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Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and Burkholderia cenocepacia are considered emerging pathogens classified as a public health problem due to extensive antimicrobial resistance. Therefore, the discovery of new therapeutic strategies has become crucial. This study aimed to evaluate the antimicrobial activity of gallic acid and methyl gallate against non-fermenting bacteria. The study included five clinical isolates of Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and Burkholderia cenocepacia. The minimum inhibitory concentrations of gallic acid and methyl gallate were determined by the broth microdilution method. Growth curves, metabolic activity, and biofilm formation of each bacterial strain in the presence or absence of phenolic compounds were performed. Finally, the therapeutic efficacy of the compounds was evaluated using an in vivo model. Gallic acid and methyl gallate showed antibacterial activity against bacterial strains in a concentration range of 64 to 256 µg/mL, both compounds reduced bacterial growth and metabolic activity of the strains, even at subinhibitory concentrations. Only, methyl gallate exhibited activity to inhibit the formation of bacterial biofilms. Moreover, gallic acid and methyl gallate increased larval survival by up to 60% compared to 30% survival of untreated larvae in a bacterial infection model in Galleria mellonella. Our results highlight the potential of gallic acid and methyl gallate as therapeutic alternatives for infections by emerging non-fermentative bacteria.

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Serratia marcescens is a global opportunistic pathogen. In vitro cytotoxicity of this bacterium is mainly related to metalloprotease serralysin (PrtS) activity. Proteolytic capability varies among the different isolates. Here, we characterized protease production and transcriptional regulators at 37°C of two S. marcescens isolates from bronchial expectorations, HU1848 and SmUNAM836. As a reference strain the insect pathogen S. marcescens Db10 was included. Zymography of supernatant cultures revealed a single (SmUNAM836) or double proteolytic zones (HU1848 and Db10). Mass spectrometry confirmed the identity of PrtS and the serralysin-like protease SlpB from supernatant samples. Elevated proteolytic activity and prtS expression were evidenced in the HU1848 strain through azocasein degradation and qRT-PCR, respectively. Evaluation of transcriptional regulators revealed higher eepR expression in HU1848, whereas cpxR and hexS transcriptional levels were similar between studied strains. Higher eepR expression in HU1848 was further confirmed through an in vivo transcriptional assay. Moreover, two putative CpxR binding motifs were identified within the eepR regulatory region. EMSA validated the interaction of CpxR with both motifs. The evaluation of eepR transcription in a cpxR deletion strain indicated that CpxR negatively regulates eepR. Sequence conservation suggests that regulation of eepR by CpxR is common along S. marcescens species. Overall, our data incorporates CpxR to the complex regulatory mechanisms governing eepR expression and associates the increased proteolytic activity of the HU1848 strain with higher eepR transcription. Based on the global impact of EepR in secondary metabolites production, our work contributes to understanding virulence factors variances across S. marcescens isolates.

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La ruptura de ligamento cruzado anterior (LCA) es la lesión de rodilla más común que se trata de una reconstruc- ción quirúrgica. El objetivo principal de esta revisión fue analizar la importancia de la función de los músculos isquiosurales como factor de riesgo de posibles lesiones de LCA. Se llevó a cabo siguiendo las normas Preferred Reported Item for Systematic Reviews and Meta- analyses (PRISMA). La fuente de la recopilación de datos fue la consulta directa de las bases de datos Pubmed, Sportdiscus, Web of Science y Scopus. Para la recuperación documental, se utilizaron varias palabras clave y se evaluó la calidad de los estudios que fueron seleccionados mediante la escala PEDro. Los déficits de la función muscular en los músculos isquiosurales se han relacionado con una mayor translación tibial anterior y, como consecuencia, un incremento del estrés tensional sobre el LCA entre los 10° y 45° de flexión de rodilla. Una co-activación de los músculos isquiosurales con el músculo cuádriceps femoral puede ser de gran ayuda para reducir los factores de riesgo de la lesión LCA.

