Investigating genetic diversity in sapucaia using inter simple sequence repeat markers.

Sapucaia is a tree species originating from the Brazilian Amazon and is widely distributed in Brazil, especially in the mid-north region (Piauí and Maranhão states). Its seeds are rich in calories and proteins, and possess great potential for commercialization. Little is known about the genetic variability in the germplasm of most Lecythis species. Here, 11 inter-simple sequence repeat primers were used to estimate the genetic variability among 17 accessions, and to determine the levels of genetic variation and the standards of population structure in sapucaia. The accessions were obtained from the active germplasm bank (AGB) of Embrapa Meio-Norte, Teresina, PI, Brazil, and corresponded to four occurrence areas. Ninety-six loci were analyzed among the studied individuals. High variation was found at the species level, where the percentage of polymorphic bands was 94.79%, Nei's genetic diversity (h) was 0.3110, and Shannon's index (I) was 0.4732. In the analyzed populations, the percentage polymorphism ranged from 20.83 to 94.79%, Nei's genetic diversity ranged from 0.0863 to 0.2969, and Shannon's index ranged from 0.1260 to 0.4457. Significant genetic differentiation was detected among the populations (ΦST = 10.66%); however, the greatest genetic differentiation was found within the populations (89.34%), between which there was an intermediate level of gene flow (Nm = 1.10). Accessions BGS 2 and BGS 4 were the most divergent, whereas accessions BGS 14 and BGS 15 were the most similar. Therefore, sapucaia analyzed from the AGB present an elevated level of genetic diversity and may have potential use in genetic breeding programs.

TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity.

There are two known mechanisms by which natural killer (NK) cells recognize and kill diseased targets: (i) direct killing and (ii) antibody-dependent cell-mediated cytotoxicity (ADCC). We investigated an indirect NK cell activation strategy for the enhancement of human NK cell killing function. We did this by leveraging the fact that toll-like receptor 9 (TLR9) agonism within pools of human peripheral blood mononuclear cells (PBMCs) results in a robust interferon signaling cascade that leads to NK cell activation. After TLR9 agonist stimulation, NK cells were enriched and incorporated into assays to assess their ability to kill tumor cell line targets. Notably, differential impacts of TLR9 agonism were observed-direct killing was enhanced while ADCC was not increased. To ensure that the observed differential effects were not attributable to differences between human donors, we recapitulated the observation using our Natural Killer-Simultaneous ADCC and Direct Killing Assay (NK-SADKA) that controls for human-to-human differences. Next, we observed a treatment-induced decrease in NK cell surface CD16-known to be shed by NK cells post-activation. Given the essential role of CD16 in ADCC, such shedding could account for the observed differential impact of TLR9 agonism on NK cell-mediated killing capacity.

Development and implementation of natural killer cell simultaneous ADCC and direct killing assay.

Assays to quantify natural killer (NK) cell killing efficacy have traditionally focused on assessing either direct killing or antibody dependent cell-mediated cytotoxicity (ADCC) independently. Due to the probability that immunotherapeutic interventions affect NK cell-mediated direct killing and NK cell-mediated ADCC differently, we developed an assay with the capacity to measure NK cell-mediated direct killing and ADCC simultaneously with cells from the same human donor. Specifically, this design allows for a single NK cell population to be split into several experimental conditions (e.g., direct killing, ADCC), thus controlling for potential confounders associated with human-to-human variation when assessing immunotherapy impacts. Our Natural Killer cell Simultaneous ADCC and Direct Killing Assay (NK-SADKA) allows researchers to reproducibly quantify both direct killing and ADCC by human NK cells. Furthermore, this optimized experimental design allows for concurrent analysis of the NK cells via flow cytometric immunophenotyping of NK cell populations which will facilitate the identification of relationships between NK cell phenotype and the subsequent killing potential. This assay will be valuable for assessing the broader impact(s) of immunotherapy strategies on both modes of NK cell killing.

Emerging Role of miR-345 and Its Effective Delivery as a Potential Therapeutic Candidate in Pancreatic Cancer and Other Cancers.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with high mortality, poor prognosis, and palliative treatments, due to the rapid upregulation of alternative compensatory pathways and desmoplastic reaction. miRNAs, small non-coding RNAs, have been recently identified as key players regulating cancer pathogenesis. Dysregulated miRNAs are associated with molecular pathways involved in tumor development, metastasis, and chemoresistance in PDAC, as well as other cancers. Targeted treatment strategies that alter miRNA levels in cancers have promising potential as therapeutic interventions. miRNA-345 (miR-345) plays a critical role in tumor suppression and is differentially expressed in various cancers, including pancreatic cancer (PC). The underlying mechanism(s) and delivery strategies of miR-345 have been investigated by us previously. Here, we summarize the potential therapeutic roles of miR-345 in different cancers, with emphasis on PDAC, for miRNA drug discovery, development, status, and implications. Further, we focus on miRNA nanodelivery system(s), based on different materials and nanoformulations, specifically for the delivery of miR-345.

Y chromosome microdeletions in Brazilian fertility clinic patients.

Microdeletions in Yq are associated with defects in spermatogenesis, while those in the AZF region are considered critical for germ cell development. We examined microdeletions in the Y chromosomes of patients attended at the Laboratory of Human Reproduction of the Clinical Hospital of the Federal University of Goiás as part of a screening of patients who plan to undergo assisted reproduction. Analysis was made of the AZF region of the Y chromosome in men who had altered spermograms to detect possible microdeletions in Yq. Twenty-three patients with azoospermia and 40 with severe oligozoospermia were analyzed by PCR for the detection of six sequence-tagged sites: sY84 and sY86 for AZFa, sY127 and sY134 for AZFb, and sY254 and sY255 for AZFc. Microdeletions were detected in 28 patients, including 10 azoospermics and 18 severe oligozoospermics. The patients with azoospermia had 43.4% of their microdeletions in the AZFa region, 8.6% in the AZFb region and 17.4% in the AZFc region. In the severe oligozoospermics, 40% were in the AZFa region, 5% in the AZFb region and 5% in the AZFc region. We conclude that microdeletions can be the cause of idiopathic male infertility, supporting conclusions from previous studies.

