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Introduction: Coronavirus disease 2019 (COVID-19) has impacted researchers' ability to continue to deliver HIV prevention and treatment interventions face to face. Although telehealth has been an important strategy to maintain research operations during the current pandemic, participants at increased risk of or living with HIV are often at higher risk of also experiencing poverty, housing instability, and other challenges that may present obstacles to successful remote delivery. Methods: We provide descriptions of remote adaptations to two randomized controlled efficacy trials of behavioral interventions for primary and secondary HIV prevention with descriptive enrollment and retention data. Results and Conclusions: Best practices for implementing telemedicine and e-health procedures are discussed, including procedures for addressing remote participation barriers (economic, health literacy, etc.) and other challenges, such as building rapport and staff support (NCT03092531 and NCT03175159).

OBJECTIVES: Acceptance and Commitment Therapy (ACT) is a third-wave behavioral and cognitive therapy that increases psychological flexibility through mindfulness, acceptance, and value-driven behavior change. ACT has been successfully used to inform a variety of health interventions. Using non-therapists to deliver ACT-based behavioral health interventions offers an opportunity to provide cost efficient and integrated care, particularly among underserved populations experiencing barriers to mental health care, such as inadequate insurance, mental health stigma, and provider shortages. This systematic review aims to: 1) identify ACT-informed behavioral health interventions delivered by laypeople and 2) review the specific characteristics of each intervention including number and duration of sessions, delivery modality, interventionist training, and intervention outcomes. METHODS: Two databases (PubMed and PsycINFO) were systematically searched for relevant literature. To further identify relevant studies, references of included manuscripts were checked, the Association for Contextual Behavioral Science’s webpage was examined, and an email was sent to the ACBS Health Special Interest Group listserv. Study abstracts and full texts (in English) were screened, resulting in 23 eligible articles describing 19 different interventions. RESULTS: A total of 1,781 abstracts were screened, 76 were eligible for full-text review, and 23 were included in a narrative synthesis. There were 19 unique interventions identified and delivered by the following: general healthcare workers (n= 7), trained researchers (n = 5), women/mothers (n= 2), municipal workers (n= 2), and teachers (n = 3). Eleven studies were RCTs and eight utilized alternative study designs. Study quality varied, with two rated as high risk for bias and eight rated to have some concerns. Target populations included clinical and non-clinical samples. There was some consistency in the effects reported in the studies: increases in pain tolerance, acceptance, and identifying and engaging in value driven behavior, improvements in cognitive flexibility, and reductions in psychological distress. CONCLUSIONS: Findings suggest that ACT interventions can be successfully delivered by a variety of laypeople and effectively address psychological distress and increase health behaviors.

Heat stress affects oocyte developmental competence and is a major cause of reduced fertility in heat stressed cattle. Negative effects of heat stress on the oocyte have been observed at morphological, biochemical and developmental levels. However, the mechanisms by which heat stress affects the oocyte at the transcriptional and epigenetic levels remain to be further elucidated. Here we aimed to investigate the effect of heat stress on oocyte quality, transcriptomic profiles and DNA methylation of oocytes collected through the transition from spring to summer under Louisiana conditions. Summer season resulted in a lower number of high quality oocytes obtained compared to the spring season. There was no difference in in vitro maturation rates of oocytes collected during spring as compared to summer. RNA sequencing analysis showed that a total of 211 and 92 genes were differentially expressed as a result of heat stress in GV and MII oocytes, respectively. Five common genes (E2F8, GATAD2B, BHLHE41, FBXO44, and RAB39B) were significantly affected by heat in both GV and MII oocytes. A number of pathways were also influenced by heat stress including glucocorticoid biosynthesis, apoptosis signaling, and HIPPO signaling in GV oocytes, and Oct4 pluripotency, Wnt/beta-catenin signaling, and melatonin degradation I in MII oocytes. In addition, fluorescent immunocytochemistry analysis showed no difference in global levels of DNA methylation and DNA hydroxymethylation at either the GV or MII stage between spring and summer oocytes. The results of this study contribute to a better understanding of the effect of heat stress on the molecular mechanisms altered in bovine oocytes.

Capacity-oriented approaches to health interventions seek to empower the target population or community to manage the health issue themselves using resources they can control. Positive deviance, resilience and asset-based approaches are three such methods of developing and implementing health interventions. This study aimed to review the efficacy of interventions explicitly applying these methods in addressing childhood obesity using adiposity as the primary outcome, measured by standardized body mass index. The search strategy was developed and implemented across four electronic databases. Of the 181 records identified and screened, 11 studies were identified as using a capacity-oriented approach overall. Asset-based approaches (n = 8 studies) consisted of 47 880 participants, positive deviance (n = 2 studies) consisted of 781 participants, and resilience-based interventions (n = 1 study) consisted of 35 participants. The asset-based approaches were mixed, with three of the eight studies showing a significant reduction in adiposity, while the other five did not find a difference. The positive deviance and resilience-based studies showed signs of efficacy in reducing adiposity. There was significant design heterogeneity across studies, and varied interpretations and definitions of the approaches were used. Further work should attempt to achieve some consensus on the use of these approaches to facilitate comparison and advance the science of capacity-oriented interventions for childhood obesity.

Embryo vitrification involves exposure to high concentrations of cryoprotectants and osmotic stress during cooling and warming in the cryopreservation process. Many of these factors can potentially affect gene expression. In this study, invitro-produced bovine embryos at the blastocyst stage were subjected to vitrification. Four recipients each were used for transferring non-vitrified (n=80) and vitrified (n=80) embryos. A total of 12 non-vitrified and 9 vitrified viable day-14 (D14) embryos were recovered by uterine flushing. RNA-seq analysis of the whole embryo or isolated trophectoderm (TE) from vitrified and fresh recovered D14 embryos revealed a total of 927 and 4376 genes with changed expression in embryos and TE isolates, respectively, as a result of vitrification. In addition, we found 671 and 61 genes commonly up- or downregulated in both vitrified whole embryos and TE. Commonly upregulated pathways by vitrification included epithelial adherens junctions, sirtuin signalling, germ cell–sertoli cell junction, ATM signalling, NER and protein ubiquitination pathways. The commonly downregulated pathways included EIF2 signalling, oxidative phosphorylation, mitochondrial dysfunction, regulation of eIF4 and p70S6K signalling and mTOR signalling pathways. Our analysis identified specific pathways and implicated specific gene expression patterns affecting embryo developmental competence that are important to cryopreservation.

