First Record of Culicoides Occidentalis in Sinaloa, Mexico.

Culicoides species adults were collected in light traps located on the coast of Elota, Sinaloa, Mexico, in February 2022. All specimens were females, and it was determined based on their morphology that they belonged to the Culicoides variipennis complex. The identification of the species was carried out by means of a comparative analysis of genetic sequences of the cytochrome c oxidase subunit I gene, which resulted in C. occidentalis, this being its first report in Sinaloa and its third nationwide.

Replication of Dengue Virus 4 in Plodia interpunctella (Lepidoptera: Pyralidae) Larvae Under Laboratory Conditions.

Background: Developing methods for the isolation and replication of dengue virus (DENV), based on nonhematophagous insect models to assess virus-host interaction, would contribute, for instance, to the creation of drugs or vaccines and eventually to the control of the disease. In this regard, nonhematophagous mosquitoes have been used as biological hosts for the isolation of DENV because they are specific and sensitive to a low viral load and viral particles with low infectivity. However, implementation of these models is mainly affected by the complexity of the establishment of the entomological colonies. Materials and Methods: In this study, the susceptibility of DENV-4 infection in Plodia interpunctella larvae was evaluated. Ten larvae, previously inoculated with supernatant from DENV-4-infected C6/36 cells, were processed to determine viral replication by the optical density and 2-ΔΔCt methods at different time intervals (1 and 7 days postinoculation). Results: A prospective increase in viral replication was observed, which did not influence the survival and development of P. interpunctella. Conclusion: These results demonstrate the infectivity of DENV-4 in P. interpunctella, thus becoming an option as a biological model for the study of this etiological agent.

Evaluation of biocompatibility and angiogenic potential of extracellular matrix hydrogel biofunctionalized with the LL-37 peptide.

BACKGROUND: Biomaterials must allow revascularization for a successful tissue regeneration process. Biomaterials formulated from the extracellular matrix (ECM) have gained popularity in tissue engineering because of their superior biocompatibility, and due to their rheological properties, ECM-hydrogels can be easily applied in damaged areas, allowing cell colonization and integration into the host tissue. Porcine urinary bladder ECM (pUBM) retains functional signaling and structural proteins, being an excellent option in regenerative medicine. Even some small molecules, such as the antimicrobial cathelicidin-derived LL-37 peptide have proven angiogenic properties. OBJECTIVE: The objective of this study was to evaluate the biocompatibility and angiogenic potential of an ECM-hydrogel derived from the porcine urinary bladder (pUBMh) biofunctionalized with the LL-37 peptide (pUBMh/LL37). METHODS: Macrophages, fibroblasts, and adipose tissue-derived mesenchymal stem cells (AD-MSC) were exposed pUBMh/LL37, and the effect on cell proliferation was evaluated by MTT assay, cytotoxicity by quantification of lactate dehydrogenase release and the Live/Dead Cell Imaging assays. Moreover, macrophage production of IL-6, IL-10, IL-12p70, MCP-1, INF-γ, and TNF-α cytokines was quantified using a bead-based cytometric array. pUBMh/LL37 was implanted directly by dorsal subcutaneous injection in Wistar rats for 24 h to evaluate biocompatibility, and pUBMh/LL37-loaded angioreactors were implanted for 21 days for evaluation of angiogenesis. RESULTS: We found that pUBMh/LL37 did not affect cell proliferation and is cytocompatible to all tested cell lines but induces the production of TNF-α and MCP-1 in macrophages. In vivo, this ECM-hydrogel induces fibroblast-like cell recruitment within the material, without tissue damage or inflammation at 48 h. Interestingly, tissue remodeling with vasculature inside angioreactors was seen at 21 days. CONCLUSIONS: Our results showed that pUBMh/LL37 is cytologically compatible, and induces angiogenesis in vivo, showing potential for tissue regeneration therapies.

