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The anthelmintic ivermectin has been reported to possess anticancer and antiviral efficacy. However, the effective concentrations reported in vitro are near the predicted aqueous solubility limit for this hydrophobic drug. We observed that ivermectin-induced cell death in two cervical cancer cell lines correlated with the formation of solid ivermectin aggregates in both serum-free and serum-supplemented culture media. Filtration of ivermectin particles0.2 μm abolished these cytolytic effects in both cell lines. An inhibitory effect on cell proliferation persisted for filtered solutions, but only for ivermectin concentrations higher than reported to be clinically attainable in humans. In addition to the importance of distinguishing between free and bound drug in solution, our data emphasize the importance of acknowledging the likely solubility limit of hydrophobic drugs when assessing their in vitro cytotoxicity.

Background Conventional radiologic modalities perform poorly in the radiated rectum and are often unable to differentiate residual cancer from treatment scarring. Purpose To report the development and initial patient study of an imaging system comprising an endorectal coregistered photoacoustic (PA) microscopy (PAM) and US system paired with a convolution neural network (CNN) to assess the rectal cancer treatment response. Materials and Methods In this prospective study (ClinicalTrials.gov identifier NCT04339374), participants completed radiation and chemotherapy from September 2019 to September 2020 and images were obtained with the PAM/US system prior to surgery. Another group's colorectal specimens were studied ex vivo. The PAM/US system consisted of an endorectal imaging probe, a 1064-nm laser, and one US ring transducer. The PAM CNN and US CNN models were trained and validated to distinguish normal from malignant colorectal tissue using ex vivo and in vivo patient data. The PAM CNN and US CNN were then tested using additional in vivo patient data that had not been seen by the CNNs during training and validation. Results Twenty-two patients' ex vivo specimens and five patients' in vivo images (a total of 2693 US regions of interest [ROIs] and 2208 PA ROIs) were used for CNN training and validation. Data from five additional patients were used for testing. A total of 32 participants (mean age, 60 years; range, 35-89 years) were evaluated. Unique PAM imaging markers of the complete tumor response were found, specifically including recovery of normal submucosal vascular architecture within the treated tumor bed. The PAM CNN model captured this recovery process and correctly differentiated these changes from the residual tumor. The imaging system remained highly capable of differentiating tumor from normal tissue, achieving an area under the receiver operating characteristic curve of 0.98 (95% CI: 0.98, 0.99) for data from five participants. By comparison, the US CNN had an area under the receiver operating characteristic curve of 0.71 (95% CI: 0.70, 0.73). Conclusion An endorectal coregistered photoacoustic microscopy/US system paired with a convolutional neural network model showed high diagnostic performance in assessing the rectal cancer treatment response and demonstrated potential for optimizing posttreatment management. © RSNA, 2021

Introduction: Duodenal eosinophilia is a key feature of functional dyspepsia, particularly in those with early satiety. Duodenal eosinophilia is also recognised in coeliac disease, although its rel...

* Informal leaders are present in healthcare organizations. They exercise influence over their peers, which can impact the effectiveness and efficiency of the organization. The purpose of this study was to explore formal leaders' perceptions of informal leaders in their organizations in order to further the knowledge base and permit managers to better develop positive informal leader strategies. A total of 322 respondents (AHRA members) returned valid surveys in response to the study. Questions contained in the survey assessed the following factors: Professional Competency, Supporting the Mission, Influence of Informal Leaders, and Future of Informal Leaders. " The results of the survey suggest that personal demographics and facility characteristics do not account for significant variation in formal leaders' perception of informal leaders in their organizations.

Women considering risk reduction surgery after a diagnosis of breast/ovarian cancer and/or inherited cancer gene mutation face difficult decisions. The safety of combined breast and gynecologic surgery has not been well studied; therefore, we evaluated the outcomes for patients who have undergone coordinated multispecialty surgery.We conducted a retrospective review of patients undergoing simultaneous breast and gynecologic surgery for newly or previously diagnosed breast cancer and/or an inherited cancer gene mutation during the same anesthetic at a single institution from 1999 to 2013.Seventy-three patients with a mean age of 50 years (range 27-88) were identified. Most patients had newly diagnosed breast cancer or ductal carcinoma in situ (62 %) and 28 patients (38 %) had an identified BRCA mutation. Almost all gynecologic procedures were for risk reduction or benign gynecologic conditions (97 %). Mastectomy was performed in 39 patients (53 %), the majority of whom (79 %) underwent immediate reconstruction. The most common gynecologic procedure involved bilateral salpingo-oophorectomy, which was performed alone in 18 patients (25 %) and combined with hysterectomy in 40 patients (55 %). A total of 32 patients (44 %) developed postoperative complications, most of which were minor and did not require surgical intervention or hospitalization. Two of the 19 patients who underwent implant reconstruction (11 %; 3 % of the entire cohort) had major infectious complications requiring explantation.Combined breast and gynecologic procedures for a breast cancer diagnosis and/or risk reduction in patients can be accomplished with acceptable morbidity. Concurrent operations, including reconstruction, can be offered to patients without negatively impacting their outcome.

Positive attitudes toward a new communication technology tend to be a significant motivator in subsequent adoption and use. The recent spurt in the adoption of social media tools such as social networking sites (SNSs) demands the examination of attitudinal variables on motives to use these Web sites. This study explicated a multidimensional measure of attitudes toward SNSs and tested a theoretical model to examine the effect of attitudes on motives to use SNSs and SNS activity. Participants (N=674) completed a cross-sectional survey consisting of measures of attitudes toward SNSs, motives of SNS use, and level of activity. Results showed support for a revised model in which attitudinal variables-ease of use, self-disclosure, and social connection-strongly predicted motives of SNS use such as passing time, information/entertainment, social conformity, and, most importantly, socialization. The motive of using SNSs as a social tool superseded the direct effect of other motives on SNS activity, suggesting that users' primary activity on SNSs was for socialization and for relational development and maintenance.

Objective To determine if maternal major depression (MD), antidepressant use, or paternal MD are associated with pregnancy outcomes after non-IVF fertility treatments. Design Cohort study. Setting Clinics. Patient(s) Participants in two randomized trials: PPCOS II (clomiphene citrate versus letrozole for polycystic ovary syndrome), and AMIGOS (gonadotropins versus clomiphene citrate versus letrozole for unexplained infertility). Intervention(s) Female and male partners completed the Patient Health Questionnaire (PHQ-9). Female medication use was collected. PHQ-9 score ≥10 was used to define currently active MD. Main Outcome Measure(s) Primary outcome: live birth. Secondary outcomes: pregnancy, first-trimester miscarriage. Poisson regression models were used to determine relative risks after adjusting for age, race, income, months trying to conceive, smoking, and study (PPCOS II versus AMIGOS). Result(s) Data for 1,650 women and 1,608 men were included. Among women not using an antidepressant, the presence of currently active MD was not associated with poorer fertility outcomes (live birth, miscarriage), but rather was associated with a slightly increased likelihood of pregnancy. Maternal antidepressant use (n = 90) was associated with increased risk of miscarriage, and male partners with currently active MD were less likely to achieve conception. Conclusion(s) Currently active MD in the female partner does not negatively affect non-IVF treatment outcomes; however, currently active MD in the male partner may lower the likelihood of pregnancy. Maternal antidepressant use is associated with first-trimester pregnancy loss, which may depend upon the type of antidepressant. Clinical Trial Registration Numbers NCT00719186 and NCT01044862.

