World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonization Project: V. Physical examination standards in endometriosis research.

OBJECTIVE: The World Endometriosis Research Foundation established the Endometriosis Phenome and Biobanking Harmonisation Project (EPHect) to create standardized documentation tools (with common data elements) to facilitate the comparison and combination of data across different research sites and studies. In 2014, 4 data research standards were published: clinician-reported surgical data, patient-reported clinical data, and fluid and tissue biospecimen collection. Our current objective is to create an EPHect standard for the clinician-reported physical examination (EPHect-PE) for research studies. DESIGN: An international consortium involving 26 clinical and academic experts and patient partners from 11 countries representing 25 institutions and organizations. Two virtual workshops, followed by the development of the physical examination standards underwent multiple rounds of iterations and revisions. SUBJECTS: N/A MAIN OUTCOME MEASURE(S): N/A RESULT(S): The EPHect-PE tool provides standardized assessment of physical examination characteristics and pain phenotyping. Data elements involve examination of back and pelvic girdle; abdomen including allodynia and trigger points; vulva including provoked vestibulodynia; pelvic floor muscle tone and tenderness; tenderness on unidigital pelvic examination; presence of pelvic nodularity; uterine size and mobility; presence of adnexal masses; presence of incisional masses; speculum examination; tenderness and allodynia at an extra-pelvic site (e.g., forearm); and recording of anthropometrics. CONCLUSION(S): The EPHect-PE standards will facilitate the standardized documentation of the physical examination, including the assessment and documentation of examination phenotyping of endometriosis-associated pelvic pain.

Comparative Effectiveness of Interventions in Initial Management of Spontaneous Pneumothorax: A Systematic Review and a Bayesian Network Meta-analysis.

STUDY OBJECTIVE: The best initial strategy for nontension symptomatic spontaneous pneumothorax is unclear. We performed a systematic review and meta-analysis to identify the most efficacious, safe, and efficient initial intervention in adults with nontension spontaneous pneumothorax. METHODS: MEDLINE, Scopus, Web of Science, and ClinicalTrials.gov were searched from January 1950 through December 2019 (print and electronic publications). Randomized controlled trials evaluating needle aspiration, narrow-bore chest tube (<14 F) with or without Heimlich valve insertion, and large-bore chest tube (≥14 F) insertion in spontaneous pneumothorax were included. Network meta-analyses were performed with a Bayesian random-effects model. RESULTS: Twelve studies were included in this review (n=781 patients). Analyses of efficacy (n=12 trials) revealed no significant differences between the interventions studied: narrow- versus large-bore chest tubes, odds ratio (OR) 1.05 (95% credible interval [CrI] 0.38 to 2.87); large-bore chest tube versus needle aspiration, OR 1.25 (95% CrI 0.65 to 2.62); and narrow-bore chest tube versus needle aspiration, OR 1.32 (95% CrI 0.54 to 3.42). Analyses of safety (n=10 trials) revealed a significant difference between needle aspiration and large-bore chest tube interventions: OR 0.10 (95% CrI 0.03 to 0.40). No differences were observed in needle aspiration versus narrow-bore chest tube (OR 0.29 [95% CrI 0.05 to 1.82]), and narrow- versus large-bore chest tube comparisons (OR 0.35 [95% CrI 0.07 to 1.67]). Analyses of efficiency were not pursued because of variation in reporting the length of stay (n=12 trials). Narrow-bore chest tube (<14 F) had the highest likelihood of top ranking in terms of immediate success (surface under the cumulative ranking curve=64%). Needle aspiration had the highest likelihood of top ranking in terms of safety (surface under the cumulative ranking curve=95.8%). CONCLUSION: In the initial management of nontension spontaneous pneumothorax, the optimal strategy between the choices of a narrow-bore chest tube (<14 F, top ranked in efficacy) and needle aspiration (top ranked in safety) is unclear. Complications were more common in large-bore chest tube (≥14 F, including 14-F tube) insertions compared with needle aspiration.

Inhibition by zinc of deoxycholate-induced apoptosis in HCT-116 cells.

The bile acid, deoxycholate, can induce apoptosis although the effect of trace elements on such cell death is unknown. The aim of this study was to determine if deoxycholate-induced apoptosis is influenced by zinc. HCT-116 colon epithelial cells were pre-treated with zinc and then exposed to deoxycholate. Membrane blebbing, formation of apoptotic bodies, and greater overall production of reactive oxygen species (ROS) occurred in cells exposed to deoxycholate, but zinc inhibited the occurrence of these three events caused by deoxycholate. Upon finer analysis, stimulation of mitochondrial superoxide production, mitochondrial dysfunction, and cytochrome c release were detected in cells exposed to deoxycholate, but zinc did not inhibit any of these three effects caused by deoxycholate. Additionally, caspase-3 activation, plasma membrane phospholipid translocation, and also chromatin condensation and fragmentation were observed in cells exposed to deoxycholate, but all of these effects of deoxycholate, including the greater overall ROS production, were all inhibited by zinc. Because zinc did not prevent the three mitochondrial effects caused by deoxycholate, the last set of findings suggested that zinc hampered activation of an initiator caspase upstream of effector caspase-3, in inhibiting deoxycholate-induced HCT-116 cell death. In examining this possibility, it was found that caspase-8 activation caused by deoxycholate was blocked by zinc. Collectively, the results suggest that zinc can inhibit deoxycholate-induced apoptotic cell death mediated by caspases.

