Differences in clinical outcomes between men with mosaic Klinefelter syndrome and those with non-mosaic Klinefelter syndrome.

Purpose: This study compared the clinical outcomes of men with Klinfelter syndrome based on karyotype. Methods: The authors analyzed the outcomes of microdissection testicular sperm extraction (micro-TESE) performed on 57 patients with Klinfelter syndrome (KS) at our clinic. Results: The average ages of the non-mosaic and mosaic KS groups were 32.2 ± 4.8 and 45.9 ± 13.1 years, respectively. The sperm retrieval rates of the non-mosaic and mosaic KS groups were 46.5% (20/43) and 50.0% (7/14), respectively. The fertilization rates after intracytoplasmic sperm injection did not significantly differ between the non-mosaic and mosaic KS groups. The mosaic KS group had higher cleavage and blastocyst development rates than the non-mosaic KS group (72.2% vs. 96.2% and 30.5% vs. 44.7%, respectively). The group using motile sperm had better outcomes than the group using immotile sperm. The embryo transfer outcomes of the non-mosaic and mosaic KS groups did not significantly differ (clinical pregnancy rate: 28.0% vs. 20.7%, miscarriage rate: 14.3% vs. 33.3%, production rate per transfer: 22.0% vs. 13.8%, and production rate per case: 58.8% vs. 57.1%). Conclusions: Compared with the non-mosaic KS group, the mosaic KS group had significantly better intracytoplasmic sperm injection outcomes because of the higher utilization rate of motile sperm.

Clinical outcome of intrauterine infusion of platelet-rich plasma in patients with recurrent implantation failure.

PURPOSE: This study aimed to evaluate the effectiveness of intrauterine infusion of platelet-rich plasma (PRP) before embryo transfer (ET) in recurrent implantation failure (RIF) cases. METHODS: The authors retrospectively analyzed 54 ET cycles involving frozen and thawed high-quality blastocysts after intrauterine PRP infusion between September 2019 and November 2020. All patients had a history of at least two times of implantation failure on ET. A total of 54 patients were categorized into two groups: thin endometrium (39 patients) and unexplained implantation failure (15 patients). In the thin-endometrium group, the endometrial thickness (EMT) was <8.0 mm at cycle days 12-14 in the prior ET cycle. RESULTS: Among the 54 ET cycles after PRP infusion, 31 (57.4%) were positive for human chorionic gonadotropin (hCG) and 27 (50%) achieved clinical pregnancy, which was significantly better than that in prior ET cycles without PRP infusion (27.2% and 9.6%, respectively). The EMT was not increased at ET date on the PRP cycle compared with that in the prior ET cycle in both patient groups. Moreover, EMT was not different between the hCG-positive and hCG-negative groups. CONCLUSION: Although intrauterine PRP infusion had no superior effect on increasing the EMT than conventional therapeutic agents, it resulted in high pregnancy rates in patients experiencing RIF with or without thin endometrium.

Involvement of HGF/MET signaling in appendicular muscle development in cartilaginous fish.

In amniotes, limb muscle precursors de-epithelialize from the ventral dermomyotome and individually migrate into limb buds. In catsharks, Scyliorhinus, fin muscle precursors are also derived from the ventral dermomyotome, but shortly after de-epithelialization, they reaggregate within the pectoral fin bud and differentiate into fin muscles. Delamination of muscle precursors has been suggested to be controlled by hepatocyte growth factor (HGF) and its tyrosine kinase receptor (MET) in amniotes. Here, we explore the possibility that HGF/MET signaling regulates the delamination of appendicular muscle precursors in embryos of the catshark Scyliorhinus canicula. Our analysis reveals that Hgf is expressed in pectoral fin buds, whereas c-Met expression in fin muscle precursors is rapidly downregulated. We propose that alteration of the duration of c-Met expression in appendicular muscle precursors might underlie the evolution of individually migrating muscle precursors, which leads to the emergence of complex appendicular muscular systems in amniotes.

Modification of pectoral fins occurs during the larva-to-juvenile transition in the mudskipper (Periophthalmus modestus).

