Resumo A produção de plantas transgênicas com características agronômicas e hortícolas desejadas vem ganhando grande importância por atender às demandas da crescente população. A transformação genética também é um passo fundamental para estudar os fundamentos das ciências das plantas. Foram desenvolvidos e utilizados diferentes protocolos de transformação, que são confiáveis e eficientes. Esses protocolos usaram genes de resistência a antibióticos ou herbicidas incorporados ao gene de interesse para identificar plantas transformadas e não transformadas. Esses genes marcadores podem representar uma ameaça ao ser humano e ao meio ambiente. O uso de marcadores visuais permite a observação direta e fácil de plantas transformadas com mais precisão. No estudo atual, foi projetado um cassete de genes com o gene da hidroxilase de produção de pigmentos (PPH) sob promotor específico de fibra (GhSCFP) e terminador NOS a jusante. Após verificar a eficiência estrutural e funcional do gene otimizado por códons usando ferramentas de bioinformática, o cassete foi enviado para síntese química a partir de fonte comercial. O cassete do gene do pigmento (PPH_CEMB), clonado em pCAMBIA-1301, foi transformado em Agrobacterium por eletroporação. O método de imersão floral mediado por Agrobacterium foi usado para transformar a inflorescência de Camelina sativa. Após a formação de sementes, foi observado um total de 600 sementes com mudança de cor, das quais 19 desenvolveram uma coloração marrom-avermelhada, enquanto as 581 restantes permaneceram amarelas. A eficiência de transformação calculada com base na mudança de cor foi de 1%. A análise de PCR das folhas obtidas após a semeadura de sementes avermelhadas confirmou a transformação do gene produtor de pigmentos, enquanto não foi observada amplificação por PCR em folhas de plantas de sementes do tipo selvagem. A partir dos resultados, é evidente que a transformação da inflorescência de C. sativa mediada por Agrobacterium é uma técnica muito eficiente e favorável ao ambiente não só para a detecção de plantas transformadas, mas também para estudar processos celulares básicos.

Abstract Production of transgenic plants with desired agronomic and horticultural traits has gained great importance to fulfill demands of the growing population. Genetic transformation is also a fundamental step to study basics of plant sciences. Different transformation protocols have been developed and used which are reliable and efficient. These protocols used antibiotic or herbicide resistance genes incorporated along with gene of interest to identify transformed plants from non-transformed ones. These marker genes may pose a threat to human and environment. Use of visual markers enables direct and easier observation of transformed plants with more precision. In current study a gene cassette with ‘pigment production hydroxylase (PPH) gene under fiber specific promoter (GhSCFP) and downstream Nos-terminator was designed. After checking the structural and functional efficiency of codon optimized gene using bioinformatics tools, the cassette was sent for chemical synthesis from commercial source. The pigment gene cassette (PPH_CEMB), cloned in pCAMBIA-1301, was transformed into Agrobacterium through electroporation. Agrobacterium-mediated floral dip method was used to transform Camelina sativa inflorescence. After seed setting a total of 600 seed were observed for change in color and out of these, 19 seeds developed a reddish-brown coloration, while the remaining 581 seeds remained yellow. The transformation efficiency calculated on basis of color change was 1.0%. PCR analysis of leaves obtained after sowing reddish seeds confirmed the transformation of pigment production gene, while no PCR amplification was observed in leaves of plants from wild type seeds. From the results it is evident that Agrobacterium-mediated transformation of C. sativa inflorescence is very efficient and environment friendly technique not only for detection of transformed plants but also to study basic cellular processes.

