bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of TGF‐β1, TGF‐α, and EGF on Cell Proliferation and Cell Death in Rat Ventral Prostatic Epithelial Cells in Culture
Authors: ILIO KENNETH Y. , SENSIBAR JULIA A. , LEE CHUNG
Published: October 2023

ABSTRACT: A tissue culture system for rat prostatic epithelial cells was developed, and the effect of epidermal growth factor (EGF), transforming growth factor alpha (TGF‐α), and transforming growth factor beta 1 (TGF‐β1) on these cells was evaluated. The primary culture was prepared by DNAse/collagenase dissociation of minced ventral prostates. Cells were initially plated in RPMI‐1640 medium containing 10% fetal bovine serum to allow the preferential attachment of stromal cells. Twenty‐four hours later, the unattached epithelial cells were replated in WAJC‐404 medium supplemented with insulin (5 μg/ml), transferrin (5 μg/ml), and selenious acid (5 ng/ml). Bovine pituitary extract (BPE) (30 μg/ml), EGF (10 ng/ml), and TGF‐β1 (0, 0.1, and 1.0 ng/ml) were added either alone or in combination according to experimental requirements. The rate of cell proliferation was assessed by counting the total cell number and by [9H]thymidine incorporation. Prostatic epithelial cells exhibited a bell‐shaped growth curve in a span of 7–8 days, with a growth peak at day 3 or 4 of culture. Treatment of cells with EGF or TGF‐α resulted in a concentration‐dependent increase in cell growth, whereas addition of TGF‐β1 into the culture resulted in an inhibition of cell proliferation that could be reversed with increasing concentrations of EGF. Cell death was assessed using the terminal deoxynudeotidyl transferase (TdT)‐mediated immunoperoxidase‐digoxigenin nick end labeling technique and the trypan blue exclusion test. Epithelial cells cultured in media containing EGF had the lowest incidence of cell death. Cells cultured in the absence of EGF demonstrated a marked increase in cells undergoing cell death. The addition of TGF‐β1 into the EGF‐depleted medium caused a further increase of cell death. These results indicated that cell proliferation and cell death in rat prostatic epithelial cells in culture could be modulated by EGF and TGF‐β1. The former stimulated cell proliferation and prevented cell death, whereas the latter inhibited proliferation in the presence or absence of EGF and induced cell death.
Androgen Receptor Immunoexpression in the Testes of Subfertile Men
Authors: VAN ROIJEN J. HERMAN , VAN ASSEN SANDER , VAN DER KWAST THEODORUS H. , de ROOIJ DICK G. , BOERSMA WIM J. A. , VREEBURG JAN T. M. , WEBER ROBERTUS F. A.
Published: October 2023

ABSTRACT: The localization and intensity of androgen receptor immunostaining was studied in the testes of 37 subfertile men with oligozoospermia and normal serum gonadotropin levels using a polyclonal antibody raised against a synthetic peptide corresponding to the first 20 N‐terminal amino add residues of the androgen receptor (AR). Furthermore, we investigated whether or not the immunoexpression of the AR in human Sertoli cells, in histologically normal testis tissue, is dependent on the stage of the spermatogenic cycle, as has been found in the rat. In the human testis, AR immunoexpression was observed in Sertoli cells, peritubular myoid cells, Leydig cells, and periarteriolar cells, but not in germinal cells. We found no evidence for a stage‐dependent immunoexpression of AR in Sertoli cells. The intensity of AR immunoexpression varied substantially between biopsy specimens of different patients. There was, however, no correlation of the intensity of AR immunoexpression in either Sertoli cells or peritubular myoid cells with spermatogenic adequacy as measured by the method of Johnsen. When, in this study, the intensity of peritubular myoid cell staining was used as a standard to evaluate the intensity of Sertoli cell staining, no correlation was detected as well. Furthermore, serum gonadotropin levels were not correlated with AR immunoexpression levels in Sertoli cells and peritubular myoid cells. These results indicate that immunodetectability of the AR is not related to the condition of the spermatogenic epithelium in patients with oligozoospermia. Inappropriate expression of the AR is neither a cause nor a consequence of idiopathic infertility in the present group of patients.
The mRNAs for the Steroid 5α‐Reductase Isozymes, Types 1 and 2, Are Differentially Regulated in the Rat Epididymis
Authors: VIGER ROBERT S. , ROBAIRE BERNARD
Published: October 2023

ABSTRACT: The enzyme steroid 5α‐reductase (EC 1.3.1.22) catalyzes the conversion of testosterone to its more potent form, dihydrotestosterone (DHT), in many androgen‐sensitive target tissues. In the epididymis, the 5α‐reduced metabolites of testosterone, DHT and 5α‐androstan‐3α, 17β‐diol (3α‐diol), are considered the primary regulators of epididymal structure and function. Two rat 5α‐reductase transcripts, designated types 1 and 2, have been identified. Our laboratory has previously characterized the endocrine and developmental regulation of the 5α‐reductase type 1 mRNA in the rat epididymis. However, regulation of the type 2 mRNA has not been investigated. Thus, we undertook to characterize the longitudinal distribution of the steady state 5α‐reductase type 2 mRNA as well as the effects of development and of unilateral efferent duct ligation on its expression in the rat epididymis. To contrast the regulation of the type 2 5α‐reductase isozyme mRNAs in the rat epididymis, these data have been compared with those previously obtained for the type 1 mRNA and enzyme activity. An analysis of the longitudinal distribution of the type 2 transcript along the epididymis revealed that its mRNA was expressed predominantly in the proximal caput region of the tissue. This regional distribution pattern differed markedly from the patterns previously described for the type 1 mRNA and enzyme activity. Surprisingly, a segment‐by‐segment analysis showed that epididymal 5α‐reductase type 2 mRNA levels were not altered as a function of increasing postnatal age. Again, this result was in marked contrast to the important changes in 5α‐reductase type 1 mRNA and enzyme activity that have been reported to occur during puberty in the rat. In the final experiment, the effect of unilateral efferent duct ligation revealed that 5α‐reductase type 2 mRNA levels increased in the initial segment of the ligated side but remained unchanged in the rest of the tissue; this was in marked contrast to the dramatic decrease (>60%) in type 1 mRNA levels observed specifically in the initial segment of the epididymis. Taken together, these experiments demonstrate that 5α‐reductase type 1 and type 2 mRNAs are differentially regulated in the rat epididymis. Moreover, this is the first report of a tissue in which the mRNAs for the steroid 5α‐reductase isozymes are regulated differently.
Secretion of Bioactive Interleukin 1 by Rat Testicular Macrophages In Vitro
Authors: Hayes R. , Chalmers S. A. , Nikolic‐Paterson D. J. , Atkins R. C. , Hedger M. P.
Published: October 2023