Insectos asociados a productos alimenticios almacenados en el área urbana de Culiacán, Sinaloa, México / Insects associated with food products stored in the urban area of Culiacán Sinaloa, Mexico

Resumen Los insectos asociados a productos almacenados dañan a una amplia variedad de alimentos en hogares y expendios, provocando baja calidad nutricional e incluso riesgo a la salud de los consumidores. El objetivo de este estudio fue determinar las especies y abundancia de insectos asociados a alimentos secos almacenados en casas y tiendas de abarrotes de la ciudad de Culiacán, Sinaloa. Se obtuvieron productos alimenticios, en casas o tiendas de abarrotes, con indicios de daños por insectos o presencia de los mismos, los cuales fueron trasladados al laboratorio, donde se aislaron los imagos. Los organismos inmaduros y las muestras de alimento seco se depositaron en cámaras de emergencia de adultos debido a que la identificación se llevó a cabo por morfología en la fase de imago. Se recolectaron 181 ejemplares de insectos pertenecientes a 8 especies; de ellas, Oryzaephilus mercator (Fauvel), Necrobia rufipes (De Greer), Plodia interpunctella (Hubner) y Cadra cautella (Walker) son nuevos registros para Sinaloa. En las casas se encontró la mayor cantidad de insectos, principalmente del orden coleoptera (U = 96.000, n = 154, P = 0.004). Las especies N. rufipes, T. castaneum (Herbst) y P. interpunctella fueron las más comunes en los sitios de estudio y en las muestras recolectadas. Por primera vez se caracterizó y se determinó la abundancia de insectos plaga de productos alimenticios almacenados en una zona urbana de Sinaloa, entidad federativa con importancia agraria en México. Ubicar taxonómicamente las especies que infestan productos almacenados ayuda a determinar el riesgo económico y de salud que representan para las zonas donde fueron encontrados. También es útil para desarrollar y aplicar medidas adecuadas de control entomológico, en caso de presentarse una plaga en la zona, lo que mantendrá la calidad e integridad de los productos que se comercializan.

Abstract Insects associated with stored produce damage a wide variety of foods in homes and grocery stores, causing poor nutritional quality and even health risk in consumers. The objective of this study was to determine the species and abundance of insects associated with dry food products stored in homes and grocery stores of the city of Culiacán, Sinaloa. Food products were obtained, in houses or grocery stores, with signs of damage by insectes or their presence, which were transferred to the laboratory, where the imagos were isolated immature and dry food samples were deposited in adult emergency chambers because the identification was carried out by morphology in the imago stage. 181 insects belonging to 8 species were collected, of which Oryzaephilus mercator (Fauvel), Necrobia rufipes (De Greer), Plodia interpunctella (Hubner) and Cadra cautella (Walker) represent new records for the state of Sinaloa. The highest number of insects that belong mainly to the order coleoptera were collected in homes (U = 96.000, n = 154, P = 0.004). N. rufipes, T. castaneum (Herbst) and P. interpunctella were the most common species at the study sites and of the food samples collected, dog food and flour were the most affected. For the first time, the abundance of insect pests of stored food products was characterized and determined in an urban area of Sinaloa, an agriculturally important state in Mexico. Identifying taxonomically the species that infest stored products will allow to determine the economic and health risk that they represent for the areas where they were found. It is also useful in developing and applying the appropriate entomological control that will maintain the quality and integrity of the products.

Extracellular matrix hydrogel derived from bovine bone is biocompatible in vitro and in vivo.

BACKGROUND: Nowadays, biomaterials used as a scaffold must be easy to deliver in the bone defect area. Extracellular matrix (ECM) hydrogels are highly hydrated polymers that can fill irregular shapes and act as bioactive materials. OBJECTIVE: This work aims to show the effects of ECM hydrogels derived from bovine bone (bECMh) on proliferation, cytotoxicity and expression of pro-inflammatory cytokines in three cells types involved in tissue regeneration, as well as biocompatibility in vivo. METHODS: In vitro, we used an extract of bECMh to test it on macrophages, fibroblasts, and adipose-derived mesenchymal stem cells (AD-MCSs). Cell proliferation was measured using the MTT assay, cytotoxicity was measured by quantifying lactate dehydrogenase release and the Live/Dead Cell Imaging assays. Concentrations of IL-6, IL-10, IL-12p70, MCP-1 and TNF-α were quantified in the supernatants using a microsphere-based cytometric bead array. For in vivo analysis, Wistar rats were inoculated into the dorsal sub-dermis with bECMh, taking as reference the midline of the back. The specimens were sacrificed at 24 h for histological study. RESULTS: In vitro, this hydrogel behaves as a dynamic biomaterial that increases fibroblast proliferation, induces the production of pro-inflammatory cytokines in macrophages, among which MCP-1 and TNF-α stand out. In vivo, bECMh allows the colonization of host fibroblast-like and polymorphonuclear cells, without tissue damage or inflammation. CONCLUSIONS: The results indicate that bECMh is a biocompatible material that could be used as a scaffold, alone or in conjunction with cells or functional biomolecules, enhancing proliferation and allowing the filling of bone defects to its further regeneration.

