Radioligand Assays for Oestradiol and Progesterone Conjugated to Protein Reveal Evidence for a Common Membrane Binding Site in the Medial Preoptic Area-Anterior Hypothalamus and Differential Modulation by Cholera Toxin and GTPγS

In this study membrane oestradiol (E) binding sites in the medial preoptic area-anterior hypothalamus (MPOA-AH) of ovariectomized (OVX) rats were characterized using standard radioligand binding techniques employing E conjugated to bovine serum albumin (BSA) at position 6 and radiolabeled with 125I (E-6-[125I-BSA]). In previous studies binding of a radioactive conjugate of progesterone (P) and BSA (P-3-[125I-BSA]) was examined using the same membrane preparation. E-6-[125I-BSA] binding was linear across a tissue concentration range of 0.005-0.02 mg protein/0.1 ml of membrane suspension. An association T1/2 of 9.5 min and a dissociation T1/2 of 52.1 min for E-6-[125I-BSA] were derived from kinetic experiments. Competition binding experiments revealed high (Ki=0.63+/-(0.50 nM) and low (Ki=161.5(96.5 nM) affinity binding sites for E-6-[125I-BSA], demonstrating different binding parameters than shown in our previous work for P-3-[125I-BSA] binding. Further studies on MPOA-AH membranes treated with cholera toxin (CTX) and GTPgammaS suggested that E-6-BSA binding sites are associated with G proteins. E-6-[125I-BSA] binding demonstrated both high-and low-affinity sites. GTPgammaS added to the assay reduced both E-6-[125I-BSA] and P-3-[125I-BSA] binding suggesting that G proteins are associated with both binding sites. Extensive analysis of both E-6-[125I-BSA] and P-3-[125I-BSA] binding sites demonstrated a reciprocal relationship such that high-affinity E-6-[125I-BSA] binding sites exhibit low affinity for P-3-[125I-BSA] and low-affinity E-6-[125I-BSA] binding sites exhibit high affinity for P-3-[125I-BSA]. Preincubating membranes with CTX or GTPgammaS reduced high-affinity E-6-[125I-BSA] binding and enhanced high-affinity P-3-[125I-BSA] binding. These results suggest that, in the MPOA-AH, membrane steroid binding sites exist in two interconvertible conformations that preferentially bind either E-6-BSA or P-3-BSA, depending on their association with a G protein. Additional studies with free steroids revealed that: (1) oestrogens (17beta-oestradiol, diethylstilbestrol) as well as synthetic oestrogen antagonists tamoxifen and ICI 182 780 displaced P-3-[125I-BSA] further suggesting a relationship between membrane binding sites for E and P-3-[125I-BSA] binding sites; and (2) treatment of OVX rats with E decreased displacement by P-3-BSA and increased displacement by ICI 182,780 and tamoxifen suggesting these antagonists affect membrane P-3-[125I-BSA] binding sites after in-vivo E treatment. The membrane binding sites for E and P demonstrate interrelationships not demonstrated by their nuclear receptors.     

Treatment options for advanced urothelial cancer after progression on chemotherapy and immune checkpoint inhibitors: a literature review

ObjectiveTo describe the current treatment landscape in advanced urothelial cancer (aUC)/metastatic urothelial cancer and in particular to review the relevant literature highlighting recent advances in the treatment of patients with aUC after progression on chemotherapy and immune checkpoint inhibitor (ICI).BackgroundaUC is a very aggressive disease with poor outcomes. Over the past several years, its treatment landscape has seen significant advances with the approval of ICI and targeted agents, which have led to improved outcomes. The current standard of care for most patients with aUC involves platinum-based chemotherapy followed by ICI after progression or as switch maintenance therapy (if no progression after chemotherapy). Treatment of patients following progression on ICI is more challenging, but novel therapies have been approved, such as erdafitinib for tumors with fibroblast growth factor receptor 2 (FGFR2) or FGFR3 activating mutation or fusion (can also be used following progression on platinum-based chemotherapy), enfortumab vedotin (EV) and sacituzumab govitecan (SG) in an unselected patient population. Many other trials in this space are currently ongoing and other promising agents may also potentially become available in the future.MethodsNarrative overview of the recent literature relevant to the treatment of advanced/metastatic urothelial cancer following progression on chemotherapy and ICI was undertaken. Relevant literature was obtained from review of computerized databases including pubmed.gov and proceedings of major conferences including American Society of clinical Oncology (ASCO) Meetings, GU ASCO Symposia and European Society of Medical Oncology (ESMO) Meetings.ConclusionsIn this narrative review, we highlight the current dynamic treatment landscape in aUC, emphasizing the recent important developments and a few examples of ongoing clinical trials. In particular, we focus on therapy options available following progression on platinum-based chemotherapy and ICI, a treatment space where until recently there had been no FDA-approved treatment options. The recent pivotal trials of antibody drug conjugates (ADCs) that led to FDA approvals in this space are highlighted, as are other agents currently in development. We conclude by discussing future directions and ongoing challenges in this evolving disease space.     

Spontaneous recovery of immune checkpoint inhibitor‐induced thyroiditis with high free T4 level: A case report

We describe a case of severe hyperthyroidism with high free thyroxine and C‐reactive protein levels, wherein thyroid function rapidly improved without treatment. In a similar case, conservative management with imaging follow‐up can be considered.     

Can the International Index of Erectile Function (IIEF-5) be used as a diagnostic tool to the severity of vasculogenic erectile dysfunction?