Aims Autologous CD34+ (auto-CD34+) cells represent an attractive option for the treatment of refractory angina. Three double-blinded randomized trials (n = 304) compared intramyocardial (IM) auto-CD34+ cells with IM placebo injections to affect total exercise time (TET), angina frequency (AF), and major adverse cardiac events (MACE). Patient-level data were pooled from the Phase I, Phase II ACT-34, ACT-34 extension, and Phase III RENEW trials to determine the efficacy and safety of auto-CD34+ cells. Methods and results Treatment effects for TET were analysed using an analysis of covariance mixed-effects model and for AF using Poisson regression in a log linear model with repeated measures. The Kaplan-Meier rate estimates for MACE were compared using the log-rank test. Autologous CD34+ cell therapy improved TET by 46.6 s [3 months, 95% confidence interval (CI) 13.0 s-80.3 s; P = 0.007], 49.5 s (6 months, 95% CI 9.3-89.7; P = 0.016), and 44.7 s (12 months, 95% CI - 2.7 s-92.1 s; P = 0.065). The relative frequency of angina was 0.78 (95% CI 0.63-0.98; P = 0.032), 0.66 (0.48-0.91; P = 0.012), and 0.58 (0.38-0.88; P = 0.011) at 3-, 6- and 12-months in auto-CD34+ compared with placebo patients. Results remained concordant when analysed by treatment received and when confined to the Phase III dose of 1 × 105 cells/kg. Autologous CD34 + cell therapy significantly decreased mortality (12.1% vs. 2.5%; P = 0.0025) and numerically reduced MACE (38.9% vs. 30.0; P = 0.14) at 24 months. Conclusion Treatment with auto-CD34+ cells resulted in clinically meaningful durable improvements in TET and AF at 3-, 6- and 12-months, as well as a reduction in 24-month mortality in this patient-level meta-analysis.

Objective To explore the self-expressed desire for, envisioned reaction to, and basic understanding of presymptomatic Alzheimer disease (AD)–related genetic and biomarker tests. Patients and Methods The Alzheimer's Prevention Registry is an online community of people at least 18 years of age who are interested in AD prevention research for purely informational purposes or to be considered for possible research participation in future studies. Information about presymptomatic testing and an online multiple choice format survey were posted from November 1, 2012, through June 20, 2013, on the registry website. Results Of 4036 respondents, 80.8% (3195/3952) wanted genetic testing if paid by insurance and 58.7% (2261/3851) if it would cost them at least $100. A total of 80.2% (3112/3879) wanted biomarker testing. If at high risk for AD, 90.5% (3478/3841) endorsed that they would "pursue a healthier lifestyle," but 11.6% (427/3706) endorsed "seriously consider suicide." The implication of a positive genetic test result was incorrectly understood by 13.1% (500/3812) and 32.6% (1255/3848) failed to view a positive biomarker test result as evidence of increased risk for or the presence of AD. Conclusion Despite efforts to increase public awareness of AD, our survey results suggest that greater education of the public is needed. Interested patients should probably undergo psychological screening to identify those at high risk of adverse psychological outcomes, and disclosure of presymptomatic test results should be anchored to tangible constructive action plans, such as healthy lifestyle changes, long-term care planning, and, when available and appropriate, participation in research trials.

Several studies using rodent subjects have now shown that extra dietary choline may prevent or even reverse the deleterious effects of pre- and early post-natal ethanol administration. Choline supplementation has been shown to attenuate many, although not all, of ethanol's effects on brain development and behavior. Our laboratory has consistently reported impaired habituation of the heart rate orienting response to a novel olfactory stimulus in animals exposed to ethanol on postnatal days (PD) 4–9. Here we examine whether supplemental choline given both during and after ethanol administration could alleviate these ethanol-induced deficits. Subjects were given 5 g/kg/day ethanol or sham intubations on PD 4–9. Half of the subjects in each group were given a single daily s.c. injection of choline chloride on PD 4–20, while the other half were injected daily with saline. Pups were tested for heart rate orienting and response habituation in a single test session on PD 23. Results replicated the ethanol-induced impairment in response habituation. However, choline supplementation had no effect on orienting or habituation in either neonatal treatment group. These findings indicate that habituation deficits induced by ethanol are not alleviated by extra dietary choline using these parameters. Choline holds great promise as a treatment for some fetal alcohol effects, but is not an effective treatment for all ethanol-related deficits.

The semiallogenic fetus is tolerated by the maternal immune system through control of innate and adaptive immune responses. Trophoblast cells secrete nanometer scale membranous particles called exosomes, which have been implicated in modulation of the local and systemic maternal immune system. Here we investigate the possibility that exosomes secreted from the first trimester and term placenta carry HLA-G and B7 family immunomodulators. Confocal microscopy of placental sections revealed intracellular colocalization of B7-H1 with CD63, suggesting that B7-H1 associates with subcellular vesicles that give rise to exosomes. First trimester and term placental explants were then cultured for 24 hours. B7H-1 (CD274), B7-H3 (CD276) and HLA-G5 were abundant in pelleted supernatants of these cultures that contained microparticles and exosomes; the latter, however, was observed only in first trimester pellets and was nearly undetectable in term explant-derived pellets. Further purification of exosomes by sucrose density fractionation confirmed the association of these proteins specifically with exosomes. Finally, culture of purified trophoblast cells in the presence or absence of EGF suggested that despite the absence of HLA-G5 association with term explant-derived exosomes, it is present in exosomes secreted from mononuclear cytotrophoblast cells. Further, differentiation of cytotrophoblast cells reduced the presence of HLA-G5 in secreted exosomes. Together, the results suggest that the immunomodulatory proteins HLA-G5, B7-H1 and B7-H3, are secreted from early and term placenta, and have important implications in the mechanisms by which trophoblast immunomodulators modify the maternal immunological environment.

This study tested whether intramyocardial (IM) administration of mobilized, purified autologous CD34(+) cells would improve total exercise time (TET) and angina frequency in patients with refractory angina.IM administration of autologous CD34(+) cells has been associated consistently with improvements in functional capacity and angina symptoms in early phase clinical trials.RENEW (Efficacy and Safety of Targeted Intramyocardial Delivery of Auto CD34+ Stem Cells for Improving Exercise Capacity in Subjects With Refractory Angina) was a randomized, double-blind, multicenter trial comparing IM CD34(+) administration with no intervention (open-label standard of care) or IM placebo injections (active control). The primary efficacy endpoint was change in TET at 12 months. Key secondary endpoints include changes in angina frequency at 3, 6, and 12 months, and TET at 3 and 6 months. The key safety analysis was the incidence of major adverse cardiovascular events through 24 months.The sponsor terminated the study for strategic considerations after enrollment of 112 of planned 444 patients. The difference in TET between patients treated with cell therapy versus placebo was 61.0 s at 3 months (95% confidence interval (CI): -2.9 to 124.8; p = 0.06), 46.2 s at 6 months (95% CI: -28.0 to 120.4; p = 0.22), and 36.6 s at 12 months (95% CI: -56.1 to 129.2; p = 0.43); angina frequency was improved at 6 months (relative risk: 0.63; p = 0.05). Autologous CD34(+) cell therapy seemed to be safe compared with both open-label standard of care and active control (major adverse cardiovascular events 67.9% [standard of care], 42.9% (active control), 46.0% [CD34(+)]).Due to early termination, RENEW was an incomplete experiment; however, the results were consistent with observations from earlier phase studies. These findings underscore the need for a definitive trial. (Efficacy and Safety of Targeted Intramyocardial Delivery of Auto CD34(+) Stem Cells for Improving Exercise Capacity in Subjects With Refractory Angina [RENEW]: NCT01508910).

