Recent changes in reproductive phenology of a K-selected aquatic insect predator, Belostoma flumineum Say (Heteroptera, Belostomatidae).

The timing of critical events like mating, migration, and development has noticeably and recently shifted in many populations of diverse organisms. Here, we report a change in the breeding phenology of giant waterbugs, Belostoma flumineum Say (Heteroptera, Belostomatidae), in the northeastern United States. Waterbugs collected in 2005 and 2006 exhibited previously typical patterns of mating and reproduction: two annual reproductive peaks in which overwintered adults mated in the spring and young adults from a new generation mated in the fall. In 2012 and 2015, despite similar sampling effort, we detected no fall breeding activity in the study area. Reproductive behaviour under controlled laboratory conditions was also different between the earlier (2005 and 2006) and recent (2012 and 2015) years: waterbugs collected in recent years exhibited significant delays in reproduction (>30 days) under similar photoperiod and thermal conditions. We discuss potential causes of this dramatic change in reproductive behaviour, such as climate change, as well as possible negative impacts of the absence of fall reproduction on populations of B. flumineum in the study region.

Pre-BCR signaling in precursor B-cell acute lymphoblastic leukemia regulates PI3K/AKT, FOXO1 and MYC, and can be targeted by SYK inhibition.

Precursor-B-cell receptor (pre-BCR) signaling and spleen tyrosine kinase (SYK) recently were introduced as therapeutic targets for patients with B-cell acute lymphoblastic leukemia (B-ALL), but the importance of this pathway in B-ALL subsets and mechanism of downstream signaling have not fully been elucidated. Here, we provide new detailed insight into the mechanism of pre-BCR signaling in B-ALL. We compared the effects of pharmacological and genetic disruption of pre-BCR signaling in vitro and in mouse models for B-ALL, demonstrating exquisite dependency of pre-BCR(+) B-ALL, but not other B-ALL subsets, on this signaling pathway. We demonstrate that SYK, PI3K/AKT, FOXO1 and MYC are important downstream mediators of pre-BCR signaling in B-ALL. Furthermore, we define a characteristic immune phenotype and gene expression signature of pre-BCR(+) ALL to distinguish them from other B-ALL subsets. These data provide comprehensive new insight into pre-BCR signaling in B-ALL and corroborate pre-BCR signaling and SYK as promising new therapeutic targets in pre-BCR(+) B-ALL.

Selective, novel spleen tyrosine kinase (Syk) inhibitors suppress chronic lymphocytic leukemia B-cell activation and migration.

Syk is a protein tyrosine kinase that couples B-cell receptor (BCR) activation with downstream signaling pathways, affecting cell survival and proliferation. Moreover, Syk is involved in BCR-independent functions, such as B-cell migration and adhesion. In chronic lymphocytic leukemia (CLL), Syk becomes activated by external signals from the tissue microenvironment, and was targeted in a first clinical trial with R788 (fostamatinib), a relatively nonspecific Syk inhibitor. Here, we characterize the activity of two novel, highly selective Syk inhibitors, PRT318 and P505-15, in assays that model CLL interactions with the microenvironment. PRT318 and P505-15 effectively antagonize CLL cell survival after BCR triggering and in nurse-like cell-co-cultures. Moreover, they inhibit BCR-dependent secretion of the chemokines CCL3 and CCL4 by CLL cells, and leukemia cell migration toward the tissue homing chemokines CXCL12, CXCL13, and beneath stromal cells. PRT318 and P505-15 furthermore inhibit Syk and extracellular signal-regulated kinase phosphorylation after BCR triggering. These findings demonstrate that the selective Syk inhibitors PRT318 and P505-15 are highly effective for inhibition of CLL survival and tissue homing circuits, and support the therapeutic development of these agents in patients with CLL, other B-cell malignancies and autoimmune disorders.

In vitro internalization, intracellular transport, and clearance of an anti-CD11a antibody (Raptiva) by human T-cells.

Efalizumab (Raptiva) is a humanized CD11a-specific monoclonal antibody that was recently approved for the treatment of moderate to severe psoriasis. In psoriasis patients, the rate of efalizumab clearance from serum is related to T-cell surface expression of CD11a, suggesting a receptor-mediated clearance model for efalizumab (Bauer et al., 1999). However, limited experimental data are available to explain how the interaction with CD11a results in the systemic clearance of efalizumab. The following studies were designed to test the hypothesis that one mechanism of anti-CD11a antibody clearance is mediated in part by cellular internalization. This was tested in vitro using purified mouse and human T-cells as a model to study the cellular uptake and clearance of anti-CD11a antibodies. Data from these studies suggest that anti-CD11a antibodies are internalized by purified T-cells. Upon internalization, the antibodies appeared to be targeted to lysosomes and were cleared from within the cells in a time-dependent manner. CD11a-mediated internalization and lysosomal targeting of efalizumab may constitute one pathway by which this antibody is cleared in vivo.