BACKGROUND: Mudskippers are amphibious fishes that use their pectoral fins to move on land. Their pectoral fins are specifically modified for terrestrial locomotion. Studies of the anatomy and kinematics of adult mudskippers suggest that modifications of the pectoral fins, such as their protrusion and elongation of the proximal radials, may provide greater control and flexibility in pectoral fin-based locomotion. However, it is unknown when and how the unique features of these pectoral fins form during the development of mudskippers, which begin life as a planktonic organism. RESULTS: Here we examined the developmental process of the pectoral fins of the mudskipper Periophthalmus modestus to address these questions. We also observed other developmental characteristics to provide clarified descriptions, including indicative morphological changes that occur during metamorphosis. CONCLUSION: Our results show that the localized cell division of the proximal part of the endoskeletal disc-the primordium of the proximal radials-and subsequent cell division along the proximal-distal axis, which is restricted to the distal part of the disc during the larva-to-juvenile transition (metamorphosis), lead to the elongation of the proximal radials.

Is human chorionic gonadotropin supplementation beneficial for frozen and thawed embryo transfer in estrogen/progesterone replacement cycles?: A randomized clinical trial.

Aim: Human chorionic gonadotropin (hCG) is used frequently for luteal support in fresh in vitro fertilization cycles as it induces progesterone secretion from the ovaries after oocyte retrieval and modulates the endometrium for implantation in fresh cycles. In contrast, hCG is not usually used for the transfer of cryopreserved-thawed embryos in estrogen/progesterone replacement cycles because ovulation is suppressed. However, several studies have shown that luteinizing hormone and hCG receptors are present in the human endometrium and that hCG can directly induce the decidualization of endometrial stromal cells in vitro. Thus, this study evaluated whether hCG supplementation can be beneficial for cryopreserved-thawed embryo transfer in estrogen/progesterone replacement cycles. Methods: One-hundred-and seventy-three cryopreserved-thawed embryo transfer cycles with estrogen/progesterone replacement were divided randomly into two groups. Transdermal oestradiol was used in combination with vaginal progesterone suppositories for HR. The embryo transfer was performed on day 17 and/or day 20 of the HR therapy cycle in both groups. In Group A, 3000 IU of hCG was administered on days 17, 20, and 23. In Group B, hCG was not used. Results: There was no significant difference in the average age of the patients, the average number of previous assisted reproductive technology cycles, or the average number of embryo transfers between the two groups. The rates of pregnancy and implantation per embryo were 37.2% and 25.3%, respectively, in Group A and 35.6% and 21.7%, respectively, in Group B. The pregnancy and implantation rates were similar in both groups. Conclusion: Supplementation with hCG is not beneficial for cryopreserved-thawed embryo transfer in estrogen/progesterone replacement cycles.

Migratory appendicular muscles precursor cells in the common ancestor to all vertebrates.

In amniote embryos, skeletal muscles in the trunk are derived from epithelial dermomyotomes, the ventral margin of which extends ventrally to form body wall muscles. At limb levels, ventral dermomyotomes also generate limb-muscle precursors, an Lbx1-positive cell population that originates from the dermomyotome and migrates distally into the limb bud. In elasmobranchs, however, muscles in the paired fins were believed to be formed by direct somitic extension, a developmental pattern used by the amniote body wall muscles. Here we re-examined the development of pectoral fin muscles in catsharks, Scyliorhinus, and found that chondrichthyan fin muscles are indeed formed from Lbx-positive muscle precursors. Furthermore, these precursors originate from the ventral edge of the dermomyotome, the rest of which extends towards the ventral midline to form body wall muscles. Therefore, the Lbx1-positive, de-epithelialized appendicular muscle precursors appear to have been established in the body plan before the divergence of Chondrichthyes and Osteichthyes.

Publisher Correction: Migratory appendicular muscles precursor cells in the common ancestor to all vertebrates.

In Fig. 2 of this Article originally published, some erroneous lines appeared on the left side of the images in panels c, e and g. The figure should have appeared as shown below. These errors have now been corrected in all versions of the Article.

Frequency of Sperm DNA Fragmentation According to Selection Method: Comparison and Relevance of a Microfluidic Device and a Swim-up Procedure.

INTRODUCTION: Multiple rounds of centrifugation or washing spermatozoa can cause sperm DNA fragmentation (SDF); however, a microfluidic approach to select spermatozoa does not require centrifugation. Reports have suggested that sperm sorting using a microfluidic device is an effective method to select good quality spermatozoa, however, it is not known whether it reduces sperm DNA damage. We investigated whether the frequency of SDF was affected by selection method during sperm processing. MATERIALS AND METHODS: Semen samples from ten men with normal, oligozoospermia and asthenozoospermia were split into two groups and sorted using a microfluidic device or by a swim-up method. Subsequently, semen parameters and SDF were measured and analyzed using paired or non-paired Student's t-tests. RESULTS: For samples sorted by the microfluidic device (Sperm Sorter Qualis(®); Menicon, Kasugai, Japan) or the swim-up method, both showed a decrease in SDF. However, the decrease was more significant when the microfluidic device was used. CONCLUSION: Sorting using the microfluidic device resulted in less SDF than did the swim-up method.

