS R.
Published: October 2023

ABSTRACT: The low density lipoprotein receptor‐related protein (LRP‐1) is a multiligand receptor capable of mediating endocytosis of a wide array of ligands that relate to both lipoprotein metabolism and proteinase regulation. Many of its ligands are proteinases, proteinase—inhibitor complexes, and lipoproteins known to be contained within the luminal fluid of the seminiferous tubules or in the interstitial spaces of the testis. Immunocytochemical analysis was performed to characterize the pattern of expression of LRP‐1 in cells of the rat testis. Immunoperoxidase staining localized LRP‐1 to the cytoplasm of Sertoli cells. The distribution and intensity of the Sertoli cell staining was found to vary according to the stages of the cycle of the seminiferous epithelium. Staining was weak in the basal cytoplasm of Sertoli cells during stages II—VIII and strong and granular in the supranuclear cytoplasm during stages XII—XIV and stage I of the cycle. Immunogold labeling showed gold particles associated with the basal and adluminal plasma membranes, with endocytic vesicles, and with endosome membranes. Labeling was also observed on the plasma membrane and membranes of the endocytic apparatus in macrophages and Leydig cells in the interstitial space. Infusion of 1251‐Labeled LRP‐1 antibody into seminiferous tubules followed by radioautography showed silver grains overlaying the adluminal plasma membrane of Sertoli cells at time 0 and in endocytic vesicles and endosomes in the supranuclear region of Sertoli cells 10‐minutes postinjection. When the 1251‐Labeled LRP‐1 antibody was injected into the interstitial space, silver grains overlayed the basal plasma membrane and coated endocytic pits of Sertoli cells at time 0 and, at 10 minutes, the grains labeled endosomes located in the basal pole of Sertoli cells. 1251‐Labeled LRP‐1 antibody also labeled the plasma membrane and the endocytic apparatus of macrophages and Leydig cells. The absence of immunogold labeling and radioautographic silver grains within lysosomes of Sertoli cells, Leydig cells, and macrophages suggests that internalized LRP‐1 is recycled back to the cell surface. The presence of LRP‐1 in the endocytic compartment of these cells is consistent with its functioning in the clearance of proteases involved in seminiferous tubule remodeling and/or the uptake of cholesterol‐bound lipoproteins necessary for the biosynthesis of testosterone. In conclusion, the results of these studies demonstrated for the first time the presence of LRP‐1 receptor in the endocytic compartments of Sertoli cells and interstitial cells of the rat testis.
Is Leydig Cell Steroidogenic Function Affected by the Germ Cell Content of the Seminiferous Tubules?
Authors: SPRANDO ROBERT L. , ZIRKIN BARRY R.
Published: October 2023

ABSTRACT: The effect of testicular germ cell content on Leydig cell steroidogenic function in vivo in adult rats was examined. Three experimental paradigms were used to effect germ cell changes. First, a vitamin A‐depletion/repletion regimen was used to achieve synchrony at different stages of the cycle of the seminiferous epithelium and thus produce testes with widely differing germ cell contents. Second, long term vitamin A depletion was used to effect germ cell, but not Leydig cell, loss. Third, Leydig cells and germ cells first were eliminated from the testes of adult rats by the administration of ethane 1,2‐dimethane sulfonate (EDS) along with testosterone‐ and estradiol‐filfed Silastic capsules; Leydig cells were then restored to the germ cell‐depleted testes by removal of the luteiniging hormone (LH)‐suppressing capsules. Serum, interstitial fluid, and seminiferous tubule fluid testosterone concentrations did not differ between rats in which at least 70% of the seminiferous tubules contained germ cells at stages VII—VIII or at stages XII—III of the cycle. The capacity of the testes of these rats to produce testosterone, assessed by their in vitro perfusion with maximally stimulating LH, also showed no differences despite the differences in germ cell content. Elimination of germ cells throughout the testes by long term vitamin A depletion also did not affect the steroidogenic function of the testes. Finally, the steroidogenic function of Leydig cells restored to germ cell‐depleted testes was indistinguishable from that of germ cell‐containing controls. These results, taken together, provide evidence supporting the contention that the germ cell content of the testis has little or no effect on testicular steroidogenic function.
The Effect of Follicle‐Stimulating Hormone Therapy on Sperm Quality: An Ultrastructural Mathematical Evaluation
Authors: STREHLER ERWIN , STERZIK KARL , de SANTO MARIELLA , ABT MARKUS , WDEDEMANN RAINER , BELLATI UMBERTO , COLLODEL GIULIA , PIOMBONI PAOLA , BACCETTI BACCIO
Published: October 2023