An alternate form of Ku80 is required for DNA end-binding activity in mammalian mitochondria.

Mammalian mitochondrial DNA end-binding activity is nearly indistinguishable from that of nuclear Ku. This observation led to the hypothesis that mitochondrial DNA end-binding activity is in part dependent upon Ku80 gene expression. To test this hypothesis, we assayed for Ku activity in mitochondrial extracts prepared from the xrs-5 hamster cell line that lacks Ku80 mRNA expression. Mitochondrial protein extracts prepared from this cell line lacked the DNA end-binding activity found in similar extracts prepared from wild-type cells. Azacytidine-reverted xrs-5 cells that acquired nuclear DNA end-binding activity also acquired mitochondrial DNA end-binding activity. Western blot analysis of human mitochondrial protein extracts using a monoclonal antibody specific for an N-terminal epitope of Ku80 identified a protein with an apparent molecular weight of 68 kDa. This mitochondrial protein was not detected by a monoclonal antibody specific for an epitope at the C-terminal end of Ku80. Consistently, while both the N- and C-terminal Ku80 monoclonal antibodies supershifted the nuclear DNA end-binding complex on an electrophoretic mobility shift assay, only the N-terminal monoclonal antibody supershifted the mitochondrial DNA end-binding complex. To confirm that the 68 kDa Ku protein was not a consequence of nuclear protein contamination of mitochondrial preparations, highly purified intact nuclei and mitochondria were treated with proteinase K which traverses the pores of intact nuclei but gains limited access into intact mitochondria. Ku80 in purified intact nuclei was sensitive to treatment with this protease, while the 68 kDa Ku protein characteristic of purified intact mitochondria was resistant. Further, immunocytochemical analysis revealed the co-localization of the N-terminal specific Ku80 monoclonal antibody with a mitochondrial-targeted green fluorescence protein. Mitochondrial localization of the C-terminal Ku80 monoclonal antibody was not observed. These data are consistent with the hypothesis that a C-terminally truncated form of Ku80 is localized in mammalian mitochondria where it functions in a DNA end-binding activity.

Human chromosomal localization, tissue/tumor expression, and regulatory function of the ets family gene EHF.

Ets factors are members of an ancient multigene family of transcription factors including oncoproteins and possibly tumor suppressors. We previously characterized a novel divergent ets gene, Ehf (ets homologous factor) in mice. Here we report the cDNA sequence, chromosomal location, and tissue/tumor expression patterns of the human EHF gene and the regulatory activity of the EHF protein. EHF maps to 11p12, which is deleted in many prostate, breast, and lung carcinomas and is a hot spot for inherited deletion- or amplification-associated developmental defects. EHF is differentially expressed in normal tissues and carcinomas and between tumor stages and is most highly expressed in the organs known to form carcinomas upon 11p12 deletion. EHF protein represses the ETS-2 induced activity of both stromelysin-1 and collagenase-1 promoters. These data suggest that EHF may contribute to human development and carcinogenesis and is a candidate for the 11p12 tumor suppressor gene.

Mammalian mitochondrial extracts possess DNA end-binding activity.

Mammalian mitochondrial protein extracts possess DNA end-binding (DEB) activity. Protein binding to a 394 bp double-stranded DNA molecule was measured using an electrophoretic mobility shift assay. Mitochondrial DEB activity was highly specific for linear DNA. Inclusion of a vast excess of non-radioactive circular DNA did not disrupt binding to radioactive f394. In contrast, binding was abolished by the inclusion of linear competitor DNA. In mammals, nuclear DEB activity is due to Ku, a hetero-dimer composed of the Ku70 and Ku86 proteins. To determine whether mitochondrial DEB activity was also due to Ku, protein extracts were prepared from the Chinese hamster XR-V15B cell line, which lacks this protein. As anticipated, nuclear extracts prepared from these cells lacked DEB activity. In contrast, mitochondrial extracts prepared from these cells had wild-type levels of DEB activity, demonstrating that this latter activity is not a consequence of nuclear contamination. Although the nuclear and mitochondrial DEB activities are independent of each other, they are nevertheless closely related, since mitochondrial DEB activity was 'supershifted' by both anti-Ku70 and anti-Ku86 antisera. The nuclear DEB protein Ku plays an essential role in nuclear DNA double-strand break repair. The DEB activity described herein may therefore play a similar role in mitochondrial DNA repair.