Expression of heparanase: a possible role in invasiveness and aggressive clinical behavior of ameloblastomas.

Heparanase is an endoglycosidase that cleaves heparan sulfate (HS), thus participating in degradation and remodeling of the extracellular matrix (ECM). Heparanase up-regulation is correlated with lymph node and distant metastasis, microvessel density and reduced postoperative survival of cancer patients. In the present study, we carried out an immunohistochemical investigation of heparanase to extend and confirm present knowledge regarding its expression in ameloblastomas (AMs), which are characterized by locally aggressive behavior. Paraffin-embedded tissue specimens of 53 AMs were stained using an antibody against heparanase. Immunohistochemical reactivity for heparanase was detected in 94.3% of the AMs examined. Heparanase was expressed strongly in peripheral columnar cells and slightly in central stellate reticulum-like cells. Small tumor nests and budding epithelial branches showed a stronger staining pattern. Stromal cells were negative for heparanase, or showed diffuse expression. However, an enhanced positive immunoreaction was present specifically near osseous tissue and adjacent to the invasive front of tumor nests. Areas of cystic degeneration showed intense heparanase immunoreactivity. The enzyme may facilitate the function of HS-binding growth factors that elicit an angiogenic response and favor osteoclastogenesis in AM.

Significance of podoplanin expression in keratocystic odontogenic tumor.

BACKGROUND: The most important clinical features of the keratocystic odontogenic tumor (KCOT) are its potential for locally destructive behavior, a tendency to recur, and its origin in the odontogenic epithelium. The clinical features of KCOT are similar to those of ameloblastoma (AM). Histologically, KCOT is distinguished from jaw cyst with keratinization (orthokeratinized odontogenic cyst; OOC). However, current scientifically based clinical parameters cannot predict any potential for neoplastic behavior, or aggressive and localized invasiveness, in patients with KCOT. We have shown that podoplanin, a lymphatic endothelial marker, is highly expressed in AM. The purpose of this study was to determine the usefulness of podoplanin for reclassification of the odontogenic keratocyst (OKC) from cyst to tumor status. METHODS: Paraffin-embedded tissue specimens of 57 OKCs (46 KCOTs and 11 OOCs) and 15 dentigerous cysts (DCs) were immunohistochemically examined using antibody against podoplanin. RESULTS: Immunohistochemical reactivity for podoplanin was detected in the cell membrane and cytoplasm of most of the basal and suprabasal layer, areas of budding basal cell proliferation, epithelial nests and peripheral cells of daughter cysts in the stromal connective tissue in KCOTs. In the case of OOC and DC, only cases associated with inflammation were positive for podoplanin. CONCLUSION: Podoplanin is strongly expressed in KCOTs in comparison with OOCs. The pattern of staining for podoplanin in KCOT could be related to its neoplastic nature, and suggests a role of the protein in tumor invasiveness.

Enhanced expression of podoplanin in ameloblastomas.

OBJECTIVE: Podoplanin, a mucin-type transmembrane glycoprotein, is specifically expressed by lymphatic but not blood vascular endothelial cells, and is also widely expressed in various specialized cell types throughout the body. Recent studies have demonstrated that it mediates a pathway leading to collective cell migration and invasion in vivo and in vitro. In the present study, we carried out an immunohistochemical investigation of podoplanin to clarify whether it is expressed in human ameloblastomas (AMs), which are characterized by locally aggressive behavior with a high rate of recurrence. In addition, we examined the localization of the epithelial marker E-cadherin and the mesenchymal marker vimentin to clarify whether AMs show epithelial-mesenchymal transition (EMT). METHODS: Paraffin-embedded tissue specimens of 38 AMs were examined immunohistochemically using antibodies against podoplanin, E-cadherin, and vimentin. RESULTS: Immunohistochemical reactivity for podoplanin was detected in the cell membrane and cytoplasm of most odontogenic tumor epithelial cells in AMs. Podoplanin was expressed strongly in peripheral columnar cells and slightly in central stellate reticulum-like cells. E-cadherin was expressed in central stellate reticulum-like cells and showed decreased expression in peripheral columnar cells. Immunoreactivity for E-cadherin was weak or negative in keratinizing cells of acanthomatous AMs, suggesting terminal differentiation of the tumor cells. Immunohistochemical reactivity for vimentin was found in stromal cells, but partial or no reaction was observed in neoplastic cells. CONCLUSION: Expression of podoplanin in AMs is considered to be associated with neoplastic odontogenic tissues; this molecule might play a role in the collective cell migration of tumor nests in AMs. The pattern of expression of E-cadherin and vimentin suggests that invasion in AMs occurs in the absence of EMT. The migration and invasion mediated by podoplanin in AMs could be related to cytoskeletal reorganization.