Introduction In developed countries, cardiovascular disease (CVD) is the leading cause of death. Identification and mitigation of CVD risk factors has been the chief strategy for prevention of CVD. To this end, the American Heart Association (AHA) and American College of Cardiology (ACC) suggest metrics of ideal cardiovascular (CV) health in seven domains and have published guidelines encouraging use of the “Simple Seven” to improve CV health. Sleep is notably absent from the Simple Seven despite substantial epidemiological evidence of its importance for CV health. We sought to measure the correlation of sleep parameters with CVD risk estimate by Framingham risk score (FRS). Methods Subjects prospectively enrolled in a CV Health Registry provided data for FRS, the seven domains of the “Simple Seven” including blood pressure, physical activity, total cholesterol, diet, weight, smoking, and glucose level, and completed validated questionnaires: Pittsburgh Sleep Quality Index (PSQI) and Berlin Questionnaire (BQ) for sleep apnea. Pearson correlations were analyzed between FRS and the “Simple Seven” as well as for the sleep questionnaires for comparison. Results Among the 646 subjects (mean age 55.4 ±9.9y, 43% men, 399W, 190B, 26H, 12A, 19other), FRS correlated substantially with blood glucose (R=0.398, p Conclusion This study suggests that sleep parameters, specifically BQ and PSQI, may contribute to risk assessment of CVD while also offering actionable information for improving CV health. Support

Capacity-oriented approaches to health interventions seek to empower the target population or community to manage the health issue themselves using resources they can control. Positive deviance, resilience and asset-based approaches are three such methods of developing and implementing health interventions. This study aimed to review the efficacy of interventions explicitly applying these methods in addressing childhood obesity using adiposity as the primary outcome, measured by standardized body mass index. The search strategy was developed and implemented across four electronic databases. Of the 181 records identified and screened, 11 studies were identified as using a capacity-oriented approach overall. Asset-based approaches (n = 8 studies) consisted of 47 880 participants, positive deviance (n = 2 studies) consisted of 781 participants, and resilience-based interventions (n = 1 study) consisted of 35 participants. The asset-based approaches were mixed, with three of the eight studies showing a significant reduction in adiposity, while the other five did not find a difference. The positive deviance and resilience-based studies showed signs of efficacy in reducing adiposity. There was significant design heterogeneity across studies, and varied interpretations and definitions of the approaches were used. Further work should attempt to achieve some consensus on the use of these approaches to facilitate comparison and advance the science of capacity-oriented interventions for childhood obesity.

While vitrification has become a valuable system used in oocyte and embryo preservation, there is still much to be learned in optimizing this protocol. Both mature and immature oocytes can be vitrified but each presents challenging aspects. Mature oocytes have microfilaments that are not yet developed in immature oocytes, which are fragile and may be disrupted by ice crystal formation during freezing. Further, currently many different cryoprotectants are used in different concentrations, most being combinations of dimethyl sulfoxide (DMSO), glycerol, and ethylene glycol. This study aimed to determine if vitrification solutions composed of ethylene glycol and either dimethyl sulfoxide or glycerol resulted in more-competent post-thaw oocytes, and to determine if maturation stage affected optimal vitrification solution. As validation of the IVF protocol, fresh mature oocytes from a commercial source were fertilized and proportion, with pronuclei formation 48 h post-IVF was recorded. Two experiments evaluated 2 cryoprotectant solutions by analysing post-vitrification and thaw competence of in vitro-fertilized oocytes to form pronuclei. Oocytes in both studies were exposed to 2 sequential vitrification solutions containing 10% DMSO or glycerol, 10% ethylene glycol and 0.5 M sucrose, and then 20% DMSO/glycerol and ethylene glycol and 0.5 M sucrose, before vitrification on cryolocks. In the first study, immature bovine oocytes (n = 200) were vitrified. Following thawing and IVM, they were analysed for pronuclei formation, with 8.49% and 0% fertilization following vitrification in DMSO and glycerol, respectively (P

BACKGROUND--The effect on subsequent respiratory function of spinal stabilisation for scoliosis in Duchenne muscular dystrophy is unclear. In order to clarify this clinical problem, changes in the forced vital capacity of a group of children with Duchenne muscular dystrophy who had undergone spinal surgery were measured and compared with a group of children with Duchenne muscular dystrophy who had not had surgery. METHODS--In this retrospective study 17 boys with Duchenne muscular dystrophy who underwent spinal stabilisation at a mean age of 14.9 years (surgical group) were compared with 21 boys with Duchenne muscular dystrophy who had not had surgery (non-surgical group). The mean (SD) Cobb angle of the surgical group at 14.9 years was 57 (16.4) degrees, and of the non-surgical group at 15 years was 45 (29.9) degrees. Forced vital capacity expressed as percentage predicted (% FVC) was measured in total over a seven year period in the surgical group and over 6.5 years in the non-surgical group, and regression equations were calculated. Survival curves for both groups were also constructed. RESULTS--No difference was found between spinal stabilisation (surgical group) and the non-surgical group in the rate of deterioration of % FVC which was 3-5% per year. There was no difference in survival in either group. CONCLUSIONS--Spinal stabilisation in Duchenne muscular dystrophy does not alter the decline in pulmonary function, nor does it improve survival.