First Evidence of Horizontal Transmission by Fecal Shedding of Dengue Virus 4 Among Aedes aegypti Larvae (Diptera: Culicidae) Under Laboratory Conditions.

The transmission pathways of dengue virus (DENV) among mosquitoes are a topic that has gained relevance in recent years because they could explain the maintenance of the virus in the wild independently of the human-mosquito horizontal transmission cycle. In this regard, Aedes aegypti larvae exposed to supernatants of C6/36 cells infected with DENV-4 were evaluated for virus excretion in feces and viability of infection in immature stages (larvae). The results demonstrate that larvae excrete DENV-4 in their feces with the potential to at least infect immature individuals of the same species. A horizontal transmission pathway of larvae-larvae DENV-4 under laboratory conditions is suggested.

First Record of Toxorhynchites moctezuma in Sinaloa, Mexico: Notes about its Morphology.

Mosquito larvae were collected in the urban area of the city of Culiacan, Sinaloa, in September of 2020. The immature stages were placed in emergence containers and fed with Aedes aegypti larvae. The adults that emerged from the immature stages were mounted on insect pins and characterized based on their morphology. The species corresponded morphologically to Toxorhynchites moctezuma, making this the first report of the species for the state of Sinaloa, Mexico. Similarities and morphological variations are discussed with previous analysis for this species.

Natural Vertical Transmission of Dengue Virus Serotype 4 in Aedes aegypti Larvae from Urban Areas in Sinaloa, Mexico.

Dengue virus (DENV) is transmitted to humans by the bite of the vector Aedes aegypti. Several researchers have suggested that the mechanism of vertical transmission of DENV in the vector is a key aspect for the prevalence of the virus in the environment and the potentiation of epidemic outbreaks of the disease. In this context and as part of an integrated study of DENV serotypes in mosquitoes of urban areas in Sinaloa, Mexico, the presence of DENV-4 in larval stages of Ae. aegypti was evaluated to demonstrate the vertical transmission of this serotype. In total, 672 larvae of Ae. aegypti were collected in 16 sectors and were grouped into 36 pools, of which 41.66% (15/36 pools) tested positive for DENV-4, with a minimum infection rate = 22.32. The analysis of the obtained sequences showed a 98% similarity to the DENV-4 with sequences previously reported in GenBank. These results show that Ae. aegypti acts as a natural reservoir for DENV-4 in this region.

Molecular Detection of Leishmania (L.) mexicana (Kinetoplastida: Trypanostomatidae) DNA in Culicoides furens (Diptera: Ceratopogonidae) from an Area with Autochthonous Canine Leishmaniasis in Northwestern Mexico.

