Modulation of peptidases by 2,4-diamine-quinazoline derivative induces cell death in the amitochondriate parasite Trichomonas vaginalis

Trichomonas vaginalis is an amitochondriate protozoan and the agent of human trichomoniasis, the most prevalent non-viral sexually transmitted infection (STI) in the world. In this study we showed that 2,4-diamine-quinazoline derivative compound (PH100) kills T. vaginalis. PH100 showed activity against fresh clinical and American Type Culture Collection (ATCC) T. vaginalis isolates with no cytotoxicity against cells (HMVI, 3T3-C1 and VERO) and erythrocytes. In addition, PH100 showed synergistic action with metronidazole, indicating that these compounds act by different mechanisms. When investigating the mechanism of action of PH100 to ATCC 30236, apoptosis-like characteristics were observed, such as phosphatidylserine exposure, membrane alterations, and modulation of gene expression and activity of peptidases related to apoptosis. The apoptosis-like cell death features were not observed for the fresh clinical isolate treated with PH100 revealing distinct profiles. Our data revealed the heterogeneity among T. vaginalis isolates and contribute with the understanding of mechanisms of cell death in pathogenic eukaryotic organisms without mitochondria

A chloroacetamide derivative as a potent candidate for fusariosis treatment

Fusariosis has presented a significant increase in their incidence in the last years. This epidemiological panorama probably is due to the increasing profile of refractory susceptibility of Fusarium spp. to available drugs, especially in immunocompromised individuals. Thus, the development of new compounds with effectiveness on these organisms is a necessity. This study evaluated the antifungal potential of a chloroacetamide derivative (4-BFCA) against resistant Fusarium strains. As a result, the compound was effective against all strains (MIC range 12.5–50 μg/mL). The time kill assay demonstrated that 4-BFCA presents a concentration-dependent fungicidal action. Although its action mechanism has not yet been elucidated, it was possible to observe its efficacy through damages and alterations provoked along the hyphae of Fusarium spp. 4-BFCA maintained a high survival rate of Tenebrio molitor larvae, suggesting that it does not cause acute systemic toxicity on this host at the concentration evaluated. In addition, 4-BFCA was 83.33% effective in combating a fungal infection in vivo on the chorioallantoid membrane of embryonated eggs. Our results are very promising and arouse interest to investigate the action of 4-BFCA on Fusarium strains since it acts as a possible candidate for the development of new therapies for the treatment of fusariosis

Management of a Congenital Flexural Deformity in a Calf - Surgical and Pathological Aspects

Background: Flexural deformities are anatomical deviations, in varying degrees, of one or more joints, and may have a congenital or acquired origin. Congenital contracture of the deep digital flexor tendon (DDFT) affects newborn calves and results in hyperflexion of the metacarpal-phalangeal joint, which in severe cases means that the animal must support its own weight on its fetlock joints. The aim of this study is to report the rapid and successful result of applying bilateral total tenotomy technique on a newborn bovine that had been diagnosed with severe bilateral DDFT contracture in the thoracic limbs.Case: A 3-week-old male bovine with a history of difficulty in maintaining a quadrupedal position was attended at the Veterinary Hospital of the Federal University of Tocantins. The animal had severe bilateral locomotor alterations in the thoracic members to the point that he walked on his fetlock joints. The physical examination revealed clinical parameters within the normal range. However, the locomotor system examination showed severe flexor deformity in the bilateral metacarpal-phalangeal joints, and the limbs were being supported on the dorsal face of the fetlock joints, which presented ulcerations, and was suggestive of a shortening of the DDFT. Based on the patient's history and clinical examination, as well as the severity of the tendon contractures, surgical treatment using the DDFT bilateral total tenotomy technique was decided upon. Postoperative treatment consisted of 2.5 mg/kg of enrofloxacin intramuscularly (IM), SID, for 5 days and 0.5 mg/kg meloxicam via IM, SID, for 3 days; as well as a dressing (cotton, medical bandage and a PVC mold) on the thoracic limbs to provide support and allow the animal to walk, until its complete recovery. One day after the surgery, the animal was able to walk with difficulty and some trembling; however, on the third day after the surgical procedure, it could stand up by itself, walk and graze normally. The surgical stitches were removed seven days after the operation, as the surgical wounds had healed adequately. The animal was followed-up for a one-month period after the total tenotomy, with no recurrence of any of the clinical signs, and the thoracic limbs had regained normal biomechanics.Discussion: Congenital flexural deformities are common in newborn calves of different breeds and their incidence is mainly in the thoracic limbs, especially in the carpal, distal interphalangeal and metacarpal-phalangeal joints as was reported in the animal in this study. The history and anamnesis of the animal, along with a thorough physical examination, in order to rule out any other congenital alterations, was extremely important in the diagnosis, and the classification of the severity of the deformity along with the choice of appropriate treatment in the present study. Although total tenotomy is rarely reported in the literature, and there are controversial results with calves, the surgical treatment was chosen in the present case due to the severity of the case. The DDFT bilateral total tenotomy surgical technique gave satisfactory results for the correction of severe bilateral flexor deformity in the thoracic limbs of the newborn crossbred calf. Complete recovery of the limb biomechanics was rapid, and there was no post-surgical complications, thus ensuring the animal could have a good and healthy life. Total DDFT tenotomy is considered a viable surgical procedure for calves with severe congenital flexor deformities; however, post-operative care is also important to ensure good final results

