Nilotinib: A Tyrosine Kinase Inhibitor Mediates Resistance to Intracellular Mycobacterium Via Regulating Autophagy.

Nilotinib, a tyrosine kinase inhibitor, has been studied extensively in various tumor models; however, no information exists about the pharmacological action of nilotinib in bacterial infections. Mycobacterium bovis (M. bovis) and Mycobacterium avium subspecies paratuberculosis (MAP) are the etiological agents of bovine tuberculosis and Johne's disease, respectively. Although M. bovis and MAP cause distinct tissue tropism, both of them infect, reside, and replicate in mononuclear phagocytic cells of the infected host. Autophagy is an innate immune defense mechanism for the control of intracellular bacteria, regulated by diverse signaling pathways. Here we demonstrated that nilotinib significantly inhibited the intracellular survival and growth of M. bovis and MAP in macrophages by modulating host immune responses. We showed that nilotinib induced autophagic degradation of intracellular mycobacterium occurred via the inhibition of PI3k/Akt/mTOR axis mediated by abelson (c-ABL) tyrosine kinase. In addition, we observed that nilotinib promoted ubiquitin accumulation around M. bovis through activation of E3 ubiquitin ligase parkin. From in-vivo experiments, we found that nilotinib effectively controlled M. bovis growth and survival through enhanced parkin activity in infected mice. Altogether, our data showed that nilotinib regulates protective innate immune responses against intracellular mycobacterium, both in-vitro and in-vivo, and can be exploited as a novel therapeutic remedy for the control of M. bovis and MAP infections.

Periprosthetic knee infection by Mycobacterium bovis and Candida guilliermondii in the context of a zoonosis: a case report and review of the literature.

INTRODUCTION: Periprosthetic joint infections are a major challenge for treating physicians. Musculoskeletal infections with Mycobacterium bovis are extremely rare, with an assumed incidence of 0.08-0.1%. Consequently, periprosthetic joint infections with Mycobacterium bovis are even less frequent. Fungal periprosthetic joint infections are very rare. No cases of Candida guilliermondii infection of implanted prostheses are described in the literature. CASE PRESENTATION: An 87-year-old Swiss man with German ethnic origin suffered from symptoms of osteoarthritis of the knee. We present the first described case of periprosthetic joint infection after total knee arthroplasty by both Mycobacterium bovis and Candida guilliermondii in the context of a zoonosis with 14 months of follow-up. The infection was presumed to originate more than 55 years earlier, when these infectious agents were still present in cattle in Switzerland. After diagnosis of the pathogens, our patient was successfully treated with tuberculostatic and mycocide medication, and a two-stage revision knee arthroplasty was performed. The medication was given for 1 year. The postoperative course was normal and he achieved ambulant musculoskeletal rehabilitation. After 14 months of follow-up no further complication emerged. At all routine consultations, there were no indications for joint inflammation, wound healing was normal, and the range of motion was flexion/extension 110/0/0°. CONCLUSIONS: We found no comparable cases in our literature search. Only a few joint infections by Mycobacterium bovis after intravesical instillation of Bacillus Calmette-Guérin are described. Primary infections without previous Bacillus Calmette-Guérin injection appear to be even less frequent. In cases where mycobacterial infection cannot be ruled out, we recommend cultivating mycobacteria cultures for weeks. In addition, a histological examination of the tissue should be carried out. After diagnosis, the concept of a two-stage reimplantation of total knee arthroplasty with mycostatic therapy for 1 year and antimycotic therapy appears to be effective.

Calcitriol increases nitric oxide production and modulates microbicidal capacity against Mycobacterium bovis in bovine macrophages.

Bovine tuberculosis, a re-emerging infectious disease caused by Mycobacterium bovis, can be transmitted to humans. Global prevalence of M. bovis in humans is underestimated and represents a serious public health risk in developing countries. In light of this situation, it is important to note that our understanding of the immunopathogenesis of human tuberculosis can be improved by studying this disease in the bovine model. Stimulation of the bovine innate immune system with calcitriol (1,25(OH)2D3) leads to an increase in bactericidal molecules involved in macrophage antimicrobial activity. It is unknown, however, if calcitriol´s effect on bovine macrophages impacts intracellular bacterial replication. With these considerations in mind, this study sought to investigate the specific role of calcitriol in tuberculosis control in bovine macrophages, in the hopes of uncovering information applicable to human tuberculosis. As such, infection with M. bovis was shown to induce expression of CYP27B1 and VDR genes in macrophages. Moreover, addition of 1,25(OH)2D3 to cultures of macrophages previously infected with mycobacteria and/or activated by LPS triggered cellular expression of nitric oxide synthase (NOS2) and increased nitrite concentrations, both indicators of nitric oxide (NO) production. By means of a microbicidal assay, addition of 1,25(OH)2D3 was seen to increase macrophage phagocytosis and to decrease mycobacterial intracellular replication. Thus, taken together, our results show that calcitriol can help stimulate the innate immune system of bovines by increasing phagocytosis and decreasing intracellular replication of microorganisms, such as M. bovis, in macrophages, through the VDR pathway.