ABSTRACT: We investigated the effect of follicle‐stimulating hormone (FSH) administration on the infrastructure of spermatozoa in order to evaluate the potential of FSH therapy for improving sperm quality. Forty‐six patients exhibiting idiopathic oligoasthenoterato‐zoospermia who attended the intrauterine insemination (IU), in vitro fertilization (IVF), or intracytoplasmic sperm injection (ICSI) program at our clinic received FSH in daily dosages of 150 IU over a period of 12 weeks. Using transmission electron microscopy, the ultrastructural analysis of spermatozoa was performed prior to the start of FSH therapy, after the treatment had been finished, and 6 weeks posttherapy. Applying a mathematical formula based on submicroscopic characteristics, we calculated the number of morphologically normal spermatozoa. After the FSH treatment, the examined subcellular organelles achieved a higher percentage of integrity. Follicle‐stimulating hormone treatment leads to a higher number of morphologically normal spermatozoa. The electron microscopic findings indicate that treatment with pure FSH may be an effective way to improve sperm quality in cases with oligoasthenoteratozoospermia. Applying the mathematical analysis based on the whole complex of the selected sperm characteristics, we obtained a way to evaluate the success of therapy for the first time.
Comparison of Responses to Adrenomedullin and Calcitonin Gene‐Related Peptide in the Feline Erection Model
Authors: CHAMPION HUNTER C. , WANG RUN , SANTIAGO JOSé A. , MURPHY WILLIAM A. , COY DAVID H. , KADOWITZ PHILIP J. , HELLSTROM WAYNE J. G.
Published: October 2023

ABSTRACT: The purpose of the present study was to investigate the effects of intracavernosal injections of adrenomedullin (ADM) and calcitonin gene‐related peptide (CGRP), two structurally similar peptides, on penile erection in the anesthetized cat. Erectile responses to ADM and CGRP were compared with responses to a standard drug combination (1.65 mg papaverine, 25 μg phentolamine, and 0.5 μg prostaglandin E1 [PGE1]). Intracavernosal injections of ADM (0.1–3 nmol) and CGRP (0.01–0.3 nmol) induced erection in a dose‐dependent manner. The maximal increase in intracavernosal pressure in response to ADM was a 75% increase, while the maximal response to CGRP was comparable to that induced by the reference combination, and the maximal increase in penile length was comparable with ADM, CGRP, and the standard drug combination. The duration of the maximal pressure increase and the total duration of the response to ADM and CGRP were more abbreviated than with the control combination, and systemic blood pressure was reduced significantly after administration of CGRP, the control combination, and the higher doses of ADM. The nitric oxide synthase inhibitor, L‐NAME, and the K+ATP‐channel antagonist, gly‐benclamide, had no effect on the erectile response to CGRP or ADM. The CGRP receptor antagonist CGRP(8–37) attenuated the erectile response to CGRP but not to ADM. These data suggest that the erectile responses to ADM and CGRP are not mediated by nitric oxide release or the opening of K+ATP channels, two mechanisms reported to be involved in penile erection, and that CGRP and ADM induce penile erection by activating different receptors.
Morphometric and Immunocytochemical Study of the Fetal, Infant, and Adult Human Vas Deferens
Authors: REGADERA JAVIER , ESPAÑA GABRIEL , ROIAS MARIANA A. , RECIO JUAN A. , NISTAL MANUEL , SUÁREZ‐QUIAN CARLOS A.
Published: October 2023

ABSTRACT: The human vas deferens (VD) is often considered simply as a conduit to transfer mature sperm from the epididymis to the ejaculatory duct. The cells that make up the epithelium of the VD, however, exhibit many characteristics of cells found in more complex epithelia, which are involved in absorption and/or secretion. In the present investigation, morphometry was utilized to characterize in detail the changes incurred by the human VD during its development, growth, and aging and to determine if these changes correlate with testicular maturation. In addition, the specific types of keratins present in the epithelial cells were defined, as well as desmin distribution in the muscular layers, during the various phases of the development, growth, and involution of the human VD. Results of the morphometric study are consistent with the interpretation that the development, growth, and aging of the VD are delayed, but parallel to, the identical phases exhibited by the human testis. Further, a differential expression of distinct keratin types was observed in the VD during the various phases examined in this study. Taken together, these two correlations may suggest that the VD is unlikely to function solely as a conduit for sperm. The rationale for this interpretation is as follows: 1) the complex developmental and maturational changes measured in the present investigation in the human VD are common to other absorptive and/or secretory epithelia; and 2) these changes parallel developmental changes observed in other androgen‐dependent epithelia of the male reproductive tract, which also function to contribute components to seminal fluid as well as to provide a conduit for sperm.
Spontaneous Degeneration of Testicular Germ Cells in Congenitally Athymic Nude Mice of Four Strains: A Light Microscopical Observation
Authors: ITOH MASAHIRO , KOKUDO YASUTAKA , SAKAMOTO YASUKI , TAKEUCHI YOSHIKI
Published: October 2023