SUMMARY: Anterior cruciate ligament (ACL) rupture is the most common knee injury to undergo surgery in the sports setting. The main objective of this review was to analyze the importance of hamstring function as a risk factor for potential ACL injury. It was conducted following the Preferred Reported Item for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The source of data collection was direct consultation of the Pubmed, Sportdiscus, Web of Science and Scopus databases. For documentary retrieval, several key words were used and the quality of the studies that were selected was assessed using the PEDro scale. Muscle function deficits in the hamstrings have been related to increased anterior tibial translation and, as a consequence, increased tensional stress on the ACL between 10° and 45° of knee flexion. A co-activation of the hamstrings with the quadriceps may be helpful in reducing the risk factors for ACL injury.

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Neurocandidiasis is a fungal infection that primarily affects neonates, which is associated with 70% case fatality rates, while pediatric patients who survive infection often have long-term neurological sequelae, making it a clinical requirement to understand the pathogenesis of neonatal neurocandidiasis. Currently, the brain regions to Candida albicans invasion during the neonatal period are not characterized. In this study, 0-day-old mice were infected with C. albicans intravenously to determine dissemination and invasion into the brain at different times post-infection by fungal burden assay and histopathological analysis, additionally cellular death and microglial activation were evaluated by flow cytometry. The results evidenced the dissemination of C. albicans within the first hour of infection in the brain. The meninges were the initial site of invasion during the first 6 hours post infection and then filamentous structures into the brain parenchyma increases during infection, the anatomic regions most susceptible to invasion being the cerebral cortex, thalamus, hypothalamus, midbrain, pons, and medulla oblongata. Furthermore, C. albicans invasion of brain tissue results in cell necrosis and activation of microglia as a consequence of fungal invasion.

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Scedosporium apiospermum is an opportunistic pathogen that can cause pulmonary infections in both immunosuppressive and immunocompetent patients. Cytokines are molecules that mediate the immune response to promote or eliminate fungal infections. In this work, we evaluated the cytokines profile in the lung and serum of mice infected with Scedosporium apiospermum. We found early production of IL-6, IL-1ß and TNF-α cytokines in the lung of infected mice during the first 5 days of infection. We suggest that release of pro-inflammatory cytokines could play a role in the control of fungal invasion.

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Candida auris is an emerging global public health threat. It is an opportunistic yeast that usually affects critically ill patients in healthcare settings and is characterized by reduced susceptibility to multiple antifungal classes. Combination therapy with antifungals and repurposed drugs is a feasible alternative to overcome this problem. The aim of this study was to examine the in vitro interactions and potential synergy of micafungin (MFG) and voriconazole (VRC) plus the antidepressant sertraline (SRT) against clinical isolates of C. auris. Conventional antifungal testing was first performed with the three drugs according to the CLSI methodology. Drug interactions were determined by the checkerboard microdilution assay using the fractional inhibitory concentration (FIC) index. Synergistic interactions were noted with the combination of MFG and SRT plus VRC with FIC values of 0.37 to 0.49 for some strains. Indifferent interactions were observed when MFG was combined with SRT with just one exception (FIC 0.53). No antagonism was observed for any combination. The combination of VRC with MCF or SRT may be relevant for treating C. auris infections.

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INTRODUCTION: The dynamics of COVID-19 transmission occurring in familial clusters may be related to sociodemographic and epidemiological characteristics of cases and contacts. The aim of this study was to identify the dynamics of COVID-19 transmission in families with more than one documented case. METHODOLOGY: Data of about 58 familiar clusters of COVID-19 was gathered and followed up clinically and by telephonic interview. Age, gender, social security plan, comorbidities, occupation, incubation, and symptoms were analyzed using Students' t-test and Chi squared test. RESULTS: The contacts were younger and healthier than cases, and students were predominant (28%). Among the symptomatic contacts, reverse transcription polymerase chain reaction yielded a positive rate of 69%. There were 2.93 contacts per case. Families with clustered cases had more family members when compared to families without clustered cases (4.2 vs. 3.3; p = 0.022). Mean age of contacts in families with clustered cases compared to families without clustered cases also showed differences (29.5 vs. 35.7; p = 0.047). CONCLUSIONS: Characterization of cases and contacts amidst a pandemic is essential for the effective implementation of health policies and research perspectives.