ABSTRACT: The relative capacities for testicular macrophages and resident peritoneal macrophages to secrete the pro‐inflammatory cytokine, interleukin 1 (lL‐1), in response to stimulation by bacterial lipopolysaccharide (LPS), were compared in vitro. Macrophages were isolated from adult male rat testicular interstitial cells or peritoneal lavage by adherence to glass coverslips or plastic culture dishes. The macrophages were immediately cultured, with or without a maximal dose of LPS (1 μg/ml), over 24 hours at 32°C. Bioactive IL‐1 production was measured by a sensitive thymocyte proliferation bioassay, employing recombinant human IL‐1β as the reference standard. In comparison with the peritoneal macrophages, testicular macrophages displayed only a very small response to LPS, producing 2.8% of the amount of IL‐1 per cell secreted by peritoneal macrophages cultured under identical conditions. Production of authentic IL‐1 was confirmed by inhibition of the bioassay response in the presence of human recombinant IL‐1 receptor antagonist. A small molecular mass (< 10 kDa based on ultrafiltration) inhibitor of IL‐1 bioactivity was also present in the medium collected from both cultures, but this inhibitory activity did not account for the differences in activity observed. In cultures of total peritoneal cells under similar conditions, addition of testosterone (10‐1,000 ng/ml) did not affect IL‐1 production in response to LPS. These data indicate that testicular macrophages have a reduced ability to secrete bioactive IL‐1, and they provide further evidence for an altered capacity for immune responses within the testis.
Difference in Glycohistochemical Lectin Staining of Collagen Fibers in the Corpora Cavernosa of Normal and Impotent Men
Authors: RAVIV GIL , WESPES ERIC , VANEGAS JUAN PABLO , PETEIN MICHEL , SCHULMAN CLAUDE C. , KISS ROBERT , DANGUY ANDRé
Published: October 2023

ABSTRACT: The objectives of this study were to investigate the value of glycohistochemical staining with three lectin types specific to a particular glycan structure (Arachis hypogaea [PNA], Triticum vulgare [WGA], and concanavalin A [Con A]) as a method of defining possible changes in the collagen structure in the corpora cavernosa in potent and impotent men. The study group consisted of 4 normal potent men and 22 men with various etiologies of impotence. The quantitative histochemical measurements were performed by means of a cell image processor. Two variables for each of the three types of lectins were studied. These were the mean optical density (MOD), which relates to glycohistochemical staining intensity, and the labeling index (LI), which is positively related to the percentage of immunostaining. Only WGA staining made it possible to discriminate significantly between the normal and pathological groups under study. The two parameters (LI, MOD) were significantly higher in the case of WGA staining in the normal group (P = 0.004 and 0.013, respectively). In contrast, only the mean LI value, in the case of the psychogenic and venogenic patients, reached a level of statistical significance (P = 0.005 and 0.001, respectively), when it increased from PNA through WGA to Con A histochemical staining. The two variables (LI, MOD) changed markedly from PNA through WGA to Con A in the arteriogenic patients (P = 0.003 and P < 0.001, respectively). WGA is of diagnostic value in distinguishing between normal and abnormal collagen in the corpora cavernosa. The difference in the lectin staining of the other groups, particularly the arteriogenic group, may be attributed to alterations in the glycosylation of the procollagen that are probably due to changes in the partial pressure of oxygen (PO2) level, an important cofactor in normal glycosylation. WGA staining may therefore be used as a marker to distinguish true psychogenic patients from those with organic diseases. Moreover, it may be used as an additional parameter in selecting the best candidates for penile revascularization.
Apical and Narrow Cells Are Distinct Cell Types Differing in Their Structure, Distribution, and Functions in the Adult Rat Epididymis
Authors: ADAMALI HUZAIFA I. , HERMO LOUIS
Published: October 2023