ABSTRACT: Many hormonal studies have demonstrated that the presence of thymic factors affects testicular function. However, morphological studies on the testis in the absence of thymic factors are very limited. In the present study, seminiferous tubules of nude mice of four strains were light microscopically observed in order to investigate the effect of congenital athymia on germinal cell components. Testes obtained from congenitally athymic nude mice of CD1, BALB/c‐nu/nu, LASAT, and KSN strains were fixed in Bouin's solution, dehydrated, and embedded in plastic. The samples were cut and their sections were stained with hematoxylin and eosin. The results showed that degeneration of testicular germ cells was focally observed in all four strains of athymic mice. The degenerating morphology was characterized by karyopyknosis and karyolysis of immature germ cells, multinuclear giant cell formation, and severe depletion of epithelial components with remaining Sertoli cells and/or some spermatogonia. KSN nude mice showed the most severe degenerative changes among the four strains of nude mice examined. Their significant degenerative changes first appeared at 2 weeks of age. In contrast, CD1 nude mice showed very slight degenerative features even at 20 weeks of age. BALB/c‐nu/nu and LASAT nude mice had more degenerated epithelia than did CD1 nude mice; however, their significant degenerative changes were apparent at 20 but not at 10 weeks of age. Therefore, the severity and the onset of spontaneous degeneration and depletion of testicular germ cells in congenitally athymic mice were different among the four strains examined.
Enhanced Susceptibility of Follicle‐Stimulating‐Hormone‐Deprived Infertile Bonnet Monkey (Macaca radiata) Spermatozoa to Dithiothreitol‐Induced DNA Decondensation In Situ
Authors: ARAVINDAN G. ROLANDS , KRISHNAMURTHY HANUMANTHAPPA , MOUDGAL N. RAGHUVEER
Published: October 2023

ABSTRACT: Immunoneutralization of endogenous follicle‐stimulating hormone (FSH) of adult male monkeys leads to oligospermia and infertility despite unchanged testosterone levels. The inability of these monkeys to impregnate despite repeated exposures to cycling females appeared to be due to abnormal alterations in the kinetics of germ cell transformations and deficient spermiogenesis. Here we investigated the stability of sperm chromatin in oFSH‐immunized monkeys as a marker for spermiogenesis. The susceptibility of spermatozoa to in vitro decondensation induced by dithiothreitol (DTT, 0.05–50 mM) was studied by measuring the nuclear fluorescence of DTT‐treated, ethidium bromide (EB)‐stained sperm using flow cytometry. Changes in sperm morphology and binding of thiol‐specific 14C‐iodoacetamide (14C‐IA) were also monitored under the same conditions. Sperm from the immunized monkeys decondensed at a lower concentration of DTT, bound more EB, and decondensed more extensively than those from control animals. The difference was apparent in sperm from all regions of the epididymis. Immunized monkey sperm also bound significantly more 14C‐IA at all concentrations of DTT. Overall, the effective concentration of DTT required to elicit 50% of maximal decondensation (ED50) of epididymal and ejaculated sperm was significantly lower for the immunized monkeys than even the caput sperm of controls. These results suggest that FSH deprivation in monkeys results in production of sperm with limited potential for disulfide formation and reduced chromatin stability.
Extragonadal Sperm Reserves, Sperm‐Depletion Rates, Numbers of Sperm per Mating, and Fertility With Successive Matings by Intact or Unilaterally Vasectomized Rats
Authors: JUDD JENNIFER E. , BERNDTSON WILLIAM E. , CASTRO ANTONIO C. S.
Published: October 2023