BACKGROUND: Despite the breadth of data supporting evidence-based practice for sepsis care in high-resource settings, there are relatively few data to guide the management of sepsis in low-resource settings, particularly in areas where HIV and tuberculosis (TB) are prevalent. Furthermore, few studies had broadened sepsis parameters to include all patients with acute infectious illness or followed patients up after hospital discharge. Understanding the epidemiology and outcomes of acute infections in a local context is the critical first step to developing locally informed targeted management strategies.OBJECTIVES: To quantify and describe the incidence of and risk factors for mortality in a cohort of patients with undifferentiated acute infectious illnesses who presented to an emergency department (ED) in the Eastern Cape region of South Africa (SA).METHODS: In this prospective cohort study, patients with suspected acute infectious illness were enrolled at a district casualty ward in Mthatha, SA, between 1 July and 1 September 2017. Demographic data, interventions, diagnostic studies and disposition were prospectively collected during the initial encounter and during the hospital stay. Follow-up was conducted both in hospital and via phone interviews 30 days after the index visit.RESULTS: A total of 301 patients presented to the ED with acute infectious illness during the study period, of whom 54.8% had complete 30-day follow-up. Of the study population, only 5.7% had a complete set of vital signs (heart rate, respiratory rate, blood pressure and temperature) documented. Of the cohort, 51.8% had HIV and 32.9% active or treated TB; 25.2% of patients died within 30 days. Accounting for medical history, diagnosis and ED interventions, risk of mortality was independently associated with age (odds ratio (OR) 1.03; 95% confidence interval (CI) 1.00 - 1.06), HIV-positive status (OR 4.10; 95% CI 1.44 - 11.67) and Quick Sequential (Sepsis-Related) Organ Failure Assessment (qSOFA) score (OR 1.90; 95% CI 1.14 - 3.19) in an adjusted model. No ED interventions were protective for mortality, with intravenous fluid administration associated with increased 30-day mortality in this cohort (OR 3.65; 95% CI 1.38 - 9.62).CONCLUSIONS: Among adults with suspected acute infectious illness in Mthatha, SA, 30-day mortality was concerningly high. Mortality was highest in patients with concomitant HIV infection. In particular, vital sign assessment to identify possible sepsis in this cohort is crucial, as it affects mortality to a meaningful extent, yet is often unavailable. Future research is needed on the management of sepsis in low-resource settings, particularly in HIV-positive individuals.

BACKGROUND. South Africa (SA) has the highest burden of HIV in the world. This study sought to evaluate the impact of high HIV prevalence on the burden of disease in an emergency department (ED).OBJECTIVES. To determine the burden of comorbidities in HIV-positive emergency care patients, their demographic profiles and severity of illness were compared with the general ED population in order to make recommendations for resource allocation and training in EDs in SA.METHODS. A prospective cross-sectional observational study was conducted from June 2017 to July 2018 in three EDs in Eastern Cape Province. All eligible patients (aged >18 years, fully conscious and clinically stable) presenting to the ED during the 6-week study period were approached and asked to give consent for a point-of-care HIV test and collection of demographic information. Simple descriptive statistics were used to analyse data. Log binomial and Poisson models were fitted to estimate prevalence ratios (PRs).RESULTS. Over the total study period, 8 000 patients presented to the ED for care across all sites and 3 537 patients were enrolled. The HIV status of 2 901 individuals (82.0%) was determined. Of those who were screened, 811 (28.0%) were identified as HIV-positive. Medical complaints were more common in HIV-positive patients (n=586, 72.3%) than in trauma patients (n=225, 27.7%). In comparison, HIV-negative patients reported fewer medical complaints (n=1 137, 54.4%) and more trauma (n=953, 45.6%) (p<0.001). HIV-positive patients were more likely to have a life-threatening emergency (n=192, 23.7%) (p=0.004), to be critically ill by triage score (p<0.001) and to be admitted to the hospital (p<0.001) than those who were HIV-negative. Despite high acuity overall, people living with HIV/AIDS were significantly less likely to be deemed critically ill according to vital signs (adjusted PR 0.94; p=0.046).CONCLUSIONS. While EDs in SA provide care to high volumes of patients with trauma-related injuries, in areas where HIV prevalence is highest, patients are more likely to present with acute medical emergencies. Providers of emergency care in SA need to be well versed in the management of HIV and associated complications.

BACKGROUND. Emergency medicine accounts for a large proportion of medical care in many low- and middle-income countries. a better understanding of the burden of disease will guide training and resource allocation priorities, but lack of electronic medical records and standardised data collection systems makes it difficult to obtain this information.OBJECTIVES. To draw attention to the proportionally large burden of trauma in emergency centres (ECs) throughout Eastern Cape Province, South Africa (SA), in the hope of influencing resource allocation and medical provider training protocols accordingly.METHODS. A secondary data analysis was performed from information gathered in HIV testing studies in two large tertiary care centres and one regional hospital in the Eastern Cape region of SA. All patients presenting to the ECs during the 6-week study period who met the inclusion criteria were approached and requested to provide consent for point-of-care HIV testing and collection of demographic information. Information collected included patient demographics, presenting complaints and final diagnoses. Simple descriptive statistics were used to analyse the data.RESULTS. Data were collected from 4 271 patients across three study sites: Frere Hospital (n=2 391), Nelson Mandela Academic Hospital (n=622) and Mthatha Regional Hospital (n=l 258). At the two tertiary care centres, most patients were between the ages of 18 and 30 years (41.2% and 32.6%, respectively) and male (57.8% and 60.2%), and 70.4% and 41.5% had traumatic injuries. The most common complaints were stab/gunshot wounds (18.3% and 20.2%). At the district hospital, the majority of patients were female (57.2%), 40.1% were between 18 and 30 years old, and 27.3% presented with traumatic injuries. Stab/gunshot wounds were the second most common complaint (7.2%) after lower respiratory tract infections (8.7%).CONCLUSIONS. From the proportion of presenting individuals sampled, we can conclude that a large proportion of care delivered in ECs in the Eastern Cape is for trauma. Local clinical capacitation efforts must focus on trauma training.