Interest in global health (GH) education is increasing across disciplines. To assess exposure to and perception of GH training among gastroenterology fellows and program directors across the USA. Design: Electronic survey study. Setting: The questionnaire was circulated to accredited US gastroenterology fellowship programs, with the assistance of the American Gastroenterological Association. Participants: Gastroenterology program directors and fellows. The questionnaire was returned by 127 respondents (47 program directors, 78 fellows) from 55 training programs (36 % of all training programs). 61 % of respondents had prior experience in GH. 17 % of programs offered GH curriculum with international elective (13 %), didactic (9 %), and research activity (7 %) being the most common. Fellows had adequate experience managing hepatitis B (93 %), cholangiocarcinoma (84 %), and intrahepatic duct stones (84 %). 74, 69 and 68 % reported having little to no experience managing hepatitis E, tuberculosis mesenteritis, or epidemic infectious enteritis, respectively. Most fellows would participate in an elective in an underserved area locally (81 %) or a 4-week elective abroad (71 %), if available. 44 % of fellows planned on working or volunteering abroad after fellowship. Barriers to establishing GH curriculum included funding (94 %), scheduling (88 %), and a lack of standardized objectives (78 %). Lack of interest, however, was not a concern. Fellows (49 %), more than faculty (29 %) (χ 2 = 21.9; p = 0.03), believed that GH education should be included in fellowship curriculum. Program directors and trainees recognize the importance of GH education. However, only 17 % of ACGME-approved fellowship programs offer the opportunity. Global health curriculum may enhance gastroenterology training.

Supplemental choline during early stages of development can result in long-lasting improvements to memory function. In addition, pre- or postnatal choline has been shown to be protective against some of the adverse effects of early alcohol exposure. The present experiment examined whether supplemental choline given to rats would protect against the effects of posttraining alcohol administration on trace fear conditioning. Posttraining alcohol exposure in adolescent rats results in poor performance in this hippocampus-dependent task, although delay conditioning is unaffected. Here, rats were given an s.c. injection of either saline or choline chloride daily on postnatal days (PD) 15-26. On PD 30 subjects were trained in a trace fear conditioning procedure. For the next 3 days animals were administered 2.5 g/kg ethanol or water control, and conditional stimulus (CS)-elicited freezing was measured on PD 34. Results indicated that posttraining alcohol disrupted the expression of trace conditioning and that supplemental choline on PD 15-26 was protective against this effect. That is, choline-treated animals subsequently given posttraining ethanol performed as well as animals not given ethanol. These results indicate that supplemental choline given during the periweaning period protects against ethanol-induced impairments in a hippocampus-dependent learning task. Findings contribute to the growing literature showing improvements in learning and memory in subjects given extra dietary choline during critical periods of brain development.

In rodents, voluntary exercise and environmental complexity increases hippocampal neurogenesis and reverses spatial learning and long-term potentiation deficits in animals prenatally exposed to alcohol. The present experiment extended these findings to neonatal alcohol exposure and to delay, trace, and contextual fear conditioning. Rats were administered either 5.25 g/kg/day alcohol via gastric intubation or received sham-intubations (SI) between Postnatal Day (PD) 4 and 9 followed by either free access to a running wheel on PD 30–41 and housing in a complex environment on PD 42–72 (wheel-running plus environmental complexity; WREC) or conventional social housing (SHSH) from PD 30 to 72. Adult rats (PD 80 ± 5) received 5 trials/day of a 10-s flashing-light conditioned stimulus (CS) paired with .8 mA footshock either immediately (delay conditioning) or after a 10-s trace interval (trace conditioning) for 2 days. Neonatal alcohol exposure impaired context and trace conditioning, but not short-delay conditioning. The WREC intervention did not reverse these deficits, despite increasing context-related freezing in ethanol-exposed and SI animals.

Founder mutations are an important cause of Lynch syndrome and facilitate genetic testing in specific ethnic populations. Two putative founder mutations in MSH6 were analyzed in 2685 colorectal cancer (CRC) cases, 337 endometrial cancer (EnCa) cases and 3310 healthy controls of Ashkenazi Jewish (AJ) descent from population-based and hospital-based case–control studies in Israel, Canada and the United States. The carriers were haplotyped and the age of the mutations was estimated. MSH6*c.3984_3987dupGTCA was found in 8/2685 CRC cases, 2/337 EnCa cases, and 1/3310 controls, consistent with a high risk of CRC (odds ratio (OR) = 9.9, 95% confidence interval (CI) = 1.2–78.9, p = 0.0079) and a very high risk of EnCa (OR = 19.6, 95% CI = 1.8–217.2, p = 0.0006). MSH6*c.3959_3962delCAAG was identified in 3/2685 CRC cases, 2/337 EnCa cases and no controls. Each mutation was observed on separate conserved haplotypes. MSH6*c.3984_3987dupGTCA and MSH6*c.3959_3962delCAAG probably arose around 585 CE and 685 CE, respectively. No carriers were identified in Sephardi Jews (450 cases and 490 controls). Truncating mutations MSH6*c.3984_3987dupGTCA and MSH6*c.3959_3962delCAAG cause Lynch syndrome and are founder mutations in Ashkenazi Jews. Together with other AJ founder mutations, they contribute substantially to the incidence of CRC and EnCa and are important tools for the early diagnosis and appropriate management of AJ Lynch syndrome patients.

A number of models have been developed to predict the probability that a person carries a detectable germline mutation in the BRCA1 or BRCA2 genes. Their relative performance in a clinical setting is variable. To compare the performance characteristics of a web-based BRCA1/BRCA2 gene mutation prediction model: the PENNII model ( www.afcri.upenn.edu/itacc/penn2 ), with studies done previously at our institution using four other models including LAMBDA, BRCAPRO, modified PENNI (Couch) tables, and Myriad II tables collated by Myriad Genetics Laboratories. Proband and family cancer history data were analyzed from 285 probands from unique families (27 Ashkenazi Jewish; 277 female) seen for genetic risk assessment in a multispecialty tertiary care group practice. All probands had clinical testing for BR.CA1 and BRCA2 mutations conducted in the same single commercial laboratory. The performance for PENNII results were assessed by the area under the receiver operating characteristic curve (AUC) of sensitivity versus 1-specificity, as a measure of ranking. The AUCs of the PENNII model were higher for predicting BRCA1 than for BRCA2 (81 versus 72%). The overall AUC was 78.7%. PENN II model for BRCA1/2 prediction performed well in this population with higher AUC compared with our experience using four other models. The ease of use of the PENNII model is compatible with busy clinical practices.