ABSTRACT: Because of the high rates of sperm production and large extragonadal sperm reserves of sexually rested male rats, mating trials are insensitive for detecting test‐induced alterations in sperm production rates. Mating trials might be more sensitive if, independently of any experimental treatments under study, the number of sperm per mating was closer to the minimum requirements for normal fertility. The present study was undertaken to assess the impact of unilateral vasectomy and/or matings with up to three females in succession, for 1 hour each, on the number of sperm per mating and fertility, in comparison to corresponding values for males allowed unlimited matings with a receptive female overnight. Unilateral vasectomy did not affect sperm production, extragonadal sperm reserves, or removal of contralateral sperm during ejaculation (P > 0.05) but caused a 50% decrease in sperm numbers per mating. Sperm output, judged from numbers of residual extragonadal sperm in unmated and mated males, was excessive (290 × 106) during conventional overnight mating with intact males and during the first and second hours of restricted mating (105 and 184 × 106 respectively, for intact males; one‐half of these amounts for unilaterally vasectomized males). In contrast, sperm output during the third successive mating was minimal (nonmeasurable) but adequate, since pregnancy rates were similar for females mated first, second, or third in succession (P > 0.05). Since successive matings reduce the number of sperm per mating by natural methods, this approach may enhance the sensitivity of mating tests when applied for assessing the potential effects of experimental treatments on sperm production.
Sperm Motion Predicts Fertility in Male Hamsters Treated with α‐Chlorohydrin
Authors: SLOTT VALERIE L. , JEFFAY SUSAN C. , DYER CHERYL J. , BARBEE RANDY R. , PERREAULT SALLY D.
Published: October 2023

ABSTRACT: An understanding of the relationship between altered sperm motion and sperm function (fertility) is important when interpreting the biological significance of toxicant‐induced changes in sperm velocity in rodent test species. Previous studies showed that a brief (4‐day) exposure of male hamsters to the model chemical α‐chlorohydrln (ACH) results in significant deficits in epididymal and uterine sperm velocity, which are associated with both a delay and a failure of fertilization in vivo. To characterize this effect in terms of fertility, similarly treated male hamsters were bred to untreated females and pups were counted the day before parturition. ACH treatment resulted in a dose‐dependent decline in the percentage of sperm‐positive females that were pregnant at the end of gestation (100, 78, 67, 22, and 0 where males were treated with 0, 33, 49, 66, and 83 mg ACH/kg/day, respectively). Cauda epididymal sperm from the same males were assayed for motion characteristics using computer‐assisted sperm analysis (CASA), and for fertilizing ability in vitro. While the percentage of motile sperm was unaffected by ACH treatment, sperm velocity declined in a dose‐dependent manner at all ACH treatment levels. Futhermore, the velocity of sperm from infertile males was shifted downward consistently across the entire velocity distribution. Since treated males tended to either be infertile (no pups) or have near normal litter size, the correlation between sperm velocity and litter size was nonlinear. Therefore, logistic regression models using velocity cut‐off values were the most useful models for predicting fertility. These results support the contention that fertility relies on there being a sufficient number of sperm that exceed a velocity threshold. Sperm from treated males were also less likely to support in vitro fertilization (IVF), providing further evidence of impaired sperm function associated with acute exposure to ACH.
Brown adipose tissue evaluation using water and triglyceride as indices by diffuse reflectance spectroscopy
Authors: Iida Tomomi , Ueda Yukio , Tsukada Hideo , Fukumoto Dai , Hamaoka Takafumi
Published: October 2023

Brown adipose tissue (BAT) is related to lipid and glucose metabolism, and BAT evaluation is expected to contribute to disease prevention and treatment. We aimed to establish a BAT evaluation method using simple and non‐invasive diffuse reflectance spectroscopy (DRS). We acquired diffuse reflectance spectra of BAT using DRS from rats with cold stimulation and analyzed the second‐derivative spectra. To predict the amount of triglyceride in BAT from the second‐derivative spectra, partial least‐squares regression analysis was performed, and we examined whether BAT weight can be predicted from the amount of triglyceride by single regression analysis. By focusing on changes in the amount of triglyceride in BAT with cold stimulation, it was suggested that this amount could be predicted spectroscopically, and the predicted amount of triglyceride could be used to estimate the BAT weight with cold stimulation. If these results can be translated into humans, they may contribute to preventing metabolic disorders.
A multilayered, dynamic model of refugee resilience: A grounded theory
Authors: Kondili Elvita , Interiano‐Shiverdecker Claudia G. , Sabah May
Published: October 2023

The goal of this study was to explore the individual, cultural–relational–communal, and systemic factors associated with refugee resilience. Using constructivist grounded theory, we explored the concept of resilience through individual and focus group interviews with 21 refugees and 25 staff members working with this population in the United States. Our findings illustrate a multilayered and dynamic model that presents resilience in refugees as a fluid and complex phenomenon involving (a) individual, (b) community, and (c) societal conditions that either facilitate or hinder refugees’ ability to bounce back from the trauma(s) they have experienced. The authors discuss the implications of this model for developing a range of intervention strategies that aid in fostering refugee resilience at individual, community, and societal levels.
Water‐free two‐phase systems: phase equilibria and partitioning of vitamins from tomatoes (Solanum lycopersicum)
Authors: Lima Thailan Souza Pereira , Buarque Filipe Smith , Pinto Ronnie Emanuel Pereira , Vasconcelos Ana Luiza dos Santos , Soares Cleide Mara Faria , de Souza Ranyere Lucena , Lima Álvaro Silva
Published: October 2023

