Pathogenesis of an experimental mycobacteriosis in an apple snail.

In this work, we aimed at investigating cell and tissue responses of the apple snail Pomacea canaliculata, following the inoculation of the zoonotic pathogen Mycobacterium marinum. Different doses were tested (10, 20, 65, and 100 M CFU) and the mortality rate was negligible. The histopathogenesis was followed at 4, 9, and 28 days after inoculation. Overt histopathological lesions were consistently observed after the two largest doses only. In the lung, marked hemocyte aggregations, including intravascular nodule formation, were observed within the large blood veins that run along the floor and roof of this organ. Hemocyte aggregations were found occluding many of the radial sinuses supplying the respiratory lamina. Acid-fast bacilli were contained in the different hemocyte aggregations. In addition, hemocytes were observed infiltrating the storage tissue, which makes up most of the lung wall, and the connective tissue of the mantle edge. Additionally, signs of degradation in the storage tissue were observed in the lung wall on day 28. In the kidney, nodules were formed associated with the constitutive hemocyte islets and with the subpallial hemocoelic space, in whose hemocytes the acid-fast structures were found. Electron microscopy analysis revealed the presence of bacteria-containing phagosomes within hemocytes located in the surface zone of the islets. Additionally, electron-dense spheroidal structures, which are likely remnants of digested mycobacteria, were observed in close proximity to the hemocytes' nuclei. The size attained by the hemocyte nodules varied during the observation period, but there was no clear dependence on dose or time after inoculation. Nodules were also formed subpallially. Some of these nodules showed 2-3 layers with different cellular composition, suggesting they may also form through successive waves of circulating cells reaching them. Nodular cores, including those formed intravascularly in the lung, would exhibit signs of hemocyte dedifferentiation, possibly proliferation, and death. Hemocyte congestion was observed in the hemocoelic spaces surrounding the pallial ends of the renal crypts, and the renal crypts themselves showed de-epithelization, particularly on day 28. The diverse cellular responses of P. canaliculata to M. marinum inoculation and the high resilience of this snail to the pathogen make it a suitable species for studying mycobacterial infections and their effects on cellular and physiological processes.

Influencia de la castración en la composición química de la carne de alpacas (Vicugna pacos)

RESUMEN Objetivo. Determinar la influencia de la castración de alpacas jóvenes sobre la composición centesimal y contenido de colesterol de su carne. Materiales y métodos. Con 20 animales machos, de 1.5 años de edad, de raza Huacaya se formaron grupos de 10 animales: experimental y control. Los del experimental, fueron sometidos a castración bilateral y los del control permanecieron "enteros". La alimentación de los animales fue en base a pastos naturales de la zona, bajo un sistema de crianza extensiva. Luego de seis meses, los animales fueron beneficiados y se tomaron muestras de tejido muscular de cinco regiones anatómicas: cuello, brazo, costillar, lomo y pierna. La humedad se determinó por desecación en estufa hasta peso constante, la grasa total por el método de Soxhlet, la proteína bruta por el método Kjeldahl, las cenizas por calcinación en mufla y el colesterol por el método colorimétrico, previa extracción de grasas con cloroformo/metanol (2:1). Resultados. El contenido de grasa es superior en los castrados (2.28%) que en los enteros (1.82%) (p≤0.01) y varía entre las distintas regiones anatómicas (p≤0.05); la humedad es mayor en enteros (76.14%) que en castrados (75.45%) (p≤0.01), pero similar entre las distintas las regiones anatómicas (p>0.05); no hay diferencia estadística en el contenido de proteínas, cenizas y colesterol (p>0.05), siendo el promedio general de 20.39%, 1.13% y 55.56 mg/100 g, respectivamente. Conclusiones. Se evidenció efecto de la castración sobre la composición centesimal de la carne de alpaca de machos jóvenes al incrementar el contenido graso y disminuir la humedad.