The significance of podoplanin expression in human inflamed gingiva.

Podoplanin, a transmembrane glycoprotein, has been considered to be expressed specifically by lymphatic endothelial cells. However, recent studies have shown that the protein is expressed in a variety of normal as well as neoplastic tissues, and that its expression might be related to cell migration and invasion. In this study, we examined podoplanin expression in inflamed gingival tissues using an immunohistochemical method. Positive immunoreactivity for podoplanin was found in the cell membrane and cytoplasm of basal cells of oral gingival epithelium when severe inflammatory cell infiltration was present in the connective tissue just under the epithelium. When inflammatory changes were weak or absent, little or no reactivity for podoplanin in the basal cells was observed. Positive reactivity for podoplanin was also detected in basal cell extensions. Surprisingly, strong immunoreactivity for podoplanin was observed in all layers of oral sulcular and junctional epithelia associated with severe inflammatory reaction in the connective tissue. These findings suggest that increased expression of podoplanin in gingival epithelium is related to the progression of chronic periodontitis.

Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in uterine endometrial carcinoma and a correlation between expression of matrix metalloproteinase-7 and prognosis.

Matrix metalloproteinases (MMPs) are associated with invasion and metastasis of several human malignant tumors, in particular MMP-7, which is mainly produced by the cancer cell itself. We examined the expression of MMP-2, 7 and 9, and tissue inhibitors of metalloproteinase (TIMP)-1 and 2 in uterine endometrial carcinoma, and compared the expression with clinicopathological characteristics in uterine endometrial carcinoma (UEC). A group of 256 patients with UEC received surgery at the Osaka City University Medical School Hospital, and 196 tumor samples were immunohistochemically stained to examine the expression of MMP-2, 7 and 9, and TIMP-1 and 2. Additionally, the invasion ability of cell stain established from UEC was examined using an in vitro invasion assay. The expression of MMP-2, 7 and 9, and TIMP-1 and 2 was observed in the cytoplasm, and the expression of MMP-2 and 7, and TIMP-1 and 2 was observed in stromal cells around the tumor cells. The expression of MMP-7 was significantly stronger in higher-grade than lower-grade tumors (P<0.05). The invasion assay showed that the invasion of cells derived from UECs was significantly inhibited by TIMP-1 and 2. The disease-free interval was significantly shorter when MMP-7 expression was intense. This increased expression of MMP-7 in high grade UECs may be associated with tumor invasion and metastasis, and MMP-7 could serve as a prognostic maker in UEC.

Expression of apoptosis-related proteins in advanced uterine cervical cancer after balloon-occluded arterial infusion chemotherapy as an indicator of the efficiency of this therapy.

We previously reported satisfactory therapeutic results of cisplatin-based cyclic balloon-occluded arterial infusion chemotherapy (BOAI) as neoadjuvant chemotherapy, which enabled treatment by hysterectomy in patients with advanced cervical cancer. We also reported expression of apoptosis among these patients and determined that the bax gene is related to this apoptosis. In the present study, we investigated the relationship between the effectiveness of BOAI therapy and expression of apoptosis regulatory genes and proteins in these cases. The subjects were 27 women with advanced cervical cancer classified as FIGO (International Federation of Gynecology and Obstetrics) stage III or higher who were admitted to the Department of Gynecology, Osaka City University Medical School Hospital between 2000 and 2003. All patients were treated by BOAI, and expression of cancer cell apoptosis was examined by the TUNEL method, expression of bax, bcl-2 and bcl-xL proteins were examined by immunohistochemistry, and expression of bax, bcl-2 and bcl-xL mRNA was examined by quantitative RT-PCR before and 3 days after BOAI. The effectiveness of BOAI therapy was thus determined. The 20 patients in whom BOAI was effective showed significantly higher expression of the bax protein and gene after BOAI, and cancer cell apoptosis was accelerated. On the other hand, the 7 patients in whom BOAI was ineffective showed significantly higher expression of the bcl-xL protein and gene after BOAI. These results suggest that bax/bcl-xL expression can be used as an indicator of the effectiveness of BOAI therapy.

Association of HPV infection with prognosis after neoadjuvant chemotherapy in advanced uterine cervical cancer.