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Karyotypes are characterized by traits such as chromosome number, which can change through whole-genome duplication and dysploidy. In the parasitic plant genus Cuscuta (Convolvulaceae), chromosome numbers vary more than 18-fold. In addition, species of this group show the highest diversity in terms of genome size among angiosperms, as well as a wide variation in the number and distribution of 5S and 35S ribosomal DNA (rDNA) sites. To understand its karyotypic evolution, ancestral character state reconstructions were performed for chromosome number, genome size, and position of 5S and 35S rDNA sites. Previous cytogenetic data were reviewed and complemented with original chromosome counts, genome size estimates, and rDNA distribution assessed via fluorescence in situ hybridization (FISH), for two, seven, and 10 species, respectively. Starting from an ancestral chromosome number of x = 15, duplications were inferred as the prevalent evolutionary process. However, in holocentric clade (subgenus Cuscuta), dysploidy was identified as the main evolutionary mechanism, typical of holocentric karyotypes. The ancestral genome size of Cuscuta was inferred as approximately 1C = 12 Gbp, with an average genome size of 1C = 2.8 Gbp. This indicates an expansion of the genome size relative to other Convolvulaceae, which may be linked to the parasitic lifestyle of Cuscuta. Finally, the position of rDNA sites varied mostly in species with multiple sites in the same karyotype. This feature may be related to the amplification of rDNA sites in association to other repeats present in the heterochromatin. The data suggest that different mechanisms acted in different subgenera, generating the exceptional diversity of karyotypes in Cuscuta.

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Cuscuta is a cytogenetically diverse genus, with karyotypes varying 18-fold in chromosome number and 127-fold in genome size. Each of its four subgenera also presents particular chromosomal features, such as bimodal karyotypes in Pachystigma. We used low coverage sequencing of the Cuscuta nitida genome (subgenus Pachystigma), as well as chromosome banding and molecular cytogenetics of three subgenus representatives, to understand the origin of bimodal karyotypes. All three species, C. nitida, C. africana (2n = 28) and C. angulata (2n = 30), showed heterochromatic bands mainly in the largest chromosome pairs. Eighteen satellite DNAs were identified in C. nitida genome, two showing similarity to mobile elements. The most abundant were present at the largest pairs, as well as the highly abundant ribosomal DNAs. The most abundant Ty1/Copia and Ty3/Gypsy elements were also highly enriched in the largest pairs, except for the Ty3/Gypsy CRM, which also labelled the pericentromeric regions of the smallest chromosomes. This accumulation of repetitive DNA in the larger pairs indicates that these sequences are largely responsible for the formation of bimodal karyotypes in the subgenus Pachystigma. The repetitive DNA fraction is directly linked to karyotype evolution in Cuscuta.

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With the advent of expanded STR (short tandem repeats) typing kits, it was necessary to determine allele frequencies and other appropriate population data parameters for El Salvador. Samples were collected from the central, east, and west regions of the country and typed for 21 forensically relevant STR loci. The data indicate that all loci are highly polymorphic, the three regions are genetically similar, and the population data are similar to those of US Hispanics. The results of this study support that the allele frequency data described herein can be used for statistical calculations for human identity testing in El Salvador.

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Candida auris is an emerging multidrug resistant fungal pathogen, which represents a major challenge for newborns systemic infections worldwide. Management of C. auris infections is complicated due to its intrinsic antifungal resistance and the limited information available on its pathogenesis, particularly during neonatal period. In this study, we developed a murine model of C. auris neonatal invasive infection. C. auris dissemination was evaluated by fungal burden and histopathological analysis of lung, brain, liver, kidney, and spleen at different time intervals. We found fungal cells in all the analyzed tissues, neonatal liver and brain were the most susceptible tissues to fungal invasion. This model will help to better understand pathogenesis mechanisms and facilitate strategies for control and prevention of C. auris infections in newborns.