ABSTRACT Objective. Determine the influence of bilateral castration in young alpacas over the centesimal chemical composition and content in their meet. Materials and methods. A total of 20 male Huacaya breed animals aged about 1.5 years old, were studied in two groups: experimental and control with 10 animals each. The experimental one was subjected to bilateral castration and the control one remained "intact". The animals' feeding was based on natural pastures from the surrounding area under an extensive rearing system. After six months, the animals were slaughtered and muscle tissue samples were taken from five anatomical regions: neck, arm, ribs, back, and leg. Moisture was determined by oven drying to constant weight, total fat by Soxhlet method, crude protein by Kjeldahl method, ashes by muffle calcination, and cholesterol by the colorimetric method after fat extraction with chloroform/ methanol (2:1). Results. Fat content was higher in castrated animals (2.28%) than in the intact ones (1.82%) (p≤0.01) and varies among different anatomical regions (p≤0.05); humidity is higher in intact animals (76.14%) than in castrated ones (75.45%) (p≤0.01), but it was similar among different anatomical regions (p>0.05); there is no statistical difference in protein, ash, and cholesterol content (p>0.05) with a general average of 20.39%, 1.13%, and 55.56 mg/100 g, respectively. Conclusion Castration effect was evidenced on the centesimal composition of alpaca meat from young males by its fat content increase and humidity decrease.

Guía de práctica clínica para el diagnóstico y manejo inicial de artritis reumatoide en el Seguro Social del Perú (EsSalud) / Clinical Practice Guideline for the diagnosis and initial management of Rheumatoid Arthritis in the Social Security of Peru (EsSalud)

La Artritis Reumatoide (AR) tiene adversas consecuencias en la salud su diagnóstico temprano y manejo óptimo requiere recomendaciones basadas en evidencia de alta calidad adaptadas a cada sistema de salud. Objetivo: Proveer recomendaciones clínicas basadas en evidencia para el diagnóstico y manejo inicial de la AR. Material y Métodos: Se conformó un grupo elaborador de la guía (GEG) que incluyó médicos especialistas y metodólogos. El GEG formuló 10 preguntas clínicas a ser respondidas por la presente GPC. Se realizaron búsquedas sistemáticas de la evidencia publicada en PubMed y CENTRAL entre diciembre 2017 a julio 2019 (revisiones sistemáticas y ­cuando fue considerado pertinente­ estudios primarios). Se seleccionó la evidencia de mayor calidad para responder cada una de las preguntas clínicas planteadas. La certeza de la evidencia fue evaluada usando la metodología Grading of Recommendations Assessment, Development, and Evaluation (GRADE) y en reuniones de trabajo periódicas, el GEG usó dicha metodología para revisar la evidencia y formular las recomendaciones, los puntos de buena práctica clínica y los flujogramas. Finalmente, la GPC fue aprobada con Resolución N° 132 ­ IETSI ­ ESSALUD ­ 2019. Resultados: La presente GPC abordó 10 preguntas clínicas, divididas en dos temas: diagnóstico y manejo. En base a dichas preguntas se formularon 16 recomendaciones (5 fuertes y 11 condicionales), 45 puntos de buena práctica clínica, y 3 flujogramas. Conclusión: El presente artículo resume la metodología y las conclusiones basadas en evidencias de la GPC para el diagnóstico y manejo inicial de la AR en EsSalud.

Rheumatoid arthritis (RA) has adverse health consequences its early diagnosis and optimal management requires high quality evidence-based recommendations tailored to each health system. Objective: To provide evidence-based clinical recommendations for the diagnosis and initial management of RA. Material and Methods: A guideline development group (GDG) was formed that included medical specialists and methodologists. The GEG formulated 10 clinical questions to be answered by this CPG. Systematic searches of the evidence published in PubMed and CENTRAL were performed between December 2017 and July 2019 (systematic reviews and -when considered relevant- primary studies). The highest quality evidence was selected to answer each of the clinical questions posed. The certainty of the evidence was evaluated using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology, and in periodic working meetings, the GEG used this methodology to review the evidence and formulate the recommendations, good clinical practice points, and flowcharts. Finally, the CPG was approved by Resolution No. 132 - IETSI - ESSALUD - 2019. Results: This CPG addressed 10 clinical questions, divided into two topics: diagnosis and management. Based on these questions, 16 recommendations (5 strong and 11 conditional), 45 points of good clinical practice, and 3 flowcharts were formulated. Conclusion: This article summarizes the methodology and evidence-based conclusions of the CPG for the diagnosis and initial management of RA in EsSalud.

Systemic L-Buthionine -S-R-Sulfoximine Treatment Increases Plasma NGF and Upregulates L-cys/L-cys2 Transporter and γ-Glutamylcysteine Ligase mRNAs Through the NGF/TrkA/Akt/Nrf2 Pathway in the Striatum.