PURPOSE: Mexico is considered endemic for Leishmania; recent reports indicate autochthonous human and canine leishmaniasis caused by Leishmania mexicana in Sinaloa state. Lutzomyia sand fly are the primary vector of the parasite, although no records of phlebotomine vectors of Leishmania exist from Sinaloa. Other hematophagous dipterans, like Culicoides, could represent possible vectors of Leishmania in absence of phlebotomines. The known distribution of Culicoides includes the southern portion of Sinaloa state, in northwestern Mexico, with records of Culicoides furens. However, no studies have demonstrated the presence of Leishmania in C. furens or its possible participation in the parasite's life cycle in Mexico. This study, therefore, sought to detect DNA of Leishmania in C. furens captured in an endemic area of autochthonous canine leishmaniasis in northwestern Mexico. METHODS: Culicoides were captured with CDC light traps, identified morphologically, and organized in pools. DNA was extracted, and used to amplify the ribosomal ITS1 region of Leishmania. PCR products were digested with HaeIII endonuclease; the banding patterns obtained were compared to reference strains. RESULTS: Leishmania mexicana DNA was detected in five out of nine pools (55%) of female C. furens. CONCLUSION: This study offers the first evidence of L. mexicana DNA in C. furens, in an endemic area of canine leishmaniasis in northwestern Mexico, where no evidence exists of the presence of phlebotomine sand fly.

First report of autochthonous canine leishmaniasis caused by Leishmania (L.) mexicana in Sinaloa, Mexico.

Leishmaniasis is a neglected tropical disease caused by different species of protozoan parasites of the genus Leishmania. Dogs have been proven as primary hosts of the parasite. Cases of cutaneous leishmaniasis in humans caused by Leishmania mexicana have been reported in Sinaloa; however, the vectors and hosts involved in the epidemiology of the parasite in northwestern Mexico are still unknown. Given the public health implications of this parasite's domestic hosts regarding the permanence and transmission of the disease to humans, the objective of the present study was to detect and determine the species of Leishmania that caused the first three cases of autochthonous canine leishmaniasis in the state of Sinaloa, Mexico. Three domestic dogs showing symptoms similar to canine leishmaniasis were identified, including chronic eye inflammation, corneal opacity, ocular exudate, emaciation and hyporexia. DNA was extracted from venous blood of the infected animals using a commercial kit. The internal transcribed spacer (ITS-1) of ribosomal DNA (rDNA) was amplified by specific primers for Leishmania from the extracted DNA, and the PCR products were digested with the restriction enzyme HaeIII. In addition, PCR products were subjected to automated sequencing. Molecular analysis showed that the infecting species was L. mexicana. This is the first report of autochthonous canine leishmaniasis caused by L. mexicana in Sinaloa, Mexico. Further studies are required to identify the species that serve as vectors and other wild and domestic hosts of the parasite, as well as to determine if there are more species of Leishmania circulating in Sinaloa.

High frequency of HPV16 European variant E350G among Mexican women from Sinaloa.

BACKGROUND & OBJECTIVES: Human papillomavirus (HPV) infections play a crucial role in the aetiology of cervical cancer (CC), and HPV16 is the primary viral genotype associated with CC. A number of variants of the HPV16 E6 gene are involved in the progression of CC, differing in their prevalence and biological and biochemical properties. This study was designed to determine the frequency of HPV types 16/18 and to identify the presence of HPV16 E6-variants in asymptomatic Mexican women. METHODS: A total of 189 cervical Pap smears were collected from women attending public health services in three different cities in Sinaloa, Mexico. Viral DNA was identified by amplification of E6 viral gene fragments using polymerase chain reaction (PCR). Identification of variants was done by sequencing a DNA fragment (321bp) of the HPV16 E6 gene. RESULTS: More than half of the women tested were HPV-positive (52.38%), with HPV16 being the most frequent genotype (21.16%), followed by HPV18 (8.99%). Sequence analysis of the E6-HPV16 PCR products showed that in all cases, the viruses corresponded to European variants. It was further observed that the E350G intra-variant was the most common (>76%). INTERPRETATION & CONCLUSIONS: This study showed a predominance of European lineage variants of HPV16 among asymptomatic women from Sinaloa, Mexico, predominantly with of the E350G variant. This variant has been shown to be associated with an increased risk of early development of CC. The use of molecular identification of carcinogenic HPV and Pap test screening may be a good strategy for monitoring women to prevent CC.

First Evidence of Vertical Infection of Dengue Virus 2 in Aedes aegypti Mosquitoes from Sinaloa, Mexico.

Fourteen pools of Aedes aegypti larvae collected within the urban area of Culiacán, Sinaloa, were analyzed by RT-PCR. The results demonstrate, for the first time, the vertical infection of serotype-2 dengue virus (DENV-2) in Sinaloa, Mexico, suggesting that Ae. aegypti acts as a natural reservoir of DENV-2 in this region.

