Defining the role of NatC dependent N-terminal acetylation in Drosophila fertility

Dissertação de Mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Levels of dioxin-like PCBs in low-volume serum samples of male patients attending fertility clinics

An accurate and easy method for the extraction, cleanup, and HRGC-HRMS analysis of dioxin-like PCBs (DL-PCBs) in low-volume serum samples (1 mL) was developed. Serum samples were extracted several times using n-hexane and purified by acid washing. Recovery rates of labeled congeners ranged from 70 to 110 % and the limits of detection were below 1 pg/g on lipid basis. Although human studies are limited and contradictory, several studies have shown that DL-PCBs can have adverse effects on the male reproductive system. In this way, the present method was applied to 21 serum samples of male patients attending fertility clinics. The total levels obtained for the patients ranged from 6.90 to 84.1 pg WHO-TEQ/g lipid, with a mean value of 20.3 pg WHO-TEQ/g lipid. The predominant PCBs (the sum of PCB 118, 156, and 105) contributed 67 % to the mean concentration of total DL-PCBs in the samples analyzed.

Optical redox ratio and endogenous porphyrins in the detection of urinary bladder cancer:a patient biopsy analysis

Bladder cancer is among the most common cancers in the UK and conventional detection techniques suffer from low sensitivity, low specificity, or both. Recent attempts to address the disparity have led to progress in the field of autofluorescence as a means to diagnose the disease with high efficiency, however there is still a lot not known about autofluorescence profiles in the disease. The multi-functional diagnostic system "LAKK-M" was used to assess autofluorescence profiles of healthy and cancerous bladder tissue to identify novel biomarkers of the disease. Statistically significant differences were observed in the optical redox ratio (a measure of tissue metabolic activity), the amplitude of endogenous porphyrins and the NADH/porphyrin ratio between tissue types. These findings could advance understanding of bladder cancer and aid in the development of new techniques for detection and surveillance.

Effects of charcoal-enriched goat manure on soil fertility parameters and growth of pearl millet (Pennisetum glaucum L.) in a sandy soil from northern Oman

The effect of charcoal feeding on manure quality and its subsequent application to enhance soil productivity has received little attention. The objectives of the present study therefore were to investigate the effects of (i) charcoal feeding on manure composition, and (ii) charcoal-enriched manure application on soil fertility parameters and growth of millet (Pennisetum glaucum L.). To this end, two experiments were conducted: First, a goat feeding trial where goats were fed increasing levels of activated charcoal (AC; 0, 3, 5, 7, and 9% of total ration); second, a greenhouse pot experiment using the manure from the feeding trial as an amendment for a sandy soil from northern Oman. We measured manure C, N, P, and K concentrations, soil fertility parameters and microbial biomass indices, as well as plant yield and nutrient concentrations. Manure C concentration increased significantly (P<0.001) from 45.2% (0% AC) to 60.2% (9% AC) with increasing dietary AC, whereas manure N, P, and K concentrations decreased (P<0.001) from 0% AC (N: 2.5%, P: 1.5%, K: 0.8%) to 9% AC (N: 1.7%, P: 0.8%, K: 0.4%). Soil organic carbon, pH, and microbial biomass N showed a response to AC-enriched manure. Yield of millet decreased slightly with AC enrichment, whereas K uptake was improved with increasing AC. We conclude that AC effects on manure quality and soil productivity depend on dosage of manure and AC, properties of AC, trial duration, and soil type.

Integrated soil fertility management in eastern and western Africa: The role of knowledge and innovation systems, its adoption and impact

