Antimicrobial Susceptibility Testing for Staphylococcus lugdunensis.

Evaluation of penicillin and oxacillin susceptibility testing was conducted on 200 Staphylococcus lugdunensis isolates. Disc diffusion with penicillin 1 IU (P1, EUCAST) and penicillin 10 IU (P10, CLSI) was compared with nitrocefin discs (Cefinase) and automated broth microdilution (Vitek 2). Oxacillin susceptibility was extrapolated from cefoxitin (FOX; 30 µg) disc diffusion and compared with Vitek 2 results. The reference methods were blaZ and mecA PCR. Penicillin zone diameter and zone edge correlated with blaZ PCR results in all except two P10-susceptible isolates (very major error [VME]) and one P1-resistant isolate (major error [ME]). A total of 148 isolates were blaZ negative, of which 146 and 149 isolates were susceptible by P1 and P10, respectively. A total of 127 were penicillin susceptible by Vitek 2. Vitek 2 overcalled resistance in 21 blaZ-negative, 20 P1-susceptible, and 22 P10-susceptible isolates (Vitek 2 ME rate, 14.2%). Two mecA-positive isolates were oxacillin resistant by FOX disc and Vitek 2 methods (categorical agreement). However, 18 FOX-susceptible mecA-negative isolates tested resistant by Vitek 2. In conclusion, Vitek 2 overestimated penicillin and oxacillin resistance compared with disc diffusion and PCR results. In our study, disc diffusion with zone edge interpretation was more accurate and specific than automated broth microdilution for S. lugdunensis.

Toward the democratization of knowledge: Using photovoice, social biography, and the "five whys" in YPAR with children.

Objectives: We assessed if using a biographical method, social biography, alongside photovoice, and the Five Whys could facilitate critical dialogue in a youth participatory action research (YPAR) context. Method: In a YPAR program, we added social biography to photovoice and the Five Whys during the problem definition phase. We coded ethnographic fieldnotes to examine the quality of critical discourse. Participants were six 10-11 year old Latinx children, some from mixed-status families. Results: Social biography enlivened critical dialogue when children defined the problem for their project: the accountability of la migra, or Immigration and Customs Enforcement (ICE). Conclusions: Social biography is a valuable tool for democratizing knowledge production with children through a liberatory framework. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Detectable Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in Human Breast Milk of a Mildly Symptomatic Patient With Coronavirus Disease 2019 (COVID-19).

SARS-CoV-2 is a novel coronavirus and causative pathogen to the pandemic illness COVID-19. Although RNA has been detected in various clinical samples, no reports to date have documented SARS-CoV-2 in human milk. This case report describes an actively breastfeeding patient with COVID-19 infection with detectable viral RNA in human milk.

Changes in complement alternative pathway components, factor B and factor H during dengue virus infection in the AG129 mouse.

The complement alternative pathway (AP) is tightly regulated and changes in two important AP components, factor B (FB) and factor H (FH) are linked to severe dengue in humans. Here, a mouse model of dengue was investigated to define the changes in FB and FH and assess the utility of this model to study the role of the AP in severe dengue. Throughout the period of viremia in the AG129 IFN signalling-deficient mouse, an increase in FB and a decrease in FH was observed following dengue virus (DENV) infection, with the former only seen in a model of more severe disease associated with antibody-dependent enhancement (ADE). Terminal disease was associated with a decrease in FB and FH, with greater changes during ADE, and accompanied by increased C3 degradation consistent with complement activation. In silico analysis of NFκΒ, signal transducer and activator of transcription (STAT) and IFN-driven FB and FH promoter elements to reflect the likely impact of the lack of IFN-responses in AG129 mice, demonstrated that these elements differed markedly between human and mouse, notably with mouse FH lacking NFκΒ and key IFN-stimulated response elements (ISRE), and FB with many more NFκΒ and STAT-responsive elements than human FB. Thus, the AG129 mouse offers utility in demonstrating changes in FB and FH that, similar to humans, are associated with severe disease, but lack predicted important human-specific and IFN-dependent responses of FB and FH to DENV-infection that are likely to regulate the subtleties of the overall AP response during dengue disease in humans.

Conventional myelosuppressive chemotherapy for non-haematological malignancy disrupts the intestinal microbiome.

BACKGROUND: The gut microbiota influences many aspects of host physiology, including immune regulation, and is predictive of outcomes in cancer patients. However, whether conventional myelosuppressive chemotherapy affects the gut microbiota in humans with non-haematological malignancy, independent of antibiotic exposure, is unknown. METHODS: Faecal samples from 19 participants with non-haematological malignancy, who were receiving conventional chemotherapy regimens but not antibiotics, were examined prior to chemotherapy, 7-12 days after chemotherapy, and at the end of the first cycle of treatment. Gut microbiota diversity and composition was determined by 16S rRNA gene amplicon sequencing. RESULTS: Compared to pre-chemotherapy samples, samples collected 7-12 days following chemotherapy exhibited increased richness (mean 120 observed species ± SD 38 vs 134 ± 40; p = 0.007) and diversity (Shannon diversity: mean 6.4 ± 0.43 vs 6.6 ± 0.41; p = 0.02). Composition was significantly altered, with a significant decrease in the relative abundance of gram-positive bacteria in the phylum Firmicutes (pre-chemotherapy median relative abundance [IQR] 0.78 [0.11] vs 0.75 [0.11]; p = 0.003), and an increase in the relative abundance of gram-negative bacteria (Bacteroidetes: median [IQR] 0.16 [0.13] vs 0.21 [0.13]; p = 0.01 and Proteobacteria: 0.015 [0.018] vs 0.03 [0.03]; p = 0.02). Differences in microbiota characteristics from baseline were no longer significant at the end of the chemotherapy cycle. CONCLUSIONS: Conventional chemotherapy results in significant changes in gut microbiota characteristics during the period of predicted myelosuppression post-chemotherapy. Further study is indicated to link microbiome changes during chemotherapy to clinical outcomes.