Cognitive outcomes associated with prenatal exposure to alcohol include low IQ, impaired short-term memory, problems with executive function, poor spatial abilities, and attention deficits. Longitudinal studies in humans prenatally exposed to alcohol have documented the persistence of such effects. Early exposure to alcohol can lead to developmental delays or more permanent impairments in neurocognitive functioning (Connor et al., 1999; Kerns et al., 1997; Korkman et al., 2003; Riley, 1990). Studies have found that although the physical abnormalities associated with prenatal alcohol exposure may normalize with maturation, impairments in attention, memory and executive function can persist into adulthood (Connor and Streissguth, 1999; Kodituwakku, 2007; Streissguth, 2007). Disturbances in attentional processes especially have been consistently reported across the lifespan of individuals with known prenatal exposure to alcohol. Such disruptions in attention have been noted as early as the first day of life (Streissguth et al., 1983), throughout childhood (Coles et al., 1997; Nanson and Hiscock, 1990; Streissguth et al., 1984), during adolescence (Brown et al., 1991; Coles et al., 2002; Korkman et al., 2003), and into adulthood (Connor et al., 1999; Kerns et al., 1997; Streissguth, 2007). Attentional problems are often noted as decreased focus, poor sustained attention and vigilance, and distractibility. Kerns et al. (1997) reported that adults with a diagnosis of fetal alcohol syndrome (FAS) were more distractible during a vigilance task. Distractibility may in part result from deficits in habituation to sensory stimuli outside the realm of the task, and such disturbances have often been described as a lack of ability to tune-out redundant and task-irrelevant information (Connor and Streissguth, 1999; Streissguth et al., 1989). Measurements of stimulus orienting have been powerful tools for examining attention, perception, learning and memory. The orienting response (OR) consists of a collection of central, autonomic, and behavioral reactions to the detection of a novel stimulus (e.g. Graham and Clifton, 1966; Sokolov, 1963). The most commonly used indices of orienting include behavioral orientation toward the source of stimulation and autonomic responses, such as changes in heart rate (Campbell et al., 1992; Lang et al., 1997). The heart rate component of the OR is defined as a decrease in rate (bradycardia), mediated primarily by activation of the parasympathetic nervous system (Graham, 1992; Hunt et al., 1994). Parasympathetic activation is functional in that it serves to reduce background noise, redistribute blood flow away from skeletal muscles to brain areas required for attention and vigilance, and can support faster reaction times (Lacey and Lacey, 1970). A fundamental property of the OR is its ability to undergo rapid habituation, which makes the OR suitable to the study of stimulus encoding and both short- and long-term recognition memory processes (e.g. Richardson and Campbell, 1991). The rate of response habituation has been considered by many as an index of cognitive development in humans and other animals, and rate of habituation assessed in infancy has been regarded as a predictor of later intelligence (e.g. Bornstein, 1989; Kavsek, 2004; McCall and Carriger, 1993). According to comparator theories (e.g. Sokolov, 1963), habituation of the orienting response reflects short-term nonassociative memory processes, as it involves the recognition that a stimulus has been encountered previously and is not biologically significant. Infant recognition memory performance also predicts later IQ (Fagan and Singer, 1983; Rose and Feldman, 1995). Indeed, McCall (1994) has proposed that inhibition is a common feature in habituation and recognition memory paradigms and that it is the propensity for inhibition that accounts for the predictive relationship between these measures and later intelligence test scores. Thus, habituation may be useful in identifying alcohol-induced cognitive deficits soon after birth that could be predictive of more complex and persistent problems that span at least into adolescence and early adulthood (Carmichael Olson et al., 1998; Kerns et al., 1997). Orienting responses have been examined in infants with a history of prenatal alcohol exposure. Coles et al. (1987) for example, reported that moderate gestational exposure to ethanol was correlated with reduced orienting and altered motor development in infants less than one month of age. Furthermore, these researchers found that the initial differences in orienting and motor competence were highly predictive of mental and motor performance assessed at 6 months. Streissguth et al. (1983) reported that infants exposed to moderate doses of ethanol during gestation exhibited reduced rates of response habituation to both auditory and visual stimuli when tested as early as 24 h after birth, with no obvious differences in initial orienting. This same cohort has been assessed repeatedly and deficits in attention and memory domains have consistently been reported (for review see Streissguth, 2007). Previous studies from our laboratory, using young rats, support the findings of Streissguth et al. (1983) in that ethanol-treated animals exhibit impaired response habituation with no observable change in stimulus orienting. Specifically, rats given ethanol on postnatal days (PD) 4-9, to model third-trimester gestational exposure in humans (Dobbing and Sands, 1979; Goodlett and Johnson, 1999), exhibit deficits in habituation of the heart rate orienting response to a novel olfactory stimulus (Hunt and Morasch, 2004; Hunt and Phillips, 2004). However, in these experiments the subjects were tested at only one age, shortly after the termination of the ethanol administration period, on PD 16. Given the well-documented persistence of attention and memory impairments observed in humans with prenatal exposure to ethanol, the purpose of the present experiment was to examine whether ethanol-induced attenuation in rate of response habituation would persist beyond the period of infancy.

Purpose To determine the impact of a physiciandirected, multifaceted health information technology (HIT) system on diabetes outcomes. Methods A pre/post-interventional study. Setting and participants The setting was Providence Primary Care Research Network in Oregon, with approximately 71 physicians caring for 117 369 patients in 13 clinic locations. The study covered Network patients with diabetes age 18 years and older. Intervention The study intervention included implementation of the CareManagerTM HIT system which augments an electronic medical record (EMR) by automating physician driven quality improvement interventions, including point-of-care decision support and care reminders, diabetes registry with care prompts, performance feedback with benchmarking and access to published evidence and patient educational materials. Measures The primary clinical measures included the change in mean value for low density lipoprotein (LDL) target Results A total of 6072 patients were identified at baseline, 70% of whom were continuously enrolled during the 24-month study. Significant improvements were observed in all diabetes related outcomes except mean HbA1c. LDL goal attainment improved from 32% to 56% (P=0.002), while mean LDL decreased by 13 mg/dL (0.33 mmol/l, P=0.002). BP goal attainment increased significantly from 30% to 52%, with significant decreases in both mean systolic and diastolic BP. The proportion of patients with an HbA1c below 7% was higher at the end of the study (P=0.008). Mean patient satisfaction remained high, with no significant difference between baseline and follow-up. Total Relative Value Units per patient per year significantly increased as a result of an increase in the number of visits in year one and the coding complexity throughout. Conclusion Implementation of a physician-directed, multifaceted HIT system in primary care was associated with significantly improved diabetes process and outcome measures.

The efficacy of current anti-cancer gene therapies is limited by the inability of gene vectors to penetrate the poorly vascularized, hypoxic regions of tumors, leaving these sites untreated. We describe a new approach for targeting gene therapy to these sites, which employs an attenuated strain of the non-pathogenic bacterium, Salmonella typhimurium, carrying an exogenous (that is, reporter or therapeutic) gene under the regulation of a new, highly hypoxia-inducible promoter (FF+20(*)). This bacterial vector was seen to rapidly migrate into, and thrive in, hypoxic areas of both mammary tumor spheroids grown in vitro and orthotopic mammary tumors after systemic injection. Using the reporter gene construct, FF+20(*)-lacZ, we show that bacterial expression of high levels of beta-galactosidase occurred only in hypoxic/necrotic sites of spheroids and tumors. We then replaced the reporter gene with one encoding a novel cytotoxic protein (HlyE) and showed that this was also expressed by bacteria only in hypoxic regions of murine mammary tumors. This resulted in a marked increase in tumor necrosis and reduced tumor growth. Our system represents a promising new strategy for delivering gene therapy to poorly vascularized regions of tumors and shows, for the first time, the efficacy of HlyE as an anti-tumor agent.

In a prospective study from 1972 to 1982, 38 patients with bleeding oesophageal varices and relatively good liver function were treated by semi-elective or emergent shunt 2-4 weeks after admission. There were no deaths within 3 months of surgery. This constituted 30% of the total 125 patients admitted with bleeding varices during that period. No referred case for treatment of portal hypertension was included in this study. The procedures performed were end-to-side portacaval shunt in 30 cases, lieno renal shunt in seven cases and interposition mesenterico-caval shunt in one case. All patients were admitted to a special unit with a two stage policy of management. Immediate endoscopic diagnosis and balloon tamponade in those with continued bleeding was followed by shunt in selected cases. Follow-up of the 38 patients showed a cumulative survival at 1 year of 89%, at 3 years of 75% and 5 years of 65%. In four cases (13%) a clinical diagnosis of portal systemic encephalopathy was made, all were controlled by medical management. In four of ten follow-up deaths, liver failure was the cause, in none of these cases was encephalopathy a problem. It is concluded that with a policy of early diagnosis and control of haemorrhage, 30% of a typical series of prospectively studied patients admitted with bleeding varices can be treated without mortality by definitive surgery. There was low incidence of encephalopathy and no cases of incapacitating mental confusion. Centralization of treatment and prospective study is essential for the implementation of such a policy of management.

Eight patients with haemorrhage from smooth muscle tumours of the upper gastro-intestinal tract were treated during a 10 year period from 1973 to 1982. Seven of the tumours were benign and one a malignant leiomyoblastoma. Seven tumours were sited in the stomach and there was one duodenal lesion. Endoscopy was performed in all eight cases and made the diagnosis definitively in four. In two cases the diagnosis was confirmed on barium meal and in the other two, diagnosis was eventually made at laparotomy. Two patients were shocked on admission and required emergency surgery. In one case a diameter greater than 10 cm suggested malignant potential and wide local resection was performed. In one other case with a tumour in the antrum, a distal partial gastrectomy was performed, and in the case with leiomyoblastoma a proximal gastrectomy was performed. One case was lost to follow-up. The mean follow-up in six cases free of disease with benign tumour was 2.6 years. The patient with a tumour of greater than 10 cm in diameter remains well at 18 months follow-up and the patient with a malignant leiomyoblastoma died 2 years after surgery.