The rise in population growth, as well as nutrient mining, has contributed to low agricultural productivity in Sub-Saharan Africa (SSA). A plethora of technologies to boost agricultural production have been developed but the dissemination of these agricultural innovations and subsequent uptake by smallholder farmers has remained a challenge. Scientists and philanthropists have adopted the Integrated Soil Fertility Management (ISFM) paradigm as a means to promote sustainable intensification of African farming systems. This comparative study aimed: 1) To assess the efficacy of Agricultural Knowledge and Innovation Systems (AKIS) in East (Kenya) and West (Ghana) Africa in the communication and dissemination of ISFM (Study I); 2) To investigate how specifically soil quality, and more broadly socio-economic status and institutional factors, influence farmer adoption of ISFM (Study II); and 3) To assess the effect of ISFM on maize yield and total household income of smallholder farmers (Study III). To address these aims, a mixed methodology approach was employed for study I. AKIS actors were subjected to social network analysis methods and in-depth interviews. Structured questionnaires were administered to 285 farming households in Tamale and 300 households in Kakamega selected using a stratified random sampling approach. There was a positive relationship between complete ISFM awareness among farmers and weak knowledge ties to both formal and informal actors at both research locations. The Kakamega AKIS revealed a relationship between complete ISFM awareness among farmers and them having strong knowledge ties to formal actors implying that further integration of formal actors with farmers’ local knowledge is crucial for the agricultural development progress. The structured questionnaire was also utilized to answer the query pertaining to study II. Soil samples (0-20 cm depth) were drawn from 322 (Tamale, Ghana) and 459 (Kakamega, Kenya) maize plots and analysed non-destructively for various soil fertility indicators. Ordinal regression modeling was applied to assess the cumulative adoption of ISFM. According to model estimates, soil carbon seemed to preclude farmers from intensifying input use in Tamale, whereas in Kakamega it spurred complete adoption. This varied response by farmers to soil quality conditions is multifaceted. From the Tamale perspective, it is consistent with farmers’ tendency to judiciously allocate scarce resources. Viewed from the Kakamega perspective, it points to a need for farmers here to intensify agricultural production in order to foster food security. In Kakamega, farmers with more acidic soils were more likely to adopt ISFM. Other household and farm-level factors necessary for ISFM adoption included off-farm income, livestock ownership, farmer associations, and market inter-linkages. Finally, in study III a counterfactual model was used to calculate the difference in outcomes (yield and household income) of the treatment (ISFM adoption) in order to estimate causal effects of ISFM adoption. Adoption of ISFM contributed to a yield increase of 16% in both Tamale and Kakamega. The innovation affected total household income only in Tamale, where ISFM adopters had an income gain of 20%. This may be attributable to the different policy contexts under which the two sets of farmers operate. The main recommendations underscored the need to: (1) improve the functioning of AKIS, (2) enhance farmer access to hybrid maize seed and credit, (3) and conduct additional multi-locational studies as farmers operate under varying contexts.

Local Understandings of Soil Fertility, Rainfall and Conservation Agriculture in Laikipia, Kenya: A Qualitative Analysis.

Conservation Agriculture (CA) is mostly referred to in the literature as having three principles at the core of its identity: minimum soil disturbance, permanent organic soil cover and crop diversity. This farming package has been described as suitable to improve yields and livelihoods of smallholders in semi-arid regions of Kenya, which since the colonial period have been heavily subjected to tillage. Our study is based on a qualitative approach that followed local meanings and understandings of soil fertility, rainfall and CA in Ethi and Umande located in the semi-arid region of Laikipia, Kenya. Farm visits, 53 semistructured interviews, informal talks were carried out from April to June 2015. Ethi and Umande locations were part of a resettlement programme after the independence of Kenya that joined together people coming from different farming contexts. Since the 1970–80s, state and NGOs have been promoting several approaches to control erosion and boost soil fertility. In this context, CA has also been promoted preferentially since 2007. Interviewees were well acquainted with soil erosion and the methods to control it. Today, rainfall amount and distribution are identified as major constraints to crop performance. Soil fertility is understood as being under control since farmers use several methods to boost it (inorganic fertilisers, manure, terraces, agroforestry, vegetation barriers). CA is recognised to deliver better yields but it is not able to perform well under severe drought and does not provide yields as high as ‘promised’ in promotion campaigns. Moreover, CA is mainly understood as “cultivating with chemicals”, “kulima na dawa”, in kiswahili. A dominant view is that CA is about minimum tillage and use of pre-emergence herbicides. It is relevant to reflect about what kind of CA is being promoted and if elements like soil cover and crop rotation are given due attention. CA based on these two ideas, minimum tillage and use of herbicides, is hard to stand as a programme to be promoted and up-scaled. Therefore CA appears not to be recognised as a convincing approach to improve the livelihoods in Laikipia.

Recovery of reproductive activity and fertility of Saanen goats affected by hydrometra after cloprostenol treatment and estrus induction during the non-breeding season (preliminary data).

