Degenerative process and cell death in salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged female exposed to the acaricide permethrin

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Cell Death in Salivary Glands of Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae) Females at Semi-engorged Feeding Stage

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A function to predict the risk for death in the neonatal sepsis

This was a prospective study of 43 septic neonates at the NICU of the School of Medicine of Botucatu, São Paulo State University. Clinical and laboratory data of sepsis were analyzed based on outcome divided into two groups, survival and death. We calculated the discriminatory power of the relevant variables for the diagnosis of sepsis in each group, and using software for Discriminant Analysis, a function was proposed. There were 43 septic cases with 31 survivals and 12 deaths. The variables that had the highest discriminatory power were: n(o) of compromised systems, the SNAP, FiO2, and (A-a)O2. The study of these and others variables, such as birth weight, n(o) of risk factors, and pH using a Linear Discriminant Function(LDF) allowed us to identify the high-risk neonates for death with a low error rate (8.33%). The LDF was: F = 0.00043 (birth weight) + 0.30367 (n(o) of risk factors) - 0.1171 (n(o) of compromised systems) + 0.33223 (SNAP) + 2.27972 (pH) - 14.96511 (FiO2) + 0.01814 ((A-a)O2). If F > 22.77 there was high risk of death. This study suggests that the LDF at the onset of sepsis is useful for the early identification of the high-risk neonates that need special clinical and laboratory surveillance.

Prospective Evaluation of Dietary Treatment in Childhood Constipation: High Dietary Fiber and Wheat Bran Intake Are Associated With Constipation Amelioration

Objectives: The aim of the study was to evaluate, over 24 months, the intake of dietary fiber (DF) and the bowel habit (BH) of constipated children advised a DF-rich diet containing wheat bran.Patients and Methods: BH and dietary data of 28 children with functional constipation defined by the Boston criteria were obtained at visit 1 (V1, n = 28) and at 4 follow-up visits (V2-V5, n = 80). At each visit the BH was rated BAD (worse/unaltered; improved but still complications) or RECOVERY (REC) (improved, no complications; asymptomatic), and a food intake questionnaire was applied. DF intake was calculated according to age (year)+5 to 10 g/day and bran intake according to international tables. Nonparametric statistics were used.Results: Median age (range) was 7.25 years (0.25-15.6 years); 21 children underwent bowel washout (most before V1/V2), and 14 had the last visit at V3/V4. DF intake, bran intake, and the BH rate significantly increased at V2 and remained higher than at V1 through V2 to V5. At V1, median DF intake was 29.9% below the minimum recommended and at the last visit 49.9% above it. Twenty-four children accepted bran at 60 visits, at which median bran intake was 20 g/day and median proportion of DF due to bran 26.9%. Children had significantly higher DF and higher bran intake at V2 to V5 at which they had REC than at those at which they presented BAD BH. DF intake > age+10 g/day was associated with bran acceptance and REC. At the last visit 21 children presented REC (75%); 20 of them were asymptomatic and 18 were off washout/laxatives.Conclusions: High DF and bran intake are feasible in constipated children and contribute to amelioration of constipation.

Preliminary Criteria for Global Flares in Childhood-Onset Systemic Lupus Erythematosus

Objective. To develop widely acceptable preliminary criteria of global flare for childhood-onset systemic lupus erythematosus (cSLE).Methods. Pediatric rheumatologists (n = 138) rated a total of 358 unique patient profiles with information about the cSLE flare descriptors from 2 consecutive visits: patient global assessment of well-being, physician global assessment of disease activity (MD-global), health-related quality of life, anti-double-stranded DNA antibodies, disease activity index scores, protein: creatinine (P:C) ratio, complement levels, and erythrocyte sedimentation rate (ESR). Based on 2,996 rater responses about the course of cSLE (baseline versus followup), the accuracy (sensitivity, specificity, and area under the receiver operating characteristic curve) of candidate flare criteria was assessed. An international consensus conference was held to rank these candidate flare criteria as per the American College of Rheumatology recommendations for the development and validation of criteria sets.Results. The highest-ranked candidate criteria considered absolute changes (Delta) of the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) or British Isles Lupus Assessment Group (BILAG), MD-global, P:C ratio, and ESR; flare scores can be calculated (0.5 x Delta SLEDAI + 0.45 x Delta P:C ratio + 0.5 x Delta MD-global + 0.02 x Delta ESR), where values of >= 1.04 are reflective of a flare. Similarly, BILAG-based flare scores (0.4 x Delta BILAG + Delta 0.65 x Delta P:C ratio + 0.5 + Delta MD-global + 0.02 x Delta ESR) of >= 1.15 were diagnostic of a flare. Flare scores increased with flare severity.Conclusion. Consensus has been reached on preliminary criteria for global flares in cSLE. Further validation studies are needed to confirm the usefulness of the cSLE flare criteria in research and for clinical care.