Abstract: Present work describes the failure analysis of AISI 304 stainless steel consisting of 7x19 construction lanyard wire rope which has failed during service. The microstructures and properties of failed wire rope have been investigated and compared with unused wire rope. Both the periphery and fracture surface of the wire rope display the presence of corrosión debris enriched with O and Cl. The fracture surfaces of the failed and unused wire ropes display intergranular and dimples, respectively. The lanyard wire rope has been exposed in corrosive atmosphere and failed in intergranular mode due to enrichment of O and Cl along the grain boundaries.

Um novo método isocrático, rápido e simples da cromatografia líquida de alta performance (HPLC) de fase normal quiral foi desenvolvido e validado para a separação enantiomérica do ácido (S)-10-hidroxicamptotecin (10 HCTN), ((4S)-4-etil-4,9-dihidro-1H-pirano [3',4':6,7] indolizina [1,2-b] quinolina-3, 14 (4H, 12H)-diona), um fármaco anti-cancerígeno. Os enantiômeros de 10 HCTN foram separados em uma coluna Chiralpak IC (fase estacionária quiral derivada de polissacarídeo imobilizado), utilizando uma fase móvel que consiste de n-hexano:etanol (50:50 v/v) em uma vazão de 1,0 mL min-1. A resolução entre os dois enantiômeros foi superior a 3 usando o método otimizado. O método desenvolvido foi validado e mostrou serrobusto, exato, enantiosseletivo, preciso e adequado para a determinação quantitativa do enantiômero-(R) tanto na matéria prima quanto no fármaco.

A new and simple, rapid, isocratic, normal phase chiral high performance liquid chromatography (HPLC) method was developed and validated for the enantiomeric separation of (S)-10-hydroxycamptothecin (10-HCTN), ((4S)-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b] quinoline-3,14(4H,12H)-dione), an anti-cancer drug substance. The enantiomers of 10-HCTN were resolved on a Chiralpak IC (immobilized polysaccharide chiral stationary phase) column using a mobile phase consisting of n-hexane:ethanol (50:50 v/v) at a flow rate of 1.0 mL min-1. The resolution between both enantiomers was greater than 3 in the optimized method. The developed method was extensively validated and proved to be robust, enantioselective, accurate, precise, and suitable for quantitative determination of (R)-enantiomer in bulk drug substance and product.

The more common lodger bee occurring in the dry forest of Costa Rica, Centris bicornuta Muscáry), has been observed nesting in new nest cavities drilled into wooden blocks placed next to cavities used by another female within 2-3 days. In contrast, new nest cavities placed in similar areas with no nesting Centris nearby were not used for weeks. These observations suggest that the presence of nesting bees may play a role in nest site selection. To confirm our observations, new nest cavities were placed in areas with or without nesting. We found nest initiation in newly placed nest cavities only in areas where bees were actively nesting. To examine the possibility that nesting locations are not unique, we placed new nest cavities in new locations either with (a) a number of completed nest cavities or (b) placed alone. Within three days we only found bees nesting in the newly placed nest cavities in situation "a". The results suggested that odor might be involved. We next compared nesting in new cavities placed alone with cavities contaminated with either (a) nest entrance plug material, (b) nest nectar, (c) nest pollen or (d) a combination of pollen and nectar. Nesting was significantly low in cavities placed next to cavities with nest entrance plug material (a), and high in cavities placed next to cavities "b, c, or d". The results suggest that pollen and /or nectar odor play a role in the location of potential nest sites.