Models have been developed to predict the probability that a person carries a detectable germline mutation in the BRCA1 or BRCA2 genes. Their relative performance in a clinical setting is unclear.To compare the performance characteristics of four BRCA1/BRCA2 gene mutation prediction models: LAMBDA, based on a checklist and scores developed from data on Ashkenazi Jewish (AJ) women; BRCAPRO, a Bayesian computer program; modified Couch tables based on regression analyses; and Myriad II tables collated by Myriad Genetics Laboratories.Family cancer history data were analyzed from 200 probands from the Mayo Clinic Familial Cancer Program, in a multispecialty tertiary care group practice. All probands had clinical testing for BRCA1 and BRCA2 mutations conducted in a single laboratory.For each model, performance was assessed by the area under the receiver operator characteristic curve (ROC) and by tests of accuracy and dispersion. Cases "missed" by one or more models (model predicted less than 10% probability of mutation when a mutation was actually found) were compared across models.All models gave similar areas under the ROC curve of 0.71 to 0.76. All models except LAMBDA substantially under-predicted the numbers of carriers. All models were too dispersed.In terms of ranking, all prediction models performed reasonably well with similar performance characteristics. Model predictions were widely discrepant for some families. Review of cancer family histories by an experienced clinician continues to be vital to ensure that critical elements are not missed and that the most appropriate risk prediction figures are provided.

Knowledge about eating disorders influences lay people's ability to recognize individuals with anorexia nervosa (AN) and bulimia nervosa (BN) and refer them to professional treatment. We assessed mental models (stored knowledge) of AN and BN in 106 college students. Results indicated that most students have general, but not specific, information about AN and BN's symptoms, consequences, causes, duration, and cures. They also believe that people with eating disorders tend to be young, White women. These findings suggest that lay recognition of eating disorders may be based primarily on observations of dysfunctional eating behaviors and therefore facilitated by additional knowledge.

HLA-G is an HLA class Ib gene that is highly expressed in human trophoblast cells. The single HLA-G mRNA is alternatively spliced to generate at least seven transcripts, three of which encode soluble isoforms. Many studies have shown that high levels of soluble antigens are associated with successful implantation and graft acceptance. To study expression, regulation and functions of two of the soluble isoforms, HLA-G5 and HLA-G6, we generated recombinant proteins in eukaryotic cells and developed monoclonal antibodies specific for each of the two proteins. In addition, we investigated the olive baboon Paan-AG gene as a potential functional correlate of HLA-G. Here, we present summaries of the studies that have been conducted in our laboratory using these tools and discuss the results within the context of the research on this topic that is ongoing in ours and other laboratories worldwide. Collectively, the data indicate that soluble HLA-G is a critical contributor to immune privilege in pregnancy and imply that this placenta-derived substance may impact other pathways leading to successful reproduction.

The electronic health record (EHR) may be an effective tool to help clinicians address tobacco use more consistently. To evaluate the impact of EHR-generated practice feedback on rates of referral to a state-level tobacco quitline, we conducted a cluster randomized clinical trial (feedback versus no feedback) within 19 primary care clinics in Oregon. Intervention clinics received provider-specific monthly feedback reports generated from EHR data. The reports rated provider performance in asking, advising, assessing, and assisting with tobacco cessation compared with a clinic average and an achievable benchmark of care. During 12 months of follow-up, EHR-documented rates of advising, assessing, and assisting were significantly improved in the intervention clinics compared with the control clinics (p

Experience-produced deficits in trace conditioning and context conditioning have been useful tools for examining the role of the hippocampus in learning. It has also been suggested that learning in these tasks is especially vulnerable to neurotoxic effects of alcohol during key developmental periods such as adolescence. In five experiments we systematically examined the presence and source of age-dependent vulnerability to the memory-disrupting effects of acute ethanol in trace conditioning and contextual fear conditioning. In Experiment 1a pre-training ethanol disrupted trace conditioning more strongly in adolescent (postnatal day, PD30-35) than adult rats (PD65-75). In Experiment 1b when pre-training ethanol was accompanied by pre-test ethanol no deficit in trace conditioning was observed in adolescents, suggesting that state-dependent retrieval failure mediated ethanol's disruption of trace conditioning at this age. Experiments 2a and 2b examined the effect of ethanol pretreatment on context conditioning. Here, adult but not adolescent rats were impaired in conditioned freezing to context cues. Experiment 2c explored state-dependency of this effect. Pre-training ethanol continued to disrupt context conditioning in adults even when ethanol was also administered prior to test. Collectively these findings reveal clear age-dependent and task-dependent vulnerabilities in ethanol's disruptive effects on hippocampus-dependent memory. Adolescents were more disrupted by ethanol in trace conditioning than adults, and adults were more disrupted by ethanol in context conditioning than adolescents. We suggest that adolescents may be more susceptible to changes in internal state (state-dependent retrieval failure) than adults and that ethanol disrupted performance in trace and context conditioning through different mechanisms. Relevance of these findings to theories of hippocampus function is discussed.

Study Objectives. To examine potential threats to internal and external study validity caused by differential patient withdrawal from a randomized controlled trial evaluating pharmacist management of hypertension, to compare the characteristics of patients who withdrew with those of patients who completed the study, and to identify characteristics that predispose patients to withdraw from hypertension management. Design. Prospective, randomized, comparative study. Setting. Network of primary care clinics. Patients. Four hundred sixty-three patients with a diagnosis of hypertension and a last documented systolic blood pressure of 160 mm Hg or greater and/or diastolic blood pressure of 100 mm Hg or greater. Intervention. Patients were randomly allocated to the pharmacist intervention or usual-care (control) group. Those in the pharmacist intervention group were collaboratively managed by a primary care clinical pharmacy specialist and their primary care provider. Patients in the control group received usual care from only their primary care provider. Measurements and Main Results. Of the 463 patients, 191 (41%) withdrew from the study after randomization and 272 (59%) completed the study. Patients who withdrew from the pharmacist intervention group were similar to patients who withdrew from the usual-care group with respect to age, sex, insurance status, and chronic conditions. Patients who smoked or had commercial insurance were more likely to withdraw from the study than the other participants. However, multivariate analysis of all variables, when adjusted for the effect of the intervention, revealed that insurance status was the only variable associated with a heightened probability of withdrawal (p=0.002). Conclusion. Although this study had a high withdrawal rate, between-group patient characteristics remained balanced. Therefore, internal validity was preserved, and outcomes from the study groups could be reliably compared. A lack of significant differences between patients who withdrew versus those who completed, with the exception of insurance status, suggests that external validity was not jeopardized.

The apoptosis cascade that plays a central role in normal and pathological processes is strictly controlled, in part by FLIP (Fas-associated death domain-like interleukin-1beta-converting enzyme-inhibitory protein), an inhibitor of caspase-8. Here, we report the expression of long and short isoforms of FLIP mRNAs and proteins in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the FLIP gene in all samples. Analysis by immunoblotting revealed that both long and short forms of FLIP proteins are present in early and late gestation human placentas with increasing levels over gestation and that FLIP proteins are present in normal and transformed trophoblast cells. Immunohistochemical experiments performed on paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy demonstrated that FLIP proteins are present in caspase-8-expressing cells and that expression patterns of FLIP differed according to cell lineage and stage of cell differentiation. The results of this study are consistent with the postulate that FLIP proteins have critical roles in placental cell survival and suggest that FLIP may protect normal and transformed trophoblast cells from cell death.