ABSTRACT: Apical and narrow cells of the initial segment and intermediate zone of the adult rat epididymis were glutaraldehyde fixed and Epon embedded for routine light (LM) and electron (EM) microscopic analysis and Bouin fixed and paraffin embedded for LM immunocytochemical analysis in order to examine their structural features, distribution, and functions. The goblet‐shaped apical cells comprised 10.7 ± 1.0% of the total epithelial population in the proximal initial segment but only 1.3 ± 0.5% in the intermediate zone. In the EM, these cells presented numerous mitochondria, few C‐shaped vesicles, and a pale round or oblong nucleus located in the upper half of their cytoplasm. The slender elongated narrow cells increased from 2.8 ± 0.3% in the proximal initial segment to 6.3 ± 0.4% in the intermediate zone. In an EM analysis, these cells presented numerous C‐shaped vesicles and mitochondria and a small flattened nucleus located in the upper half of their cytoplasm. The structural features of both these cell types differed not only from each other but also from the neighboring principal and basal cells of each region. Of the various antibodies examined to lysosomal proteins, narrow and apical cells expressed high levels of cathepsin D, while β‐hexosaminidase A was expressed at high levels in narrow cells but only moderately in apical cells. Apical cells were intensely reactive for the Yf subunit of glutathione S‐transferase (GST)‐P, whereas no reaction was seen in narrow cells; the Yo subunit of GST was localized within both cell types but only in the proximal initial segment. Narrow cells exclusively expressed carbonic anhydrase II. Selective differences in the immunolocalization of these various proteins were also noted between these two cell types and principal and basal cells. The localization of cathepsin D and β‐hexosaminidase A within narrow and apical cells suggests these cells may be involved in the degradation of specific proteins within their lysosomes, whereas the presence of GSTs may aid in protecting spermatozoa from a changing environment of harmful electrophiles. Localization of carbonic anhydrase II exclusively within narrow cells suggests that these cells may modify the pH of the lumen resulting in the quiescence of sperm motility in the proximal end of the epididymis. Together, the data indicate that apical and narrow cells differ not only from each other but also from principal and basal cells in their structure and relative distribution. They also express different proteins within the distinct epididymal regions, indicating that they perform different functions.
The Neural Cell Adhesion Molecule (NCAM) Provides Clues to the Development of Testicular Leydig Cells
Authors: MAYERHOFER ARTUR , LAHR GEORGIA , SEIDL KLAUS , EUSTERSCHULTE BEATE , CHRISTOPH ANNETTE , GRATZL MANFRED
Published: October 2023

ABSTRACT: As previously shown, Leydig cells in culture dramatically up‐regulate the expression of the neural cell adhesion molecule (NCAM) gene and express alternatively spliced forms. Because the family of NCAM adhesion molecules is known to be involved in cell migration and differentiation, we examined the potential involvement of NCAMs in Leydig cell differentiation in the developing testis. We detected NCAM‐immunoreactive cells in the rat and mouse at embryonic day (ED) 13–14 in epithelia of mesonephric tubules and cell clusters between the mesonephros and testis. At about ED 17–18, strongly NCAM‐immunoreactive cells were demonstrated extending from the mesonephros/mesonephric tubules into the region occupied by the forming rete testis and spreading into the testis itself. Within the testis, interstitial fetal Leydig cells, identified with an antiserum directed against P450‐side‐chain cleavage enzyme, were also NCAM immunoreactive, although to a lesser degree, and in part expressed NCAM‐associated polysialic acid. In situ hybridization histochemistry demonstrated the presence of NCAM mRNA, mainly in the cell population corresponding to the strongly immunoreactive cells. NCAM forms with molecular weights of 140 and 180 kDa, the latter polysialylated, predominate in immunoblots of fetal testes. Because testicular development occurs along the mesonephros, from which precursor cells are thought to migrate into the developing gonad and then differentiate into the various testicular cell types, our results may suggest that the expression of NCAMs and NCAM modification are associated with cells that appear to migrate into the developing testis. Although it remains to be proven whether these cells could be precursor cells of Leydig cells, this assumption is supported by the fact that within the developing testis, NCAMs and NCAM modifications are expressed during differentiation of testicular P450‐side‐chain cleavage enzyme‐positive Leydig cells. Thus, the adhesion molecule NCAM and its variants and modifications are expressed in the developing testis and may be of functional, “morphogenic” importance. Because Leydig cells in vitro and during fetal development express NCAMs, these molecules may prove to be a suitable specific marker for the study of Leydig cell development and differentiation.
Purification, cDNA Cloning, and Developmental Changes in the Steady‐State mRNA Level of Rat Testicular Tissue Inhibitor of Metalloproteases‐2 (TIMP‐2)
Authors: Grima Josephine , Calcagno Karen , Cheng C. Yan
Published: October 2023

ABSTRACT: Using multiple high‐performance liquid chromatography (HPLC) steps and high‐performance electrophoresis chromatography (HPEC) in conjunction with an [125l]collagen film assay to identify inhibitors of metalloproteases, we have purified a 22‐kDa polypeptide to apparent homogeneity from primary Sertoli cell‐enriched culture medium. Partial N‐terminal amino acid sequence analysis revealed that this protein is similar to the human tissue inhibitor of metalloproteases‐2 (TIMP‐2). To determine the similarity of rat testicular TIMP‐2 to the human homolog, a full‐length cDNA coding for rat testicular TIMP‐2 was isolated from a rat Sertoli cell cDNA expression library and sequenced. Analysis of the nucleotide sequence and the deduced amino acid sequence of the rat testicular TIMP‐2 cDNA revealed an 84 and 98% homology with the human TIMP‐2 nucleotide and amino acid sequences, respectively. A survey of its mRNA transcripts in different tissues by northern blots revealed the presence of two mRNA species of 3.7 and 1.3 kb in the testis and brain but not in the kidney, spleen, epididymis, and liver in adult male rats. Studies using polymerase chain reaction (PCR) and Southern blot to detect the TIMP‐2 mRNA using total RNA isolated from germ cells, Sertoli cells, and Leydig cells have shown that only Sertoli and Leydig cells expressed TIMP‐2 mRNA. These results indicate that Sertoli cells are the major source of TIMP‐2 in the testis behind the blood‐testis barrier (seminiferous tubule barrier). During testicular development from 3 to 60 days of age, the testicular steady‐state TIMP‐2 mRNA level increased steadily with an advancement of age. Such an increase in the steady‐state testicular TIMP‐2 mRNA level apparently is not the result of an up‐regulation by germ cells because germ cells cocultured with Sertoli cells failed to elicit an increase in the Sertoli cell steady‐state TIMP‐2 mRNA level. The results of this study suggest that TIMP‐2 secreted by Sertoli cells may play a role in tissue restructuring and germ cell migration during spermatogenesis.
Evaluation of Computer‐Assisted Semen Analysis (CASA) With IDENT Stain to Determine Sperm Concentration
Authors: ZINAMAN MICHAEL J. , UHLER MEIKE L. , VERTUNO ELIZABETH , FISHER SUSAN G. , CLEGG ERIC D.
Published: October 2023