2016

Glutathione and abscisic acid supplementation influences somatic embryo maturation and hormone endogenous levels during somatic embryogenesis in Podocarpus lambertii Klotzsch ex Endl.

Here we propose a protocol for embryogenic cultures induction, proliferation and maturation for the Brazilian conifer Podocarpus lambertii, and investigated the effect of abscisic acid (ABA) and glutathione (GSH) supplementation on the maturation phase. ABA, zeatin (Z) and salicylic acid (SA) endogenous levels were quantified. Number of somatic embryos obtained in ABA-supplemented treatment was signifi- cant higher than in ABA-free treatment, showing the relevance of ABA supplementation during somatic embryos maturation. Histological analysis showed the stereotyped sequence of developmental stages in conifer somatic embryos, reaching the late torpedo-staged embryo. GSH supplementation in maturation culture medium improved the somatic embryos number and morphological features. GSH 0 mM and GSH 0.1 mM treatments correlated with a decreased ABA endogenous level during maturation, while GSH 0.5 mM treatment showed constantlevels. Alltreatments resulted in decreased Z endogenous levels, supporting the concept that cytokinins are important during the initial cell division but not for the later stages of embryo development. The lowest SA levels found in GSH 0.5 mM treatment were coincident with early embryonic development, and this treatment resulted in the highest development of somatic embryos. Thus, a correlation between lower SA levels and improved somatic embryo formation can be hypothesized

The effect of cross-linker on endogenous enzymatic activity within radicular dentin and bond strength between intraradicular post and radicular dentin

The primary aim was to evaluate the effect of 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (EDC) on endogenous enzymatic activity within radicular dentin and push-out bond strength of adhesively luted fiber posts, at baseline and after artificial aging. Additionally, the effect of different cementation strategies on endogenous enzymatic activity and fiber post retention was evaluated. The experiment was carried out on extracted human teeth, following endodontic treatment and fiber post cementation. Three cementation strategies were performed: resin cement in combination with etch-and-rinse (EAR) adhesive system, with self-etch (SE) system and self-adhesive (SE) cement. Each of the mentioned strategies had a control and experimental (EDC) group in which root canal was irrigated with 0.3M EDC for 1 minute. The push-out bond strength test was performed 24h after cementation and after 40.000 thermocycles. In order to investigate the effect of EDC and different cementation strategies, in situ zymography analyses of the resin-dentin interfaces were conducted. Statistical analyses were conducted with the software Stata 12.0 (Stata Corp, College Station, Texas, USA) and the significance was set for p<0.05. The results of statistical analysis (ANOVA) showed that the variables “EDC”, “root region” and “artificial aging” significantly influenced fiber posts’ retention to root canal (p<0.05). The highest values were observed in coronal third. The mean values observed after artificial aging were lower when compared to baseline, however EDC was effective in preserving bond strength. The level of enzymatic activity varied between the groups and EDC had a beneficial effect on silencing the enzymatic activity. Within the limitations of the study, it was concluded that the choice of cementation strategy did not influence posts’ retention, while EDC contributed to the preservation of bond strength after artificial aging and reduced enzymatic activity within radicular dentin. In vivo trials are necessary to confirm the results of this in vitro study.

Distraction by task-irrelevant stimuli: the effects of endogenous spatial attention and visual working memory load

Salient stimuli, like sudden changes in the environment or emotional stimuli, generate a priority signal that captures attention even if they are task-irrelevant. However, to achieve goal-driven behavior, we need to ignore them and to avoid being distracted. It is generally agreed that top-down factors can help us to filter out distractors. A fundamental question is how and at which stage of processing the rejection of distractors is achieved. Two circumstances under which the allocation of attention to distractors is supposed to be prevented are represented by the case in which distractors occur at an unattended location (as determined by the deployment of endogenous spatial attention) and when the amount of visual working memory resources is reduced by an ongoing task. The present thesis is focused on the impact of these factors on three sources of distraction, namely auditory and visual onsets (Experiments 1 and 2, respectively) and pleasant scenes (Experiment 3). In the first two studies we recorded neural correlates of distractor processing (i.e., Event-Related Potentials), whereas in the last study we used interference effects on behavior (i.e., a slowing down of response times on a simultaneous task) to index distraction. Endogenous spatial attention reduced distraction by auditory stimuli and eliminated distraction by visual onsets. Differently, visual working memory load only affected the processing of visual onsets. Emotional interference persisted even when scenes occurred always at unattended locations and when visual working memory was loaded. Altogether, these findings indicate that the ability to detect the location of salient task-irrelevant sounds and identify the affective significance of natural scenes is preserved even when the amount of visual working memory resources is reduced by an ongoing task and when endogenous attention is elsewhere directed. However, these results also indicate that the processing of auditory and visual distractors is not entirely automatic.