Atresia Ani (Imperforated Anus) in Calves: Clinical, Surgical and Pathological Aspects

Background: Anal atresia is a congenital malformation, which often affects calves, and is related to the imperforation of the membrane that separates the endoderm of the posterior intestine from the ectodermal anal membrane. It is commonly associated with other congenital malformations and skeletal anomalies. The clinical signs generally appear in the first days of the animal's life, due to a retention of feces. The diagnosis is clinical and is based on observation, anamnesis and a physical examination of the animal. The only viable treatment is surgical. This paper aims to report 6 cases of anal atresia in bovine calves, 4 males and 2 females that were successfully treated surgically. Cases: This work reports 6 cases of anal atresia in 4 male calves and 2 female calves. Three presented total atresia (type II), one partial (type I) and in both female calves, anal atresia and rectovaginal fistula (type IV) were observed. The animals were all of undefined race. Five of the cases were from northern Tocantins, 4 males and 1 female (anal atresia with rectovaginal fistula), and 1 female (anal atresia with rectovaginal fistula) was from Valença, RJ. All animals were born active, by eutocic/natural birth, and assumed a quadrupedal position followed by the first feeding as normal. They were aged between 2 days and 6 months, and had a clinical history of abdominal distention and difficulty or inability defecating, and the females both also had a rectovaginal fistula, all cases compatible with anal atresia. Based on the patient's history and clinical examination, surgical treatment for anal reconstruction was decided upon. Postoperative treatment consisted of enrofloxacin [2.5 mg/kg - intramuscularly (IM), once daily (SID), for 5 days] and fluxinin meglumine [1.1 mg/kg - IM, SID, for 3 days] ; as well as a healing ointment which was applied to the area of the surgical wound, every 12 h, for 7 days. There were no trans-surgical complications. The animals showed progressive recovery after anal reconstruction and the stitches were removed in all cases on the 10th postoperative day, with no postoperative complications and no recurrence of any clinical signs from that moment on.Discussion: The study of congenital and hereditary changes enables the identification of their origins, can help prevent new cases and, some of them, are open to economically viable treatment and/or correction that can improve the well-being of the animal and prevent economic losses due to death or animal sacrifice, as reported in the present study. Anal atresia is the most common congenital defect of the lower gastrointestinal tract in calves, being an isolated abnormality, or associated with other malformations, especially of the distal spinal column such as the absence of a tail (perosumus acaudato), as one of the animals in this study. The clinical signs and physical examination are sufficient to establish the diagnosis, as demonstrated in this report, which is usually made in newborn animals, due to the lack or difficulty in defecation associated with no anal orifice and/or swelling in the perineal region. The treatment of choice for anal atresia is surgical, in order to construct an anal neo-orifice and thus avoid endotoxemic shock as well as providing relief and well-being for the animals. As observed in this study, when anal atresia is diagnosed early, and surgical treatment is properly instituted, the prognosis is favorable. The surgery is considered of low complexity, quick and it can be carried out in the field. Thus, from a commercial point of view, considering the costs of the procedures and the value of the calf at the end of weaning, such treatments are beneficial to the owners. In addition, the surgical treatment is essential for animal health and welfare in cases of anal atresia.Keywords: cattle, congenital defects, hereditary pathology, perosomus acaudato, surgery

Caracterização morfoagronômica de acessos de bucha vegetal (Luffa spp.)