Glutathione (GSH) is the most abundant intracellular antioxidant. GSH depletion leads to oxidative stress and neuronal damage in the central nervous system (CNS). In mice, the acute systemic inhibition of GSH synthesis by L-buthionine-S-R-sulfoximine (BSO) triggers a protective response and a subsequent increase in the CNS GSH content. This response might be modulated by a peripheral increment of circulating nerve growth factor (NGF). NGF is an important activator of antioxidant pathways mediated by tropomyosin-related kinase receptor A (TrkA). Here, we report that peripheral administration of BSO increased plasma NGF levels. Additionally, BSO increased NGF levels and activated the NGF/TrkA/Akt pathway in striatal neurons. Moreover, the response in the striatum included an increased transcription of nrf2, gclm, lat1, eaac1, and xct, all of which are involved in antioxidant responses, and L-cys/L-cys2 and glutamate transporters. Using antibody against NGF confirmed that peripheral NGF activated the NGF/TrkA/Akt/Nrf2 pathway in the striatum and subsequently increased the transcription of gclm, nrf2, lat1, eaac1, and xct. These results provide evidence that the reduction of peripheral GSH pools increases peripheral NGF circulation that orchestrates a neuroprotective response in the CNS, at least in the striatum, through the NGF/TrkA/Akt/Nrf2 pathway.

Comparison of Biological Properties of Two Medicinal Extracts of the Tehuacan-Cuicatlan Valley.

In the Valley of Tehuacan-Cuicatlan, Cyrtocarpa procera and Bursera morelensis are located and are used in traditional medicine. In this research, several biological properties were evaluated. The methanol extracts of C. procera (MeCp) and B. morelensis (MeBm) were obtained by maceration. The antibacterial activities of the extracts were evaluated by the Kirby-Baüer disc-diffusion method. The wound healing activity was evaluated by histopathological analysis. Both extracts had a bacteriostatic effect in the Staphylococcus aureus (MeCp MIC = 0.25 mg/mL and MeBm MIC = 1 mg/mL) and the Vibrio cholerae (MeCp MIC = 1 mg/mL and MeBm MIC = 4 mg/mL). Both extracts demonstrated a wound healing efficacy similar to the reference standard (Recoveron). They also showed a high antioxidant capacity (MeCp SC50 = 5.75 µg/mL and MeBm SC50 = 4.27 µg/mL). These results are related to the concentration of phenols (MeCp = 166 and MeBm = 236.6 mg GAe/g) and flavonoids of MeCp = 16 and MeBm = 22 µg Qe/g. Both extracts, acting in a similar way in microorganisms that cause infection thanks to their antioxidant activity, favor the healing of wounds. This is the first study in which the biological properties of these two species are compared.

Boron and Coumaphos Residues in Hive Materials Following Treatments for the Control of Aethina tumida Murray.

In the search of alternatives for controlling Aethina tumida Murray, we recently proposed the BAA trap which uses boric acid and an attractant which mimics the process of fermentation caused by Kodamaea ohmeri in the hive. This yeast is excreted in the feces of A. tumida causing the fermentation of pollen and honey of infested hives and releasing compounds that function as aggregation pheromones to A. tumida. Since the boron is the toxic element in boric acid, the aim of this article is to assess the amount of boron residues in honey and beeswax from hives treated with the BAA trap. For this aim, the amount of bioaccumulated boron in products of untreated hives was first determined and then compared with the amount of boron of products from hives treated with the BAA trap in two distinct climatic and soil conditions. The study was conducted in the cities of Padilla, Tamaulipas, and Valladolid, Yucatan (Mexico) from August 2014 to March 2015. The quantity of boron in honey was significantly less in Yucatan than in Tamaulipas; this agrees with the boron deficiency among Luvisol and Leptosol soils found in Yucatan compared to the Vertisol soil found in Tamaulipas. In fact, the honey from Yucatan has lower boron levels than those reported in the literature. The BAA treatment was applied for four months, results show that the BAA trap does not have any residual effect in either honey or wax; i.e., there is no significant difference in boron content before and after treatment. On the other hand, the organophosphate pesticide coumaphos was found in 100% of wax samples and in 64% of honey samples collected from Yucatan. The concentration of coumaphos in honey ranges from 0.005 to 0.040 mg/kg, which are below Maximum Residue Limit (MRL) allowed in the European Union (0.1 mg/kg) but 7.14% of samples exceeded the MRL allowed in Canada (0.02 mg/kg).