Cytogenetic And Morphologic Approaches Of Hybrids From Experimental Crosses Between Triatoma Lenti Sherlock & Serafim, 1967 And T. Sherlocki Papa Et Al., 2002 (hemiptera: Reduviidae).

The reproductive capacity between Triatoma lenti and Triatoma sherlocki was observed in order to verify the fertility and viability of the offspring. Cytogenetic, morphological and morphometric approaches were used to analyze the differences that were inherited. Experimental crosses were performed in both directions. The fertility rate of the eggs in crosses involving T. sherlocki females was 65% and 90% in F1 and F2 offspring, respectively. In reciprocal crosses, it was 7% and 25% in F1 and F2 offspring, respectively. The cytogenetic analyses of the male meiotic process of the hybrids were performed using lacto-acetic orcein, C-banding and Feulgen techniques. The male F1 offspring presented normal chromosome behavior, a finding that was similar to those reported in parental species. However, cytogenetic analysis of F2 offspring showed errors in chromosome pairing. This post-zygotic isolation, which prevents hybrids in nature, may represent the collapse of the hybrid. This phenomenon is due to a genetic dysregulation that occurs in the chromosomes of F1. The results were similar in the hybrids from both crosses. Morphological features, such as color and size of connexive and the presence of red-orange rings on the femora, were similar to T. sherlocki, while wins size was similar to T. lenti in F1 offspring. The eggshells showed characteristics that were similar to species of origin, whereas the median process of the pygophore resulted in intermediate characteristics in the F1 and a segregating pattern in F2 offspring. Geometric morphometric techniques used on the wings showed that both F1 and F2 offspring were similar to T. lenti. These studies on the reproductive capacity between T. lenti and T. sherlocki confirm that both species are evolutionarily closed; hence, they are included in the brasiliensis subcomplex. The extremely reduced fertility observed in the F2 hybrids confirmed the specific status of the species that were analyzed.

Fatty Acid Synthase Inhibitors Induce Apoptosis In Non-tumorigenic Melan-a Cells Associated With Inhibition Of Mitochondrial Respiration.

The metabolic enzyme fatty acid synthase (FASN) is responsible for the endogenous synthesis of palmitate, a saturated long-chain fatty acid. In contrast to most normal tissues, a variety of human cancers overexpress FASN. One such cancer is cutaneous melanoma, in which the level of FASN expression is associated with tumor invasion and poor prognosis. We previously reported that two FASN inhibitors, cerulenin and orlistat, induce apoptosis in B16-F10 mouse melanoma cells via the intrinsic apoptosis pathway. Here, we investigated the effects of these inhibitors on non-tumorigenic melan-a cells. Cerulenin and orlistat treatments were found to induce apoptosis and decrease cell proliferation, in addition to inducing the release of mitochondrial cytochrome c and activating caspases-9 and -3. Transfection with FASN siRNA did not result in apoptosis. Mass spectrometry analysis demonstrated that treatment with the FASN inhibitors did not alter either the mitochondrial free fatty acid content or composition. This result suggests that cerulenin- and orlistat-induced apoptosis events are independent of FASN inhibition. Analysis of the energy-linked functions of melan-a mitochondria demonstrated the inhibition of respiration, followed by a significant decrease in mitochondrial membrane potential (ΔΨm) and the stimulation of superoxide anion generation. The inhibition of NADH-linked substrate oxidation was approximately 40% and 61% for cerulenin and orlistat treatments, respectively, and the inhibition of succinate oxidation was approximately 46% and 52%, respectively. In contrast, no significant inhibition occurred when respiration was supported by the complex IV substrate N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD). The protection conferred by the free radical scavenger N-acetyl-cysteine indicates that the FASN inhibitors induced apoptosis through an oxidative stress-associated mechanism. In combination, the present results demonstrate that cerulenin and orlistat induce apoptosis in non-tumorigenic cells via mitochondrial dysfunction, independent of FASN inhibition.

