AIM:Sex-specific immune responses to intra-amniotic infections remain poorly understood despite their key role in preterm birth. METHODS:We infected male and female amniotic fluid cells (AFCs) with M. hominis to explore the potential impact of sex-specific innate immune responses, evaluating the gene and protein expression levels of IL-1β, IL-6, IL-8, TNF-α, TLR2 and TLR4, the production of nitrites, and the levels of selected miRNAs. KEY FINDINGS:The gene expression of IL-1β, IL-6, IL-8, TNF-α, and TLR2 were elevated in infected females AFCs, while only IL-6, IL-8 and TLR4 expression were up-regulated in infected males. Protein levels of IL-6, TNF-α and TLR2 were up-regulated exclusively in infected females. Furthermore, infected female AFCs produced higher levels of nitrites. MiRNA expression revealed an up-regulation of miR-29a-3p in infected females, and miR-223-3p in infected males, with miR-29b-3p showing up-regulation in both sexes upon infection. SIGNIFICANCE:The response to intrauterine infections differs between males and females. Female foetuses may possess a greater capacity to manage the infection and inflammation, underscoring the importance of personalized prenatal care.

(1) Background: It has been previously shown that sourdough bread, compared to commercial yeast bread, elicits a lower postprandial glycemic and insulinemic response in patients with impaired glucose tolerance (IGT). Aims: Our aim was to evaluate the following aspects in persons with type 2 diabetes (T2DM): (1) glucose variations induced by three different doughs: X = bread prepared with functional alkaline biocrystal water, Y = sourdough-leavened bread, and W = bakery yeast bread; (2) the utility of flash glucose monitoring (FGM) to measure GL. (2) Methods: Twelve T2DM following diets (six males, diabetes duration 10.9 ± 1.3 years with no complications, Hba1c < 7.0%), after 12 h of fasting, consumed 180 g of the study breads leavened/matured for 48 (X), 8 (Y), and 4 h (W) at room temperature with 200 mL of water, in a random order, in single-blind conditions, on three different days. All patients had FGM running for the entire period of the experiments. Insulin was determined by capillary blood obtained for the basal and peak glucose concentrations. (3) Results: The peak glucose and peak insulin concentrations were significantly (p < 0.05) higher for W versus both X and Y, without significant differences between X and Y. The area under the curve of glucose variations for over 240 min was significantly higher in W than X (p < 0.01) and Y (p < 0.05), without significant differences between X and Y. (4) Conclusions: (1) Bread prepared with biocrystal water has the same lower GL of sourdough bread compared to bakery yeast bread, and it is easier to manage its leavening/maturation period; (2) FGM is a reliable method for determining rapid glucose changes in response to a carbohydrate meal in persons with type 2 diabetes.

The cultivation of cyanobacteria by exploiting available in situ resources represents a possible way to supply food and oxygen to astronauts during long-term crewed missions on Mars. Here, we evaluated the possibility of cultivating the extremophile cyanobacterium Chroococcidiopsis thermalis CCALA 050 under operating conditions that should occur within a dome hosting a recently patented process to produce nutrients and oxygen on Mars. The medium adopted to cultivate this cyanobacterium, named Martian medium, was obtained using a mixture of regolith leachate and astronauts’ urine simulants that would be available in situ resources whose exploitation could reduce the mission payload. The results demonstrated that C. thermalis can grow in such a medium. For producing high biomass, the best medium consisted of specific percentages (40%vol) of Martian medium and a standard medium (60%vol). Biomass produced in such a medium exhibits excellent antioxidant properties and contains significant amounts of pigments. Lipidomic analysis demonstrated that biomass contains strategic lipid classes able to help the astronauts facing the oxidative stress and inflammatory phenomena taking place on Mars. These characteristics suggest that this strain could serve as a valuable nutritional resource for astronauts.

We report the use of a new multiplex Real-Time PCR platform to simultaneously identify 24 pathogens and 3 antimicrobial-resistance genes directly from respiratory samples of COVID-19 patients. Results were compared to culture-based diagnosis.Secondary infections were detected in 60% of COVID-19 patients by molecular analysis and 73% by microbiological assays, with no significant differences in accuracy, indicating Gram-negative bacteria as the predominant species. Among fungal superinfections, Aspergillus spp. were detected by both methods in more than 7% of COVID-19 patients. Oxacillin-resistant S. aureus and carbapenem-resistant K. pneumoniae were highlighted by both methods.Secondary microbial infections in SARS-CoV-2 patients are associated with poor outcomes and an increased risk of death. Since PCR-based tests significantly reduce the turnaround time to 4 hours and 30 minutes (compared to 48 hours for microbial culture), we strongly support the routine use of molecular techniques, in conjunction with microbiological analysis, to identify co/secondary infections.