ABSTRACT: This study was undertaken to compare a new fluorescent stain‐based computer‐assisted semen analysis (CASA) system (IDENT) for determining human sperm concentration to the manual hemacytometric method and to conventional CASA (CASA‐CONV). Normal healthy semen donors as well as patients provided samples that were evaluated for sperm concentration with the CASA‐IDENT method, the hemacytometer method, and CASA‐CONV. Each field was examined visually to determine the sources of overcounting and undercounting for the two CASA methods. Four ranges of sperm concentration were examined: 0–10, >10–30, >30–100, and >100 × 106/ml. The main outcome measures were sperm concentration, debris counted as sperm, and missed sperm. Our results showed that significantly more debris was counted as sperm and more sperm were missed with CASA‐CONV than CASA‐IDENT. As the sperm density increased, so did the number of counting errors for the CASA‐CONV system. The error rate was much greater using CASA‐CONV (12.1 ± 42.2%) than with CASA‐IDENT (0.4 ± 0.7%) when compared to hemacytometer counts(P = 0.068). We conclude that the CASA‐IDENT method of sperm counting is highly accurate and less time‐consuming when compared to the hemacytometer method. There are significant differences in the amount of debris counted as sperm and number of missed sperm between CASA‐CONV and CASA‐IDENT with varying sperm density. With both parameters, the counts are more accurate using the CASA‐IDENT method.
Media and Dilution Procedures Tested to Minimize Handling Effects on Human, Rabbit, and Bull Sperm for Computer‐Assisted Sperm Analysis (CASA)
Authors: FARRELL PAUL B. , FOOTE ROBERT H. , McARDLE MARGARET M. , TROUERN‐TREND VICTORIA L. , TARDIF ANGELA L.
Published: October 2023

ABSTRACT: Proper handling of semen prior to computer‐assisted sperm analysis (CASA) is critical if the analysis is to be representative of the fresh sample. The effects of diluting medium or dilution and holding time before CASA on multiple sperm characteristics were studied. Four replicates of unselected semen samples from each of eight human donors were diluted with phosphate‐buffered saline (PBS)‐glucose plus bovine serum albumin (BSA), with Tyrode's albumen lactate pyruvate (TALP), and with high‐potassium TALP (K‐TALP) to a concentration of approximately 25 × 106 sperm/ml. The diluted semen was held for 0, 1, and 2 hours at approximately 30°C before CASA, with little difference between the three diluents in all 12 variables measured. There was a decline of 3–6% in the proportion of motile sperm over a 2‐hour period (P < 0.05). Donors were the largest source of differences (P < 0.05). Rabbit sperm (five bucks, four ejaculates per buck) were processed in a manner similar to that of the human sperm. There was a major effect of media. The average percentages of motile sperm over 2 hours in TALP, K‐TALP, and PBS were 76, 42, and 29%, respectively (P < 0.05), with a decline of only 3% in TALP during the 2 hours. Hyperactivity and other characteristics were affected by treatment. Donors were a large source of variation. Bull semen (10 bulls, two ejaculates per bull) either was not diluted or diluted with TALP 2x or 4x and held for 0, 1, and 2 hours at 30°C. It was then diluted to 25 × 106 sperm/ml with TALP. There was little change in most sperm characteristics in any treatment during the first hour, although many of the changes were statistically significant. The percentage of motile sperm in undiluted semen declined from 87% to 82% over 2 hours. Modified TALP was a suitable medium for sperm from all three species, and a simple PBS‐glucose‐BSA medium can be used for human sperm.
An evaluation of video‐prompting procedures via telehealth to teach first aid skills to children with intellectual and developmental disabilities
Authors: Sureshkumar Brittney M. , Zonneveld Kimberley L. M.
Published: October 2023

Unintentional injuries are one of the leading causes of morbidity and mortality among children with intellectual and developmental disabilities (IDD). First aid training involves teaching critical first aid skills, some of which are designed to treat unintentional injuries. To date, no study has (a) evaluated the effects of a video‐prompting procedure to teach first aid skills to children with IDD or (b) attempted to teach these skills to children by using a telehealth delivery format. We used a concurrent multiple‐baseline‐across‐skills design to evaluate the efficacy of a video‐prompting procedure via telehealth to teach five children with IDD to perform first aid on themselves for insect stings, minor cuts, and minor burns under simulated conditions. For all participants, our procedure produced large improvements that maintained for a minimum of 4 weeks. Furthermore, the effects of the training generalized to novel confederates for all participants, and these effects maintained for a minimum of 4 weeks.
Pirh2 modulates amyloid‐β aggregation through the regulation of glucose‐regulated protein 78 and chaperone‐mediated signaling
Authors: Singh Abhishek , Tiwari Shubhangini , Singh Sarika
Published: October 2023

