Experimental Trypanosoma cruzi biclonal infection in Triatoma infestans: detection of distinct clonal genotypes using kinetoplast DNA probes

Monitored biclonal densities of parasites were offered to third-stage larvae of Triatoma infestans via an artificial feeding device and 30 days later, the gut contents of the insects were processed for microscopic examination and polymerase chain reaction (PCR) detection of Trypanosoma cruzi kinetoplast DNA [kDNA]). A total of 15 mixtures involving nine different stocks attributed to the 19/20, 32 and 39 major clonal genotypes of Trypanosoma cruzi were used. The presence of each T. cruzi clonal genotype after completion of the cycle through the insects was investigated by hybridising the PCR amplification products with genotype-specific minicircle kDNA probes. Sixty-five out of 90 examined insects (72.2%) were positive for parasites by microscopic examination and 85 (94.4%) were positive by PCR. The results show that almost half of the biclonal infections are not detectable after completion of the cycle, and that there are important differences in detection of such biclonal infections according to the clonal genotypes considered. Moreover, elimination of a clonal genotype by another is a frequent, but not constant, pattern in biclonal infections of T. infestans. The use of PCR and kDNA probes makes it possible to avoid the culture phase, which makes detection of mixed infections much easier in epidemiological surveys. Moreover, the fact that T. infestansdoes not transmit different T. cruzi clonal genotypes with the same efficiency has strong implications for the reliability of xenogiagnosis.     

Long-term inhibition by auxin of leaf blade expansion in bean and Arabidopsis

The role of auxin in controlling leaf expansion remains unclear. Experimental increases to normal auxin levels in expanding leaves have shown conflicting results, with both increases and decreases in leaf growth having been measured. Therefore, the effects of both auxin application and adjustment of endogenous leaf auxin levels on midrib elongation and final leaf size (fresh weight and area) were examined in attached primary monofoliate leaves of the common bean (Phaseolus vulgaris) and in early Arabidopsis rosette leaves. Aqueous auxin application inhibited long-term leaf blade elongation. Bean leaves, initially 40 to 50 mm in length, treated once with alpha-naphthalene acetic acid (1.0 mm), were, after 6 d, approximately 80% the length and weight of controls. When applied at 1.0 and 0.1 mm, alpha-naphthalene acetic acid significantly inhibited long-term leaf growth. The weak auxin, beta-naphthalene acetic acid, was effective at 1.0 mm; and a weak acid control, benzoic acid, was ineffective. Indole-3-acetic acid (1 microm, 10 microm, 0.1 mm, and 1 mm) required daily application to be effective at any concentration. Application of the auxin transport inhibitor, 1-N-naphthylphthalamic acid (1% [w/w] in lanolin), to petioles also inhibited long-term leaf growth. This treatment also was found to lead to a sustained elevation of leaf free indole-3-acetic acid content relative to untreated control leaves. Auxin-induced inhibition of leaf growth appeared not to be mediated by auxin-induced ethylene synthesis because growth inhibition was not rescued by inhibition of ethylene synthesis. Also, petiole treatment of Arabidopsis with 1-N-naphthylphthalamic acid similarly inhibited leaf growth of both wild-type plants and ethylene-insensitive ein4 mutants.     

Implementation of first Croatian Law on Protection of Persons with Mental Disorders

The aim of this article was to examine Croatian psychiatric practice regarding involuntary hospitalization, after the Law on Protection of Persons with Mental Disorders became effective, on January 1, 1998. Data on the practice of involuntary hospitalizations of patients with mental disorders in Vrapce Psychiatric Hospital were collected from the medical records, for the years 1998 and 1999. Data regarding involuntary hospitalizations from other Croatian hospitals and departments were obtained from heads of psychiatric hospitals and departments for the first five months of 1998. The rate of involuntarily hospitalized patients in Vrapce Psychiatric Hospital rose significantly from 1998 to 1999 (p < 0.01). The rate of patients involuntarily hospitalized under section 21, subsection 3 rose significantly from 1998 to 1999 (p < 0.01), while rate of patients involuntarily hospitalized under section 22, subsection 1 decreased significantly in the same period (p < 0.01) in Vrapce Psychiatric Hospital. The implementation of the Law on protection of persons with mental disorders was not applied uniformly in all Croatian psychiatric institutions during first five months of 1998. Further analyses on this subject are necessary in order to investigate the influence of changes and supplements to the Law on the protection of persons with mental disorders on the practice of involuntary hospitalizations.     