Whether the human papillomavirus (HPV) status of the tumor affects the sensitivity to neoadjuvant chemotherapy, and the prognosis in advanced uterine cervical cancer (FIGO stage III or higher) remains unknown. We examined the HPV status of 43 patients who had received CDDP therapy by balloon-occluded arterial infusion (BOAI), as neoadjuvant chemotherapy for advanced uterine cervical cancer (squamous cell carcinoma) stage III or higher. DNA was extracted from formalin-fixed, paraffin-embedded tumor samples obtained by punch biopsy before the neoadjuvant chemotherapy. The detection of HPV and its typing were analyzed by a polymerase chain reaction (PCR)-based assay using consensus primers for the L1 consensus regions. HPV DNA was detected in all 43 patients (100%): 29 cases with HPV 16 (67.4%), 5 cases with HPV 33 (11.6%), 4 cases with HPV 31 (9.3%), 3 cases with HPV 35 (7.0%), 1 case with HPV 18 (2.3%) and 1 case with HPV 58 (2.3%). The HPV types were divided into 3 groups, HPV 16, HPV 33 and other HPV types (HPV 18, 31, 35, 58), and comparisons and examinations were performed among the 3 groups. Although the rates of tumor reduction and operation accomplishment after 3 courses of BOAI showed no significant differences among the 3 groups, there were significant differences in the survival rates. The survival rate of advanced uterine cervical cancer patients with HPV 33 infection was the highest, followed by that of patients with HPV 16 infection. The survival rates of patients with the other types of HPV infection were the worst among the 3 groups and significantly lower than those of patients with HPV 16 or HPV 33 infection. The differences in the curative effect after BOAI may depend on the different characters of the HPV types.

VEGF mRNA in adipocytes increase with rebound weight-gain after diet-restriction.

The mechanism of rebound body weight-gain after a restricted-diet state is unclear. We investigated the expression of angiogenic factors in human adipocytes with a changing nutritional state in culture medium, and attempted to ascertain the mechanisms involved in rebound weight-gain. Adipocytes were divided into three groups; the first group (control group) was cultured in medium with 10% fetal calf serum (FCS), the second (DR3% group) was cultured in medium with 3% FCS, and the third (DR6% group) was cultured in medium with 6% FCS. After being cultured for 48 h, each was next cultured with 12% FCS for a further 48 h. When made to change from a low nutrition state to a higher one, adipocytes changed from hypotrophic to hypertrophic. Simultaneously, vascular endothelial growth factor (VEGF) in the culture medium increased significantly. When investigated immunohistochemically, the expression of VEGF was similarly shown in the cytoplasm of adipocytes. The same tendency with the same quantity of mRNA was shown by RT-PCR. These results show that VEGF produced and secreted from adipocytes increases, when the cultivation state of adipocytes is changed from a low nutritional state to a higher one. VEGF produced and secreted from adipocytes may be related to rebound weight-gain.

Relaxant mechanisms of parathyroid hormone in rat mesenteric artery.

The effects of parathyroid hormone (PTH) on tension and intracellular Ca level ([Ca ] ) were examined in ring preparations of rat mesenteric artery using isometric tension recording and the fura-2 method, respectively. The PTH (30 n ) elicited relaxation in arterial rings precontracted by phenylephrine regardless of the presence or absence of endothelium. In the endothelium-denuded arterial rings precontracted by 3 micro M of phenylephrine or 60 m of potassium chloride (KCl), PTH-related protein and PTH produced concentration-dependent relaxation to the same extent, but inhibited contraction induced by phenylephrine more effectively than that induced by KCl. Phenylephrine-induced tonic contraction was changed to a phasic one with decreased peak tension in the presence of PTH. Similar changes were observed with extracellular Ca removal or methoxyverapamil plus SK&F96365, respective of voltage-gated and receptor-operated Ca channel inhibitors. Phenylephrine evoked a concentration-dependent contraction concomitant with an increase in [Ca ]. PTH reduced both responses to the same extent. In a Ca -free solution, PTH inhibited a phasic contraction and a transient increase in [Ca ] in response to phenylephrine but not caffeine. Reverse transcriptase-polymerase chain reaction showed that PTH and PTH receptors were expressed in the rat mesenteric artery. In this tissue, PTH increased cyclic adenosine monophosphate (cAMP) levels. These results suggest that the inhibitory effect of PTH on alpha -adrenoceptor-mediated contraction results from the inhibition of Ca influx through receptor-operated and voltage-gated Ca channels, and Ca release from Ca stores, probably via increased cAMP in the rat mesenteric artery.