Amyloid‐β (Aβ) protein aggregation in the brain is a pathological hallmark of Alzheimer's disease (AD) however, the underlying molecular mechanisms regulating amyloid aggregation are not well understood. Here, we studied the propitious role of E3 ubiquitin ligase Pirh2 in Aβ protein aggregation in view of its regulatory ligase activity in the ubiquitin‐proteasome system employing both cellular and sporadic rodent models of AD. Pirh2 protein abundance was significantly increased during Streptozotocin (STZ) induced AD conditions, and transient silencing of Pirh2 significantly inhibited the Aβ aggregation and modified the dendrite morphology along with the substantial decrease in choline level in the differentiated neurons. MALDI‐TOF/TOF, coimmunoprecipitation, and UbcH7‐linked in vitro ubiquitylation analysis confirmed the high interaction of Pirh2 with chaperone GRP78. Furthermore, Pirh2 silencing inhibits the STZ induced altered level of endoplasmic reticulum stress and intracellular Ca2+ levels in neuronal N2a cells. Pirh2 silencing also inhibited the AD conditions related to the altered protein abundance of HSP90 and its co‐chaperones which may collectively involve in the reduced burden of amyloid aggregates in neuronal cells. Pirh2 silencing further stabilized the nuclear translocation of phospho‐Nrf2 and inhibited the altered level of autophagy factors. Taken together, our data indicated that Pirh2 is critically involved in STZ induced AD pathogenesis through its interaction with ER‐chaperone GRP78, improves the neuronal connectivity, affects the altered level of chaperones, co‐chaperones, & autophagic markers, and collectively inhibits the Aβ aggregation.
Support effect of Mn/Na2WO4‐based catalysts in oxidative coupling of methane
Authors: Torshizi Hasan Oliaei , Nakhaei Pour Ali , Salimi Alireza , Nezhadalibaghan Zahra
Published: October 2023

BACKGROUND

In this work, the supported Mn‐Na2WO4/γ‐Al2O3, Mn‐Na2WO4/γ‐Al2O3‐TiO2 and Mn‐Na2WO4/ γ‐Al2O3‐SiO2 catalysts were prepared using the dry impregnation procedure and tested for suitability to the oxidative coupling of methane (OCM) reaction. The synthesized catalyst was characterized by various techniques such as X‐ray diffraction (XRD), Fourier transform infrared (FTIR), X‐ray photon spectroscopy (XPS), temperature‐programmed reduction (TPR), Brunauer–Emmett–Teller (BET) surface area and field emission scanning electron microscopy (FESEM), and the support effect of the prepared catalysts in the OCM reaction was evaluated.

RESULTS

The XPS results showed that the tetrahedral WO42− phase is presented in all prepared catalysts and it is expected that the presence of this phase is correlated with high OCM activity. The XPS results show that the amount of O2− in the treated Mn‐Na2WO4/γ‐Al2O3‐TiO2 catalyst at 800 °C is higher than that in the Mn‐Na2WO4/γ‐Al2O3 and Mn‐Na2WO4/γ‐Al2O3‐SiO2 catalysts, but is reduced with increasing temperature. At 850 °C, the amount of O2− in the treated Mn‐Na2WO4/γ‐Al2O3 catalyst is higher than in the Mn‐Na2WO4/γ‐Al2O3‐TiO2 and Mn‐Na2WO4/γ‐Al2O3‐SiO2catalysts.

CONCLUSION

The results show that the MnSiO3 and MnTiO3 sites may deliver a much easier Mn2+↔Mn3+ cycle than the MnAl2O4 and MnWO4 sites at low temperatures, and may play a principal role in enhancing the catalyst activity in the OCM reaction at 800 °C. © 2023 Society of Chemical Industry (SCI).
False‐positive supraclavicular lymph node detected on chest computed tomography in oncology patients: Clinical implication based on subsequent neck ultrasonography and ultrasonography‐guided tissue sampling
Authors: Jeong Yujin , Cho Eun , Baek Hye Jin , Jang Jeong Yoon , Choi Kwang Ho
Published: October 2023

Purpose

The purpose of this study was to assess the prevalence and clinical implications of false‐positive supraclavicular lymph node (LN) detected on chest computed tomography (CT), using subsequent neck ultrasonography (US) and US‐guided tissue sampling.

Methods

Among 172 patients with suspected supraclavicular LNs identified on CT, 87 underwent neck US or US‐guided tissue sampling. Receiver operating characteristic curve and logistic regression analyses were performed to determine the diagnostic performance of US and independent predictors of false‐positive LNs.

Results

Among 87 patients, 49 (56.3%) were pathologically confirmed as metastases, 26 (29.9%) were negative for malignancy, and 12 (13.8%) had pseudolesions or schwannomas. The diagnostic indices were as follows: sensitivity, 91.8%; specificity, 92.3%; PPV, 95.7%; NPV, 85.7%; and accuracy, 92.0% (AUC = 0.921; 95% CI: 0.832–0.970, p < 0.001). The false‐positive group had a higher mean age than the true‐positive group (mean age, 69.8 ± 9.2 vs. 63.9 ± 9.8, p = 0.003). Logistic regression analyses revealed that age ≥ 65 years was the only independent predictor of false‐positive LNs (OR = 4.391; 95% CI: 1.037–18.582; p = 0.044).