Trichomonas vaginalis and Mycoplasma hominis, two microorganisms causing infections of the urogenital tract, are closely associated in that they establish an endosymbiosis relationship, the only case among human pathogens. As a result, the presence of one microorganism may be considered a sign that the other is present as well. Identification of the two pathogens in clinical samples is based on cultivation techniques on specific media, even though in recent years, new sensitive and rapid molecular techniques have become. Here, we demonstrate that the concomitant presence of T.vaginalis in urogenital swabs may lead to a delay in the identification of M.hominis, and thus to an underestimation of bacterial infections when cultural techniques are used.

Abstract Background It has been previously showed that sourdough bread, compared to commercial yeast bread, elicits a lower postprandial glycemic and insulinemic response patients with Impaired Glucose Tolerance (IGT). Aims To evaluate in type 2 diabetic patients (T2DM): 1) Glucose load (GL) to three different doughs: X = prepared with functional alkaline biocrystal water, Y = Sourdough leavened bread, and W = baker yeast bread; 2) the utility of flash glucose monitoring (FGM) to measure GL. Methods Ten T2DM on diet (6 Males, diabetes duration 10.8 ± 1.2 years with no complications, Hba1c < 7.0%), after 12 hours fasting, consumed 180 grams study bread leavened/matured for 48 (X),8 (Y), and 4 hours (W) at room temperature with 200 ml water, in random order, single-blind, in three different days. All patients had a FGM running for the entire period of the experiments. Insulin was determined by capillary blood obtained at basal and at the peak glucose concentration. Results Peak glucose and peak insulin concentration were significantly (p < 0.05) higher for W versus both X and Y, without significant differences between X and Y. Glucose Load over 240 minutes was significantly higher in W than X (p < 0.01) and Y (p < 0.05), without significant differences between X and Y. Conclusions 1) bread prepared with biocrystal water has the same lower GL of Sourdough Bread compared to baker yeast bread with an easier management of the leavening/maturation period; 2) FGM is a reliable method for determining the GL in response to a carbohydrate meal in type 2 diabetic patients.

Abstract Background It has been previously showed that sourdough bread, compared to commercial yeast bread, elicits a lower postprandial glycemic and insulinemic response patients with Impaired Glucose Tolerance (IGT). Aims To evaluate in type 2 diabetic patients (T2DM): 1) Glucose load (GL) to three different doughs: X = prepared with functional alkaline biocrystal water, Y = Sourdough leavened bread, and W = baker yeast bread; 2) the utility of flash glucose monitoring (FGM) to measure GL. Methods Ten T2DM on diet (6 Males, diabetes duration 10.8 ± 1.2 years with no complications, Hba1c < 7.0%), after 12 hours fasting, consumed 180 grams study bread leavened/matured for 48 (X),8 (Y), and 4 hours (W) at room temperature with 200 ml water, in random order, single-blind, in three different days. All patients had a FGM running for the entire period of the experiments. Insulin was determined by capillary blood obtained at basal and at the peak glucose concentration. Results Peak glucose and peak insulin concentration were significantly (p < 0.05) higher for W versus both X and Y, without significant differences between X and Y. Glucose Load over 240 minutes was significantly higher in W than X (p < 0.01) and Y (p < 0.05), without significant differences between X and Y. Conclusions 1) bread prepared with biocrystal water has the same lower GL of Sourdough Bread compared to baker yeast bread with an easier management of the leavening/maturation period; 2) FGM is a reliable method for determining the GL in response to a carbohydrate meal in type 2 diabetic patients.

Trichomonas vaginalis is a pathogenic protozoan diffused worldwide capable of infecting the urogenital tract in humans, causing trichomoniasis. One of its most intriguing aspects is the ability to establish a close relationship with endosymbiotic microorganisms: the unique association of T. vaginalis with the bacterium Mycoplasma hominis represents, to date, the only example of an endosymbiosis involving two true human pathogens. Since its discovery, several aspects of the symbiosis between T. vaginalis and M. hominis have been characterized, demonstrating that the presence of the intracellular guest strongly influences the pathogenic characteristics of the protozoon, making it more aggressive towards host cells and capable of stimulating a stronger proinflammatory response. The recent description of a further symbiont of the protozoon, the newly discovered non-cultivable mycoplasma Candidatus Mycoplasma girerdii, makes the picture even more complex. This review provides an overview of the main aspects of this complex microbial consortium, with particular emphasis on its effect on protozoan pathobiology and on the interplays among the symbionts.

