Society of Radiologists in Ultrasound Consensus on Routine Pelvic US for Endometriosis.

Endometriosis is a prevalent and potentially debilitating condition that mostly affects individuals of reproductive age, and often has a substantial diagnostic delay. US is usually the first-line imaging modality used when patients report chronic pelvic pain or have issues of infertility, both common symptoms of endometriosis. Other than the visualization of an endometrioma, sonologists frequently do not appreciate endometriosis on routine transvaginal US images.

CFD-based bioreactor model with proportional-integral-derivative controller functionality for dissolved oxygen and pH.

A physics-based model for predicting cell culture fluid properties inside a stirred tank bioreactor with embedded PID controller logic is presented. The model evokes a time-accurate solution to the fluid velocity field and overall volumetric mass transfer coefficient, as well as the ongoing effects of interfacial mass transfer, species mixing, and aqueous chemical reactions. The modeled system also includes a direct coupling between process variables and system control variables via embedded controller logic.

Untargeted proteomics reveals upregulation of stress response pathways during CHO-based monoclonal antibody manufacturing process leading to disulfide bond reduction.

Monoclonal antibody (mAb) interchain disulfide bond reduction can cause a loss of function and negatively impact the therapeutic's efficacy and safety. Disulfide bond reduction has been observed at various stages during the manufacturing process, including processing of the harvested material. The factors and mechanisms driving this phenomenon are not fully understood.

Allosteric regulation within the highly interconnected structural scaffold of AraC/XylS homologs tolerates a wide range of amino acid changes.

To create bacterial transcription "circuits" for biotechnology, one approach is to recombine natural transcription factors, promoters, and operators. Additional novel functions can be engineered from existing transcription factors such as the E. coli AraC transcriptional activator, for which binding to DNA is modulated by binding L-arabinose.

Predictors for Discontinuation of Intrauterine Devices That Underwent Ultrasound-Guided Placement in a Long-term Follow-up Cohort.

Objectives: To investigate long-term outcomes of ultrasound-guided intrauterine device (IUD) insertion. The rate of and reasons for IUD discontinuation were reviewed, including the prevalence of uterine fibroids.

Methods: A retrospective cohort of patients who underwent transabdominal ultrasound (TAUS)-guided IUD insertion was reviewed.

Metabolic understanding of disulfide reduction during monoclonal antibody production.

The disulfide reduction of intact monoclonal antibodies (mAbs) and subsequent formation of low molecular weight (LMW) species pose a direct risk to product stability, potency, and patient safety. Although enzymatic mechanisms of reduction are well established, an understanding of the cellular mechanisms during the bioreactor process leading to increased risk of disulfide reduction after harvest remains elusive. In this study, we examined bench, pilot, and manufacturing-scale batches of two mAbs expressed in Chinese hamster ovary (CHO) cells, where harvested cell culture fluid (HCCF) occasionally demonstrated disulfide reduction.

Increased MSX level improves biological productivity and production stability in multiple recombinant GS CHO cell lines.

Increasing cell culture productivity of recombinant proteins via process improvements is the primary focus for research groups within biologics manufacturing. Any recommendations to improve a manufacturing process obviously must be effective, but also be robust, scalable, and with product quality comparable to the original process. In this study, we report that three different GS CHO cell lines developed in media containing a standard concentration of the selection agent methionine sulfoximine (MSX), but then exposed to increased MSX concentrations during seed train expansion, achieved titer increases of 10-19%.

Dyspareunia Related to GSM: Association of Total Vaginal Thickness via Transabdominal Ultrasound.

Introduction: It has previously been suggested in the literature that ultrasound measurement of total vaginal wall thickness (TVT) differs significantly between pre- and postmenopausal women, indicating that it may be a useful and noninvasive objective assessment to correlate the degree of vaginal atrophy to patient-reported symptoms.

Aim: The purpose of this cross-sectional pilot study was to determine whether TVT in postmenopausal women, as measured with transabdominal ultrasound, is associated with patient-reported dyspareunia and symptoms related to genitourinary symptomatology.

Methods: Postmenopausal women presenting for pelvic ultrasound had TVT and total mucosal thickness (TMT) measured via transabdominal ultrasound.

Sonographically Guided Office Endometrial Sampling: Indications and Results.