Conclusion

Subsequent US can be helpful for evaluating suspicious supraclavicular LNs detected on CT to establish appropriate management, especially in older patients.
A multi‐institutional study designed by members of Tokyo Guidelines (TG) Core Meeting to elucidate the clinical characteristics and pathogenesis of acute cholangitis after bilioenteric anastomosis and biliary stent insertion with a focus on biliary obstruction: Role of transient hepatic attenuation difference (THAD) and pneumobilia in improving TG diagnostic performance
Authors: Kato Hiroyuki , Takada Tadahiro , Strasberg Steven , Isaji Shuji , Sano Keiji , Yoshida Masahiro , Itoi Takao , Okamoto Kohji , Kiriyama Seiki , Yagi Shintaro , Matsubara Takashi , Higuchi Ryota , Ohyama Tetsuji , Misawa Takeyuki , Mukai Shuntaro , Mori Yasuhisa , Asai Koji , Mizuno Shugo , Abe Yuta , Suzuki Kenji , Homma Yuki , Hata Jiro , Tsukiyama Kana , Kumamoto Yusuke , Tsuyuguchi Toshio , Maruo Hirotoshi , Asano Yukio , Hori Shutaro , Shibuya Makoto , Mayumi Toshihiko , Toyota Naoyuki , Umezawa Akiko , Gomi Harumi , Horiguchi Akihiko
Published: October 2023

Background/Purpose

The aim of this study was to clarify the clinical characteristics of acute cholangitis (AC) after bilioenteric anastomosis and stent‐related AC in a multi‐institutional retrospective study, and validate the TG18 diagnostic performance for various type of cholangitis.

Methods

We retrospectively reviewed 1079 AC patients during 2020, at 16 Tokyo Guidelines 18 (TG 18) Core Meeting institutions. Of these, the post‐biliary reconstruction associated AC (PBR‐AC), stent‐associated AC (S‐AC) and common AC (C‐AC) were 228, 307, and 544, respectively. The characteristics of each AC were compared, and the TG18 diagnostic performance of each was evaluated.

Results

The PBR‐AC group showed significantly milder biliary stasis compared to the C‐AC group. Using TG18 criteria, definitive diagnosis rate in the PBR‐AC group was significantly lower than that in the C‐AC group (59.6% vs. 79.6%, p < .001) because of significantly lower prevalence of TG 18 imaging findings and milder bile stasis. In the S‐AC group, the bile stasis was also milder, but definitive‐diagnostic rate was significantly higher (95.1%) compared to the C‐AC group. The incidence of transient hepatic attenuation difference (THAD) and pneumobilia were more frequent in PBR‐AC than that in C‐AC. The definitive‐diagnostic rate of PBR‐AC (59.6%–78.1%) and total cohort (79.6%–85.3%) were significantly improved when newly adding these items to TG18 diagnostic imaging findings.

Conclusions

The diagnostic rate of PBR‐AC using TG18 is low, but adding THAD and pneumobilia to TG imaging criteria may improve TG diagnostic performance.
Identifying and locating carotenoids in supra‐orbital combs of male black grouse (Lyurus tetrix) using Raman and transmission electron microscopy: A histological study using rehydrated tissue samples
Authors: Dobson Jessica L. , Pike Thomas W. , Gonzalez‐Rodriguez Jose , Soulsbury Carl D.
Published: October 2023

Colourful signals have long been implicated as indicators of individual quality in animals. Bare‐skin signals are an understudied aspect of avian colouration compared with plumage studies, despite displaying rapid changes in size and colour in response to different environmental or physiological stressors. Even fewer studies have focused on the underlying histology of these structures and the importance this plays in the resulting skin colour. Using the Black Grouse (Lyrurus tetrix), we identified the underlying structure of individual dermal spikes, which make up the red supra‐orbital comb (a known integumentary signal of male quality), and highlight visual structural differences between combs of different sizes. In addition, we used Raman spectroscopy to indicate the presence of carotenoids within the tissue, something that had previously only been inferred through characteristic reflectance patterns. An increased understanding of the structural basis of colour of featherless parts of the skin opens up exciting new avenues for interpreting the information content of integumentary signals.
Multi‐Parametric MRI for Evaluating Variations in Renal Structure, Function, and Endogenous Metabolites in an Animal Model With Acute Kidney Injury Induced by Ischemia Reperfusion
Authors: Tao Quan , Zhang Qianqian , An Ziqi , Chen Zelong , Feng Yanqiu
Published: October 2023

Background

Ischemia reperfusion injury (IRI)‐induced acute kidney injury (AKI) may occur after renal ischemic injury. There is a lack of an accurate and comprehensive detection technique for IRI‐AKI.

Purpose

To longitudinally evaluate IRI‐AKI in rats by renal structure, function, and metabolites using multi‐parametric MRI (mpMRI).

Study Type

Prospective.

Animal Model

Forty‐eight rats undergoing IRI‐AKI.

Field Strength/Sequence

7‐T, T1 mapping, and arterial spin labeling (ASL): echo planar imaging (EPI) sequence; blood oxygen level‐dependent (BOLD): gradient recalled echo (GRE) sequence; T2 mapping, quantitative magnetization transfer (qMT), and chemical exchange saturation transfer (CEST): rapid acquisition with relaxation enhancement (RARE) sequence.

Assessment

The mpMRI for IRI‐AKI was conducted at 0 (control), 1, 3, 7, 14, and 28 days, all included eight rats. The longitudinal mpMRI signal of manually outlined cortex, outer stripe of the outer medulla (OSOM), inner stripe of the outer medulla, and medulla plus pelvis were calculated and compared, their diagnosis performance for IRI‐AKI also been evaluated.

Statistical Tests

Pearson correlations analysis for correlation between mpMRI signal and renal injury, unpaired t‐tests for comparing the signal changes, and receiver operating characteristics (ROC) analysis was used to identify most sensitive indicator of mpMRI. A P‐value <0.05 was considered statistically significant.

Results

Compared with control kidneys, the T1 and T2 values of the cortex and medulla in IRI kidneys increased and reached their highest values on day 14, and the kidneys also showed the most severe edema and segments blurred. The RBF in the cortex and OSOM showed a significant decline after day 3. The BOLD signal in the OSOM largest increased on day 28. The cortical PSR and the amine‐CEST both decreased with IRI‐AKI progression, and amine‐CEST achieved the highest AUC for the diagnosis (0.899).