Labor Saved,Calories Lost: The Energetic Impact of Domestic Labor‐saving Devices

Objective: As the prevalence of obesity has increased, so has sedentariness. Progressive sedentariness has been attributed to greater use of labor saving devices, such as washing machines, and less nonexercise walking (e.g., walking to work). However, there is a paucity of data to support this conclusion. In this study, we address the hypothesis that domestic mechanization of daily tasks has resulted in less energy expenditure compared with performing the same tasks manually. Research Methods and Procedures: Energy expenditure was measured in four groups of subjects (122 healthy adult men and women total) from Rochester, Minnesota. Energy expenditure was measured using indirect calorimetry while subjects performed structured tasks such as cleaning dishes and clothes, stair climbing, and work‐associated transportation, and these values were compared with the respective mechanized activity. Results: Energy expenditure was significantly greater and numerically substantial when daily domestic tasks were performed without the aid of machines or equipment (clothes washing: 45 ± 14 vs. 27 ± 9 kcal/d; dish washing: 80 ± 28 vs. 54 ± 19 kcal/d; transportation to work: 83 ± 17 vs. 25 ± 3 kcal/d; stair climbing: 11 ± 7 vs. 3 ± 1 kcal/d; p < 0.05). The combined impact of domestic mechanization was substantial and equaled 111 kcal/d. Discussion: The magnitude of the energetic impact of the mechanized tasks we studied was sufficiently great to contribute to the positive energy balance associated with weight gain. Efforts focused on reversing sedentariness have the potential to impact obesity.     

Defining the Pharmacodynamic Profile and Therapeutic Index of NHS-IL12 Immunocytokine in Dogs with Malignant Melanoma

Background

Interleukin (IL)-12 is a pro-inflammatory cytokine that mediates T-helper type 1 responses and cytotoxic T-cell activation, contributing to its utility as anti-cancer agent. Systemic administration of IL-12 often results in unacceptable toxicity; therefore, strategies to direct delivery of IL-12 to tumors are under investigation. The objective of this study was to assist the preclinical development of NHS-IL12, an immunocytokine consisting of an antibody, which targets necrotic tumor regions, linked to IL-12. Specifically this study sought to evaluate the safety, serum pharmacokinetics, anti-tumor activity, and immune modulation of NHS-IL12 in dogs with naturally occurring cancers.

Methodology/Principal Findings

A rapid dose-escalation study of NHS-IL12 administered subcutaneously to dogs with melanoma was conducted through the Comparative Oncology Trials Consortium (COTC). Eleven dogs were enrolled in four dose-escalation cohorts; thereafter, an additional seven dogs were treated at the defined tolerable dose of 0.8 mg/m². The expanded cohort at this fixed dose (ten dogs in total) was accrued for further pharmacokinetics and pharmacodynamics assessment. NHS-IL12 levels, serum cytokine concentrations, and peripheral blood mononuclear cell characterization (post-treatment) and draining lymph node immune profiling, and tumor biopsies (pre- and post-treatment) were collected. Adverse events included thrombocytopenia, liver enzymopathies, fever, and vasculitis. Correlation between interferon (IFN)-γ induction, adverse events, and NHS-IL12 exposure (maximum concentration and area under the concentration-time curve) were dose-dependent. Serum IL-10 levels and intratumoral CD8⁺ populations increased after treatment. Partial responses, according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria, were observed in two dogs treated with NHS-IL12 0.8 mg/m² and 1.6 mg/m².

Conclusions/Significance

NHS-IL12 was administered safely to dogs with melanoma and both immunologic and clinical activity was observed. This study successfully defined a narrow therapeutic window for systemic delivery of NHS-IL12 via the subcutaneous route. Results will inform the design and implementation of first-in-human clinical trials of NHS-IL12 in cancer patients.     

Community-Based Ecotourism and Primate Watching as a Conservation Tool in the Amazon Rainforest

International Journal of Primatology - Community-based tourism is a participatory approach implemented to support the economic and social development of local communities. Here, we present the...     

Sex-specific responses in placental fatty acid oxidation,esterification and transfer capacity to maternal obesity

Fatty acid metabolism and oxidation capacity in the placenta, which likely affects the rate and composition of lipid delivered to the fetus remains poorly understood. Long chain polyunsaturated fatty acids, such as docosahexaenoic acid (DHA), are critical for fetal growth and brain development. We determined the impact of maternal obesity on placental fatty acid oxidation, esterification and transport capacity by measuring PhosphatidylCholine (PC) and LysoPhosphatidylCholine (LPC) containing DHA by mass spectrometry in mother-placenta-baby triads as well as placental free carnitine and acylcarnitine metabolites in women with normal and obese pre-pregnancy BMI. Placental protein expression of enzymes involved in beta-oxidation and esterification pathways, MFSD2a (lysophosphatidylcholine transporter) and OCTN2 (carnitine transporter) expression in syncytiotrophoblast microvillous (MVM) and basal (BM) membranes were determined by Western Blot. Maternal obesity was associated with decreased umbilical cord plasma DHA in LPC and PC fractions in male, but not female, fetuses. Basal membrane MFSD2a protein expression was increased in placenta of males of obese mothers. In female placentas, despite an increased MVM OCTN2 expression, maternal obesity was associated with a reduced MUFA-carnitine levels and increased esterification enzymes. We speculate that lower DHA-PL in fetal circulation of male offspring of obese mothers, despite a significant increase in transporter expression for LPC-DHA, may lead to low DHA needed for brain development contributing to neurological consequences that are more prevalent in male children. Female placentas likely have reduced beta-oxidation capacity and appear to store FA through greater placental esterification, suggesting impaired placenta function and lipid transfer in female placentas of obese mothers.