ObjectiveThe objective of this observational study was to compare the International Index of Erectile Function (IIEF-5) with penile duplex in the diagnosis of vasculogenic erectile dysfunction severity. 150 subjects complaining of erectile dysfunction for >6 months have been divided into two groups according to their response to intracavernous injection (ICI) test; 50 patients who showed good response ? (E4) and one hundred patients who showed poor response ? (E4) up to maximum dose of 1 cc Quadmix with abnormal penile duplex. The results of the duplex are correlated to the IIEF-5 score of the patients.FindingsThere is statistically significant difference between mean value of IIEF-5 in both good and poor responders (P-value = 0.0000), significant difference between mean value of (age, duration, PSV, EDV, diameter of artery after injection, percent of increase in arterial diameters and RI) between the good and poor responders. There was no evidence of statistically significant difference between mean value of IIEF-5 in both arteriogenic and venogenic subgroups of poor responders group. There was neither evidence of statistically significant correlations between IIEF-5 and penile duplex results in both good and poor responders groups, nor between IIEF-5 and penile duplex results in arteriogenic and venogenic and combined subgroups.ConclusionIIEF-5 might not be a suitable diagnostic tool of the severity of vascular affection in ED.     

宫颈管内与宫腔内供精人工授精妊娠结局的比较

目的比较分析宫颈管内和宫腔内供精人工授精(AID)妊娠结局的影响。方法对548个AID治疗周期进行回顾性分析,其中宫颈管内人工授精(ICI)200例360个周期,宫腔内人工授精(IUI)125例188个周期,对2种授精部位的AID妊娠结局进行比较。结果 ICI组的周期妊娠率为14.74%(51/346),其中自然周期妊娠率为18.54%(33/178),促排卵周期妊娠率为10.71%(18/168),流产率为11.76%(6/51);IUI组的周期妊娠率为28.49%(51/179),其中自然周期的妊娠率为32.04%(33/103),促排卵周期妊娠率为23.68%(18/76),流产率为5.88%(3/51);组间差异均有统计学意义(P0.05)。结论授精部位可能是影响AID成功妊娠的关键因素之一,采用IUI法可能会提高AID的妊娠率。     

阳痿的超声诊断分析

目的应用多普勒超声（DUS）技术观察阳痿（ED）患者阴茎血流情况,并进行诊断分型,为临床治疗提出指导性建议。方法对32例国际勃起功能评分（IIEF-5）为12．1-15．7（13．6±5．1）的ED患者,测定双侧海绵体动脉及阴茎背静脉内径、阴茎海绵体动脉收缩期最大血流速度（PSV）、舒张期最低血流速度（EDV）、阻力指数（RI）。阴茎海绵体内一次性注射（ICI）罂粟碱30mg＋酚妥拉明1mg 10min后,再次测定双侧海绵体动脉及阴茎背静脉内径、阴茎海绵体动脉PSV、EDV、RI、阴茎背静脉流速。应用SPSS12．0统计软件进行统计学分析。结果非血管性ED8例阴茎海绵体动脉PSV均≥35cm／s,RI≥1．0;阴茎背静脉常为间断低速静脉血流;15例为动脉性ED,阴茎海绵体动脉PSV均〈25cm/s,阴茎背静脉见间断低速静脉血流;6例为静脉性ED,阴茎海绵体动脉EDV均〉0cm／s,阴茎背静脉见持续静脉血流;3例为动静脉混合性ED,阴茎海绵体动脉PSV均〈25cm／s,阴茎背静脉见持续静脉血流。结论DUS检测阴茎海绵体动脉和阴茎背静脉血流并结合国际勃起功能评分（IIEF-5）是诊断阳痿疾病及其分型的有效方法。     

Metachronous Esophageal Ulcers after Immune-mediated Colitis Due to Immune Checkpoint Inhibitor Therapy: A Case Report and Literature Review

Although cases of gastrointestinal toxicity of pembrolizumab have been reported, cases of acute immune-mediated colitis accompanied with metachronous esophageal disorders (esophagitis and ulcer) are rare. We herein report a case of acute colitis and metachronous esophageal ulcers due to an immune-related adverse event following concomitant pembrolizumab chemotherapy for lung adenocarcinoma. To our knowledge, there have so far been no reports of cases in which both acute immune-mediated colitis and metachronous esophageal ulcers developed. We therefore report the details of this case along with a review of the pertinent literature.     

Physiology: Effects of the pure anti-oestrogen ICI 182780 on oestrogen receptors, progesterone receptors and Ki67 antigen in human endometrium in vivo

The new steroidal pure anti-oestrogen ICI 182780 was studiedfor the first time in pre-menopausal women. A total of 30 patientsrequiring hysterectomy for benign gynaecological disease wererandomized to ICI 182780, 12 mg/day i.m. (n = 19) or no treatment(n = 11) for 7 days prior to surgery. Immunohistochemical measurementswere made in the snap-frozen, resected endometrium for oestrogenreceptors (ER), progesterone receptors (PgR) and Ki67, a nuclearantigen whose expression is closely related to proliferation.Five control patients ovulated prior to surgery and, as expected,the secretory endometria had lower Ki67 antigen concentrationsthan endometria from the proliferative phase. The endometriafrom patients treated with ICI 182780 had reduced Ki67 comparedwith controls. This demonstration of reduced proliferative activityindicates that the pharmacological effectiveness of the treatmentwas maintained despite increased plasma oestradiol concentrations.In contrast to results from rodents, ICI 182780 did not markedlyreduce ER expression, although there was significantly lowerER in the myometrial cells of the treated group. The lack ofeffect on PgR shows a dissociation between the drug's effecton this oestrogendependent protein and its effects on proliferation.