BACKGROUND

The present work provides diagrams for water‐free two‐phase systems based on polypropylene glycol 2000 (PPG 2000) + [C2mim][OAc] + alcohols (primary: methanol, ethanol, 1‐propanol and 1‐butanol; secondary: 2‐propanol and 2‐butanol; and tertiary: 2‐methyl‐2‐propanol) at 298.15 ± 1.00 K and 0.10 ± 0.01 MPa. As a proof of concept, the systems were applied to study the selective partitioning of vitamins, namely β‐carotene and α‐tocopherol.

RESULTS

The hydrophilic/hydrophobic balance is the driving force for phase separation. The more hydrophilic alcohols are responsible for the wider biphasic area, and branching increases hydrophilicity and the biphasic area compared to linear alcohols. Additionally, it was verified that the mathematical determination of the tie‐lines satisfactorily describes the positioning in the phase diagram when compared with the analytical determination. In the proposed systems, pure biomolecules partitioned selectively: α‐tocopherol migrated to the PPG 2000 phase (top phase) while β‐carotene to the [C2mim][OAc] phase (bottom phase). For the study with real biomolecules, solid–liquid extraction was performed using 1‐butanol (more selective alcohol) in a solid–liquid ratio of 1:10 at 200 rpm for 60 min. The partitioning trend followed the behavior of systems using pure biomolecules. The highest selectivity (5.3) was observed for the system composed of PPG 2000 + [C2mim][OAc] + 1‐butanol or 2‐butanol. The partition system did not impair the antioxidant potential expressed as DPPH inhibition.

CONCLUSIONS

Water‐free two‐phase systems can be formed and applied to partitioning vitamins from tomatoes. © 2023 Society of Chemical Industry (SCI).
Population‐level analysis of natural control of HIV infection in Zambia and South Africa: HPTN 071 (PopART)
Authors: Grant‐McAuley Wendy , Piwowar‐Manning Estelle , Clarke William , Breaud Autumn , Zewdie Kidist Belay , Moore Ayana , Ayles Helen Mary , Kosloff Barry , Shanaube Kwame , Bock Peter , Meehan Sue‐Ann , Maarman Gerald , Fidler Sarah , Hayes Richard , Donnell Deborah , Eshleman Susan H. ,   for the HPTN 071 (PopART) Study Team
Published: October 2023

Introduction

HIV controllers have low viral loads (VL) without antiretroviral treatment (ART). We evaluated viraemic control in a community‐randomized trial conducted in Zambia and South Africa that evaluated the impact of a combination prevention intervention on HIV incidence (HPTN 071 [PopART]; 2013–2018).

Methods

VL and antiretroviral (ARV) drug testing were performed using plasma samples collected 2 years after enrolment for 4072 participants who were HIV positive at the start of the study intervention. ARV drug use was assessed using a qualitative laboratory assay that detects 22 ARV drugs in five drug classes. Participants were classified as non‐controllers if they had a VL ≥2000 copies/ml with no ARV drugs detected at this visit. Additional VL and ARV drug testing was performed at a second annual study visit to confirm controller status. Participants were classified as controllers if they had VLs <2000 with no ARV drugs detected at both visits. Non‐controllers who had ARV drugs detected at either visit were excluded from the analysis to minimize potential confounders associated with ARV drug access and uptake.

Results

The final cohort included 126 viraemic controllers and 766 non‐controllers who had no ARV drugs detected. The prevalence of controllers among the 4072 persons assessed was 3.1% (95% confidence interval [CI]: 2.6%, 3.6%). This should be considered a minimum estimate, since high rates of ARV drug use in the parent study limited the ability to identify controllers. Among the 892 participants in the final cohort, controller status was associated with biological sex (female > male, p = 0.027). There was no significant association between controller status and age, study country or herpes simplex virus type 2 (HSV‐2) status at study enrolment.