Human prenatal ethanol exposure that occurs during a period of increased synaptogenesis known as the “brain growth spurt” has been associated with significant impairments in attention, learning, and memory. The present experiment assessed whether administration of ethanol during the brain growth spurt in the rat, which occurs shortly after birth, disrupts attentional performance. Rats were administered 5.25 g/kg/day ethanol via intragastric intubation from postnatal days (PD) 4–9, sham-intubation, or no intubation (naive). Beginning at PD 90, animals were trained to asymptotic performance in a two-lever attention task that required discrimination of brief visual signals from trials with no signal presentation. Finally, manipulations of background noise and inter-trial interval duration were conducted. Early postnatal ethanol administration did not differentially affect acquisition of the attention task. However, after rats were trained to asymptotic performance levels, those previously exposed to ethanol demonstrated a deficit in detection of signals but not of non-signals compared to sham-intubated and naive rats. The signal detection deficit persisted whenever these animals were re-trained in the standard task, but further task manipulations failed to interact with ethanol pretreatment. The present data support the hypothesis that early postnatal ethanol administration disrupts aspects of attentional processing in the rat.

Maternal antigen presenting cells, which are macrophages and dendritic cells, are scattered throughout human decidualized endometrium during all stages of pregnancy. These powerful, multi-functional leukocytes reside in close proximity to uterine glandular epithelium, uterine blood vessels, and HLA-G-producing invasive cytotrophoblast cells. Macrophages and dendritic cells, which express the HLA-G receptors, ILT2 and ILT4, play major roles in driving innate and adaptive immune responses, altering the behavior of local stromal cells, shaping the cytokine microenvironment, and protecting the tissue from infection. Therefore, encounters between decidual antigen presenting cells and HLA-G molecules are likely to influence uterine and placental homeostasis as well as local maternal immune responses to the fetus during pregnancy.

Context: The premise of the problem-oriented medical record is that an accurately defined problem list will directly result in more thorough and efficient patient care. However, little empirical evidence exists demonstrating improved patient outcomes as a result of an adequately structured problem list. Objective: To determine the impact of problem list documentation of heart failure on the likelihood that evidence-based pharmacotherapy has been prescribed. Design: Cross-sectional study. Setting: Community-based primary care clinics in Portland, Oregon. Subjects: Active patients in the network with a left ventricular ejection fraction of 40% or less, with and without heart failure, in their structured problem list. Main Outcome Measures: The proportion of patients prescribed medications with known benefits for systolic dysfunction. Results: In this group of patients with known systolic dysfunction, the likelihood of therapy with either an angiotensin converting enzyme inhibitor or angiotensin II receptor blocker was higher in patients who had heart failure listed on their problem list compared to patients who did not (92.2% vs 76.7%; P

Background: Recent studies have revealed that the adolescent brain may be especially vulnerable to ethanol-induced toxicity. Corticolimbic regions are more severely damaged following ethanol exposure during adolescence than during adulthood. The consequences of adolescent ethanol exposure on cognition however, have only recently begun to be explored. Methods: Male and female rats were administered 0, 1.5, 2.5 or 4.5 g/kg ethanol (20% v/v) by acute intragastric gavage during adolescence (postnatal days [PD] 28, 30, 32 and 34). On PD 40, half of the subjects in each dose group were given 5 pairings of a 10-sec flashing light (CS; conditioned stimulus) immediately followed by mild footshock (US; unconditioned stimulus), a procedure known as delay conditioning. The other half were also given 5 CS-US pairings, but the US was presented 10 sec after CS offset, a procedure known as trace conditioning. All subjects were tested for CS-elicited freezing 24 h later. Results: There was no effect of adolescent ethanol exposure on delay conditioned responding, with all subjects demonstrating comparable levels of CS-elicited freezing. In contrast, the amount of freezing in the trace conditioned subjects was negatively correlated with prior ethanol dose. Specifically, exposure to 2.5 or 4.5 g/kg during adolescence resulted in a deficit in trace conditioned responding. Conclusions: These data indicate that intermittent exposure to ethanol during adolescence results in impairment in hippocampal-dependent trace conditioning that persists beyond the period of ethanol exposure. Delay conditioning was unaffected by prior ethanol treatment, indicating that there was no difficulty in detecting the CS or US, or in the ability to engage in freezing behavior. These results suggest that the adolescent brain may be particularly vulnerable to the effects of repeated exposure to ethanol that can have consequences for nonspatial, hippocampal-dependent cognitive abilities.

The placenta utilizes both active and passive mechanisms to evade rejection by the maternal immune system. Recently, the mRNA for two newly cloned members of the B7 family of immunomodulatory cell-associated proteins have been identified in the human term placenta. In this article, we review the current knowledge of the B7 family member B7-H1, and discuss how it may participate in modulation of the maternal immune system at the maternal-fetal interface. B7-H1 has been found to possess immunostimulatory or immunoinhibitory properties, and immunohistological examination of first trimester and term placenta has revealed that this protein is abundant in the placenta. B7-H1 is highly expressed by both the syncytiotrophoblast and extravillous cytotrophoblast, both of which lie in direct contact with maternal blood and tissue. Further, treatment of the choriocarcinoma cell line, JEG-3, with recombinant human interferon (IFN)-γ resulted in a dose-dependent increase in the abundance of the message for B7-H1, suggesting that IFN-γ could regulate expression of B7-H1 by the trophoblast. These studies document that the positioning of B7-H1 at the maternal-fetal interface is such that it could participate in suppression of activated maternal leukocytes.

To identify baseline characteristics of women with unexplained infertility to determine whether treatment with an aromatase inhibitor will result in a lower rate of multiple gestations than current standard ovulation induction medications.Randomized, prospective clinical trial.Multicenter university-based clinical practices.A total of 900 couples with unexplained infertility.Collection of baseline demographics, blood samples, and ultrasonographic assessments.Demographic, laboratory, imaging, and survey characteristics.Demographic characteristics of women receiving clomiphene citrate (CC), letrozole, or gonadotropins for ovarian stimulation were very consistent. Their mean age was 32.2 ± 4.4 years and infertility duration was 34.7 ± 25.7 months, with 59% primary infertility. More than one-third of the women were current or past smokers. The mean body mass index (BMI) was 27 and mean antimüllerian hormone level was 2.6; only 11 women (1.3%) had antral follicle counts of5. Similar observations were identified for hormonal profiles, ultrasound characterization of the ovaries, semen parameters, and quality of life assessments in both male and female partners.The cause of infertility in the couples recruited to this treatment trial is elusive, as the women were regularly ovulating and had evidence of good ovarian reserve both by basal FSH, antimüllerian hormone levels, and antral follicle counts; the male partners had normal semen parameters. The three treatment groups have common baseline characteristics, thereby providing comparable patient populations for testing the hypothesis that use of letrozole for ovarian stimulation can reduce the rates of multiples from that observed with gonadotropin and CC treatment.NCT 01044862.

Organophosphorus (OP) compounds have been reported to inhibit Ca/Mg-ATPase, but the relevance of this inhibition to organophosphate-induced delayed neuropathy (OPIDN) has not been explored. To determine if inhibition of this enzyme was related to the development of OPIDN, neuropathic and nonneuropathic OP compounds were sted for their ability to inhibit Ca-stimulated ATPase activity in the P2 synaptosomal fraction from hen brain. Following in vitro exposure to 10(-3) to 10(-5) M OP compounds, Ca-stimulated ATPase activity was inhibited by chlorpyrifos, chlorpyrifos-oxon, phenyl saligenin phosphate (PSP), and tri-o-tolyl phosphate (TOTP), but not by parathion, paraoxon, or diisopropyl fluorophosphate (DFP). Further investigation of inhibition induced by chlorpyrifos determined that inhibition was noncompetitive with respect to calcium and ATP. OP compound hydrophobicity was well correlated with in vitro inhibition of Ca-stimulated ATPase, suggesting that OP compounds interact with membrane lipids, and this interaction may contribute to the noncompetitive inhibition of Ca-stimulated ATPase that was observed. Subsequent to in vivo exposure, DFP, but not PSP, produced inhibition of Ca-stimulated ATPase activity in the hen brain P2 synaptosomal fraction. These data indicate that inhibition of Ca-stimulated ATPase activity is not correlated to neuropathic potential and demonstrate that inhibition of Ca/Mg-ATPase is not responsible for OPIDN.