Objectives: Mycoplasma hominis, an opportunistic pathogen of the human lower urogenital tract, can survive and replicate within the protozoan Trichomonas vaginalis, establishing an endosymbiotic relation-ship. The intracellular location may provide a means for the bacteria to evade the immune system and protection from antimicrobial activities. Our aim was to investigate the influence of the endosymbiotic association of M. hominis with trichomonad cells on bacterial antibiotic susceptibility.Methods: We evaluated antibiotic resistance patterns in a group of M. hominis isolated from T. vaginalis clinical specimens as well as in M. hominis isolated from patients without trichomoniasis. Using an experimental model system, we compared the minimum inhibitory concentration (MIC) and lethal con-centration (MLC) of tetracycline on M. hominis endosymbionts of T. vaginalis and extracellular bacteria.Results: The incidence rate of M. hominis strains resistant to C14 and C15 macrolide antibiotics was higher in intracellular strains associated with T. vaginalis compared with extracellular bacteria isolated from women not affected by trichomoniasis. However, sensitivity to tetracycline and quinolones was similar in both groups. In vitro experiments demonstrated that M. hominis strains, when isolated as endosymbionts from T. vaginalis, exhibited reduced sensitivity to tetracycline when cultured extracellularly for at least eight weeks.Conclusion: The intracellular localization of bacteria within trichomonad cells may affect antibiotic susceptibility.(c) 2023 The Author(s). Published by Elsevier Ltd on behalf of International Society for Antimicrobial Chemotherapy. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ )

Nowadays, fungal infections increase, and the demand of novel antifungal agents is constantly rising. In the present study, silver, titanium dioxide, cobalt (II) hydroxide and cobalt (II,III) oxide nanomaterials have been synthesized from Spirulina platensis extract. The synthesis mechanism has been studied using GCMS and FTIR thus confirming the involvement of secondary metabolites, mainly amines. The obtained products have been analysed using XRD, SEM, TGA and zeta potential techniques. The findings revealed average crystallite size of 15.22 nm with 9.72 nm for oval-shaped silver nanoparticles increasing to 26.01 nm and 24.86 nm after calcination and 4.81 nm for spherical-shaped titanium dioxide nanoparticles which decreased to 4.62 nm after calcination. Nanoflake shape has been observed for cobalt hydroxide nanomaterials and for cobalt (II, III) oxide with crystallite size of 3.52 nm and 13.28 nm, respectively. Silver nanoparticles showed the best thermal and water dispersion stability of all the prepared structures. Once subjected to three different Candida species (C. albicans, C. glabrata, and C. krusei) silver nanoparticles and cobalt (II) hydroxide nanomaterials showed strong antifungal activity at 50 μg/mL with minimum inhibitory concentration (MIC) values. After light exposition, MIC values for nanomaterials decreased (to 12.5 μg/mL) for C. krusei and increased (100 μg/mL) for C. albicans and C. glabrata.

Trichomoniasis, the most common non-viral sexually transmitted infection worldwide, is caused by the protozoon Trichomonas vaginalis. The 5- nitroimidazole drugs, of which metronidazole is the most prescribed, are the only effective drugs to treat trichomoniasis. Resistance against metronidazole is increasingly reported among T. vaginalis isolates. T. vaginalis can establish an endosymbiosis with two Mycoplasma species, Mycoplasma hominis and Candidatus Mycoplasma girerdii, whose presence has been demonstrated to influence several aspects of the protozoan pathobiology. The role of M. hominis in T. vaginalis resistance to metronidazole is controversial, while the influence of Ca. M. girerdii has never been investigated. In this work, we investigate the possible correlation between the presence of Ca. M. girerdii and/or M. hominis and the in vitro drug susceptibility in a large group of T. vaginalis isolated in Italy and in Vietnam. We also evaluated, via RNA-seq analysis, the expression of protozoan genes involved in metronidazole resistance in a set of syngenic T. vaginalis strains, differing only for the presence/absence of the two Mycoplasmas. Our results show that the presence of M. hominis significantly increases the sensitivity to metronidazole in T. vaginalis and affects gene expression. On the contrary, the symbiosis with Candidatus Mycoplasma girerdii seems to have no effect on metronidazole resistance in T. vaginalis.