Conclusions

To our knowledge, this report presents the first large‐scale, population‐level study evaluating the prevalence of viraemic control and associated factors in Africa. A key advantage of this study was that a biomedical assessment was used to assess ARV drug use (vs. self‐reported data). This study identified a large cohort of HIV controllers and non‐controllers not taking ARV drugs, providing a unique repository of longitudinal samples for additional research. This cohort may be useful for further studies investigating the mechanisms of virologic control.
A Composite of Polyether Ether Ketone and Silica‐Coated Copper Particles for Creating Tailored Conductive Tracks via Laser Printing
Authors: Schnettger Alexander , Holländer Ulrich , Maier Hans J.
Published: October 2023

Conventional substrates for optoelectronic systems include inorganic or organic carrier materials; however, these systems are typically subjected to environmentally harmful multistep processes to prepare printed circuit boards. To mitigate these issues, the present article reports a polyether ether ketone (PEEK)‐based composite densely filled with copper microparticles, prepared using a simple, cost‐effective, and sustainable synthesis method. The material exhibits high thermal conductivity but is electrically nonconductive prior to undergoing laser treatment. To prevent the composite from exhibiting electrical conductivity, the copper particles are coated with a thin silica layer through a sol–gel reaction. The thermal stability of PEEK and the Cu–PEEK composites with Cu contents of up to 70 vol%, which are prepared via heat melding, is investigated by thermogravimetric analysis, differential scanning calorimetry, and Fourier‐transform infrared spectroscopy to clarify the manner in which copper affects the chemical structure of the polymer. The developed composite exhibits a significantly higher thermal conductivity than that of the unfilled PEEK polymer. This paper also describes the effects of laser treatment on the surface morphology. Overall, this study suggests that conductive tracks with low electrical resistance can be created on electrically insulating substrates with high thermal conductivity.
Crosslinked copolymers with degradable oligo(lactide) branches
Authors: Sandner Barbara , Steurich Simone , Wartewig Siegfried
Published: October 2023

Methods for preparation of oligo(lactone) macromonomers end‐capped with methacrylate groups are summarized. The conversion of C=C double bonds during the crosslinking copolymerization of the macromonomers has been studied by means of Laser Raman spectroscopy at room temperature. Glass transition, mechanical properties and the degradation rate of composite materials prepared by copolymerization in the presence of hydroxylapatite may systematically be influenced both by the type of lactone monomer, e.g. D,L‐ or L,L‐dilactide, diglycolide, and the comonomer, e.g. 2‐hydroxyethyl methacrylate, tetrahydro‐furfuryl methacrylate, tri(ethylene glycol) dimethacrylate.

The composites should be useful as bone implant materials with lower polymerization exotherm and better biocompatibility than conventional materials based on methyl methacrylate.
A Randomized Phase 3 Study Comparing P2B001 to its Components (Low‐Dose Extended‐Release Rasagiline and Pramipexole) and to Optimized Doses of Marketed Extended‐Release Pramipexole in Early Parkinson's Disease
Authors: Olanow C. Warren , Hauser Robert A. , Burdick Daniel J. , Dhall Rohit , de Marcaida Joy Antonelle , Gil Ramon A. , Kreitzman David L. , Elmer Lawrence W. , McGarry Andrew , Kieburtz Karl ,   for the P2B Study Group
Published: October 2023

Background

There remains uncertainty as to the optimal way to initiate therapy for Parkinson's disease (PD) to maximize benefit and minimize adversity.

Objectives

The objective was to determine if P2B001 (a fixed, low‐dose, extended‐release [ER] combination of pramipexole 0.6 mg and rasagiline 0.75 mg) is superior to each of its components and compare its safety and efficacy to optimized treatment with marketed doses of pramipexole‐ER.

Methods

This was a 12‐week, double‐blind study (NCT03329508). Total of 544 untreated patients with PD were randomized (2:2:2:1) to treatment with P2B001, its individual components (pramipexole‐ER 0.6 mg or rasagiline‐ER 0.75 mg), or commercial doses of pramipexole‐ER titrated to optimal dose (1.5–4.5 mg). The primary endpoint was change from baseline to week 12 in Unified Parkinson's Disease Rating Scale (UPDRS) parts II and III. The key secondary endpoint was the change from baseline in the Epworth Sleepiness Scale (ESS) for P2B001 versus the titrated dose of pramipexole‐ER.