Objectives: The purpose of this study was to identify clinical indications in which endometrial sampling was performed under transabdominal ultrasound (US) guidance and to evaluate the outcomes of this approach in an academic practice.

Methods: We retrieved data from the electronic medical record for all patients referred to the gynecologic US unit for transabdominal US-guided endometrial sampling from January 2011 to June 2017. Data retrieved included age, parity, previous cesarean delivery or pelvic surgery, indication for endometrial biopsy, US abnormalities, clinical reasoning for US-guided sampling, and pathologic reports.

Sonographically guided insertion of intrauterine device: Indications and results.

From 2011 to 2015, a total of 67 patients were referred for IUD insertion guided with transabdominal sonography (TAS). Fifty-six of the 67 patients had successful IUD insertion under TAS guidance. The clinical indications for referral included fibroids, uterine position, previous history of IUD expulsion, and limited tolerance of pelvic examination.

Transabdominal sonography to measure the total vaginal and mucosal thicknesses.

Transabdominal and transvaginal sonography are used to measure bladder wall and detrusor thickness. Only transvaginal sonography has been used to measure the vaginal wall thickness. We describe the use of transabdominal sonography to measure the total vaginal wall thickness and total vaginal mucosal thickness at the bladder trigone.

Assessing the thickness of the vaginal wall and vaginal mucosa in pre-menopausal versus post-menopausal women by transabdominal ultrasound: A feasibility study.

Objective: As life expectancy increases, the number of women reporting adverse genito-urinary symptoms (genitourinary syndrome of menopause; GSM) from menopause, including vaginal dryness and sexual pain, also will increase. Current objective measurements of vaginal atrophy such as maturation index require vaginal swabs and are invasive; at present, no minimally invasive measurements exist. The purpose of this study was to assess whether total vaginal wall thickness (TVT) and total vaginal mucosa thickness (TMT) as measured by transabdominal ultrasound could qualify as additional objective markers of vaginal wall thinning which could be related to menopausal status.

1.65 Å resolution structure of the AraC-family transcriptional activator ToxT from Vibrio cholerae.

ToxT is an AraC-family transcriptional activator protein that controls the expression of key virulence factors in Vibrio cholerae, the causative agent of cholera. ToxT directly activates the expression of the genes that encode the toxin-coregulated pilus and cholera toxin, and also positively auto-regulates its own expression from the tcp promoter. The crystal structure of ToxT has previously been solved at 1.

3-Substituted Biquinolinium Inhibitors of AraC Family Transcriptional Activator VirF from Obtained Through In Situ Chemical Ionization of 3,4-Disubstituted Dihydroquinolines.

During a structure-activity relationship optimization campaign to develop an inhibitor of AraC family transcriptional activators, we discovered an unexpected transformation of a previously reported inhibitor that occurs under the assay conditions. Once placed in the assay media, the 3, 4-disubstituted dihydroquinoline core of the active analogue rapidly undergoes a decomposition reaction to a quaternary 3-substituted biquinolinium. Further examination established an SAR for this chemotype while also demonstrating its resilience to irreversible binding of biologically relevant nucleophiles.

Small-molecule inhibitor of the Shigella flexneri master virulence regulator VirF.

VirF is an AraC family transcriptional activator that is required for the expression of virulence genes associated with invasion and cell-to-cell spread by Shigella flexneri, including multiple components of the type three secretion system (T3SS) machinery and effectors. We tested a small-molecule compound, SE-1 (formerly designated OSSL_051168), which we had identified as an effective inhibitor of the AraC family proteins RhaS and RhaR, for its ability to inhibit VirF. Cell-based reporter gene assays with Escherichia coli and Shigella, as well as in vitro DNA binding assays with purified VirF, demonstrated that SE-1 inhibited DNA binding and transcription activation (likely by blocking DNA binding) by VirF.

Identification of a small-molecule inhibitor of bacterial AraC family activators.

Protein members of the AraC family of bacterial transcriptional activators have great promise as targets for the development of novel antibacterial agents. Here, we describe an in vivo high-throughput screen to identify inhibitors of the AraC family activator protein RhaS. The screen used two Escherichia coli reporter fusions: one to identify potential RhaS inhibitors and a second to eliminate nonspecific inhibitors from consideration.

