Control of Helicobacter pylori with engineered probiotics secreting selective guided antimicrobial peptides.

Helicobacter pylori is the primary cause of 78% of gastric cancer cases, providing an opportunity to prevent cancer by controlling a single bacterial pathogen within the complex gastric microbiota. We developed highly selective antimicrobial agents against H. pylori by fusing an H. pylori-binding guide peptide (MM1) to broad-spectrum antimicrobial peptides. The common dairy probiotic Lactococcus lactis was then engineered to secrete these guided antimicrobial peptides (gAMPs). When co-cultured in vitro with H. pylori, the gAMP probiotics lost no toxicity compared to unguided AMP probiotics against the target, H. pylori, while losing >90% of their toxicity against two tested off-target bacteria. To test binding to H. pylori, the MM1 guide was fused to green fluorescent protein (GFP), resulting in enhanced binding compared to unguided GFP as measured by flow cytometry. In contrast, MM1-GFP showed no increased binding over GFP against five different off-target bacteria. These highly selective gAMP probiotics were then tested by oral gavage in mice infected with H. pylori. As a therapy, the probiotics outperformed antibiotic treatment, effectively eliminating H. pylori in just 5 days, and also protected mice from challenge infection as a prophylactic. As expected, the gAMP probiotics were as toxic against H. pylori as the unguided AMP probiotics. However, a strong rebound in gastric species diversity was found with both the selective gAMP probiotics and the non-selective AMP probiotics. Eliminating the extreme microbial dysbiosis caused by H. pylori appeared to be the major factor in diversity recovery. IMPORTANCE Alternatives to antibiotics in the control of Helicobacter pylori and the prevention of gastric cancer are needed. The high prevalence of H. pylori in the human population, the induction of microbial dysbiosis by antibiotics, and increasing antibiotic resistance call for a more sustainable approach. By selectively eliminating the pathogen and retaining the commensal community, H. pylori control may be achieved without adverse health outcomes. Antibiotics are typically used as a therapeutic post-infection, but a more targeted, less disruptive approach could be used as a long-term prophylactic against H. pylori or, by extension, against other gastrointestinal pathogens. Furthermore, the modular nature of the guided antimicrobial peptide (gAMP) technology allows for the substitution of different guides for different pathogens and the use of a cocktail of gAMPs to avoid the development of pathogen resistance.

Rare Thrombotic Complications of COVID-19: A Case Series.

Coronavirus disease 2019 (COVID-19) is known to manifest with bilateral pneumonia and acute respiratory distress syndrome. This infection with severe acute respiratory syndrome coronavirus 2 (SAR-CoV-2) is alarming because it not only affects the respiratory system but may also cause thromboembolic events. Multiple studies have reported procoagulation/hypercoagulable complications in COVID-19. This case series is a valuable addition to the literature because it reflects unique presentations of thrombotic events in COVID-19 patients. We report two cases in which patients presented with thromboembolic complications secondary to COVID-19 infection: one with severe bowel ischemia and the other with blue toe syndrome. To formulate management strategies to prevent fatal outcomes for patients with COVID-19, physicians must be vigilant in identifying life-threatening thromboembolic complications from this disease.

Inhibition of nuclear factor κB in the lungs protect bleomycin-induced lung fibrosis in mice.

BACKGROUND: Pulmonary fibrosis is a debilitating condition with limited therapeutic avenues. The pathogenicity of pulmonary fibrosis constitutes involvement of cellular proliferation, activation, and transformational changes of fibroblast to myofibroblasts. It is a progressive lung disease and is primarily characterized by aberrant accumulation of extracellular matrix proteins in the lungs with poor prognosis. The inflammatory response in the pathogenesis of lung fibrosis is suggested because of release of several cytokines; however, the underlying mechanism remains undefined. A genetic model is the appropriate way to delineate the underlying mechanism of pulmonary fibrosis. METHODS AND RESULTS: In this report, we have used cc-10 promoter based IκBα mutant mice (IKBM, an inhibitor of NF-κB) which were challenged with bleomycin (BLM). Compared to wild-type (WT) mice, the IKBM mice showed significant reduction in several fibrotic, vascular, and inflammatory genes. Moreover, we have identified a new set of dysregulated microRNAs (miRNAs) by miRNA array analysis in BLM-induced WT mice. Among these miRNAs, let-7a-5p and miR-503-5p were further analyzed. Our data showed that these two miRNAs were upregulated in WT-BLM and were reduced in IKBM-BLM mice. Bioinformatic analyses showed that let-7a-5p and miR-503-5p target for endothelin1 and bone morphogenic receptor 1A (BMPR1A), respectively, and were downregulated in WT-BLM mice indicating a link in pulmonary fibrosis. CONCLUSION: We concluded that inhibition of NF-κB and modulation of let-7a-5p and miR-503-5p contribute a pivotal role in pulmonary fibrosis and may be considered as possible therapeutic target for the clinical management of lung fibrosis.

Effect of Rubus idaeus L. Consumption During Pregnancy on Maternal Mice and Their Offspring.