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This study aimed to assess the species distribution and antifungal susceptibility patterns of 200 strains of Aspergillus isolated from clinical specimens (n = 146) and soil samples (n = 54) in Mexico. ITS, ß-tubulin, and calmodulin DNA sequencing was performed for species identification. Broth microdilution susceptibility testing for amphotericin B, voriconazole, posaconazole, itraconazole, isavuconazole, anidulafungin, caspofungin, and micafungin was done according to CLSI for all strains. A. fumigatus was most frequently recovered from clinical specimens, while A. niger was commonly encountered in soil, both followed by A. flavus in the second place. A total of 60 (30%) cryptic species were identified, with A. tubingensis and A. tamarii being the most commonly found. The decreased susceptibility to amphotericin B and azoles was 32% for both, and were mainly led by A. fumigatus, whereas this percentage decreased to 9% for caspofungin, particularly in A. terreus. More than 75% of cryptic species were susceptible in vitro to all antifungals. Multi-azole decreased susceptibility was detected only in seven isolates. Given that antifungal resistance in Aspergillus spp. is an increasing worldwide threat that causes major challenges in the clinical management of aspergillosis, these data highlight the need for continuous epidemiological surveillance of these pathogens for the implementation of locally adequate treatment strategies. LAY SUMMARY: This is an epidemiological study in Mexico. A. fumigatus was most frequent in clinical specimens and A. niger in soil samples. A. tubingensis and A. tamarii were the most common cryptic species. Resistance to amphotericin B and azoles was 32% each, and 9% for caspofungin.

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Background: Serratia marcescens is an enteric bacterium with increasing incidence in clinical settings, attributed mainly to the opportune expression of diverse virulence determinants plus a wide intrinsic and acquired antibiotic resistance. Methods: The aim of this study was to compare the virulence factor profiles of 185 Serratia marcescens isolates from different clinical origins. In vitro proteolytic and hemolytic activities, biofilm formation, and motility were assessed in each strain. Additionally, the pathogenicity of four hypervirulent strains was analyzed in vivo in Galleria mellonella. Results: We found that bacterial isolates from wound/abscess and respiratory tract specimens exhibited the highest protease activity along with a strong biofilm production, while uropathogenic isolates showed the highest hemolytic activity. Swarming and swimming motilities were similar among all the strains. However, respiratory tract isolates showed the most efficient motility. Two hyperhemolytic and two hyperproteolytic strains were detected; the latter were more efficient killing Galleria mellonella with a 50%-60% larval mortality 48 hours after challenge. Conclusion: A correlation was found between biofilm formation and proteolytic and hemolytic activities in biopsy specimens and bloodstream isolates, respectively. Overall, it becomes critical to evaluate and compare the clinical strains virulence diversity in order to understand the underlying mechanisms that allow the establishment and persistence of opportunistic bacterial infections in the host.

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A pulmonary infection model due to Scedosporium apiospermum in immunocompetent mice was developed. BALB/c mice were infected by endotracheal intubation with 5 × 106 conidia/mouse and disease progression was evaluated on days 1, 3, 5, 7, 11, 16, 21, 30, 50 and 60 post-infection through quantitative culture and histopathological analysis of lungs, livers, spleens, brains, and kidneys. There was no extrapulmonary dissemination during the study nor shown to be a lethal infection. The fungal burden in lungs was maintained from day 1-5 and gradually decreased by day 30 post-challenge. On day 60, 30% of mice showed complete elimination of the fungus. Severe alterations in the lung tissue were observed, as well as the presence of conidia and hyphae surrounded by a cellular infiltrate composed mainly of neutrophils in the first days of the infection. The elimination of fungal cells and normal tissue morphology were recovered throughout the study.

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Low molecular weight protein tyrosine phosphatases (LMW-PTP) are ubiquitous enzymes found across a spectrum of genera from prokaryotes to higher eukaryotes. LMW-PTP belong to the Cys-based PTP class II protein family. Here, we show that LMW-PTP can be categorized into two different groups, referred as class II subdivision I (class II.I) and subdivision II (class II.II). Using BPtpA from the opportunistic pathogen Burkholderia cenocepacia, as a representative member of the LMW-PTP class II.I, we demonstrated that four conserved residues (W47, H48, D80, and F81) are required for enzyme function. Guided by an in silico model of BPtpA, we show that the conserved residues at α3-helix (D80 and F81) contribute to protein stability, while the other conserved residues in the W-loop (W47 and H48) likely play a role in substrate recognition. Overall, our results provide new information on LMW-PTP protein family and establish B. cenocepacia as a suitable model to investigate how substrates are recognized and sorted by these proteins.