Efeito do extrato aquoso de camomila (Chamomilla recutita L. ) na prenhez de ratas e no desenvolvimento dos filhotes / Effect of aqueous extract of chamomile (Chamomilla recutita L. ) on rat pregnancy and offspring development

As plantas medicinais possuem substâncias ativas e, muitas vezes, o efeito tóxico sobre o organismo é desconhecido ou ignorado pelos usuários. A camomila é muito utilizada pela população, porém, contra-indicada para gestantes por possuir indícios de atividade emenagoga e relaxante da musculatura lisa. Por esses motivos, o objetivo do presente estudo foi avaliar os efeitos do extrato aquoso de camomila na gestação e nos filhotes gerados. Foram utilizadas 9 ratas da linhagem Wistar, divididas em 3 grupos, D1 e D2 que receberam infusão de camomila a 5% e 10% respectivamente, e o grupo controle que recebeu soro fisiológico. Os tratamentos foram administrados por via oral, desde o 1º ao 7º dia após o cruzamento. Os parâmetros estudados foram: prevalência de abortos, ganho de peso materno durante a prenhez, morte fetal e materna, malformações fetais grosseiras, número de recém-nascidos, peso dos filhotes, e análise de reflexos neurológicos dos filhotes (postural, preensão e orientação) no 1º, 3º, 5º e 10º dias de vida. Houve gestação em 70% do grupo controle, 40% do D1 e 80% do D2. Não houve diferença no ganho de peso materno no 7º e 21º dia, porém, os grupos tratados obtiveram ganho de peso menor em relação ao controle no 14º dia de gestação (p=0,04). As diferenças entre o número de recém-nascidos não foram significantes. Quanto ao peso dos recém-nascidos, os animais tratados apresentaram menor ganho de peso aos dias 1, 3, 5, e 10 após o nascimento (p=0,005; p=0,001; p<0,001; p<0,001; respectivamente). Ocorreram diferenças no reflexo postural no 1º dia, ocorrendo aceleração (p=0,005); já no reflexo de preensão (p=0,006), e no reflexo de orientação (p=0,01) houve retardo no desaparecimento, sem alteração nos demais dias sobre os outros parâmetros avaliados. A camomila pode influenciar, tanto no ganho de peso materno durante a gestação, como no dos filhotes após o nascimento, e pode provocar alterações nos reflexos neurológicos. Não se observou alteração nos demais parâmetros estudados.

Medicinal plants have active substances and their toxic effect on the organism is often unknown or ignored by users. Chamomile has been widely used by the population; however, it is contraindicated for pregnant women due to evidence of its emmenagogue and relaxing activity on smooth muscles. For these reasons, the aim of this study was to evaluate the effects of aqueous extract of chamomile on pregnancy and generated offspring. Nine Wistar rats were divided into three groups, D1 and D2, which received chamomile infusion at 5% and 10%, respectively, and the control group which received saline solution. Treatments were administered from the 1st to the 7th day after breeding. The studied parameters were abortion prevalence, maternal weight gain during pregnancy, fetal and maternal death, gross fetal malformation, newborn number, weight and neurological reflexes of the offspring on the 1st, 3rd, 5th and 10th days of life. There was pregnancy in 70% of control group, 40% of D1 and 80% of D2. There was no difference in maternal weight gain on the 7th and 21st days; however, treated groups had smaller weight gain, compared to control, on the 14th day of pregnancy (p=0.04). The differences among newborn numbers were not significant. As to weight of newborns, treated animals had smaller weight gain on days 1, 3, 5 and 10 after birth (p=0.005; p=0.001; p<0.001; p<0.001; respectively). For neurological effects, there were differences in postural reflex on the 1st day, resulting in acceleration; on the other hand, for grasp (p=0.006) and orienting reflex (p=0.01), there was a delay in disappearing, and no changes occurred on the remaining days for the other assessed parameters. Chamomile may influence maternal weight gain during pregnancy, as well as offspring weight gain after birth, and may cause changes in neurological reflexes. There was no alteration in the remaining studied parameters.

Y chromosome microdeletions in Brazilian fertility clinic patients

Microdeletions in Yq are associated with defects in spermatogenesis, while those in the AZF region are considered critical for germ cell development. We examined microdeletions in the Y chromosomes of patients attended at the Laboratory of Human Reproduction of the Clinical Hospital of the Federal University of Goiás as part of a screening of patients who plan to undergo assisted reproduction. Analysis was made of the AZF region of the Y chromosome in men who had altered spermograms to detect possible microdeletions in Yq. Twenty-three patients with azoospermia and 40 with severe oligozoospermia were analyzed by PCR for the detection of six sequence-tagged sites: sY84 and sY86 for AZFa, sY127 and sY134 for AZFb, and sY254 and sY255 for AZFc. Microdeletions were detected in 28 patients, including 10 azoospermics and 18 severe oligozoospermics. The patients with azoospermia had 43.4% of their microdeletions in the AZFa region, 8.6% in the AZFb region and 17.4% in the AZFc region. In the severe oligozoospermics, 40% were in the AZFa region, 5% in the AZFb region and 5% in the AZFc region. We conclude that microdeletions can be the cause of idiopathic male infertility, supporting conclusions from previous studies.