[Mycotic aorta aneurysm treated with an autologous venous graft].

Systemic side effects, including sepsis, due to bacille Calmette-Guérin treatment for carcinoma in situ in the bladder, are observed in 15% of the patients. In rare cases, patients have developed systemic infections and mycotic aneurysms. In this case report, a 72-year-old man developed a mycotic aortic aneurysm, and the appropriate tuberculostatic drugs had no effect on his systemic infection. He was successfully treated surgically, replacing the affected aortic segment with an autologous venous graft, resulting in complete remission. A follow-up PET-CT three months later showed no sign of ongoing aortic infection.

Diagnosis of human bovine tuberculosis aided by PET/CT scanning and EBUS-TBNA.

Human bovine tuberculosis is a rare zoonotic infection in developed countries which has been achieved predominantly by effective eradication programmes in cattle. The principal modes of transmission are consumption of unpasteurised dairy products and close contact with infected cattle. The clinical and radiological presentation is indistinguishable from tuberculosis caused by Mycobacterium tuberculosis The diagnosis should be considered in individuals with relevant risk factors who present with intra/extrathoracic pathology. We describe and discuss a case of bovine tuberculosis with a synchronous primary bronchus carcinoma in an immunocompetent individual who presented with a solitary pulmonary nodule and contralateral mediastinal lymphadenopathy on CT imaging. The diagnosis of M. bovis infectionwas aided by 18F-fluorodeoxyglucose positron emission tomography/CT imaging and endobronchial ultrasound-guided mediastinal lymph node sampling.

Expanding the tetrahydroquinoline pharmacophore.

Tetrahydroquinoline is a privileged scaffold with a large number of biological applications. The tetrahydroquinoline pharmacophore has been expanded to yield 34 compounds. Biological screening of these compounds led to the identification of tetrahydroquinoline as neurotropic agents not reported earlier.

Integrated health messaging for multiple neglected zoonoses: Approaches, challenges and opportunities in Morocco.

Integrating the control of multiple neglected zoonoses at the community-level holds great potential, but critical data is missing to inform the design and implementation of different interventions. In this paper we present an evaluation of an integrated health messaging intervention, using powerpoint presentations, for five bacterial (brucellosis and bovine tuberculosis) and dog-associated (rabies, cystic echinococcosis and leishmaniasis) zoonotic diseases in Sidi Kacem Province, northwest Morocco. Conducted by veterinary and epidemiology students between 2013 and 2014, this followed a process-based approach that encouraged sequential adaptation of images, key messages, and delivery strategies using auto-evaluation and end-user feedback. We describe the challenges and opportunities of this approach, reflecting on who was targeted, how education was conducted, and what tools and approaches were used. Our results showed that: (1) replacing words with local pictures and using "hands-on" activities improved receptivity; (2) information "overload" easily occurred when disease transmission pathways did not overlap; (3) access and receptivity at schools was greater than at the community-level; and (4) piggy-backing on high-priority diseases like rabies offered an important avenue to increase knowledge of other zoonoses. We conclude by discussing the merits of incorporating our validated education approach into the school curriculum in order to influence long-term behaviour change.

Antitubercular activity of new coumarins.

The present article describes a series of 21 N '-benzylidene-2-oxo-2H-chromene-3-carbohydrazides 4a-4v, which were synthesized and evaluated for their cell viabilities in non-infected and Mycobacterium bovis Bacillus Calmette-Guerin-infected macrophages. Subsequently, the non-cytotoxic compounds 4c, 4g, 4h, 4j, 4l and 4t were assessed against Mycobacterium tuberculosis ATCC 27294 using the microplate Alamar Blue assay and the activity expressed as the minimum inhibitory concentration in µg/mL. These compounds exhibited a significant activity (50-100 µg/mL) when compared to the first-line drugs, such as pyrazinamide (PZA >100 µg/mL). These results could be considered a good starting point for further studies to develop new lead compounds to treat multidrug-resistant tuberculosis.

Selective delivery of rifampicin incorporated into poly(DL-lactic-co-glycolic) acid microspheres after phagocytotic uptake by alveolar macrophages, and the killing effect against intracellular Mycobacterium bovis Calmette-Guérin.