Modifications occur when interviewers contradict statements made by witnesses or imply that witnesses provided information that they (interviewers) did not provide. Because of their suggestive nature, modifications threaten the reliability of investigative interviews. This study investigated developmental differences in witnesses' responses to modifications during interviews as well as in inclusion of modified misinformation in subsequent answers. Preschool, elementary school, and college students were interviewed about a video presentation. In the experimental conditions, the interviewer contradicted information about the video provided by the participants. Participants then answered two sets of follow-up questions: one immediately following the interview and another 6-8 days later. Results indicated that participants were more likely to ignore modifications than to correct or agree with them. Adult participants were most likely to disagree with modifications. Preschoolers were most likely to incorporate modified misinformation into subsequent answers. Implications of these findings for investigative interviews are discussed.

The uterus and placenta of the mouse and rat produce a member of the prolactin (PRL) family referred to as decidual/trophoblast PRL-related protein (d/tPRP). This cytokine/hormone has been hypothesized to regulate decidual cell activities needed for the establishment and maintenance of gestation. An alkaline phosphatase (AP)-tagging strategy was used to identify d/tPRP target cells. AP-d/tPRP bound to virtually all cells and tissues to which it was exposed, consistent with our earlier evidence that d/tPRP binds to heparin-containing molecules. Moreover, we found that co-incubation with heparin or pretreatment with heparitinase greatly decreased the binding of AP-d/tPRP to tissue sections. In addition, we observed that the AP-d/tPRP probe bound to the surface of Chinese hamster ovary (CHO) cells but not to heparan sulfate-deficient CHO-pgsD-677 cells. Potential unique non-heparin d/tPRP binding sites within mouse and rat uteroplacental tissues were identified by consecutively incubating sections with AP-d/tPRP followed by heparin. This strategy led to the identification of d/tPRP target cells associated with the uterus and the labyrinth zone of the chorioallantoic placenta. Within the uterus, d/tPRP specifically bound to eosinophils. d/tPRP-binding and eosinophil peroxidase activity were co-localized and showed similar patterns of distribution during the estrous cycle, pregnancy, and following hormonal manipulation. d/tPRP interactions with eosinophils were further demonstrated in the lung and intestine, with eosinophils isolated from the peritoneum, and in mice with generalized tissue eosinophilia. Collectively, these findings suggest that intercellular d/tPRP targeting is mediated through associations with heparin-containing molecules which help direct d/tPRP to specific interactions with eosinophils within the uterus and with the labyrinthine compartment of the chorioallantoic placenta.

In human, mouse, and rat pregnancy, maternal NK cells accumulate and differentiate at implantation sites. These cells, termed uterine NK (uNK) cells, express NO synthase (NOS)-2 and develop cytolytic molecules such as perforin and granzymes during differentiation in situ. In this study, relationships between expression of the NOS-2 gene, uNK cell population density and tissue distribution, and synthesis of perforin were investigated. Uteri from wild-type (WT) and NOS-2−/− mice were collected at gestation days (g.d.) 8, 10, 12, 14, and 16 (n, >2/g.d.). Histochemical staining failed to reveal any differences between the population densities or tissue distributions of uNK cells in WT and NOS-2−/− uteri at any stage of gestation. By contrast, immunohistochemical staining with anti-perforin Abs demonstrated significantly fewer perforin-positive uNK cells in two uterine compartments of NOS-2−/− mice in comparison to the same compartments in WT mouse uteri. Perforin-positive uNK cells were reduced in NOS-2−/− metrial glands at g.d. 8, 10, and 12 and in decidua basalis at g.d. 12 (p < 0.05). Analysis of perforin protein by immunoblotting confirmed this observation. Northern blot hybridization studies showed that loss of perforin protein in NOS-2−/− mice was accompanied by decreased steady-state levels of perforin mRNA. These results demonstrate that migration of uNK cells into the uterus, selection of residency sites, and proliferation in situ are independent of NOS-2. By contrast, their differentiation, including transcription and translation of the cytotoxic molecule perforin, was shown to rely on normal expression of the NOS-2 gene.

Fertilization, implantation, and placentation are dynamic cellular events that require not only synchrony between the maternal environment and the embryo, but also complex cell-to-cell communication. This communication involves integrins, a large family of proteins involved in the attachment, migration, invasion, and control of cellular function. Over the past decade, investigators have learned that integrins participate in multiple reproductive events including fertilization, implantation, and placentation in many species. This review will describe: (i) the expression of integrins on gametes and during the establishment and development of the placenta; (ii) regulatory pathways for controlling expression of integrins in the uterus and developing placenta; (iii) the function of integrins as determined by null-mutations; and (iv) reproductive dysfunction in women related to inappropriate integrin expression in the uterus and/or placenta.

Objective: Lactoferrin is an iron-binding protein that has been implicated in protection against infections and allogeneic recognition reactions and in the control of cell growth. We studied the biochemical characteristics and expression of the unique lactoferrin epitopes (LF 1 ) in human placentas. Study Design: Immunohistologic studies of normal human term placentas were done by using monoclonal antibodies to LF 1 . Double-antibody experiments were done by using monoclonal antibodies to markers of inflammation (macrophages, human leukocyte antigen [HLA-DR]). LF 1 was studied immunochemically by using lactoferrin fragments produced by the reaction of lactoferrin with trypsin or N -glycanase. Results: Anti-LF 1 monoclonal antibodies reacted with most interstitial cytotrophoblasts in the basal plate and with villous cytotrophoblasts of some but not all chorionic villi. Cytotrophoblasts expressing LF 1 were associated with large numbers of HLA-DR–reactive macrophages. Anti-LF 1 monoclonal antibodies reacted with 2 distinct tryptic fragments of lactoferrin, and these reactivities were not affected by treatment with N -glycanase. Conclusion: Placental cytotrophoblasts express unique epitopes of lactoferrin (LF 1 ). Such expression is increased in the presence of activated macrophages. This expression could be an extraembryonic response to inflammation and maternal allogeneic recognition as an effort to protect trophoblastic cells. The LF 1 epitopes might represent conserved polypeptide epitopes on 2 homologous lobes of lactoferrin. (Am J Obstet Gynecol 1999;181:460-7.)

PRL-like protein A (PLP-A) is a member of the PRL family expressed in trophoblast cells coincident with establishment of the chorioallantoic placenta. The purpose of this investigation was to identify targets for PLP-A. Using an alkaline phosphatase-tagging strategy, we show that PLP-A specifically interacts with a population of natural killer (NK) lymphocytes within the mesometrial compartment of decidua from pregnant and pseudopregnant rats. These observations are supported by the codistribution of PLP-A targets with cells expressing the rat NK cell surface marker, gp42, the absence of PLP-A binding in conceptuses from NK cell-deficient tgε26 mice, and the specific interaction of PLP-A with a rat NK cell line, RNK-16. We have further demonstrated that PLP-A effectively suppresses RNK-16 cell cytolytic activities. Our results provide evidence for a new paradigm of embryonic-maternal communication involving a PLP-A signaling pathway between trophoblast cells and uterine NK lymphocytes.