Contexto sociodemográfico de los homicidios en México D.F.: un análisis espacial / Sociodemographic context of homicide in Mexico City: a spatial analysis

Objetivo. Investigar el patrón de distribución espacial de la tasa de homicidios y su relación con las características sociodemográficas en las delegaciones de Benito Juárez, Coyoacán y Cuauhtémoc de la Ciudad de México en el año 2010. Métodos. Estudio inferencial de corte transversal que usa métodos de análisis espacial para estudiar la asociación espacial de la tasa de homicidios y las características demográficas. La asociación espacial fue determinada a través del cociente de localización, análisis de regression múltiple y el uso de la regresión geográficamente ponderada. Resultados. Los homicidios muestran un patrón de localización heterogéneo con altas tasas en zonas con uso del suelo no residencial, con baja densidad de población y baja marginación. Conclusiones. El uso de herramientas de análisis espacial son instrumentos poderosos para el diseño de políticas de seguridad pública preventiva y recreativa que busquen reducir la mortalidad por causas externas como homicidios.

Objective. Investigate the spatial distribution pattern of the homicide rate and its relation to sociodemographic features in the Benito Juárez, Coyoacán, and Cuauhtémoc districts of Mexico City in 2010. Methods. Inferential cross-sectional study that uses spatial analysis methods to study the spatial association of the homicide rate and demographic features. Spatial association was determined through the location quotient, multiple regression analysis, and the use of geographically weighted regression. Results. Homicides show a heterogeneous location pattern with high rates in areas with non-residential land use, low population density, and low marginalization. Conclusions. Spatial analysis tools are powerful instruments for the design of prevention-and recreation-focused public safety policies that aim to reduce mortality from external causes such as homicides.

Contexto sociodemográfico de los homicidios en México D.F.: un análisis espacial / Sociodemographic context of homicide in Mexico City: a spatial analysis

OBJETIVO:Investigar el patrón de distribución espacial de la tasa de homicidios y su relación con las características sociodemográficas en las delegaciones de Benito Juárez, Coyoacán y Cuauhtémoc de la Ciudad de México en el año 2010. MÉTODOS: Estudio inferencial de corte transversal que usa métodos de análisis espacial para estudiar la asociación espacial de la tasa de homicidios y las características demográficas. La asociación espacial fue determinada a través del cociente de localización, análisis de regresión múltiple y el uso de la regresión geográficamente ponderada. RESULTADOS: Los homicidios muestran un patrón de localización heterogéneo con altas tasas en zonas con uso del suelo no residencial, con baja densidad de población y baja marginación. CONCLUSIONES: El uso de herramientas de análisis espacial son instrumentos poderosos para el diseño de políticas de seguridad pública preventiva y recreativa que busquen reducir la mortalidad por causas externas como homicidios.

OBJECTIVE:Investigate the spatial distribution pattern of the homicide rate and its relation to sociodemographic features in the Benito Juárez, Coyoacán, and Cuauhtémoc districts of Mexico City in 2010. METHODS: Inferential cross-sectional study that uses spatial analysis methods to study the spatial association of the homicide rate and demographic features. Spatial association was determined through the location quotient, multiple regression analysis, and the use of geographically weighted regression. RESULTS: Homicides show a heterogeneous location pattern with high rates in areas with non-residential land use, low population density, and low marginalization. CONCLUSIONS: Spatial analysis tools are powerful instruments for the design of prevention- and recreation-focused public safety policies that aim to reduce mortality from external causes such as homicides.

Evaluation of the medicinal properties of Cyrtocarpa procera Kunth fruit extracts.