Macrophages and their phagocytotic abilities play a dominant role for defense against infected organisms. However, Mycobacterium tuberculosis can survive in the phagosomes of macrophages. In this study, the effective delivery of a drug and the killing effect of tubercle bacilli within macrophages were investigated utilizing the phagocytotic uptake of rifampicin (RFP) that had been incorporated into poly(DL-lactic-co-glycolic) acid (PLGA) microspheres. The microspheres were composed of PLGA that had a monomer ratio (lactic acid/glycolic acid) of either 50/50 or 75/25. They had molecular weights from 5000 to 20,000, and diameters of 1.5, 3.5, 6.2 and 8.9 microm. The most significant factor for phagocytotic activity of macrophages was the diameter of the microspheres. By contrast, molecular weight and monomer ratio of PLGA did not influence phagocytosis. The amount of RFP delivered into cells was also investigated. RFP-PLGA microspheres composed of PLGA with a molecular weight of 20,000 and monomer ratio of 75/25 showed the highest amount of delivery (4 microg/1 x 10(6) cells). Fourteen days after infection, the survival rate of treated intracellular bacilli was 1% when compared with untreated cells. There was almost no killing effect of free RFP (4 or 15 microg/ml) on intracellular bacilli. In vivo efficacy of RFP-PLGA was also examined in rats infected with M. tuberculosis Kurono. Intratracheal administration of RFP-PLGA microspheres was shown to be superior to free RFP for killing of intracellular bacilli and preventing granuloma formation in some lobes. These results suggest that phagocytotic activity could be part of a new drug delivery system that selectively targeted macrophages.

Evaluation of susceptibility of Mycobacterium bovis to antituberculous drugs by radiometric BACTEC 460TB system.

Susceptibility of Mycobacterium bovis strains to antituberculous drugs (isoniazid and rifampin) was detected by radiometric BACTEC 460TB system. M.bovis strains were isolated from tissue samples showing tuberculous lesions collected at an abbattoir from cattle belonging to 47 tuberculosis outbreaks occurring in Northern Italy in 1995-1999. Forty-six out of 61 strains (75.4%) resulted susceptible to both isoniazid and rifampin. Thirteen strains (21.3%) were resistant to isoniazid only. No strains showed resistance to rifampin only. Two strains (3.3%) resulted resistant to both drugs, showing antituberculous multidrug-resistance. Given the compulsory eradication program of bovine tuberculosis by elimination of infected animals and the ban on antituberculous drug treatments in animals, detection of resistant M. bovis strains appears of great interest.

Swelling of hand and forearm caused by Mycobacterium bovis.

A 75-year-old native Dutch farmer presented with a painless swelling of his right hand extending into his forearm, accompanied by general malaise and low grade fever. His medical history revealed coxitis tuberculosa in 1954 and injury of the same hand in 1978. His present swelling appeared to be caused by Mycobacterium bovis, probably due to endogenous reactivation. Whether the mycobacteria reached the hand hematogenously or were directly inoculated at the time of injury of his hand in 1978 remains unclear.

Tratamento intermitente da tuberculose bovina com isoniazida / Intermittent treatment of bovine tuberculosis with isoniazid

Foi adotado um regime de tratamento intermitente da tuberculose com isoniazida em 55 bovinos reagentes positivos e 13 suspeitos. A isoniazida em forma de sal cristalizado foi fornecida aos animais misturada em raçäo concentrada, em doses de 25mg/kg p.v., nas 2ª, 4ª e 6ª feiras, durante 6 meses. A cura da tuberculose foi avaliada pela dessensibilizaçäo alérgica, após o término do tratamento através de tuberculinizaçöes comparativas realizadas de 3 em 3 meses, durante um ano. Este monitoramento revelou progressiva reduçäo da sensibilidade alérgica até o desaparecimento definitivo. Aos 90 dias pós-tratamento já havia 31 (45,5//) bovinos negativos, 19 (27,9//) suspeitos e 18 (26,4//) positivos; aos 180 dias, 47 (69,1//) negativos, 10 (14,7//) suspeitos e 11 (15,1//) positivos; aos 270 dias, 51 (75,0//) negativos, 12 (17,6//) suspeitos e 5 (7,3//) positivos e aos 360 dias, 63 (92,6//) negativos, 5 (7,3//) suspeitos e nenhum mais positivo. Como os reagentes suspeitos, no final do tratamento podem ser considerados livres da infecçäo, o percentual de curas foi de 100//, mostrando que o tratamento intermitente com 3 dosagens de isoniazida semanais foi täo eficiente quanto o regime contínuo. O regime intermitente reduziu de 120 para 80 o número de doses administradas e reduziu o custo em 40//