Fungal (Penicillium chrysogenum) and yeast (Saccharomyces cerevisiae) ATP sulfurylases were shown to have very similar kinetic and chemical properties except that the fungal enzyme (a) contains a highly reactive Cys residue (SH-1) whose modification results in sigmoidal velocity curves (Renosto, F., Martin, R. L., and Segel, I. H. (1987) J. Biol. Chem. 262, 16279-16288) and (b) is allosterically inhibited by 3'-phosphoadenosine 5'-phosphosulfate (PAPS), while the yeast enzyme displays neither of these properties. The fungal enzyme subunit (64.3 kDa, 572 amino acids) is also larger than the yeast enzyme subunit (59.3 kDa, 521 amino acids). To correlate the unique allosteric properties of the fungal enzyme with specific structural features, we cloned and sequenced the ATP sulfurylase gene (aps) from P. chrysogenum. The yeast and fungal enzymes are homologous over the first 400 amino acids and contain two regions high in basic residues which are conserved in sulfurylases from Arabidopsis and the Riftia pachyptila (hydrothermal vent tube worm) chemolithotrophic symbiont. These regions may participate in forming the binding sites for MgATP2- and SO4(2-). The fungal enzyme has no sites for MgATP2- and SO4(2-). The fungal enzyme has no significant sequence homology to the yeast enzyme in the C-terminal 172 amino acids. This C-terminal region contains SH-1 (Cys-508) and has homology to MET14 (S. cerevisiae), CYSC (E. coli), and NODQ (Rhizobium meliloti), i.e. adenosine 5'-phosphosulfate (APS) kinase. The cumulative results suggest that (a) the allosteric PAPS binding site of P. chrysogenum ATP sulfurylase is located in the C-terminal domain of the protein and (b) that this domain may have evolved from APS kinase. In spite of the homology, this C-terminal region does not account for the APS kinase activity of P. chrysogenum. Fungal ATP sulfurylase has no significant homology to (or regulatory properties in common with) CYSD or CYSN, proteins reported to comprise E. coli ATP sulfurylase (Leyh, T., Vogt, T. F., and Suo, Y. (1992) J. Biol. Chem. 267, 10405-10410).

Three experiments were conducted to determine the effects of feeding diets containing laboratory synthesized deoxynivalenol (DON), or Fusarium (naturally) contaminated wheat containing DON on performance and health of young pigs (32.8–41.1 kg body weight). Diets containing 3.4–19.1 mg DON kg−1 were fed to pigs ad libitum for 14 d (exps. 1 and 2) or 7 wk (exp. 3) after which time each pig was slaughtered and blood, tissue and organ samples taken for analysis. All pigs responded to DON-contaminated diets with an immediate reduction in feed intake and body weight gain followed by some degree of recovery that was dependent on the dietary DON concentration. Negative linear relationships were found between feed intake or body weight gain and dietary DON concentration with pure DON and naturally contaminated wheat diets. Feed intake and weight gain were, respectively, 18 and 23% lower when a diet containing naturally contaminated wheat was fed compared to a diet containing an equivalent concentration of pure DON. Effects of DON on organ weights, relative to body weight, were inconsistent among experiments. The gastric mucosa was thinner and more eroded and had a higher degree of folding in the esophageal region in most groups fed DON-contaminated diets compared to those fed the control diet. Effects of DON on the fundic and cardiac regions of the stomach were inconsistent among treatments and experiments. There was little or no effect of DON on most of the blood parameters measured. Key words: Pure deoxynivalenol, naturally contaminated wheat, swine, intake, gain, vomitoxin, mycotoxin, fusarium, trichothecene

The dose limiting toxicities of the short infusion trial of the dacarbazine analog, CB10-277, were nausea and vomiting which appeared to be related to the peak plasma level of the parent drug. In addition, based on mouse studies, these dose limiting toxicities occurred at a less than optimal level of the monomethyl metabolite, the presumed species required for antitumour activity. An alternative schedule that would avoid the parent drug peak plasma levels of short infusion, while possibly allowing an increase in the amount of monomethyl metabolite produced was considered. Thus, a 24 h continuous infusion schedule, repeated every 21 days was explored. Twenty-two patients received 42 courses with a dose range of 4,700-15,000 mg m-2. The dose limiting toxicity was myelosuppression (leucopenia and thrombocytopenia). Although nausea and vomiting also occurred, it was manageable with routine antiemetic therapy. Other toxicities included diarrhoea, hallucinations, malaise, muscle ache, headache and flushing and all were < or = WHO grade 2. Pharmacokinetic studies were performed with 13 courses which included all dose levels. The mean t1/2 of the parent drug was 178 min. Area under the concentration x time curve (AUC) at the highest dose for the parent drug and the monomethyl metabolite were 2,350 and 9 mM x minutes, respectively. This monomethyl metabolite AUC and the associated myelosuppression showed a more favourable comparison to the preclinical data determined in mice than the results from the short infusion trial of CB10-277. Therefore, the recommended Phase II dose and schedule of this drug was 12,000 mg m-2 given by 24 h continuous infusion.

Decarbazine is an imidazole dimethyltriazene with reproducible activity in patients with metastatic melanoma. CB10-277 is a phenyl dimethyltriazene which, like dacarbazine, requires metabolic activation to its corresponding monomethyl species for antitumour activity. In preclinical models (human melanoma xenografts and transplantable rodent tumours) CB10-277 showed a similar spectrum and level of activity when compared to dacarbazine. Pharmacokinetic studies were performed with CB10-277 in mice treated i.v. at the LD10 (750 mg m-2) and plasma analysed by HPLC. The parent drug area under the plasma concentration vs time curve (AUC) was 142 mM x minutes. Drug metabolism occurred as evidenced by the HPLC identification of the monomethyl species (AUC = 8 mM x minutes) as well as other metabolites. A Phase I trial using a short infusion with doses repeated every 21 days has been performed. Thirty-six patients received 80 courses over a dose range of 80-6,000 mg m-2. The dose limiting toxicity was nausea and vomiting which occurred in 80% of the evaluable courses > or = 900 mg m-2. The only other common side effect was a flushing or warm sensation, which occurred in over 75% of courses at > or = 1,350 mg m-2. There were no hemodynamic consequences. Responses occurred in patients with melanoma (one complete, two partial, one mixed/11), sarcoma (one mixed/6) and carcinoid (one partial/l). Pharmacokinetics were performed in 46 courses. The CB10-277 AUC increased linearly with dose (r = 0.9203, P < 0.001) up to 700 mM x minutes at 6,000 mg m-2). Evidence of CB10-277 metabolism was observed, as in mice, by detection of the monomethyl species and other metabolites. However, the plasma levels of the monomethyl species in patients (1.8 and 3.7 mM x minutes at 6,000 mg m-2) were less than those predicted from studies in mice. Despite this, antitumour activity in dacarbazine sensitive histologies was observed and additional studies with CB10-277 are recommended.