BACKGROUND: The fruit of Cyrtocarpa procera is used to treat stomach diseases by people living in San Rafael, Coxcatlan, Puebla. This work investigated the antibacterial, antioxidant, cytotoxic and anti-inflammatory activities of the fruit produced by this species. METHODS: Methanol extract was obtained by maceration. After obtaining the methanol extract (MeOH1), methanol subextract (MeOH2) and hexane (H) were obtained. The antibacterial activities of MeOH1, MeOH2 and H were evaluated through disc-diffusion. The quenching of free radicals was evaluated by decolorizing a methanolic DPPH solution. The cytotoxic activity of MeOH2 was evaluated by in vitro assay system of growth inhibition of human cervical carcinoma cell line (CasKi). The IL-1ß and TNF-α were determined through ELISA in the supernatants of the macrophage cell line (RAW 264.7). The MeOH2 subextract was separated by column chromatography, seventy-three fractions were collected. RESULTS: The Gram-positive and -negative bacteria examined were sensitive to MeOH1 and MeOH2; the MeOH2 was bactericidal toward Staphyloccocus aureus (MIC = 4 mg/mL) and Vibrio cholera (MIC = 4 mg/mL). The MeOH2 inhibited the DPPH radical (SC50 = 69.7 µg/mL), but a cytotoxicity assay revealed that the extract is not toxic according to the National Cancer Institute (LD50 = 22.03 µg/mL). The production of proinflammatory cytokines (IL- 1ß and TNF- α) by LPS- stimulated macrophages was reduced after the treatments. The methanol extract contained various organic acids, such as citric acid, palmitic acid and α- linoleic acid. CONCLUSIONS: The fruits of Cyrtocarpa procera are employed to treat ailments such as diarrhea, in this study were demonstrated some biological activities involved in a bacterial infection. This is the first research about of the medicinal properties of C. procera fruit.

[Sociodemographic context of homicide in Mexico City: a spatial analysis]. / Contexto sociodemográfico de los homicidios en México D.F.: un análisis espacial.

OBJECTIVE: Investigate the spatial distribution pattern of the homicide rate and its relation to sociodemographic features in the Benito Juárez, Coyoacán, and Cuauhtémoc districts of Mexico City in 2010. METHODS: Inferential cross-sectional study that uses spatial analysis methods to study the spatial association of the homicide rate and demographic features. Spatial association was determined through the location quotient, multiple regression analysis, and the use of geographically weighted regression. RESULTS: Homicides show a heterogeneous location pattern with high rates in areas with non-residential land use, low population density, and low marginalization. CONCLUSIONS: Spatial analysis tools are powerful instruments for the design of prevention- and recreation-focused public safety policies that aim to reduce mortality from external causes such as homicides.

Resistencia a la traccion de postes de fibra de vidrio cementados con resina autoadhesivo y resina de autocurado / Resistance to the traction of glass fiber post cemented with adhesive and self-curing resin

Determinar la resistencia a la tracci¢n de los cementos de resina autoadhesivo Relyx u100 y de resina autocurado Multilink en la retenci¢n de postes de fibra de vidrio. Material y m‚todos. Se realiz¢ un estudio comparativo, transversal, in vitro, prospectivo, experimental. Se utiliz¢ 40 premolares unirradiculares, se separaron en dos grupos conformados por 20 piezas dentales y fueron seccionados a nivel del techo cameral, se obtuvo as¡ la uniformidad de la muestra con una medida de 18 mm cada una. Se realiz¢ el tratamiento de conductos con la t‚cnica corona apical respectiva y se esper¢ 7 d¡as posteriores al tratamiento. Se desobtur¢ con fresas Gates, Pesso y luego con la fresa del kit de los postes, se estandariz¢ el mismo tama¤o para todos los dientes. Se procedi¢ a la cementaci¢n de los postes seg£n las especificaciones establecidas por los fabricantes Relyx u100 y Multilink. Las piezas dentales fueron colocadas en una probeta (cubo de acr¡lico r pido) por un extremo y, por el otro extremo, los postes en otro cubo. Las probetas para las pruebas de fractura se colocaron dentro de la m quina universal de ensayos hasta el momento en que perdieron su adhesi¢n, una vez obtenidos los resultados fueron archivados en la ficha de recolecci¢n de datos para, posteriormente, realizar las comparaciones respectivas. Resultados. Los postes de fibra de vidrio cementados con resina autocurado presentaron una resistencia a la tracci¢n 30,6 kg, y el cemento de resina autoadhesivo present¢ una resistencia a la tracci¢n de 24,2 kg. Conclusiones. El cemento de resina autocurado present¢ una mayor resistencia a la tracci¢n, comparada con el cemento de resina autoadhesivo, sin embargo, no hubo diferencia estad¡stica.