T. vaginalis and M. hominis form a unique case of endosymbiosis that modulates the parasite’s pathobiology. Recently, a new nonculturable mycoplasma species (“ Candidatus Mycoplasma girerdii”) has been described as closely associated with the protozoon.

Vaginal infections affect millions of women annually worldwide. Therapeutic options are limited, moreover drug-resistance increases the need to find novel antimicrobials for health promotion. Recently phytochemicals were re-discovered for medical treatment. Myrtle (Myrtus communis L.) plant extracts showed in vitro antioxidant, antiseptic and anti-inflammatory properties thanks to their bioactive compounds. The aim of the present study was to create novel nanodevices to deliver three natural extracts from leaves, seeds and fruit of myrtle, in vaginal milieu. We explored their effect on human cells (HeLa, Human Foreskin Fibroblast-1 line, and stem cells isolated from skin), resident microflora (Lactobacillus acidophilus) and on several vaginal pathogens (Trichomonas vaginalis, Escherichia coli, Staphylococcus aureus, Candida albicans, Candida kefyr, Candida glabrata, Candida parapsilosis, Candida krusei). Polycaprolactone-Gelatin nanofibers encapsulated with leaves extract and soaked with seed extracts exhibited a different capability in regard to counteracting microbial proliferation. Moreover, these nanodevices do not affect human cells and resident microflora viability. Results reveal that some of the tested nanofibers are interesting candidates for future vaginal infection treatments.

Ever since the publication of the seminal paper by Lynn Margulis in 1967 proposing the theory of the endosymbiotic origin of organelles, the study of the symbiotic relationships between unicellular eukaryotes and prokaryotes has received ever-growing attention by microbiologists and evolutionists alike. While the evolutionary significance of the endosymbiotic associations within protists has emerged and is intensively studied, the impact of these relationships on human health has been seldom taken into account. Microbial endosymbioses involving human eukaryotic pathogens are not common, and the sexually transmitted obligate parasite Trichomonas vaginalis and the free-living opportunistic pathogen Acanthamoeba represent two unique cases in this regard, to date. The reasons of this peculiarity for T. vaginalis and Acanthamoeba may be due to their lifestyles, characterized by bacteria-rich environments. However, this characteristic does not fully explain the reason why no bacterial endosymbiont has yet been detected in unicellular eukaryotic human pathogens other than in T. vaginalis and Acanthamoeba, albeit sparse and poorly investigated examples of morphological identification of bacteria-like microorganisms associated with Giardia and Entamoeba were reported in the past. In this review article we will present the body of experimental evidences revealing the profound effects of these examples of protist/bacteria symbiosis on the pathogenesis of the microbial species involved, and ultimately their impact on human health.

The protozoon Trichomonas vaginalis is responsible for trichomoniasis, a common sexually transmitted infection associated with an increased risk of HIV infection and adverse pregnancy outcomes. The protozoon has the surprising ability to establish a symbiotic relationship with other microorganisms. In fact, most T.vaginalis isolates intracellularly host the vaginal bacterium Mycoplasma hominis and can harbor up to four dsRNA viruses. Moreover, a novel Mycoplasma species named Ca. Mycoplasma girerdii has been recently described as associated with trichomonad cells. Trichomonas vaginalis colonizes the human vagina and its presence causes profound alterations of the resident microbiota, leading to dysbiosis. In healthy women, vaginal microbiota is characterized by the presence of a complex population of aerobic and anaerobic microorganisms living in a physiologically dynamic system dominated by bacteria of the genera Lactobacillus. The most common microbial vaginal imbalance is bacterial vaginosis, a polymicrobial disease associated with several adverse reproductive outcomes and increased risk of HIV infection. Here, we review the current knowledge regarding the interactions between both T.vaginalis and M.hominis and the vaginal microbiota, and we discuss the possibility of a cooperation between T.vaginalis and its symbionts in the development of vaginal dysbiosis.