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Abstract Background: Both necrotizing enterocolitis and acute kidney injury are tightly related conditions, which independently increase mortality in newborns. Necrotizing enterocolitis is an inflammatory disease with a systemic repercussion that leads to inflammatory kidney changes predisposing to renal damage. Methods: This study assessed risk factors for the development of acute kidney injury in patients diagnosed with necrotizing enterocolitis and compared mortality between patients with or without acute kidney injury. Thirty-nine patients with the diagnosis of necrotizing enterocolitis were included, regardless of the gestational age. Results: Of 39 patients, 38.5% developed acute kidney injury. Survival showed to be significantly lower in patients with acute kidney injury (54.4 days) when compared to newborns without acute kidney injury (76.22 days; p = 0.014). Mortality in patients with acute kidney injury was 46.7%, increasing up to 62.5% with severe kidney damage. The hazard ratio for mortality was 4.708 for acute kidney injury (p = 0.025). The severity of enterocolitis showed to be an independent risk factor in developing acute kidney injury and severe kidney injury (odds ratio [OR] = 1.841, p = 0.034 and OR = 1.917, p = 0.027, respectively). Conclusions: Newborns with necrotizing enterocolitis should be evaluated for early recognition of acute kidney injury. Prospective studies with a higher number of patients are needed to identify modifiable risk factors to impact in the prevention of these conditions.

Resumen Introducción: La enterocolitis necrosante y el daño renal agudo son condiciones íntimamente relacionadas que incrementan independientemente la mortalidad en recién nacidos. La enterocolitis necrosante es una enfermedad inflamatoria sistémica que desencadena cambios renales inflamatorios, predisponiendo el desarrollo de daño renal. Métodos: Se analizaron los factores de riesgo para el desarrollo de daño renal agudo en pacientes con diagnóstico de enterocolitis necrosante y se comparó la mortalidad entre los pacientes sin daño renal y los pacientes con daño renal agudo. Se incluyeron 39 pacientes con diagnóstico de enterocolitis necrosante, independientemente de la edad gestacional. Resultados: De los 39 pacientes, el 38.5% desarrolló daño renal agudo. La sobrevida de los que desarrollaron daño renal agudo (54.4 días) mostró ser significativamente menor al compararse con los recién nacidos que no presentaron daño renal (76.22 días; p = 0.014). La mortalidad en los pacientes con daño renal agudo fue del 46.7%, que se incrementó hasta el 62.5% en aquellos con daño renal grave. El riesgo de mortalidad fue de 4.708 para daño renal agudo (p = 0.025). La gravedad de la enterocolitis necrosante demostró ser un factor de riesgo independiente para el desarrollo de daño renal agudo y de daño renal agudo severo (razón de momios [RM] = 1.841; p = 0.034 y RM = 1.917; p = 0.027, respectivamente). Conclusiones: Los recién nacidos con diagnóstico de enterocolitis necrosante deben ser evaluados para reconocer de forma temprana la presencia de daño renal agudo. Se requiere de estudios prospectivos con mayor número de pacientes para identificar factores de riesgo modificables que puedan impactar en la prevención de estas patologías.

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Background: Both necrotizing enterocolitis and acute kidney injury are tightly related conditions, which independently increase mortality in newborns. Necrotizing enterocolitis is an inflammatory disease with a systemic repercussion that leads to inflammatory kidney changes predisposing to renal damage. Methods: This study assessed risk factors for the development of acute kidney injury in patients diagnosed with necrotizing enterocolitis and compared mortality between patients with or without acute kidney injury. Thirty-nine patients with the diagnosis of necrotizing enterocolitis were included, regardless of the gestational age. Results: Of 39 patients, 38.5% developed acute kidney injury. Survival showed to be significantly lower in patients with acute kidney injury (54.4 days) when compared to newborns without acute kidney injury (76.22 days; p = 0.014). Mortality in patients with acute kidney injury was 46.7%, increasing up to 62.5% with severe kidney damage. The hazard ratio for mortality was 4.708 for acute kidney injury (p = 0.025). The severity of enterocolitis showed to be an independent risk factor in developing acute kidney injury and severe kidney injury (odds ratio [OR] = 1.841, p = 0.034 and OR = 1.917, p = 0.027, respectively). Conclusions: Newborns with necrotizing enterocolitis should be evaluated for early recognition of acute kidney injury. Prospective studies with a higher number of patients are needed to identify modifiable risk factors to impact in the prevention of these conditions.