Concentrations of cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn) were measured in adult barnacles (Elminius modestus Darwin) from Waitemata and Manukau Harbours in the Auckland area, New Zealand. As in studies on sediments reported in the literature, it was possible to identify areas of likely anthropogenic influence, e.g., around the Auckland Harbour Bridge for Pb, Cu, and Zn. Groups of individuals with highest concentrations for these metals showed 19.8–23.8 mg Pb kg−1, 198–266 mg Cu kg−1, and 4460–6530 mg Zn kg−1 (95% confidence limits, dry weight basis). Cd concentrations found for all barnacles from the Auckland area ranged from 0.8 to 3.1 mg kg−1. Two samples from Omaha Beach, 60 km north of Auckland, were used as a reference. Accordingly, groups of individuals with lowest concentrations for Pb, Cu, and Zn could be allocated to this site using the Student‐Newman‐Keuls Multiple Range Test (0.5–1.3 mg Pb kg−1, 8–10 mg Cu kg−1, and 144–214 mg Zn kg−1 ; 95% confidence limits). Only Cd conc...

Garlic has been used as a flavouring agent, traditional medicine, and functional food to improve physical or mental well-being. Garlic and garlic products generally have been regarded as safe but a number of conflicting reports in the literature and confounding factors make it difficult to unequivocally establish the clinical efficacy and safety of these products either alone or in the presence of therapeutic products. A preliminary study was undertaken with fresh garlic and garlic products using the major cDNA-expressed human cytochrome P-450 isozymes associated with the metabolism of HIV/AIDS drugs, and purified P-glycoprotein (P-gp) cell membranes to ascertain the risk potential for generating interactions with therapeutic products.A broad screening was undertaken with 10 garlic products (aged, odourless, oil, freeze-dried) and 3 varieties of fresh garlic bulbs (common, Elephant and Chinese), all purchased from local outlets, to examine their potential to affect human cytochrome P-450 2C9*1, 2C9*2, 2C19, 2D6, 3A4, 3A5 and 3A7 mediated-metabolism of marker substrates using an in vitro fluorometric microtiter plate assay. Four garlic products were screened for their potential to interact with P-gp using an in vitro colourmetric ATPase assay.Extracts of fresh garlic, different brands and lots of odourless garlic and representative samples of garlic oil, freeze dried garlic, and aged garlic exhibited an inhibitory effect on cytochrome P450 2C9*1, 2C19, 3A4, 3A5 and 3A7 mediated metabolism of a marker substrate. The activity of 2D6 mediated-metabolism was generally unaffected by garlic. Extracts of the fresh garlic stimulated CYP2C9*2 metabolism of the marker substrate. With the extracts tested, garlic had very low to moderate P-gp interaction as compared with the positive control verapamil.Our in vitro findings demonstrate that garlic components can affect cytochrome P-450 2C, 2D and 3A mediated-metabolism of the isoforms studied. The safety and efficacy of conventional therapeutic products metabolized by the affected isozymes, particularly those with a narrow therapeutic index, taken concomitantly with garlic needs to be examined further under clinical settings.

In patients with localized prostate cancer, radical prostatectomy and radiation therapy, although effective in controlling localized disease, are often associated with significant side effects attributable to injury of adjacent tissues. Moreover, patients with metastatic disease eventually fail systemic hormonal or chemotherapy because of the development of progressive, refractory disease. In this study, we evaluated the safety and efficacy of a novel suicide gene therapy that could potentially spare normal tissue while bypassing molecular mechanisms of apoptosis resistance by using chemically inducible effector caspases to trigger apoptosis in prostate cancer cells. Initially, we compared the ability of a panel of inducible Fas signaling intermediates to kill human and murine prostate cancer cell lines. On the basis of the superior killing by downstream caspase-1 and caspase-3, replication-deficient adenoviral vectors expressing conditional caspase-1 (Ad-G/iCasp1) or caspase-3 (Ad-G/iCasp3), regulated by nontoxic, lipid-permeable, chemical inducers of dimerization (CID), were constructed. Upon vector transduction followed by CID administration, aggregation and activation of these recombinant caspases occur, leading to rapid apoptosis. In vitro, both human (LNCaP and PC-3) and murine (TRAMP-C2 and TRAMP-C2G) prostate cancer cell lines were efficiently transduced and killed in a CID-dependent fashion. In vivo, direct injection of Ad-G/iCasp1 into s.c. TRAMP-C2 tumors caused focal but extensive apoptosis without evidence for a bystander effect at the maximal viral dose (i.e., 2.5 x 10(10) viral particles/25 microl) in host animals that also received CID compared with control animals. Treatment with Ad-G/iCasp1 plus CID resulted in a transient, yet significant, reduction both in tumor growth and volume compared with tumors treated with vector but not CID (P0.035) or vector-diluent plus CID (P0.022), both of which grew more rapidly. These results demonstrate that CID-regulated, caspase-based suicide gene therapy is safe and can inhibit the growth of experimental prostate cancer in vitro and in vivo through potent induction of apoptosis, providing a rationale for further development.

SR233377 is a novel thioxanthenone analogue that demonstrated solid tumor selectivity in vitro with activity confirmed in vivo against several murine tumors including those of colon, pancreas, and mammary origin. Its primary preclinical dose-limiting toxicities included myelosuppression and neurological toxicity. The neurological toxicity was acute and could be ameliorated in mice when the drug was administered as a 1-h infusion instead of rapid i.v. injection. As a result of its preclinical efficacy profile, SR233377 entered Phase I clinical investigation. The compound was administered i.v. over 2 h on day 1 repeated every 28 days. The starting dose was 33 mg/m2 (one-tenth the mouse LD10). Escalations continued to 445 mg/m2 (six escalations), where dose-limiting toxicity was observed. At this dose, acute ventricular arrhythmias, including one patient with torsades de pointes and transient cardiac arrest, occurred. Because this toxicity might have been related to the plasma peak, the protocol was amended to a 24-h infusion beginning at 225 mg/m2. With this dose, prolongation of the corrected QT interval (QTc) over the pretreatment levels resulted. Because prolonged QTc is a known forerunner to acute ventricular arrhythmias, clinical development of SR233377 was stopped. However, preclinical antitumor and toxicity studies with analogues are underway with hopes of identifying a new clinical candidate with similar antitumor effects that is devoid of cardiac toxic effects.