Early embryonic death in mares: clinical and hormonal aspects

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Modes of cell death in the hypopharyngeal gland of the honey bee (Apis mellifera L)

Different modes of cell death have been revealed in the regressing hypopharyngeal glands of worker honey bees. The hypopharyngeal gland, which is well developed in young nursing bees to produce protein for larval food, was seen to regress naturally in foraging adult worker bees. A range of techniques including histology, cytochemistry, in situ TUNEL, Annexin V and Comet assays indicated that cells within the gland demonstrate progressive symptoms of apoptosis, necrosis and a vacuolar form of programmed cell death. The latter mode of cell death did not display chromatin margination, but was accompanied by an enhanced level of autophagic and hydrolytic activity in which a cytosolic source of acid phosphatase became manifest in the extra-cisternal spaces. Normal and annexin-positive cells were found to occur in the younger nursing bees, whilst necrosis and an aberrant vacuolar type of apoptosis predominated in the older foraging bees. The relevance of these results to the classification of programmed cell death is discussed. (C) 2000 Academic Press.

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy. Tissue sections were subsequently stained with haematoxylin-eosin, bromophenol blue, silver, or a variant of the critical electrolyte concentration (CEC) method. Ultrathin sections were contrasted with uranyl acetate and lead citrate. Glands were processed for the histochemical and cytochemical localization of acid phosphatase, as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed. The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and, additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen. The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed. The nuclei were pyknotic, with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

Fipronil-induced cell death in salivary glands of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females

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Markers of cell death in salivary glands of males of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari, Ixodidae)

The salivary glands of Rhipicephalus sanguineus males at stages: unfed (control), at day seven post-attachment, and at days three and seven post-detachment from the host were examined using methods of enzymatic analysis and cell viability. At these stages of feeding, different staining patterns were observed in the cells of type IV, III, II and I acini, which were affected by degeneration in this sequence. Acid phosphatase reaction was inversely proportional to that of ATPase, while ATPase reaction was proportional to membrane integrity.Salivary gland cells of unfed males exhibited intact nucleus and plasma membrane, suggesting that the acid phosphatase detected may participate in the normal physiology of acini. In males at day seven post-attachment, intact membranes were observed in almost all types of acini, as well as stronger reaction for acid phosphatase, nuclear changes, and decrease in ATPase reaction, changes associated with the degenerative process. At days three and seven post-detachment degeneration progress, being observed loss of membrane integrity, nuclear changes, prominent decrease in ATPase reaction, and an increase in acid phosphatase reaction in the first case and a decreased of it at day seven post-detachment from the host. During cell death, alterations occurred in the following sequence: a) nuclear changes, b) loss of ATPase reaction, c) loss of integrity of the plasma membrane, and d) increase of acid phosphatase. The latter might be associated with the late degradation of cytoplasmic remnants, characterizing the process of cell death in glands of R. sanguineus males as atypical or non-classic apoptosis. (C) 2008 Elsevier B.V. All rights reserved.

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera : Apidae)

The silk glands of bees are a good model for the study of cell death in insects. With the objective to detect the nuclear features during glandular regression stage, larvae at the last instar and pre-pupae were collected and their silk glands were dissected and processed for ultrastructural analysis and histologically for cytochemical and imunocytochemical analysis. The results showed that the cellular nuclei exhibited characteristics of death by atypical apoptosis as well as autophagic cell death. Among the apoptosis characteristic were: nuclear strangulation with bleb formation in some nuclei, DNA fragmentation in most of the nuclei and nucleolar fragmentation. Centripetal chromatin compaction was observed in many nuclei, forming a perichromatin halo differing from typical apoptotic nuclei. With regards to the characteristics of autophagic-programmed cell death, most relevant was the delay in the collapse of many nuclei. (C) 2006 Elsevier Ltd. All rights reserved.

Ultrastructural analysis of the effect of mating delay on cell death in the ovaries of virgin honey bee (Apis mellifera L.) queens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)