Results

P2B001 provided superior efficacy compared to each of its components; mean (95% CI) treatment differences in UPDRS II + III scores were −2.66 (95% CI, −4.33 to −1.00) versus pramipexole‐ER 0.6 mg (P = 0.0018) and − 3.30 (95% CI, −4.96 to −1.63) versus rasagiline‐ER 0.75 mg (P < 0.0001). P2B001 had comparable efficacy with the titrated dose of pramipexole‐ER (mean, 3.2 mg), but significantly less worsening in daytime‐sleepiness (ESS treatment difference: −2.66 [95% CI, −3.50 to −1.81]; P < 0.0001). P2B001 was well‐tolerated with fewer sleep‐related and dopaminergic adverse events than titrated doses of pramipexole‐ER including somnolence, orthostatic hypotension, and neuropsychiatric side effects.

Conclusions

P2B001 had superior efficacy to its individual components and was comparable with commercially used doses of pramipexole‐ER with less worsening of sleepiness and fewer dopaminergic adverse events. These findings support considering once‐daily P2B001 as initial therapy for patients with early PD. © 2023 International Parkinson and Movement Disorder Society.
Childhood cancer data initiative: Status report
Authors: Jagu Subhashini , Mardis Elaine R. , Wedekind Mary F. , Widemann Brigitte C. , Kingery Rena K. , Gonzalez Samantha L. , Shern Jack F. , Reaman Gregory H.
Published: October 2023

In March 2023, over 800 researchers, clinicians, patients, survivors, and advocates from the pediatric oncology community met to discuss the progress of the National Cancer Institute's Childhood Cancer Data Initiative. We present here the status of the initiative's efforts in building its data ecosystem and updates on key programs, especially the Molecular Characterization Initiative and the planned Coordinated National Initiative for Rare Cancers in Children and Young Adults. These activities aim to improve access to childhood cancer data, foster collaborations, facilitate integrative data analysis, and expand access to molecular characterization, ultimately leading to the development of innovative therapeutic approaches.
Polymerization‐induced self‐assembly for drug delivery: A critical appraisal
Authors: Hochreiner Eleonora G. , van Ravensteijn Bas G. P.
Published: October 2023

Polymerization‐induced self‐assembly (PISA) with (in situ) encapsulation of (therapeutic) cargo has proven to be an efficient preparation method for loaded polymeric micelles and polymersomes, thereby presenting significant opportunities in the field of drug delivery. However, despite extensive research efforts, no significant advances toward systematic in vivo studies or clinical applications have been achieved to date. In this Review, we outline the current state‐of‐the‐art of cargo encapsulation via PISA with a specific focus on developments achieved in the past 5 years. Considering the general requirements for functional drug delivery systems, we identify the major hurdles that still need to be overcome in order to push PISA‐derived systems from a promising academic exercise to viable candidates for clinical translation.
Modelling the dynamics of hydrogen synthesis from methane in nanosecond‐pulsed plasmas
Authors: Morais Eduardo , Bogaerts Annemie
Published: October 2023

A chemical kinetics model was developed to characterise the gas‐phase dynamics of H2 production in nanosecond‐pulsed CH4 plasmas. Pulsed behaviour was observed in the calculated electric field, electron temperature and species densities at all pressures. The model agrees reasonably with experimental results, showing CH4 conversion at 30% and C2H2 and H2 as major products. The underlying mechanisms in CH4 dissociation and H2 formation were analysed, highlighting the large contribution of vibrationally excited CH4 and H2 to coupling energy from the plasma into gas‐phase heating, and revealing that H2 synthesis is not affected by applied pressure, with selectivity remaining unchanged at ~42% in the 1–5 bar range.
The role of plasma concentrations and drug characteristics of beta‐blockers in fall risk of older persons
Authors: Ploegmakers K. J. , van Poelgeest E. P. , Seppala L. J. , van Dijk S. C. , de Groot L. C. P. G. M. , Oliai Araghi S. , van Schoor N. M. , Stricker B. , Swart K. M. A. , Uitterlinden A. G. , Mathôt R. A. A. , van der Velde N.
Published: October 2023

Beta‐blocker usage is inconsistently associated with increased fall risk in the literature. However, due to age‐related changes and interindividual heterogeneity in pharmacokinetics and dynamics, it is difficult to predict which older adults are more at risk for falls. Therefore, we wanted to explore whether elevated plasma concentrations of selective and nonselective beta‐blockers are associated with an increased risk of falls in older beta‐blocker users. To answer our research question, we analyzed samples of selective (metoprolol, n = 316) and nonselective beta‐blockers (sotalol, timolol, propranolol, and carvedilol, n = 179) users from the B‐PROOF cohort. The associations between the beta‐blocker concentration and time to first fall were assessed using Cox proportional hazard models. Change of concentration over time in relation to fall risk was assessed with logistic regression models. Models were adjusted for potential confounders. Our results showed that above the median concentration of metoprolol was associated with an increased fall risk (HR 1.55 [1.11–2.16], p = .01). No association was found for nonselective beta‐blocker concentrations. Also, changes in concentration over time were not associated with increased fall risk. To conclude, metoprolol plasma concentrations were associated with an increased risk of falls in metoprolol users while no associations were found for nonselective beta‐blockers users. This might be caused by a decreased β1‐selectivity in high plasma concentrations. In the future, beta‐blocker concentrations could potentially help clinicians estimate fall risk in older beta‐blockers users and personalize treatment.
Investigation of the human metabolism and disposition of the prolyl hydrolase inhibitor daprodustat using IV microtracer with Entero‐Test bile string
Authors: Tai Guoying , Xia Fangming , Chen Cathy , Pereira Adrian , Pirhalla Jill , Miao Xiusheng , Young Graeme , Beaumont Claire , Chen Liangfu
Published: October 2023