To determine the resistance to the traction of the cements of autoadhesive resin Relyx u100 and Multilink self-curing resin in retaining glass fiber posts. Material and methods. A comparative, cross, in vitro, prospective, experimental study was made. 40 monoradicular premolars were used, they were separated into two groups comprised of 20 teeth and they were sectioned on cameral roof level obtaining thereby the uniformity of the sample with a measurement of 18 mm each. The root canal with crown apical technique was performed and seven days after treatment were waited. They were desobtureted with Gates, Pesso drill and then with the drill of the posts kit, the same size was standardized for all teeth. It was proceeded to the cementation of posts according to specifications set by manufacturers of Multilink and Relyx u100. The teeth were placed in a test tube (acrylic cube fast) at one end and at the other end in another hub studs. The specimens for fracture tests were placed within the universal testing machine Amsler mark up the time they lost their accession after the results obtained were kept in the data collection sheet to then be able to perform the respective comparisons. Results. The glass fiber poles cemented with cement of self-curing resin presented a tensile strength Kg. 30,6 and the cement of autoadhesive resin presented a tensile strength of 24,2 Kg. Conclusions. The cement of self-curing resin had a higher tensile strength compared with the cement of autoadhesive resin, however we evidenced that there is no statistical difference.

[Laparoscopic drainage of liver abscess. Initial experience]. / Drenaje laparoscópico de absceso hepático. Experiencia inicial.

BACKGROUND: Radiologically guided percutaneous drainage has proven to be simple and effective and currently is considered the gold standard of care with success rates between 80 and 87%. However, not all abscesses are amenable to or will resolve with percutaneous drainage. The purpose of this study is to report our 1-year initial experience with laparoscopic drainage of liver abscesses. METHODS: Patients with diagnosis of liver abscess and who had a surgical indication and were submitted to laparoscopic drainage were studied prospectively from January to December of 2004. RESULTS: Six patients underwent laparoscopic drainage. There were three men and three women with a mean age of 50.8 years. Four patients had single lesions and two had multiple lesions. Mean size of the abscesses was 10.9 x 7.7 cm, segments primarily affected were 5 and 8. Two patients had amebic and four pyogenic abscesses. Mean operative time was 142 min. All patients were eating by the day following surgery and were ambulating between 24 and 72 h postoperatively. All patients reported to be asymptomatic at the time of their last visit. There was one recurrence that resolved with a second procedure. CONCLUSIONS: Success rate with the laparoscopic drainage of liver abscesses (83.3%) makes this procedure an effective and viable surgical alternative as a primary approach in selected patients or after a failed percutaneous drainage.

Infection of BALB/c, C57Bl/6 mice and F1 hybrid CB6F1 mice with strains of leishmania mexicana isolated from Mexican patients with localized or diffuse cutaneous leishmaniasis

Mice from the syngeneic strains BALB/c, C57Bl/6 and (BALB/cxC57Bl/6) F1 hybrids (CB6F1) were infected in the fottpad with six different stains of Leishmania mexicana mexicana isolated from Mexican patients. Three Leishmania strains were isolated from patients with localized cutaneous leishmaniasis (LCL, the benign form of the disease and three from patients with diffuse cutaneous leishmaniasis (DCL, the malignant form of the disease). In BALB/c mice, four Leishmania strains showed a sustained fast growth from 4 to 5 weeks postinfection until the end of the experiment (15 weeks), and the other two grew slowly up to 10 or 12 weeks after infection and then started to grow faster. In C57Bl/6 mice four Leismania strains showed a limited to moderate growth up to 6 to 11 weeks postinfection and then started to decrease. One strain showed a moderate growth during the entire experiment and one strain grew as fast as in BLB/c mice up to 11 weeks postinfection and then started to decrease. The CB6F1 hybrid behaved like the C57Bl/6 parent strain with five Leishmania strains but was much more resistant to one Leishmania strain than the C57Bl/6 mice. Sex of the mouse did not influence the outcome of infection. One important purpose of this work was to see if the Leishmania strains that cause DCL are intrinsically more virulent than those that cause the benign form (LCL). Although important variations in virulence among the Leishmania strains were observed, especially in BALB/c mice, they were not correlated with the type of disease caused in humans