Effects Of Partial Inhibition Of Respiratory Complex I On H2o 2 Production By Isolated Brain Mitochondria In Different Respiratory States.

The aim of this work was to characterize the effects of partial inhibition of respiratory complex I by rotenone on H2O2 production by isolated rat brain mitochondria in different respiratory states. Flow cytometric analysis of membrane potential in isolated mitochondria indicated that rotenone leads to uniform respiratory inhibition when added to a suspension of mitochondria. When mitochondria were incubated in the presence of a low concentration of rotenone (10 nm) and NADH-linked substrates, oxygen consumption was reduced from 45.9 ± 1.0 to 26.4 ± 2.6 nmol O2 mg(-1) min(-1) and from 7.8 ± 0.3 to 6.3 ± 0.3 nmol O2 mg(-1) min(-1) in respiratory states 3 (ADP-stimulated respiration) and 4 (resting respiration), respectively. Under these conditions, mitochondrial H2O2 production was stimulated from 12.2 ± 1.1 to 21.0 ± 1.2 pmol H2O2 mg(-1) min(-1) and 56.5 ± 4.7 to 95.0 ± 11.1 pmol H2O2 mg(-1) min(-1) in respiratory states 3 and 4, respectively. Similar results were observed when comparing mitochondrial preparations enriched with synaptic or nonsynaptic mitochondria or when 1-methyl-4-phenylpyridinium ion (MPP(+)) was used as a respiratory complex I inhibitor. Rotenone-stimulated H2O2 production in respiratory states 3 and 4 was associated with a high reduction state of endogenous nicotinamide nucleotides. In succinate-supported mitochondrial respiration, where most of the mitochondrial H2O2 production relies on electron backflow from complex II to complex I, low rotenone concentrations inhibited H2O2 production. Rotenone had no effect on mitochondrial elimination of micromolar concentrations of H2O2. The present results support the conclusion that partial complex I inhibition may result in mitochondrial energy crisis and oxidative stress, the former being predominant under oxidative phosphorylation and the latter under resting respiration conditions.

Long-term Follow-up Of Patients With 46,xy Partial Gonadal Dysgenesis Reared As Males.

Background/Aims. Studies on 46,XY partial gonadal dysgenesis (PGD) have focused on molecular, gonadal, genital, and hormone features; little is known about follow-up. Our aim was to analyze long-term outcomes of PGD. Methods. Retrospective longitudinal study conducted at a reference service in Brazil. Ten patients were first evaluated in the 1990s and followed up until the 2010s; follow-up ranged from 13.5 to 19.7 years. All were reared as males and had at least one scrotal testis; two bore NR5A1 mutations. Main outcomes were: associated conditions, pubertal development, and growth. Results. All patients had normal motor development but three presented cognitive impairment; five had various associated conditions. At the end of the prepubertal period, FSH was high or high-normal in 3/6 patients; LH was normal in all. At the last evaluation, FSH was high or high-normal in 8/10; LH was high or high-normal in 5/10; testosterone was decreased in one. Final height in nine cases ranged from -1.57 to 0.80 SDS. All had spontaneous puberty; only one needed androgen therapy. Conclusions. There is good prognosis for growth and spontaneous pubertal development but not for fertility. Though additional studies are required, screening for learning disabilities is advisable.

Interferentes endócrinos no ambiente

Although the hypothesis that environmental chemicals may exhibit endocrine disrupting effects is not new, the issue has been a growing level of concern due to reports of increased incidences of endocrine-related disease in humans, including declining male fertility, and more significantly, to adverse physiological effects observed in wildlife where cause and effect relationships are more evident. The list of endocrine disrupting chemicals (EDCs) includes a range of anthropogenic compounds, phytoestrogens, naturally occurring sex steroids and synthetic estrogens. Within the aquatic environment, the presence of EDCs has concerned many scientists and water quality regulators. Discharge of effluents from treatment facilities is likely to be a significant source of input of contaminants to many systems, and the potential for concentration of hydrophilic compounds and transformation products within sludges has implications for their disposal. Then, understanding the processes and the fate of EDCs on the environment, as well as the mechanisms of endocrine disruption, may facilitate controlling or limiting exposure of both humans and the environment to these compounds.