Genetic Polymorphisms of Tumor Necrosis Factor Alpha and Susceptibility to Dengue Virus Infection in a Mexican Population.

To evaluate the association of the -308 and -238 tumor necrosis factor alpha (TNF-α) gene polymorphisms with clinical manifestations of dengue and TNF-α serum levels in a northwestern Mexican population. The study populations included dengue fever (DF) and dengue hemorrhagic fever (DHF) patients, and a group of healthy controls (HCs) without history of dengue. Polymerase chain reaction-restriction fragment length polymorphism and Enzyme-Linked Immunosorbent Assay were performed to determine genotypes and serum concentration of TNF-α, respectively. There were no significant differences in alleles, genotypes, and haplotype frequencies between patients and HCs. However, when patients were separated into DF and DHF, there was an increased prevalence of the -308 GA genotype in HCs compared to DHF (odds ratio [OR] = 0.129, 95% confidence interval [CI] = 0.018-0.945, p = 0.025), as well as the GG haplotype (OR = 0.49, 95% CI = 0.273-0.880, p = 0.01757) in DF. The genotypes of both polymorphisms were not associated with hematologic manifestations. Serum TNF-α levels were significantly higher in patients than in HCs (p = 0.004). Our results suggest a minimal effect of the -308 and -238 TNF-α gene polymorphisms in dengue patients and that their increased serum levels of TNF-α are independent of genotypes.

First Record of Aedes albopictus in Sinaloa, Mexico.

We report here the discovery of Aedes albopictus for the first time in Sinaloa state, Mexico. The mosquito larvae were collected from small water containers in the urban area of Culiacan city, Sinaloa state. Identification of the species was done primarily by morphology, followed by confirmation with polymerase-chain-reaction-based molecular method.

Spontaneous cure after natural infection with Gnathostoma turgidum (Nematoda) in Virginia opossums (Didelphis virginiana).

Seasonality of the nematode Gnathostoma turgidum in Virginia opossums (Didelphis virginiana) in the wild has been reported; however, the mechanisms involved in deworming are unknown. We monitored the parasitologic and biologic changes in four Virginia opossums naturally infected with G. turgidum by coproparasitologic examination and abdominal ultrasonography. Eggs became detectable in the feces of opossums in May, peaked in July and August, and suddenly decreased in October. Adults of G. turgidum were expelled in the feces mainly in September. Ultrasonography of the liver showed slight damage during May. Lesions in the stomach appeared in April and persisted until September. The abnormalities of the liver and stomach were resolved in November. These data suggest that G. turgidum is likely expelled as a result of host immunologic mechanisms, although termination of a natural life span cannot be definitively excluded.

[A tetracycline-inducible gene system allows for regulated production of proteins with therapeutic potential]. / El sistema génico inducible por tetraciclina permite la producción regulada de proteínas con potencial terapéutico.

The adoption of a bacterial system of control of genic expression with tetracycline, combined with the advances in the identification of regulatory sequences and mechanisms of expression of eukaryotic genes, has increased the versatility and effectiveness of techniques to reintroduce and to make genes function in cells and in superior organisms by being able to control the activity of the transgene in a temporary or reversible manner. This approach has also facilitated making detailed studies of different cellular processes and diseases; for example, the study of the function of oncogenes and other genes involved in the formation and progression of cancer, the generation of cellular models for recombinant protein production with therapeutic purposes, the ex-vivo genetic manipulation of extracted cells from patients and returned to them for gene therapy procedures, as well as the modulation of transgenes to revert hereditary suffering in animal models.

[Telomerase: an enzyme with multiple applications in cancer research]. / La telomerasa: una enzima con múltiples aplicaciones en la investigación del cáncer.

The telomerase is a ribonucleoprotein enzyme to which multiple functions have been attributed, the most important of these is the maintenance of the telomere which is related with cellular immortalization and cancer. 85% of human tumors have telomerase activity, that in normal cells goes undetected. These characteristics make the telomerase an attractive target for chemotherapy.