Polymicrobial brain stem abscess due to Streptococcus anginosus and Actinomyces species.

This report describes a rare case of a pyogenic brain stem abscess. The lesion was readily identified by computed tomographic (CT) scan and magnetic resonance imaging (MRI). Streptococcus anginosus was grown from the pus and Actinomyces colonies were found on histopathology. The lesion was successfully treated with stereotactic surgery and antibiotic therapy. A review of the relevant literature including the role of stereotaxy in the treatment of the brain stem abscess is discussed. Copyright 1999 Harcourt Publishers Ltd.

Lowering the language barrier in an acute psychiatric setting.

OBJECTIVES: The aim of this paper is to investigate the effectiveness of a training and policy strategy to improve communication opportunities in an acute inpatient unit for patients of non-English-speaking background (NESB) with low English proficiency. METHOD: A pre- and post-intervention design involved: (i) a survey of the multilingual skills of 80 clinical staff; (ii) recording of patients' ethnic background and proficiency; (iii) pre- and post-intervention data collection of the main outcome measure (communications with patients in a language other than English [LOTE]); and (iv) staff training, and active encouragement, in interpreter use. English proficiency was assessed using the population census proficiency question. RESULTS: Of 257 admissions, 33% were of NESB and 19% preferred to speak a LOTE. The staff survey yielded a 49% return rate and showed that, of 11 LOTEs spoken by patients, seven were also spoken by 17 of the staff. Twenty-nine percent of staff were not clinically proficient in these languages. Compared to the NESB population, a higher proportion of NESB patients rated low on proficiency. Following the intervention, interpreter bookings and booking duration increased significantly. CONCLUSIONS: A standard training package and a policy promoting interpreter use improved communication opportunities in an acute setting where language needs are typically poorly met. Failure to ensure effective communicate raises risks of misdiagnosis and inappropriate treatment. By measuring patients' proficiency directly, the present study identified a higher level of need for interpreter services than estimated by past reports.

Time-dependent changes in cocaine-seeking behavior and extracellular dopamine levels in the amygdala during cocaine withdrawal.

Cocaine and cocaine-associated cues elicit craving in addicts and reinstate cocaine-seeking behavior in rats. Craving and cocaine-seeking behavior may be mediated by withdrawal-induced changes in dopamine (DA) neurotransmission in the amygdala. To examine whether there are concomittant changes in cocaine-seeking behavior and extracellular DA levels during withdrawal, experimental rats were trained to self-administer cocaine (0.75 mg/kg i.v.). After 14 daily 3-hour training sessions, animals underwent either a 1-day, 1-week, or 1-month withdrawal period. Extracellular DA levels were assessed during baseline, extinction, cue reinstatement, and cocaine (15 mg/kg i.p.) reinstatement of cocaine-seeking behavior (i.e., defined as the difference in nonreinforced lever presses on an active minus inactive lever). Cocaine-seeking behavior became more intense during the course of cocaine withdrawal. Additionally, basal and cocaine-induced extracellular DA levels were enhanced after the 1-month withdrawal period. We suggest that the former may reflect a persistent elevation in tonic extracellular DA levels in the amygdala, whereas the latter may reflect a persistent elevation in phasic extracellular DA levels.

Amphotericin B colloidal dispersion for treatment of candidemia in immunocompromised patients.

The efficacy and safety of amphotericin B colloidal dispersion (ABCD; Amphotec, Sequus Pharmaceuticals, Menlo Park, CA), a lipid complex of amphotericin B, were evaluated in immunocompromised patients with candidemia. These patients were recruited from five open-label clinical trials of ABCD therapy for fungal infections subsequent to bone marrow transplantation, hematologic malignancies, solid tumors, solid-organ transplantation, or other severe underlying disorders. ABCD was given intravenously in a median daily dose of 3.9 mg/kg for < or =72 days. Response rates were as follows: 53% overall (n = 88), 66% for patients with candidemia alone (n = 67), and 14% for patients with disseminated candidemia (n = 21). Nephrotoxicity occurred in 16% of patients, with either doubling of the baseline serum creatinine level or an increase of > or =1 mg/dL or a > or =50% decrease in calculated creatinine clearance. On average, there were no significant changes in the levels of serum creatinine or bilirubin from baseline to the end of treatment. In conclusion, ABCD was safe and effective for treating immunocompromised patients with candidemia.

Assessing case records using monitor criteria developed from consensus guidelines.