The AraC/XylS family activator RhaS negatively autoregulates rhaSR expression by preventing cyclic AMP receptor protein activation.

The Escherichia coli RhaR protein activates expression of the rhaSR operon in the presence of its effector, L-rhamnose. The resulting RhaS protein (plus L-rhamnose) activates expression of the L-rhamnose catabolic and transport operons, rhaBAD and rhaT, respectively. Here, we further investigated our previous finding that rhaS deletion resulted in a threefold increase in rhaSR promoter activity, suggesting RhaS negative autoregulation of rhaSR.

Differences in the mechanism of the allosteric l-rhamnose responses of the AraC/XylS family transcription activators RhaS and RhaR.

Proteins in the largest subset of AraC/XylS family transcription activators, including RhaS and RhaR, have C-terminal domains (CTDs) that mediate DNA-binding and transcription activation, and N-terminal domains (NTDs) that mediate dimerization and effector binding. The mechanism of the allosteric effector response in this family has been identified only for AraC. Here, we investigated the mechanism by which RhaS and RhaR respond to their effector, l-rhamnose.

Transcription activation by the DNA-binding domain of the AraC family protein RhaS in the absence of its effector-binding domain.

The Escherichia coli L-rhamnose-responsive transcription activators RhaS and RhaR both consist of two domains, a C-terminal DNA-binding domain and an N-terminal dimerization domain. Both function as dimers and only activate transcription in the presence of L-rhamnose. Here, we examined the ability of the DNA-binding domains of RhaS (RhaS-CTD) and RhaR (RhaR-CTD) to bind to DNA and activate transcription.

Functional consequences of exchanging domains between LacI and PurR are mediated by the intervening linker sequence.

Homologue function can be differentiated by changing residues that affect binding sites or long-range interactions. LacI and PurR are two proteins that represent the LacI/GalR family (>500 members) of bacterial transcription regulators. All members have distinct DNA-binding and regulatory domains linked by approximately 18 amino acids.

Linker regions of the RhaS and RhaR proteins.

Substitutions within the interdomain linkers of the AraC/XylS family proteins RhaS and RhaR were tested to determine whether side chain identity or linker structure was required for function. Neither was found crucial, suggesting that the linkers do not play a direct role in activation, but rather simply connect the two domains.

Cyclic AMP receptor protein and RhaR synergistically activate transcription from the L-rhamnose-responsive rhaSR promoter in Escherichia coli.

The Escherichia coli rhaSR operon encodes two AraC family transcription activator proteins, RhaS and RhaR, which regulate expression of the l-rhamnose catabolic regulon in response to l-rhamnose availability. RhaR positively regulates rhaSR in response to l-rhamnose, and RhaR activation can be enhanced by the cyclic AMP (cAMP) receptor protein (CRP) protein. CRP is a well-studied global transcription regulator that binds to DNA as a dimer and activates transcription in the presence of cAMP.

Amino acid contacts between sigma 70 domain 4 and the transcription activators RhaS and RhaR.

The RhaS and RhaR proteins are transcription activators that respond to the availability of L-rhamnose and activate transcription of the operons in the Escherichia coli L-rhamnose catabolic regulon. RhaR activates transcription of rhaSR, and RhaS activates transcription of the operon that encodes the L-rhamnose catabolic enzymes, rhaBAD, as well as the operon that encodes the L-rhamnose transport protein, rhaT. RhaS is 30% identical to RhaR at the amino acid level, and both are members of the AraC/XylS family of transcription activators.

Children who question their heterosexuality.

Many gay, lesbian, and bisexual adults report a period of childhood sexual questioning--an uneasy questioning of their heterosexuality brought on by same-sex attractions and motivating same-sex sexual exploration. This article evaluates hypotheses about the correlates, causes, and consequences of childhood sexual questioning. Participants were 182 children in the 4th through 8th grades.

Impact of high pressure freezing on DH5alpha Escherichia coli and red blood cells.

The impact of high pressure and freezing on survivability of Escherichia coli and human red blood cells was evaluated to determine the utility of high-pressure transitions for preserving living cells. Based on microscopy and survivability, high pressures did not directly impact physical damage to living cells. E.