Data Conclusion

Multi‐parametric MRI may show comprehensive variations in IRI‐AKI, and amine‐CEST may exhibit the highest accuracy for diagnosis of IRI‐AKI.

Level of Evidence

1

Technical Efficacy

Stage 2
Impact of core protein naturally selected mutants associated with HBeAg‐negative status in HBV biosynthesis
Authors: Elizalde María Mercedes , Giadans Cecilia Graciela , Campos Rodolfo Héctor , Flichman Diego Martín
Published: October 2023

Hepatitis B e antigen (HBeAg) loss represents a late stage of chronic hepatitis B virus (HBV) infection associated with a drastic decrease in HBV‐DNA, a lower risk of disease progression, and the occurrence of several mutations in the preCore/core region. However, the underlying mechanisms supporting the downregulation of viral replication have yet to be elucidated. In the present study, the analysis of the frequency of subgenotype D1 core protein (HBc) mutations associated with HBeAg status revealed a higher mutation rate in HBeAg‐negative sequences compared to HBeAg‐positive ones. Particularly, 22 amino acids exhibited a higher frequency of mutation in HBeAg‐negative sequences, while the remaining residues showed a high degree of conservation. Subsequently, the assessment of HBc mutants derived from HBeAg‐negative patients in viral structure and replicative capacity revealed that HBc mutations have the ability to modulate the subcellular localization of the protein (either when the protein was expressed alone or in the context of viral replication), capsid assembly, and, depending on specific mutation patterns, alter covalently closed circular DNA (cccDNA) recycling and up‐ or downregulate viral replication. In conclusion, HBc mutations associated with HBeAg‐negative status impact on various stages of the HBV life cycle modulating viral replication during the HBeAg‐negative stage of infection.
Epidemiological characteristics, clinical manifestations, and mental health status of human mpox cases: A multicenter cross‐sectional study in China
Authors: Fu Leiwen , Wang Bingyi , Wu Ke , Yang Liuqing , Hong Zhongsi , Wang Ziyu , Meng Xiaojun , Ma Ping , Qi Xiao , Xu Guangyong , Wang Fuxiang , Zhang Weijie , Huang Chenming , Zhang Dingyan , Lin Yuanlong , Cao Wanxian , Sun Ping , Wu Weibo , Gao Yong , Lv Fan , Qian Jun , Lu Hongzhou , Zou Huachun
Published: October 2023

Human mpox is occurring worldwide, however, evidence from the Asian Pacific Region is limited. In this multicenter cross‐sectional study, information of confirmed mpox cases diagnosed between June 1 and July 31, 2023 in China. Information included demographic and epidemiological characteristics, and clinical manifestations, laboratory results, and mental health status of mpox cases. A total of 115 confirmed mpox cases were enrolled. All cases were men. A total of 102 (90.3%) identified as homosexual. The median age was 31.0 years (interquartile range 27.0–36.5). A total of 65 (56.5%) were HIV‐positive, of whom 92.3% were receiving antiretroviral therapy (ART). A total of 19/39 (40.4%) had a CD4 cell count <500 cells/μL. Systemic features such as fever (73.0%), lymphadenopathies (49.6%), and myalgia (28.7%) were commonly observed. Skin lesions were present in all participants: 49.6% in the genital area and 27.0% in the perianal area. Vesicular rash (78.3%) and papular rash (44.3%) were the most common lesion morphologies. People living with HIV were more likely to have anxiety than those living without HIV. The majority of mpox cases had primary genital lesions and sexual activities before diagnosis, which supports the likelihood of sexual contact transmission. Guidelines on hospitalization and isolation protocols for mpox patients necessitate further confirmation.
How do socioeconomic differentiation and rural governance affect households' land transfer decisions? Evidence from China
Authors: Liu Chongchong , Wang Ying , Yang Bin , Zheng Liang
Published: October 2023

In rural areas, social and economic differentiation profoundly impact rural development patterns, which, in return, impacts rural village social and economic structures and the land transfer decisions of rural households residing within the village. Research on agricultural land transfer has rarely been discussed under the lens of the differentiated socioeconomic structure of rural villages, as well as rural governance capacities. This paper contributes to the existing body of research on the determinants of rural land transfer by examining the role of village socioeconomic differentiation and rural governance based on a quantitative analysis of factors at different scales. The results show that: (1) compared with clan‐based unity structure, kinship‐based split structure and dispersed structure have a positive effect on household land out‐transfer and a negative effect on land in‐transfer. (2) The homogeneity of residents had a negative impact on out‐transfer, while the financial revenue of the village had significant positive influences on households' out‐transfer. (3) The higher the trust in cadres and the evaluation of the local government, the greater the tendency of households to transfer land in, but none of this effect is significant. (4) The socialized agricultural services and the share of non‐farm have significant effects on land transfer. Moreover, households with better geographical conditions, such as those near the county capital and towns, are more likely to transfer their land out. These findings offer significant insights for the management of agricultural scale and the revitalization of rural areas in China, while also offering valuable ideas for the implementation of differentiated agricultural control and rural governance policies in other countries.
Advanced Design of a III‐Nitride Light‐Emitting Diode via Machine Learning
Authors: Jiang Zhuoying , Jiang Ying , Chen Mengyu , Li Jinchai , Li Penggang , Chen Binghuan , Zhao Shanshan , Wang Jie , Jiang Sijie , Cai Miaomin , Li Lin , Li Cheng , Huang Kai , Lu Weifang , Kang Junyong , Zhang Rong
Published: October 2023