OBJECTIVE: To develop criteria to enable the monitoring of general practitioner (GP) case records using consensus guidelines for conditions commonly managed in general practice and to measure how well the case records of a non random sample of GPs conformed to these criteria. METHOD: An iterative process was used to develop criteria from consensus guidelines for 19 conditions. Criteria were also developed to enable monitoring of the structure and content of the patient case record. A non random sample of GPs in Adelaide was approached to allow measurement of the content of their case records against these criteria. This measurement was undertaken by allied health professionals. An overall percentage score of conformity with the criteria was created for 10 acute, and six chronic conditions and for the patient case record review. These were rank ordered and Kendall's rank order correlation coefficients were used to compare the results in these three areas of practice. RESULTS: Criteria were successfully developed for each condition. Thirty-one GPs had their patient case records assessed. There was substantial variability between these practitioners in their conformity to the criteria. Kendall's rank order coefficients found statistically significant correlation between the results for acute and chronic conditions, and between acute conditions and the patient case record review section. CONCLUSION: It is feasible to develop criteria that enable measurement of the conformity of GP case records to these criteria. The overall level of conformity, together with the substantial variability found between practitioners suggest that there is a need for GPs to address this area of their practice.

Development of consensus guidelines for conditions commonly managed in general practice. Royal Australian College of General Practitioners.

OBJECTIVE: Initially, the project objective was to develop audit protocols for commonly managed conditions in general practice. However, as the nature of the project evolved these objectives changed to developing consensus guidelines. METHOD: An iterative process was undertaken using mail-outs to gain items for potential inclusion in an audit protocol. This resulted in a change of direction to management guidelines. Consensus, as to the validity of inclusion of items in each guideline, was then achieved by continuing to use an iterative process. RESULTS: Guidelines for 30 presenting complaints were developed. Because a number of differential diagnoses were possible for each complaint, a total of 105 guidelines were developed. A total of 263 general practitioners participated. CONCLUSION: The concept of discriminant actions was developed. It was agreed that this described the clinical decision making that occurs in the general practices of participating practitioners. It is feasible to use an iterative method undertaken by mail to develop consensus guidelines for conditions that are commonly managed by general practitioners.

Reported diabetes mellitus management among South Australian general practitioners.

Diabetes mellitus is a chronic disease commonly managed in general practice. This article reviews the findings of a questionnaire survey of 173 randomly chosen South Australian general practitioners and their reported management in the areas of detection, diagnosis, assessment, monitoring and GP knowledge. Substantial differences between respondents were found in all areas, suggesting the need for improved educational methods aimed at all general practitioners.

Knowledge of and reported asthma management among South Australian general practitioners.

AIM: This study, carried out in 1989, set out to assess general practitioners' knowledge of asthma management and their reported management practices. METHOD: Of 153 randomly selected South Australian general practitioners 127 (83%) completed a questionnaire designed to explore issues relating to the management of asthma. RESULTS: The survey revealed substantial differences between general practitioners in their knowledge and management practices including the assessment of the severity of asthma, the need for objective monitoring, that is by the use of spirometry and peak flow meters, and the use of medication. Overall, the sampled general practitioners believed that patient-related factors were the main barriers to effective treatment of asthma. CONCLUSION: The findings of this study suggest that ideal asthma management was not being attained. More research is required to ascertain why such variability among practitioners exists and how best to remedy these differences.

A pattern of decline in general practice obstetric care in Adelaide.

Some educational, training and politico-economic reasons for the decline in GP obstetrics in Adelaide, South Australia, are described. It is recommended that similar studies be done in other centres in Australia to evaluate the general nature of the findings.

The effect of prazosin on patients with symptoms of benign prostatic hypertrophy.

OBJECTIVE: To evaluate the effect of prazosin on the symptoms of benign prostatic hypertrophy for patients of general practitioners. SETTING AND PATIENTS: Eighty-two general practitioner patients from 317 who were identified as being eligible for inclusion in the trial. They were randomly allocated to receive either prazosin or placebo for the first 3 month arm then crossed-over for the second 3 months. RESULTS: Seventy-six patients completed the trial. An analogue scale of patient rating of symptoms showed a statistically significant decrease in symptoms for patients taking prazosin when compared with patients taking placebo, regardless of the order in which they had been administered. A total symptom score and an obstructive symptom score for benign prostatic hypertrophy showed a statistically significant difference between the two treatments at 4 weeks. A difference was still apparent at 8 and 12 weeks but it was not statistically significant. No difference was detected for irritative symptoms. CONCLUSION: Prazosin may have a limited place in the management of benign prostatic hypertrophy.