A bucha vegetal pode apresentar importância no mercado de produtos agroecológicos devido às fibras naturais e aplicabilidade em diversos setores da agroindústria. O objetivo do trabalho foi caracterizar oito acessos de bucha vegetal para identificar genótipos potenciais para fins de melhoramento. Foi utilizado o delineamento experimental de blocos completos ao acaso, com oito tratamentos e três repetições. Os tratamentos corresponderam a oito acessos de bucha vegetal (A1 até A8), com a parcela constituída por quatro plantas. A caracterização dos acessos foi realizada pela adoção de 16 descritores morfológicos. Os dados foram submetidos à análise de variância univariada pelo teste F de Snedecor, teste de Tukey a 5% de probabilidade, estudo da divergência genética foi feito pela matriz de distâncias Euclidiana, análise multivariada de agrupamento pelo método UPGMA e a importância relativa dos descritores para divergência genética pelo método de Singh. Foi notada uma variação considerável para doze dos descritores avaliados. Observou-se variação entre os acessos estudados para razão foliar, número de ramificações, descritores de flores masculinas e femininas e número médio de frutos por acesso. No método de agrupamento UPGMA, a caracterização pode sugerir cruzamentos entre genitores divergentes, com a formação de quatro grupos: GI (A6), GII (A1 e A8), GIII (A2, A3 e A7) e GIV (A4 e A5). Por fim, os descritores comprimentos de pétala, cálice e receptáculo das flores femininas foram os que mais contribuíram para a diversidade com, respectivamente, 87,87; 10,02 e 1,71%

Combining the pharmacophore features of coumarins and 1,4-substituted 1,2,3-triazoles to design new acetylcholinesterase inhibitors : fast and easy generation of 4-methylcoumarins/1,2,3-triazoles conjugates via Click Chemistry

Coumarins are a large class of compounds that display a range of interesting biological properties, being considered privileged structures because of the ability of their 2H-chromen-2-one nuclei to bind to multiple pharmacological targets. We hypothesized that the linkage of a second pharmacophore nucleus to the 2H-chromen-2-one core, the 1,2,3-triazole moiety, would entail more selective and pharmacologically active coumarins. Therefore, we describe the synthesis of fourteen 4-methylcoumarins/1,4-substituted 1,2,3-triazole conjugates, which were predicted by in silico methods to inhibit acetylcholinesterase (AChE) and proved to be moderate in vitro inhibitors of this enzyme. Molecular docking simulations suggest that the most active of these compounds has a putative binding mode similar to donepezil, both occupying the peripheral anionic site of AChE, which is associated with the secondary noncholinergic functions of the enzyme. This highlights the potential of this series for further optimization in the search of new coumarins for the treatment of Alzheimer’s disease

New 99mTc-Labeled Digitoxigenin Derivative for Cancer Cell Identification

In recent years, cardiac glycosides (CGs) have been investigated as potential antiviral and anticancer drugs. Digitoxigenin (DIG) and other CGs have been shown to bind and inhibit Na+/K+-adenosinetriphosphatase (ATPase). Tumor cells show a higher expression rate of the Na+/K+-ATPase protein or a stronger affinity towards the binding of CGs and are therefore more prone to CGs than non-tumor cells. Cancer imaging techniques using radiotracers targeted at specific receptors have yielded successful results. Technetium-99m (99mTc) is one of the radionuclides of choice to radiolabel pharmaceuticals because of its favorable physical and chemical properties along with reasonable costs. Herein, we describe a new Na+/K+-ATPase targeting radiotracer consisting of digitoxigenin and diethylenetriaminepentaacetic acid (DTPA), a bifunctional chelating ligand used to prepare 99mTc-labeled complexes, and its evaluation as an imaging probe. We report the synthesis and characterization of the radiolabeled compound including stability tests, blood clearance, and biodistribution in healthy mice. Additionally, we investigated the binding of the compound to A549 human non-small-cell lung cancer cells and the inhibition of the Na+/K+-ATPase by the labeled compound in vitro. The 99mTc-labeled DTPA–digitoxigenin (99mTc-DTPA–DIG) compound displayed high stability in vitro and in vivo, a fast renal excretion, and a specific binding towards A549 cancer cells in comparison to non-tumor cells. Therefore, 99mTc-DTPA–DIG could potentially be used for non-invasive visualization of tumor lesions by means of scintigraphic imaging