We have previously shown that antibodies to CTLA-4, an inhibitory receptor on T cells, can be effective at inducing regression of transplantable murine tumors. In this study, we demonstrate that an effective immune response against primary prostate tumors in transgenic (TRAMP) mice can be elicited using a strategy that combines CTLA-4 blockade and an irradiated tumor cell vaccine. Treatment of TRAMP mice at 14 weeks of age resulted in a significant reduction in tumor incidence (15% versus control, 75%), as assessed 2 months after treatment. Histopathological analysis revealed that treated mice had a lower tumor grade with significant accumulation of inflammatory cells in interductal spaces when treated with anti-CTLA-4 and a granulocyte-macrophage colony-stimulating factor-expressing vaccine. Vaccination of nontransgenic mice with this regimen resulted in marked prostatitis accompanied by destruction of epithelium, indicating that the immune response was, at least in part, directed against normal prostate antigens. These findings demonstrate that this combinatorial treatment can elicit a potent antiprostate response and suggest potential of this approach for treatment of prostate cancer.

We have characterized the temporal expression of the insulin-like growth factor (IGF) axis in the transgenic adenocarcinoma of mouse prostate (TRAMP) model as prostate cancer progression in this model closely mimics that observed in the human disease, and the model provides samples representing the earliest stages of prostate cancer that are clinically the most difficult to obtain. We report that prostate-specific IGF-I mRNA expression increased during prostate cancer progression in TRAMP mice and was elevated in the accompanying metastatic lesions, whereas prostatic IGF-I mRNA remained at nontransgenic levels in androgen-independent disease. Expression of IGF-II mRNA, however, was reduced in primary prostate cancer, metastatic lesions, and androgen-independent disease. Expression of type-1 IGF receptor (IGF1R) mRNA, encoding the cognate receptor for both IGF-I and IGF-II, as well as type-2 IGF receptor (IGF2R) mRNA was not found to be altered during primary prostate cancer progression in intact TRAMP mice but was dramatically reduced in metastatic lesions and in androgen-independent disease. Similar to reports from clinical disease, serum IGF-I levels were observed to increase precociously in TRAMP mice early in disease progression but remained at nontransgenic levels after castration. Elevated serum levels of IGF-binding protein 2 were observed to correlate with advanced prostate cancer in the TRAMP model. Together these observations implicate IGF-I as an important factor during the initiation and progression of primary prostate cancer and provide evidence that there is a strong selection against expression of IGF1R and IGF2R in metastatic and androgen-independent disease.

The studies presented herein quantitated ductal branching morphogenesis in the anterior prostate (AP) of the newborn rat. Four parameters were measured: epithelial area, epithelial perimeter, node number, and form factor. Nine natural and synthetic androgens were tested for their effectiveness in inducing postnatal prostatic development using 808 newborn rat APs in 68 dose-response experiments. Based on these studies it was shown that testosterone (T) was slightly more effective than dihydrotestosterone (DHT) in supporting ductal branching morphogenesis in the developing rat AP. Furthermore, the activity of T could not be accounted for simply by conversion of T to DHT. Synthetic androgens, 7alpha-methyl-19-nortestosterone and methyltrienolone (R1881), which cannot be 5alpha-reduced to DHT, also induced extensive ductal branching and elicited responses less than those to T and not statistically different from those to DHT. This suggests that although DHT is sufficient for prostatic development, it is not necessary for postnatal ductal branching morphogenesis and growth of the prostate. 5Alpha-androstan-3alpha,17beta-diol was particularly potent in inducing ductal branching, eliciting a response greater than or comparable to those of T and DHT. Androsterone, androstanedione, 5alpha-androstan-3beta,17beta-diol and 5beta-androstan-3alpha,17beta-diol induced ductal branching, but to a lesser extent than either T or DHT. These studies challenge the assumption that DHT is essential for prostatic development, specifically during ductal branching morphogenesis of the neonatal rat prostate.

Two new thioxanthenones, 183577 and 232759, have rekindled interest in the development of representatives from this class of structures as useful anticancer agents. Although the mechanism of action is unknown, both compounds demonstrated a similar spectrum of solid tumor selectivity. 232759 was selected for clinical development because it showed no hepatotoxicity in preliminary studies, whereas 183577 showed hepatotoxicity but only at the maximum tolerated dose (MTD). The limiting toxicity for the clinical candidate was myelosuppression in preliminary studies. Plasma and tissue drug levels, as well as protein binding, were studied in mice using optimal administration times at the MTD for each drug (for 183577, this was a 4-h infusion at 1350 mg/m2 and for 232759, it was a 5-min injection at 240 mg/m2), as well as at one-half the MTD for the clinical candidate. The drugs were 96-100% bound by plasma proteins. The peak drug concentrations, half-life, and area under the concentration-time curve in plasma for 183577 were 3483 ng/ml, 465 min, and 2018 microgram/ml. min, respectively. The peak drug concentration, half-life, and area under the concentration-time curve in plasma for 232759 were 5257 ng/ml, 44 min, and 276 microgram/ml. min, respectively, at the MTD and 2810 ng/ml, 40 min, and 110 microgram/ml. min at one-half the MTD. In all instances of simultaneous measurements, drug concentrations were equal or higher in tissues than they were in plasma. Unlike the plasma and kidney concentrations of 183577, the liver concentrations did not show a declining trend over the 8-h observation period. Declines in plasma, liver, kidney, and tumor levels of 232759 were detected over the 8-h observation period. The sustained high 183577 concentration in liver is believed to be responsible for its prolonged half-life and hepatotoxicity. Evidence for metabolism of the parent drugs was based on the finding of additional peaks on the high-pressure liquid chromatography tracings. Future studies will focus on identification and antitumor studies of these presumed metabolites in hopes of a better understanding of the solid tumor activity profiles and toxic effects of these compounds.