Interferon-γ (IFN-γ) is a potent proinflammatory cytokine that modulates hematopoietic cell maturation, differentiation, activation, and apoptosis. To evaluate the postulate that locally produced IFN-γ could influence uterine hematopoietic cells, specific protein was detected by immunohistochemistry and messenger RNA (mRNA) was identified by in situ hybridization in cycling and pregnant mouse uteri. In cycling uteri, IFN-γ was limited to luminal and glandular epithelial cells during the estrus phase of the cycle. In pregnant uteri, IFN-γ was prominent at early (gestation day 6–10) and late (gestation day 18) stages. IFN-γ-producing cells identified by in situ hybridization included uterine epithelial cells, natural killer cells, macrophages, placental trophoblast cells, and cells in the degenerating metrial gland. Collectively, the data indicate that programming of immune and other cells via autocrine and paracrine pathways could be achieved by locally produced IFN-γ, and suggest that uteroplacental IFN-γ may be most influential during early and late stages of pregnancy. J. Leukoc. Biol. 64: 393–400; 1998.

Unlike other somatic cells, human placental trophoblast cells do not express the highly polymorphic HLA-A and HLA-B human leukocyte major histocompatibility antigens that would stimulate maternal immunological rejection of the fetus. To investigate mechanisms underlying cell lineage-specific expression, cell lines were generated from homozygous matings of HLA-B27 transgenic mice. Trophoblast cell lines were generated from gestation day 10 placentas and fibroblasts were cultured from gestation day 13/14 embryos. Polymerase chain reaction (PCR) readily identified HLA-B DNA in transgenic trophoblastic cells but specific mRNA was of low abundance, being detectable by reverse transcriptase PCR but not by Northern blot hybridization. HLA-B-specific protein in/on the trophoblast cells was undetectable by cell enzyme-linked immunosorbent assay and the protein was not induced by exposing the trophoblastic cells to interferon-gamma (IFN-gamma). Restricted expression was specific for the HLA-B transgene and its antigen; IFN-gamma-inducible endogenous H-2Db class I antigens were detectable on the trophoblast cells. In contrast to the trophoblastic cells, HLA-B27 transgenic fibroblasts expressed IFN-gamma-inducible HLA class I antigens as well as H-2Db antigens. Thus, the mechanism(s) regulating expression of the polymorphic HLA-B antigen in trophoblastic cells is gene-specific, IFN-gamma-resistant and operative at the level of transcription or immediate post-transcription.

Pregancy in the human presents an “immunological paradox,” because of the unexpected willingness of mothers to accept genetically disparate tissues. The fact that the fetus can develop unharmed for nine months shows that protective mechanisms must exist to permit its survival. The conditions that permit the genetically dissimilar human fetus to evade rejection by its mother's immune system have been the subject of intense interest for several decades. As the placental cells, which are in contact with maternal blood or tissue, are devoid of HLA class II antigens, interest has focused on the expression of HLA class molecules. Recent developments in the constitutive, transcriptional, and translational expression of HLA class I molecules on anatomically and morphologically different subpopulations of trophoblast cells will form the basis of this short review.

Introduction The Indo-Guyanese population is the largest immigrant minority population in Schenectady, New York. A clinic-based study in Schenectady and surveillance reports from Guyana found high diabetes prevalence and mortality among Guyanese of Indian descent. No community-based study has focused on diabetes among Indo-Guyanese immigrants in the United States. We sought information on the prevalence of diabetes and its complications in Indo-Guyanese adults in Schenectady and compared it with the prevalence among non-Hispanic white adults in Schenectady. Methods We administered a cross-sectional health survey at community venues in Schenectady in 2011. We identified diagnosed diabetes and its complications through self-reports by using a reliability-tested questionnaire. The final data set included 313 Indo-Guyanese and 327 non-Hispanic white adults aged 18 years or older. We compared the prevalence of diagnosed diabetes and diabetes complications between Indo-Guyanese and non-Hispanic whites. Results Most Indo-Guyanese participants were born in Guyana, whereas most non-Hispanic whites were born in the United States. The crude prevalence of diagnosed diabetes among Indo-Guyanese participants and non-Hispanic whites was 30.3% and 16.1%, respectively. The age-standardized prevalence was 28.7% among Indo-Guyanese participants, significantly higher than that among non-Hispanic whites (14.5%, P < .001). Indo-Guyanese participants who had diabetes had a lower body mass index and were more likely to report poor or fair general health and eye or vision complications than non-Hispanic whites who had diabetes. Conclusion Our study confirms the higher prevalence of diabetes in Indo-Guyanese adults in Schenectady. The higher prevalence of complications suggests poor control of diabetes. Excess burden of diabetes in this population calls for further research and public health action.

Signals transduced by binding of tumour necrosis factor alpha (TNF) and lymphotoxin alpha (LT-alpha) trimers to high-affinity cell membrane receptors, TNF-RI (p55/p60) and TNF-RII (p75/p80), affect many cell functions. In this study, expression of the gene encoding TNF-RI in uteri of cyclic mice was mapped using in situ hybridization. TNF-RI hybridization signals fluctuated during the cycle. Signal intensity was highest during dioestrus-II, when mRNA encoding TNF-RI was present in endometrial epithelial and stroma cells, as well as in myometrial smooth muscle and connective tissue cells. The ability of oestradiol and progesterone to modulate steady state concentrations of mRNA encoding TNF-RI in uterine cells was assessed by using in situ and northern blot hybridization procedures. Seven days after ovariectomy, low concentrations of mRNA encoding TNF-RI were detected by northern analysis and weak in situ hybridization signals were identified in epithelia and some myometrial connective tissue cells. Administration of oestradiol, progesterone or oestradiol plus progesterone to ovariectomized animals stimulated temporal and cell type-specific changes in steady state concentrations of mRNA encoding TNF-RI that were unique to each hormonal regimen. Maximal induction of mRNA encoding TNF-RI required 24 h of oestradiol stimulation and 72 h of progesterone stimulation. In uteri treated with oestradiol plus progesterone, the oestradiol pattern predominated over the progesterone pattern. Thus, multiple cell types in cyclic mouse uteri express the gene encoding TNF-RI, and expression in specific cells is controlled by female steroid hormones.

A consecutive series of 19 patients were treated for median sternotomy dehiscence by secondary sternal closure with interosseous absorbable sutures and superimposed pectoralis major myocutaneous advancement flaps. These patients were selected for this treatment only on the basis of the quality and quantity of remaining bone stock after debridement. Using this technique there have been no failures of primary therapy with a zero 30-day mortality rate. All patients have achieved good functional and aesthetic results with mechanically stable sternums, wounds confined to the chest and elimination of sepsis. This technique has the advantages of being simple, safe and relatively quick and avoids many of the inherent complications and disadvantages of other techniques and flaps commonly used in the management of this complication.

Diffusion magnetic resonance imaging (MRI) provides a sensitive indicator of cerebral hypoxia. We investigated if apparent diffusion coefficient (ADC) and transverse relaxation (T2) predict symptoms of acute mountain sickness (AMS), or merely indicate the AMS phenotype irrespective of symptoms. Fourteen normal subjects were studied in two groups; unambiguous AMS and no-AMS at 3,800 m altitude (intermediate AMS scores were excluded). T2 relaxation was estimated from a T2 index of T2-weighted signal normalized by cerebrospinal fluid signal. Measurements were made in normoxia and repeated after 2 days sustained hypoxia (AMS group symptomatic and no-AMS group asymptomatic) and after 7 days hypoxia (both groups asymptomatic). Decreased ADC directly predicted AMS symptoms ( P < 0.05). Apparent diffusion coefficient increased in asymptomatic subjects, or as symptoms abated with acclimatization. This pattern was similar in basal ganglia, white matter, and gray matter. Corpus callosum behaved differently; restricted diffusion was absent (or rapidly reversed) in the splenium, and was sustained in the genu. In symptomatic subjects, T2,index decreased after 2 days hypoxia and further decreased after 7 days. In asymptomatic subjects, T2,index initially increased after 2 days, but decreased after 7 days. T2,index changes were not predictive of AMS symptoms. These findings indicate that restricted diffusion, an indicator of diminished cerebral energy status, directly predicts symptoms of AMS in humans at altitude.