Daprodustat is an oral small molecule hypoxia‐inducible factor (HIF) prolyl hydroxylase inhibitor (PHI) approved in Japan and the United States for the treatment of anemia associated with chronic kidney disease. This phase 1, nonrandomized, 2‐period, crossover study in 6 healthy men characterized and quantified the metabolites generated after a microtracer IV infusion of 50 μg (125 nCi) [14C]‐daprodustat administered concomitantly with a nonradiolabeled therapeutic dose of a 6‐mg daprodustat tablet, followed by a single oral solution dose of 25 mg (62.5 μCi) [14C]‐daprodustat. High‐performance liquid chromatography (HPLC) coupled with radioactivity detection (TopCount or AMS) and HPLC‐tandem mass spectrometry (HPLC‐MSn) were used for quantitative measurement and structural identification of radioactive metabolites in plasma, urine, feces, and bile. Following oral administration of [14C]‐daprodustat, unchanged daprodustat was the principal circulating drug‐related component, accounting for 40% of plasma radioactivity. Predominant oxidative metabolites M2, M3, M4, and M13 individually represented 6–8% of the plasma radioactivity and together accounted for the majority of radioactivity in urine and feces (53% in both matrices; 12% and 41% of dose, respectively). Unchanged daprodustat was not detected in urine and was only 0.7% of total radioactivity in feces (<0.5% of dose), with the remainder of the dose accounted for by oxidative metabolites. The radio‐metabolic profile of duodenal bile following IV infusion of [14C]‐daprodustat was similar to that observed in feces after oral administration. The data suggested that oral daprodustat was extensively absorbed, cleared exclusively by oxidative metabolism, and eliminated via hepatobiliary (primary) and urinary (secondary) excretion.
Thoracic SMARCA4‐deficient undifferentiated tumour: Diagnostic challenges and potential for misdiagnosis in small tissue samples
Authors: Maartens Deborah Johanna , Moolla Muhammad Saadiq , Ndaba Sibusiso , Vlok Sucari Susanna Catherina , Hendricks Firzana , Koegelenberg Coenraad Frederik Nicolaas , van Wyk Abraham Christoffel
Published: October 2023

We report a diagnostically challenging case of a SMARCA4‐deficient undifferentiated tumour to emphasize its potential to mimic other malignant tumours on histology, especially in small biopsies and where rhabdoid morphology is lacking. A 48‐year‐old man, who was known for chronic obstructive pulmonary disease and polysubstance use, presented with dyspnoea and an anterior mediastinal mass that had grown rapidly over a seven‐month period. The rapid growth and location in the anterior mediastinum raised clinical suspicion for lymphoma or a germ cell tumour. Microscopic examination of a transthoracic, ultrasound‐guided, core needle biopsy revealed relatively uniform, malignant epithelioid cells with clear cytoplasm, but lacking any rhabdoid features. Tumour necrosis was prominent. The immunohistochemistry panel was negative for lymphoma markers, but positive for SALL4 (a marker typically associated with germ cell tumours), CD34, EMA, and HepPar1, while expression of SMARCA4 and claudin‐4 was entirely lost. Only focal cytokeratin expression was demonstrated. SMARCB1 (INI1) expression was retained. The diagnosis of SMARCA4‐DUT was made based on these findings. Unfortunately, the tumour was already at an advanced stage at diagnosis (stage IVA) and the patient had a poor performance status. He was treated with palliative radiotherapy with no significant improvement in performance status and passed away 3 months after diagnosis. The case highlights the importance of considering SMARCA4‐DUT in the differential diagnosis of an undifferentiated, rapidly growing thoracic tumour and the potential for misdiagnosis on a small tissue sample, particularly as rhabdoid morphology may be absent.