The purpose of this study was to determine how family physicians in the United States treat acute bronchitis in an otherwise healthy adult.A 33-item questionnaire on the diagnosis and treatment of acute bronchitis was mailed to a random sample of 500 physicians who are members of the American Board of Family Practice.Thirty-two of the 500 sampled physicians could not be located by mail; 265 of those who received the questionnaire responded. The response rate was 57% (265/468). Sixty-three percent of responding physicians indicated that antibiotics are their first choice of treatment for the otherwise healthy, nonsmoking adult with acute bronchitis. The decision to use antibiotics as the first choice of treatment did not vary by physician's sex, age, years in practice, practice location, practice type, or percentage of HMO patients. Only 6% of responding physicians reported using beta 2 agonist bronchodilators as their first choice of treatment. Physicians in this study stated that they prescribe an antibiotic 75% of the time in treating nonsmoking patients with acute bronchitis (first choice or otherwise). If the patient is a smoker, physicians reported that they prescribe antibiotics 90% of the time (F = 110.25; df = 1; P.0001). Physicians reported that for patients who smoke it takes longer for coughs to totally resolve and longer for them to return to a normal activity level than for nonsmokers.Family physicians report that antibiotics are their most common treatment for acute bronchitis in the otherwise healthy adult. Previous clinical trials have shown only marginal improvement in symptoms when patients with this condition are treated with an antibiotic. With antibiotic resistance emerging as a major global health problem, it is essential that other methods of treatment be evaluated.

The purpose of this study was to determine how family physicians in the United States diagnose acute bronchitis in otherwise healthy adults.A 33-item questionnaire on the diagnosis and treatment of acute bronchitis was mailed to a random sample of 500 physicians who are members of the American Board of Family Practice.Two hundred sixty-five physicians responded. Of those who did not respond, 32 could not be located. Of those who did respond, 10 were either retired or were practicing in another specialty. The net response rate was 56% (255/458). Responding physicians stated that character of cough and sputum production are the most important items used in diagnosing acute bronchitis. Fifty-eight percent indicated that the cough should be productive, and 60% stated that the sputum should be purulent. Seventy-two percent of respondents did not feel that wheezing or rhonchi need to be present. Younger physicians and those who selected antibiotics as their first treatment choice were more likely to define acute bronchitis as the presence of a productive cough with purulent sputum (P.05). Physicians from an academic setting were more likely to define acute bronchitis as a productive cough (P.05). Thirty-six percent of physicians from practices serving populations withor = 60% managed care patients included wheezing or rhonchi in the definition of acute bronchitis, compared with 26% of all others (P.05).Variations in the diagnosis of acute bronchitis in otherwise healthy adults can be attributed to physician age, treatment choice, and practice setting. A significant number of family physicians did not require a productive cough as part of the diagnostic criteria for acute bronchitis. This finding needs to be considered in studies evaluating treatment. Additional qualitative studies are necessary to identify other factors involved in diagnosing acute bronchitis.

To develop a syngeneic transplantable system to study immunotherapeutic approaches for the treatment of prostate cancer, three cell lines were established from a heterogeneous 32 week tumor of the transgenic adenocarcinoma mouse prostate (TRAMP) model. TRAMP is a transgenic line of C57BL/6 mice harboring a construct comprised of the minimal -426/+28 rat probasin promoter driving prostate-specific epithelial expression of the SV40 large T antigen. TRAMP males develop histological prostatic intraepithelial neoplasia by 8-12 weeks of age that progress to adenocarcinoma with distant metastases by 24-30 weeks of age. The three cell lines (TRAMP-C1, TRAMP-C2, and TRAMP-C3) express cytokeratin, E-cadherin, and androgen receptor by immunohistochemical analysis and do not appear to have a mutated p53. Although TRAMP-C1 and TRAMP-C2 are tumorigenic when grafted into syngeneic C57BL/6 hosts, TRAMP-C3 grows readily in vitro but does not form tumors. The T antigen oncoprotein is not expressed by the cell lines in vitro or in vivo. The rationale for establishing multiple cell lines was to isolate cells representing various stages of cellular transformation and progression to androgen-independent metastatic disease that could be manipulated in vitro and, in combination with the TRAMP model, provide a system to investigate therapeutic interventions, such as immunotherapy prior to clinical trials.

The pyrazoloacridine (PZA) analogue NSC366140 (PD115934) entered clinical trial based on unique preclinical characteristics including solid tumor selectivity in vitro, marked antitumor activity in vivo against murine solid tumors, selectivity against noncycling cells, and activity against multidrug-resistant tumor cells. After identification of the pre-clinical efficacy and an acceptable toxicity profile, a Phase I study of PZA was carried out. A total of 28 patients was entered and received a total of 67 treatment courses. The drug was administered via a 1-h infusion every 21 days. The starting dose was 30 mg/m2 with 2-fold dose escalations through 480 mg/m2. The next dose escalation was 50%, to 720 mg/m2. Grade I through grade IV toxicities were observed. Since no dose-limiting toxicities were observed at 480 mg/m2, and up to grade IV toxicities were observed at 720 mg/m2, an intermediate dose, 600 mg/m2, was evaluated. Dose-limiting toxicities at 720 mg/m2 were hematological (grade III and IV neutropenia) in four of six patients and neurological (up to grade III cerebral toxicities, including restlessness, dizziness, agitation/anxiety, personality changes, and nightmares, as well as myoclonus) in three of six patients treated. The pharmacokinetic parameters which helped predict these toxicities included area under the curve and peak plasma level. Pharmacokinetic studies showed interpatient variations in all parameters studied. The mean area under the curve levels of PZA at the highest two dose levels in patients were near the level detected in mice at their maximum tolerated total dose. The recommended starting dose for Phase II trials using this schedule is 600 mg/m2.