Neste trabalho foi desenvolvido um procedimento espectrofotométrico derivativo para a determinação de etilenobisditiocarbamato de zinco(II), Zineb, com base na formação do complexo azul de molibdato de sódio em meio ácido. A lei de Beer é obedecida até 40 µg mL-1 de Zineb no comprimento de onda de 956 nm. O limite de detecção foi de 0,006 µg mL-1 de Zineb para a razão S/N = 3 considerando diversos parâmetros como o efeito da concentração de ácido. A interferência de diversos íons na determinação de Zineb foi avaliada. A maioria dos metais alcalinos e sais metálicos não causou interferências. O procedimento tem boa sensibilidade e foi empregado para a determinação de Zineb em amostras comerciais de alimentos contendo Dithane Z 78 e Hexathane 75% W.P.. Os resultados foram comparados com procedimentos previamente propostos. Zineb foi determinado com sucesso sem a ocorrência de interferências na presença de outros ditiocarbamatos, tais como Ziram, Thiram, Ferbam etc.

A derivative spectrophotometric procedure was developed for the determination of zinc(II) ethylenebisdithiocarbamate, Zineb, after formation of its blue colored complex with sodium molybdate in acidic medium. The Beer's law is obeyed up to 40 µg mL-1of Zineb at 956 nm. The detection limit was 0.006 µg mL-1 for Zineb when S/N ratio is 3 taking into account various parameters, such as the effect of acid concentration. The interference of a large number of ions on the determination of Zineb was evaluated. Most of the alkaline metals and metal salts did not interfere. The procedure presented proper sensitivity and it was applied for determining Zineb in food stuffs and commercial samples of Dithane Z 78 and Hexathane 75% W.P.. Results were compared with earlier reported methods. Zineb was successfully determined without any interferences in the presence of other dithiocarbamates like ziram, thiram, ferbam etc.

Pegmatitos graníticos são tradicionalmente conhecidos por terem intercrescimentos gráficos, pertíticos e mirmequíticos, relacionados a quartzo e feldspatos potássicos e sódicos. Também são considerados para caracterizar os tipos de pegmatitos, distinguindo-os dos granitos e outros tipos de rochas plutônicas relacionadas. Granito gráfico é aceito como sinônimo do pegmatito granítico. Estudos sistemáticos, do autor e seus colegas, sobre depósitos de pegmatitos graníticos de gemas permitiram a definição de duas províncias de gemas águas marinhas no ENE do Brasil: uma no NeoProterozóico e outra em seqüência Arqueana. Feldspatos potássicos na primeira apresentam intercrescimentos pertíticos e, na outra, o intercrescimento gráfico domina com quartzo grosseiro centimétrico anômalo ao longo das clivagens de K-feldspato. Diversos pegmatitos graníticos encontrados no complexo Arqueano, na Província de Águas Marinhas de Lages Pintadas, Santa Cruz, Estado do RN, apresentam esta textura-estrutura. Intercrescimento gráfico é atribuído a cristalização eutética, sucedida por fluidos hidrotermais com enriquecimento de sílica permitindo o crescimento através da difusão e nucleação de quartzo e ao alongo das clivagens de feldspato potássico. No terreno Arqueano, a abundancia de chert reciclado formando migmatitos metapsamíticos atravessados por numerosos veios de quartzo e granito gráfico grosseiro, contribuiu para o crescimento de berilo e da água marinha.

Granitic pegmatites are traditionally known to contain graphic, perthitic and myrmekitic intergrowths related to quartz and K- and Na- feldspars. They are further considered to characterise the pegmatite types distinguishing them from the granites and other related plutonic rock types. Graphic granite is accepted also as a synonym to granitic pegmatite. Systematic studies, by the author and colleagues, on the granitic pegmatite gem deposits have permitted the definition of two aquamarine gem provinces in ENE Brazil, one in the NeoProterozoic and the other in the Archaean sequences. Potash feldspars in the pegmatites in the former show perthitic intergrowths, whereas in the latter graphic intergrowth dominates with anomalously coarse centimetric quartz along the cleavages of K-feldspar. Several granitic pegmatites hosted in Archaean complex, in Lages Pintadas Aquamarine Province, Santa Cruz, RN State, present this texture-structure. Graphic intergrowth is attributed to the eutectic crystallization, succeeded by hydrothermal fluids with silica enrichment permitting the growth through diffusion and nucleation of quartz and along cleavages of potash feldspar. In the Archaean terrain, the abundance of recycled chert forming metapsammitic migmatites traversed by numerous quartz veins and coarse graphic granites, has contributed to the growth of beryl and also the aquamarines.