Gallium nitride (GaN)‐based light‐emitting diodes (LEDs) have obtained great market success in the past 20 years. However, the traditional research paradigm, i.e., experimental trial‐and‐error method, no longer adapts to the industry development. In this work, an efficient approach is demonstrated to design and optimize GaN‐based LED structures via machine learning (ML). By using the dataset of GaN‐based LED structures over the past decade to train four typical ML models, it is found that the convolutional neural network (CNN) provides the most accurate prediction, with a root mean square error (RMSE) of 1.03% for internal quantum efficiency (IQE) and 11.98 W cm−2 for light output power density (LOPD). Based on the CNN model, 1) the feature importance analysis is adopted to reveal the critical features for LED performance; 2) the predicted trends of IQE and LOPD match well with the physical mechanism, being consistent with the experimental and simulation results; and 3) a high‐throughput screening is demonstrated to predict the properties of over 20 000 structures within seconds to obtain high efficiency LED structures. This ML‐based LED design method enables direct guiding of the LED structure optimization in terms of key parameter selection during manufacturing and greatly accelerates the development cycle of GaN‐based LEDs.
Adhesives Based on Poly(glycidyl methacrylate‐co‐butyl acrylate) with Controlled Structure: Curing Behavior and Adhesion Properties on Metal Substrates
Authors: Cañamero Pedro , Fernández‐García Marta , de la Fuente José Luis
Published: October 2023

The adhesion properties of poly(glycidyl methacrylate (GMA)‐co‐butyl acrylate (BA)) statistical copolymers, synthesized by atom transfer radical polymerization (ATRP), are investigated employing three different curing agents or hardeners, such as diethanolamine (DEA), dicyandiamide (DICY), and 2‐cyanoacetamide (2‐CA) on copper, iron, brass, aluminum, and titanium metal surfaces. This work describes the treatment of the different surfaces, establishes the optimal curing conditions from differential scanning calorimetry (DSC) analysis of these novel adhesive systems, and evaluates the results of the single‐lap shear test for metal joints. Thus, by dynamic DSC measurements of the mixtures, a low curing temperature of 90 °C is defined when DEA is used as a curative; while systems based on DICY and 2‐CA require temperatures of 150 °C and 160 °C, respectively. In addition, the curing process of this controlled acrylic copolymer with DICY exhibits a singular behavior, possibly due to the curing reaction mechanism, where multiple epoxy‐amine ring‐opening polyaddition reactions take place between DICY's active hydrogens and epoxy groups of poly(GMA‐co‐BA). This latter curing system shows the highest adhesion features with lap‐shear strength at room temperature of 15.5 MPa, using copper as metallic substrate; however, the best results are obtained using 2‐CA as curing agent with aluminum and iron.
A potent and broad‐spectrum neutralizing nanobody for SARS‐CoV‐2 viruses, including all major Omicron strains
Authors: Yao Hebang , Wang Hongyang , Zhang Zhaoyong , Lu Yuchi , Zhang Zhiying , Zhang Yu , Xiong Xinyi , Wang Yanqun , Wang Zhizhi , Yang Haitao , Zhao Jincun , Xu Wenqing
Published: October 2023

SARS‐CoV‐2 viruses are highly transmissible and immune evasive. It is critical to develop broad‐spectrum prophylactic and therapeutic antibodies for potential future pandemics. Here, we used the phage display method to discover nanobodies (Nbs) for neutralizing SARS‐CoV‐2 viruses especially Omicron strains. The leading nanobody (Nb), namely, Nb4, with excellent physicochemical properties, can neutralize Delta and Omicron subtypes, including BA.1, BA.1.1 (BA.1 + R346K), BA.2, BA.5, BQ.1, and XBB.1. The crystal structure of Nb4 in complex with the receptor‐binding domain (RBD) of BA.1 Spike protein reveals that Nb4 interacts with an epitope on the RBD overlapping with the receptor‐binding motif, and thus competes with angiotensin‐converting enzyme 2 (ACE2) binding. Nb4 is expected to be effective for neutralizing most recent Omicron variants, since the epitopes are evolutionarily conserved among them. Indeed, trivalent Nb4 interacts with the XBB1.5 Spike protein with low nM affinity and competes for ACE2 binding. Prophylactic and therapeutic experiments in mice indicated that Nb4 could reduce the Omicron virus loads in the lung. In particular, in prophylactic experiments, intranasal administration of multivalent Nb4 completely protected mice from Omicron infection. Taken together, these results demonstrated that Nb4 could serve as a potent and broad‐spectrum prophylactic and therapeutic Nb for COVID‐19.
Long‐term storage does not affect the DNA methylation profiles of vitrified‐warmed human embryos
Authors: Zhu Ling , Sun Liwei , Liu Weiwei , Han Wei , Huang Guoning , Li Jingyu
Published: October 2023

With the widespread application of embryo cryopreservation in assisted reproductive techniques, it is necessary to assess the safety of long‐term cryopreservation of human embryos and it is unclear whether storage time has an impact on the DNA methylation profiles of human embryos. Nine women who received IVF treatment were recruited for this study. The retrieved eight‐cell human embryos were classified into three groups including fresh embryos, cryopreserved embryos stored for 3 years, and cryopreserved embryos stored for 8 years. Single‐cell whole‐genome bisulfite sequencing (scWGBS) was conducted. The genome‐wide methylation pattern of the fresh and two cryopreserved groups were similar. In addition, the methylation level in different genomic regions showed comparable patterns and no significant differences were observed in the methylation level of imprinted genes among the three groups. A total of 587 differentially methylated regions (DMRs) in the 3‐year group and 540 DMRs in the 8‐year group were identified comparing to fresh group. However, they were not enriched in promoters and had a similar genome‐wide distributions, suggesting that these DMRs may not contribute to the changes in corresponding gene expressions. Our study illustrated that long‐term cryopreservation will not affect the DNA methylation profiles of human eight‐cell embryos at single‐cell level.