Trichomonas vaginalis is an anaerobic protist, responsible for the most prevalent non-viral sexually transmitted infection in humans. One of the most intriguing aspects of T. vaginalis pathobiology is the complex relationship with intracellular microbial symbionts: a group of dsRNA viruses belonging to family of Totiviridae ( T. vaginalis virus), and eubacteria belonging to the Mycoplasma genus, in particular M ycoplasma hominis . Both microorganisms seem to strongly influence the lifestyle of T. vaginalis , suggesting a role of the symbiosis in the high variability of clinical presentation and sequelae during trichomoniasis. In the last few years many aspects of this unique symbiotic relationship have been investigated: M. hominis resides and replicates in the protozoan cell, and T. vaginalis is able to pass the bacterial infection to both mycoplasma-free protozoan isolates and human epithelial cells; M. hominis synergistically upregulates the proinflammatory response of human monocytes to T. vaginalis . Furthermore, the influence of M. hominis over T. vaginalis metabolism and physiology has been characterized. The identification of a novel species belonging to the class of Mollicutes (Candidatus Mycoplasma girerdii ) exclusively associated to T. vaginalis opens new perspectives in the research of the complex series of events taking place in the multifaceted world of the vaginal microbiota, both under normal and pathological conditions.

The flagellated protozoon Trichomonas vaginalis, responsible for trichomoniasis, can establish a symbiotic relationship with the bacterium Mycoplasma hominis and can harbor double-stranded RNA Trichomonasvirus (TVV). In this study, we investigated by real-time PCR the prevalence of the four TVVs and of M. hominis among 48 T. vaginalis strains isolated in Italy, and we evaluated a possible association with metronidazole resistance. Fifty percent of the analyzed trichomonad strains tested positive for at least one TVV T. vaginalis, with TVV2 being the most prevalent, followed by TVV1 and TVV3. Two T. vaginalis strains were infected by TVV4, detected in Europe for the first time. Interestingly, we found more than one TVV species in 75% of positive trichomonad strains. M. hominis was present in 81.25% of T. vaginalis isolates tested, and no statistically significant association was observed with the infection by any TVV. Metronidazole sensitivity of T. vaginalis isolates was evaluated in vitro, and no correlation was observed between minimal lethal concentration and the presence of TVVs. This is the first report on TVV infection of T. vaginalis in Italy. Even if no association of TVV positive isolates with the presence of the symbiont M. hominis or with metronidazole resistance was observed, further studies are needed to shed light on the effective role of infecting microorganisms on the pathophysiology of T. vaginalis.

Mycoplasma hominis is considered an opportunistic pathogen able to colonize the lower urogenital tract; in females the infection is associated with severe pregnancy and postpartum complications, including abortion, endometritis, preterm delivery, and low birth weight. Molecular mechanisms of pathogenicity and virulence effectors remain poorly characterized. A number of studies in the last decade have demonstrated that M. hominis can establish an endosymbiotic relationship with Trichomonas vaginalis, a urogenital parasitic protozoon, also associated with adverse pregnancy outcomes. Recently, two bacterial genes (alr and goiB) associated with amniotic cavity invasion and a single gene (goiC) associated with intra-amniotic infections and high risk of preterm delivery have been identified in M. hominis isolated from a group of pregnant patients. In this work we demonstrate that a high number of M. hominis intracellularly associated with T. vaginalis have goiC gene, in association with alr and goiB. In addition, we demonstrate that metronidazole treatment of M. hominis-infected T. vaginalis allows delivering viable intracellular goiC positive M. hominis from antibiotic-killed protozoa and that free M. hominis can infect human cell cultures. Results suggest that molecular diagnostic strategies to identify both pathogens and their virulence genes should be adopted to prevent severe complications during pregnancy.

The symbiosis between the parasitic protist Trichomonas vaginalis and the opportunistic bacterium Mycoplasma hominis is the only one currently described involving two obligate human mucosal symbionts with pathogenic capabilities that can cause independent diseases in the same anatomical site: the lower urogenital tract. Although several aspects of this intriguing microbial partnership have been investigated, many questions on the influence of this symbiosis on the parasite pathobiology still remain unanswered. Here, we examined with in vitro cultures how M. hominis could influence the pathobiology of T. vaginalis by investigating the influence of M. hominis on parasite replication rate, haemolytic activity and ATP production. By comparing isogenic mycoplasma-free T. vaginalis and parasites stably associated with M. hominis we could demonstrate that the latter show a higher replication rate, increased haemolytic activity and are able to produce larger amounts of ATP. In addition, we demonstrated in a T. vaginalis-macrophage co-culture system that M. hominis could modulate an aspect of the innate immuno-response to T. vaginalis infections by influencing the production of nitric oxide (NO) by human macrophages, with the parasite-bacteria symbiosis outcompeting the human cells for the key substrate arginine. These results support a model in which the symbiosis between T. vaginalis and M. hominis influences host-microbes interactions to the benefit of both microbial partners during infections and to the detriment of their host.