Inhibition of deoxycholate-induced apoptosis in iron-depleted HCT-116 cells.

The bile acid, deoxycholate (DOC), can induce apoptosis in cells containing adequate amounts of all key nutrients, but it is unknown whether DOC-induced apoptosis can occur in cells lacking a single key nutrient. The aim of this study was to determine if DOC is able to induce apoptosis in HCT-116 colon epithelial cells depleted of iron. The cells were made iron-deficient by pre-treating them with the iron chelator, deferoxamine (DFO), before subsequent exposure to DOC. Mitochondrial dysfunction was detected in control cells exposed to DOC, but not in iron-depleted cells exposed to DOC. Moreover, characteristic features of apoptosis, namely, membrane blebbing, formation of apoptotic bodies, cytochrome c release into cytosol, generation of the activated form of caspase-3, chromatin condensation and fragmentation, and also plasma membrane phospholipid translocation, were all induced by DOC in control cells but not in iron-depleted cells. Treating DFO-pretreated cells with ferrous sulfate to replenish cellular iron restored the ability of DOC to induce apoptosis. In relating these findings to oxidative stress, it was found that DOC also induced the formation of reactive oxygen species and caused DNA damage in control cells, but not in iron-depleted cells. Collectively, the results suggest that in order for HCT-116 cells to undergo apoptosis when exposed to DOC, adequate amounts of intracellular iron must be present.

Upregulation of GADD153 by butyrate: involvement of MAPK.

Butyrate inhibits the proliferation of cancer cells, but the early molecular events initiated by butyrate have not been fully identified. Herein, butyrate is shown to affect the growth arrest and DNA damage-inducible gene 153 (GADD153) in HCT-116 human colon adenocarcinoma cells. Despite absence of any detectable cellular DNA damage, the expression of GADD153 was upregulated before several features characteristic of apoptosis appeared. Butyrate-induced upregulation of GADD153 mRNA was attenuated by actinomycin D, but apparently not by cycloheximide. In investigating possible involvement of MAPK in mediating the effect of butyrate on GADD153 mRNA expression, the extracellular regulated kinase (ERK) inhibitor PD98059, but neither the JNK inhibitor SP600125 nor the p38 MAPK inhibitor SB203580, blunted the ability of butyrate to upregulate GADD153 mRNA expression. U0126, a selective inhibitor of upstream MEK, had a similar effect as PD98059 on butyrate-induced GADD153 mRNA upregulation. Collectively, these findings suggest that butyrate caused activation of the GADD153 gene at the level of transcription involving mainly the MEK/ERK branch of the MAPK signal transduction pathway. Moreover, these molecular events were not the result of any DNA damage and occurred before several features characteristic of apoptosis became evident.

Protection of human colon epithelial cells against deoxycholate by rottlerin.

The bile salt, deoxycholate (DOC), can harm cells and cause disease. Hence, there is interest in identifying compounds capable of protecting cells against DOC. In HCT-116 colon epithelial cells, DOC increased generation of reactive oxygen species and caused DNA damage and apoptosis. These effects of DOC were inhibited by rottlerin, which is a phenolic compound of plant origin. In elucidating its mechansim, rottlerin prevented the release of cytochrome c from mitochondria into cytosol, and also prevented the cleavage of caspase-3. Yet, rottlerin by itself markedly decreased mitochondrial membrane potential and increased mitochondrial superoxide production, but this did not result in cytochrome c release or in caspase-3 cleavage. At a higher test concentration, two other phenolic phytochemicals, namely, quercetin and resveratrol, were each able to largely prevent the occurrence of apoptosis in cells exposed to DOC. In contrast, epigallocatechin gallate, curcumin, and genistein were ineffective.

Paradoxical effect of diphenyleneiodonium in inducing DNA damage and apoptosis.

Diphenyleneiodonium (DPI) is often used as a molecular tool in unravelling redox-sensitive cellular events involving NADPH oxidase. However, to better understand unexpected actions of DPI, it was ascertained if DPI affects cellular DNA. DPI induced single-strand breaks in DNA of HCT-116 cells, although this only slightly increased GADD153 expression. Nevertheless, after sustaining DNA damage, the DPI-treated cells subsequently had features characteristic of apoptosis, such as translocated membrane phospholipid and nuclei containing condensed chromatin. Paradoxically, DPI attenuated the DNA damage and overall ROS production caused by sodium deoxycholate (DOC), although DPI did not inhibit DOC-induced generation of mitochondrial [image omitted] . Furthermore, DPI prevented the occurrence of apoptosis caused by DOC. However, other known chemical inhibitors of NADPH oxidase did not produce the same results as DPI in negating the effects of DOC. Collectively, these disparate findings suggest that DPI can act not in accord with conventional wisdom depending on the experimental conditions.