The trigger for human labor is a scientific mystery. This research examined Rubus idaeus (RI), commonly referred to as red raspberry, which is widely purported to be efficacious in promoting parturition processes and favorable birth outcomes. This randomized controlled trial sought to determine the influence of RI consumption during gestation on C57BL/6N Tac mice and their offspring. The aims of this study were to (1) determine differences in the length of gestation, gestational weight gain, and litter size where RI is consumed daily at varied strengths and (2) determine differences in offspring characteristics and behavior where maternal RI consumption occurred. Once paired, mice were randomly assigned to one of three groups: placebo (n = 10) receiving plain water, RI aqueous extract fluid of 1.78 mg/mL (n = 10), or RI aqueous extract fluid of 2.66 mg/mL (n = 10). All received the same standardized diet throughout gestation. Pregnant mice were weighed with chow intake and fluid consumption determined daily. Gestation length and litter size were recorded at the time of birth. Differences in offspring characteristics were also determined and included physical characteristics (weight, physical development) and neuromotor reflexes and behaviors (locomotive abilities, geotaxis reflex, cliff avoidance reflex, and swimming development). When compared with controls, high-dose RI ingestion resulted in shorter length of gestation and smaller litter size (P ≤ .05). There was also an increase in fluid consumption and a decrease in pup weights on postnatal day 4 and 5 with RI treatment (P ≤ .05). Altogether, results suggest that RI influences parturition and fecundity processes with transplacental exposure impacting offspring characteristics.

Light-activated doxorubicin-encapsulated perfluorocarbon nanodroplets for on-demand drug delivery in an in vitro angiogenesis model: Comparison between perfluoropentane and perfluorohexane.

Phase-transition perfluorocarbon (PFC) nanodroplets have been developed for on-demand drug delivery carriers with external triggers such as ultrasound or laser irradiation techniques. Although various perfluorocarbons, including perfluoropentane (C5F12) and perfluorohexane (C6F14), have been investigated for their theranostic use, comparison of the phase-transition efficiency, the drug delivery efficacy by light activation, and physical properties of the PFC nanodroplets have not been reported. We have synthesized gold nanorod-coated doxorubicin-encapsulated perfluorocarbon nanodroplets using perfluoropentane and perfluorohexane as light-activated on-demand drug delivery carriers, called PF5 and PF6, respectively. When gold nanorods on the perfluorocarbon nanodroplets resonate with a laser wavelength, plasmonic heat generated on the gold nanorods vaporizes the nanodroplets to gas bubbles (phase-transition), and releases the encapsulated drug from the nanodroplet core. Overall, the nanodroplet size, drug encapsulation efficiency, number density, and cytotoxicity were similar between PF5 and PF6. However, the long-term stability against passive phase-transition or coalescence in physiological conditions and the phase-transition efficiency were different from each other. PF6 was better in long-term stability but showed lower phase-transition than PF5. The lower phase-transition of PF6 might have led to lower drug delivery efficiency compared to PF5. This is probably because PF6 has higher temperature thresholds required for phase-transition due to its higher boiling point. The study demonstrated feasibility of the light-activated nanodroplets for on-demand targeted nanotherapy, which suppresses the development of angiogenesis.

Experimental infection of Rhipicephalus sanguineus with Ehrlichia chaffeensis.

Ehrlichia chaffeensis, the etiologic agent of human monocytic ehrlichiosis, is a tick-borne rickettsial pathogen that is infective to a wide range of mammals, including dogs and people. Amblyomma americanum, the lone star tick, is considered the primary vector of E. chaffeensis, but this pathogen has been detected in other tick species, including the brown dog tick, Rhipicephalus sanguineus. We hypothesized that the Arkansas strain of E. chaffeensis is infective to R. sanguineus, and used a novel PCR assay to test for acquisition of this pathogen by R. sanguineus and A. americanum ticks that were simultaneously fed on experimentally infected dogs. Although E. chaffeensis was not frequently detected in peripheral blood of these dogs, the pathogen was detected in both tick species and in canine lung, kidney, lymph node, bone marrow and frontal lobe samples. One dog (AFL) was maintained for several years, and ticks again acquired E. chaffeensis from this dog 566 days after intradermal inoculation with E. chaffeensis, but the pathogen was not detected in ticks fed on the same dog at 764 or 1086 days after the intradermal inoculation.

Age-related changes in the serotonin 2A receptor in the hypoglossal nucleus of male and female rats.

Hypoglossal motoneuron output to the genioglossus muscle contributes to upper airway patency. Serotonin (5HT) plays an important role in regulating hypoglossal motoneuron excitability via serotonin 2A receptors (5HT(2A)). The purpose of this study was to investigate whether there are age-associated changes in 5HT(2A) receptor expression in the hypoglossal nucleus of male and female rats. The brains of young, middle-aged and old F344 rats were sectioned, reacted immunocytochemically for the presence of 5HT(2A) receptor, and the staining density quantified. The estrus stage of female rats was determined and circulating sex hormone levels measured and correlated with 5HT(2A) levels. The results show that there was significantly greater 5HT(2A) receptor immunoreactivity in the hypoglossal nucleus of female than of male rats. With increasing age, there was an increase in 5HT(2A) receptor immunoreactivity in the hypoglossal nucleus of female rats, whereas no age-associated changes were observed in male rats. Previous studies have shown a reduction in 5HT-dependent respiratory plasticity and an age-associated decrease in 5HT in the hypoglossal nucleus in male but not female rats. Data from the present study suggest that aging male rats fail to compensate adequately for reduced 5HT in the hypoglossal nucleus by upregulating the expression of the 5HT(2A) receptor.