Introducción: La enterocolitis necrosante y el daño renal agudo son condiciones íntimamente relacionadas que incrementan independientemente la mortalidad en recién nacidos. La enterocolitis necrosante es una enfermedad inflamatoria sistémica que desencadena cambios renales inflamatorios, predisponiendo el desarrollo de daño renal. Métodos: Se analizaron los factores de riesgo para el desarrollo de daño renal agudo en pacientes con diagnóstico de enterocolitis necrosante y se comparó la mortalidad entre los pacientes sin daño renal y los pacientes con daño renal agudo. Se incluyeron 39 pacientes con diagnóstico de enterocolitis necrosante, independientemente de la edad gestacional. Resultados: De los 39 pacientes, el 38.5% desarrolló daño renal agudo. La sobrevida de los que desarrollaron daño renal agudo (54.4 días) mostró ser significativamente menor al compararse con los recién nacidos que no presentaron daño renal (76.22 días; p = 0.014). La mortalidad en los pacientes con daño renal agudo fue del 46.7%, que se incrementó hasta el 62.5% en aquellos con daño renal grave. El riesgo de mortalidad fue de 4.708 para daño renal agudo (p = 0.025). La gravedad de la enterocolitis necrosante demostró ser un factor de riesgo independiente para el desarrollo de daño renal agudo y de daño renal agudo severo (razón de momios [RM] = 1.841; p = 0.034 y RM = 1.917; p = 0.027, respectivamente). Conclusiones: Los recién nacidos con diagnóstico de enterocolitis necrosante deben ser evaluados para reconocer de forma temprana la presencia de daño renal agudo. Se requiere de estudios prospectivos con mayor número de pacientes para identificar factores de riesgo modificables que puedan impactar en la prevención de estas patologías.

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Interspecific hybridization and genome duplication to form allopolyploids are major evolutionary events in angiosperms. In the parasitic genus Cuscuta (Convolvulaceae), molecular data suggested the existence of species of hybrid origin. One of them, C. veatchii, has been proposed as a hybrid between C. denticulata and C. nevadensis, both included in sect. Denticulatae. To test this hypothesis, a cytogenetic analysis was performed with CMA/DAPI staining and fluorescent in situ hybridization using 5S and 35S rDNA and genomic probes. Chromosomes of C. denticulata were small with a well-defined centromeric region, whereas C. nevadensis had larger, densely stained chromosomes, and less CMA+ heterochromatic bands. Cuscuta veatchii had 2n = 60 chromosomes, about 30 of them similar to those of C. denticulata and the remaining to C. nevadensis. GISH analysis confirmed the presence of both subgenomes in the allotetraploid C. veatchii. However, the number of rDNA sites and the haploid karyotype length in C. veatchii were not additive. The diploid parentals had already diverged in their chromosomes structure, whereas the reduction in the number of rDNA sites more probably occurred after hybridization. As phylogenetic data suggested a recent divergence of the progenitors, these species should have a high rate of karyotype evolution.

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BACKGROUND: Invasive pulmonary aspergillosis is a life-threatening fungal disease principally caused by the ubiquitous mould Aspergillus fumigatus. This clinical entity is a major cause of morbidity and mortality (principally, but not restricted to, immunocompromised individuals). A few recent reports suggest in vitro fungicidal activity of sertraline against Aspergillus spp., but this activity has not yet been investigated in vivo. OBJECTIVES: To evaluate the antifungal activity of sertraline in two in vivo models of aspergillosis. METHODS: The antifungal activity of sertraline as monotherapy at three different doses (3, 10 and 15 mg/kg) was evaluated in Galleria mellonella and in a murine model of invasive pulmonary aspergillosis. Therapeutic efficacy parameters determined were larval survival and health index score for G. mellonella, whereas pulmonary fungal burden, galactomannan and lung histopathology were assessed in the murine model. RESULTS: Sertraline treatments improved larval survival and health index score, especially at doses of 10 and 15 mg/kg. Moreover, 10 mg/kg sertraline was able to reduce pulmonary fungal burden with an efficacy comparable with that of 3 mg/kg amphotericin B and 10 mg/kg voriconazole. CONCLUSIONS: To the best of our knowledge, this is the first in vivo study that evaluates the antifungal activity of sertraline against A. fumigatus, showing a possible promising option for the adjuvant treatment of pulmonary aspergillosis.

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