Pharmacokinetic studies that consisted of measuring the plasma drug profile, tissue drug distribution, and elimination in urine and feces were performed in female C57BL/6 x DBA/2 (hereafter called B6D2F1) and male B6D2F1A/2 and C57BL/6 x CH3 (hereafter called B6C3F1) mice following treatment with a 1-h i.v. infusion of the PZA, PD115934 (NSC 366140). This drug is the first of a new class of cytotoxic agents and was selected for clinical trials because of both its broad antitumor activity in vivo against murine solid tumors and human xenografts, and its in vivo toxicity profile that was predictable based on drug dose and schedule of administration. The pharmacokinetic results obtained here in mice have been used to facilitate the dose escalations during the Phase I trial and to determine pharmacokinetic drug exposure targets for its acute and sub-acute toxic effects. Plasma samples from three to four mice per time point were pooled, and then individual tissue samples from the same mice were collected at specified times following treatment. All samples were prepared using solid-phase extraction and assayed using high pressure liquid chromatography. The acute dose-limiting toxicity was neurological and occurred immediately after treatment at 300 mg/m2. The peak plasma level range at the acute maximum tolerated dose was 1040-1283 ng/ml. Thus, peak plasma levels1000 ng/ml were the acute toxicity target. Variations in the area under the plasma drug concentration x the time curve were observed that did not appear to be related to sex or age. The previously defined subacute dose-limiting toxicity was myelosuppression that occurred at a maximum tolerated dose of 600 mg/m2 (300 mg/m2 x 2) in B6D2F1 females. Thus, the area under the plasma drug concentration x the time curve in B6D2F1 females at this dose (1048 microg/ml x min) was the area under the plasma drug concentration x the time curve target. Drug levels were detected at 60 min following treatment in all tissues examined with a plasma:tissue ratio as high as 1:500. The organs with the highest levels were kidney, pancreas, liver, lung, and brain. Fecal excretion was low (range, 0.04-0.20% of the dose administered) and was not clearly different between males and females. Urinary excretion was higher (range, 5-28% of the dose administered) and did show evidence of sex-related differences, with male urinary drug excretion being higher than female urinary drug excretion. The drug was/=95% protein bound. Preliminary evidence for drug metabolism was found in urine and feces and will be further explored.

Fungal (Penicillium chrysogenum) and yeast (Saccharomyces cerevisiae) ATP sulfurylases were shown to have very similar kinetic and chemical properties except that the fungal enzyme (a) contains a highly reactive Cys residue (SH-1) whose modification results in sigmoidal velocity curves (Renosto, F., Martin, R. L., and Segel, I. H. (1987) J. Biol. Chem. 262, 16279-16288) and (b) is allosterically inhibited by 3'-phosphoadenosine 5'-phosphosulfate (PAPS), while the yeast enzyme displays neither of these properties. The fungal enzyme subunit (64.3 kDa, 572 amino acids) is also larger than the yeast enzyme subunit (59.3 kDa, 521 amino acids). To correlate the unique allosteric properties of the fungal enzyme with specific structural features, we cloned and sequenced the ATP sulfurylase gene (aps) from P. chrysogenum. The yeast and fungal enzymes are homologous over the first 400 amino acids and contain two regions high in basic residues which are conserved in sulfurylases from Arabidopsis and the Riftia pachyptila (hydrothermal vent tube worm) chemolithotrophic symbiont. These regions may participate in forming the binding sites for MgATP2- and SO4(2-). The fungal enzyme has no sites for MgATP2- and SO4(2-). The fungal enzyme has no significant sequence homology to the yeast enzyme in the C-terminal 172 amino acids. This C-terminal region contains SH-1 (Cys-508) and has homology to MET14 (S. cerevisiae), CYSC (E. coli), and NODQ (Rhizobium meliloti), i.e. adenosine 5'-phosphosulfate (APS) kinase. The cumulative results suggest that (a) the allosteric PAPS binding site of P. chrysogenum ATP sulfurylase is located in the C-terminal domain of the protein and (b) that this domain may have evolved from APS kinase. In spite of the homology, this C-terminal region does not account for the APS kinase activity of P. chrysogenum. Fungal ATP sulfurylase has no significant homology to (or regulatory properties in common with) CYSD or CYSN, proteins reported to comprise E. coli ATP sulfurylase (Leyh, T., Vogt, T. F., and Suo, Y. (1992) J. Biol. Chem. 267, 10405-10410).

Androgen-dependent male urogenital development occurs via mesenchymal-epithelial interactions in which mesenchyme induces epithelial morphogenesis, regulates epithelial proliferation, and evokes expression of tissue-specific secretory proteins. Mesenchymal-epithelial interactions continue to be important into adulthood. For example, mesenchyme of the urogenital sinus (UGM) and seminal vesicle (SVM) induce dramatic morphologic and functional changes in various adult epithelia. Since adult epithelial cells are unquestionably responsive to mesenchymes that can elicit expression of alternative morphologic and functional phenotypes, established carcinomas might also be influenced by their connective tissue environment. In this regard, Dunning prostatic tumor has been induced by UGM or SVM to differentiate into tall columnar secretory epithelial cells. This change in cytodifferentiation is associated with a reduction in growth rate and loss of tumorigenesis. The role of soluble growth factors in the mechanism of mesenchymal-epithelial interactions is discussed.

The pharmacokinetics of the anthrapyrazole CI-941 has been investigated in conjunction with the Phase I evaluation of the drug with the intent of applying a pharmacokinetically guided dose escalation strategy. A starting dose of 5 mg/m2 was chosen, based on one-tenth the 10% lethal dose in mice. Due to the steep dose lethality relationship and nonlinear pharmacokinetics in mice, a target area under the CI-941 plasma concentration x time curve (AUC) of 110 microM x min (i.e., 40% of the mouse 10% lethal dose AUC) was chosen. This AUC was achieved in mice at 40 mg/m2. A total of 37 patients received 74 courses of CI-941 (5 to 55 mg/m2), with 26 patients consenting to pharmacokinetic monitoring. CI-941 was rapidly cleared from plasma, and a triexponential open model could be fitted to the data (t1/2 alpha = 7.6 +/- 2 min, t1/2 beta = 65 +/- 27 min, t1/2 zeta = 21 +/- 9 h). CI-941 was subjected to only limited urinary elimination, accounting for 5.2 +/- 2.8% of the administered dose. Wide interpatient variability in plasma CI-941 clearance at the starting dose and subsequent doses precluded the implementation of a pharmacokinetically guided dose escalation scheme, and the dose was escalated in 5-mg/m2 increments until the maximally tolerated dose was achieved. A number of investigations were performed to study potential reasons for variability in CI-941 clearance. However, CI-941 plasma protein binding (95 +/- 1%) and measures of pretreatment renal (51Cr-EDTA clearance), hepatic (plasma alanine transaminase and alkaline phosphatase levels), or cardiac function (left ventricular ejection fractions) did not relate strongly to CI-941 clearance. In patients treated at 40 mg/m2, the AUC values (156 to 415 microM x min) approximated or exceeded the target AUC. Fifty mg/m2 was the Phase II recommended dose. Further prospective studies are warranted to assess the utility of pharmacokinetically guided dose escalation strategies and to determine whether or not the variability encountered in clearance is unique to CI-941.

The estrogen receptor (ER) is a nuclear protein with a hormone- and a DNA-binding domain. We examined the DNA binding of ER in MCF-7 cells and 79 primary breast cancers by gel mobility shift assay using as a probe the estrogen response element (ERE). The mobility shift assay showed saturable, specific binding of ER to ERE in crude, high molar extracts containing greater than or equal to 4 mg/ml protein. Nonspecific binding was reduced by increasing concentrations of poly(deoxyinosidylate.deoxycytidylate) and shortening of the ERE probe from 35 to 15 base pairs. In the presence of Mg2+ the ER-ERE complex formation was hormone dependent at 22 degrees C but not at 37 degrees C. In the absence of Mg2+ estradiol was not necessary for ER-ERE complex formation. Correlation of the mobility shift assay with the hormone-binding (E2) assay showed agreement in 55 of the 79 tumors. Both assays were positive (E2 +/ERE+) in 35 cases and both were negative (E2-/ERE-) in 20 cases. In 11 tumors the hormone-binding assay was positive and the mobility shift assay negative (E2+/ERE-), suggesting an alteration of the DNA-binding domain. In 13 cancers the hormone-binding assay was negative and the mobility shift assay positive (E2-/ERE+) suggesting an alteration of the hormone-binding domain. By performing both hormone- and DNA-binding assays of ER and the hormone-binding assay of progesterone receptor (PR), we found the following subgroups of breast cancer: (a) E2+/ERE+/PR+, (b) E2+/ERE+/PR-, (c) E2+/ERE-/PR+, (d) E2+/ERE-/PR-, (e) E2-/ERE+/PR+, (f) E2-/ERE+/PR-, (g) E2-/ERE-/PR-. The simultaneous determination of 17 beta-estradiol and ERE binding may provide a better definition of the ER status of individual tumors and prove useful in refining endocrine therapy of patients with breast cancer.

The optimization of a simple, sensitive procedure using a chick embryotoxicity screening test (CHEST) bioassay for detection of toxic compounds is presented. Dosing protocols of eggs, using several mycotoxins (aflatoxin B„ deoxynivalenol, T-2 toxin) and appropriate controls, were evaluated for embryonic sensitivity, overall practicality of the procedure, and consistency of results. It was found that both type of carrier solvent and volume injected could significantly affect overall embryonic mortality. The chick embryo was most sensitive to the effects of toxins and solvents after 1 or 2 days of incubation; a rapid decrease in response was observed as the age of the embryo at dosing increased. Following administration of the toxins just below the shell membrane by way of a small hole (

Fifty-four barrows (27.5 ± 0.5 kg) were given ad libitum for 7 wk one of 18 diets prepared from field corn inoculated with different strains of Fusarium graminearum, naturally contaminated corn and wheat, pure deoxynivalenol (DON), and clean corn. The effect of 4.7 mg pure DON kg−1 feed was not significantly different from that of the control, whereas pigs on several diets at a similar DON level had significantly lower feed intake and weight gain values. There was no correlation between DON concentration in the diets, amount ingested and pig performance. Key words: Vomitoxin, deoxynivalenol, pigs, inoculated corn, Fusarium, mycotoxins

Twelve species of barnacles were identified from the fouling community on the parts of the ‘Maui’ oil platform that were submerged during its tow from Japan in 1975 and after its arrival in New Zealand. The stalked barnacles Lepas anatifera L. var. (a) Darwin, L. anserifera L., Conchoderma auritum (L.), and C. virgatum (Spengler) probably settled during the tow across the tropical Pacific, and have been recorded in New Zealand waters before from ships (the Lepas spp.) and vertebrates (the Conchoderma spp.). The acorn barnacles were small, and probably settled in Japanese waters; Balanus variegatus Darwin and B. amphitrite Darwin already occur in northern New Zealand waters. The other six species—B. improvisus Darwin, B. albicostatus Pilsbry, B. reticulatus Utinomi, Megabalanus volcano (Pilsbry), M. rosa (Pilsbry), and Tetraclita squamosa japonica Pilsbry—have not been recorded from New Zealand before.

Diets containing deoxynivalenol (DON) prepared from Fusarium inoculated corn and Luprosil® NC, a propionate-based feed preservative, were offered to 34-kg barrows for 7 wk. The preservative improved overall performance of pigs fed the control or DON-containing diets, but did not appear to specifically reduce the adverse effects of DON. Key words: Deoxynivalenol, pigs, feed preservative, propionate, Fusarium graminearum

Fifteen species of scalpellid barnacles were collected between 1974 and 1979 by dredging to a maximum depth of 2515 m on the New Zealand shelf. The species are assigned to genera following recent proposals by other authors. Graviscalpellum new genus is proposed for the hermaphroditic arcoscalpellids. Graviscalpellum persona (Annandale) is a new record for New Zealand seas. A fuller sequence of ontogeny is described for GymnoscaJpellum intermedium (Hoek), with a proposal that Gymnoscalpellum and Litoscalpellum are synonymous. A key to the New Zealand scalpellid barnacles is given.

Two related ways to invariantly study higher derivatives, new-tensors and derivative strings, are compared, and the idea of suppression operator, or multiplier, is introduced. It renders the formal expression of the new objects almost as simple as that of ordinary tensors. Jacobians as a subgroup of the full second-order group are identified, and the harmonic subgroup leaving Laplace’s equation invariant is determined. The wider principle of general covariance entailed by the full group is considered. In particular, electrogravitic potentials are defined, and the exploration of their field theory begun.

Species composition and biomass of plankton samples taken on transects across an upwelling plume off Farewell Spit in February 1981 are presented. Copepods were numerically dominant, particularly Oithona similis and Paracalanus indicus. Also abundant were large phytoplankters (Chaetoceros sp. and Trichodesmium sp.), crustacean faecal pellets, and euphausiid larvae. The distribution of zooplankton species suggests mixing of inshore waters and neritic populations with upwelled waters. These data are discussed with respect to an interpretation of the upwelling system as a northwards transport of zooplankton and its enhanced productivity into the South Taranaki Bight.

Cyclosporin A was produced in a low foaming semi-synthetic media by carrageenan-entrappedTolypocladium inflatum in an airlift bioreactor with an external circulation loop. The cyclosporin A recovered by a single ethyl acetate extraction was essentially free of contaminating media and endogenously produced microbial products.