Mycoplasma hominis bursitis in a simultaneous pancreas-kidney transplant recipient: case report and literature review.

Mycoplasma hominis can be isolated frequently from the genitourinary tract of some healthy individuals. On rare occasions, it acts as a pathogen in immunocompromised patients such as transplant recipients. Here, we describe the case of a 39-year-old man with end-stage kidney disease secondary to diabetic nephropathy who received a simultaneous pancreas-kidney transplant. He developed pancreatitis and arterial thrombosis 2 weeks post-transplant and required a pancreatectomy. His kidney allograft function remained normal. He developed severe left hip pain 2 weeks post-transplant with a trochanteric bursal effusion detected on magnetic resonance imaging. The effusion grew M. hominis. The patient was treated with 100 mg of doxycycline twice daily for 9 months with full resolution of the effusion at 4 months post-treatment. We also review all previously reported M. hominis infections in transplant recipients.

Dengue Virus Induces Increased Activity of the Complement Alternative Pathway in Infected Cells.

Severe dengue virus (DENV) infection is associated with overactivity of the complement alternative pathway (AP) in patient studies. Here, the molecular changes in components of the AP during DENV infection in vitro were investigated. mRNA for factor H (FH), a major negative regulator of the AP, was significantly increased in DENV-infected endothelial cells (EC) and macrophages, but, in contrast, production of extracellular FH protein was not. This discord was not seen for the AP activator factor B (FB), with DENV induction of both FB mRNA and protein, nor was it seen with Toll-like receptor 3 or 4 stimulation of EC and macrophages, which induces both FH and FB mRNA and protein. Surface-bound and intracellular FH protein was, however, induced by DENV, but only in DENV antigen-positive cells, while in two other DENV-susceptible immortalized cell lines (ARPE-19 and human retinal endothelial cells), FH protein was induced both intracellularly and extracellularly by DENV infection. Regardless of the cell type, there was an imbalance in AP components and an increase in markers of complement AP activity associated with DENV-infected cells, with lower FH relative to FB protein, an increased ability to promote AP-mediated lytic activity, and increased deposition of complement component C3b on the surface of DENV-infected cells. For EC in particular, these changes are predicted to result in higher complement activity in the local cellular microenvironment, with the potential to induce functional changes that may result in increased vascular permeability, a hallmark of dengue disease.IMPORTANCE Dengue virus (DENV) is a significant human viral pathogen with a global medical and economic impact. DENV may cause serious and life-threatening disease, with increased vascular permeability and plasma leakage. The pathogenic mechanisms underlying these features remain unclear; however, overactivity of the complement alternative pathway has been suggested to play a role. In this study, we investigate the molecular events that may be responsible for this observed alternative pathway overactivity and provide novel findings of changes in the complement system in response to DENV infection in primary cell types that are a major target for DENV infection (macrophages) and pathogenesis (endothelial cells) in vivo Our results suggest a new dimension of cellular events that may influence endothelial cell barrier function during DENV infection that could expand strategies for developing therapeutics to prevent or control DENV-mediated vascular disease.

Sustained improvement in vancomycin dosing and monitoring post-implementation of guidelines: Results of a three-year follow-up after a multifaceted intervention in an Australian teaching hospital.

INTRODUCTION: Despite vancomycin being in use for over half-a-century, it is still not dosed or monitored appropriately in many centers around the world. The objective of this study was to determine the effectiveness of a multifaceted intervention to implement a vancomycin dosing and monitoring guideline across multiple medical and surgical units over time. METHODS: This was an observational before-and-after interventional cohort study. The pre-intervention period was August to December 2010-2011 and the post-intervention period was September to November 2012-2014. The implementation strategy comprised: face-to-face education, online continuing medical education, dissemination of pocket guideline and email reminder. Outcome measures included: appropriate prescribing of loading and maintenance doses, therapeutic drug monitoring, time to attain target range and nephrotoxicity. RESULTS: Post-implementation prescribing of loading doses increased (10.4%-43.6%, P=<0.001), guideline adherent first maintenance dose (44%-68.4% P = 0.04), correct dose adjustment from (53.1%-72.2%, P = 0.009). Beneficial effects pre and post-implementation were observed for adherent timing of initial concentration (43.2%-51.9%, P = 0.01), concentrations in target range (32.6%-44.1%, P = 0.001), time to target range (median 6-4 days, P=<0.001), potentially nephrotoxic concentrations (30.7%-20.9%, P=<0.001) and nephrotoxicity (10.4%-6.8%, P=<0.001). CONCLUSIONS: A multifaceted intervention to implement a vancomycin dosing and monitoring guideline significantly improved prescribing, monitoring, pharmacokinetic and safety outcomes for patients treated with vancomycin over an extended period. However, increased guideline adoption by clinicians is required to maximize and prolong the utility of this important agent.

FUT2 genotype influences lung function, exacerbation frequency and airway microbiota in non-CF bronchiectasis.

OBJECTIVE: To assess whether FUT2 (secretor) genotype affects disease severity and airway infection in patients with non-cystic fibrosis bronchiectasis. PARTICIPANTS: Induced sputum samples were obtained from 112 adult patients with high-resolution CT scan-proven bronchiectasis and at least two exacerbations in the previous year, as part of an unrelated randomised control trial. OUTCOME MEASURES: Presence of null FUT2 polymorphisms were determined by gene sequencing and verified by endobronchial biopsy histochemical staining. Outcome measures were FEV1% predicted, exacerbation frequency, and bacterial, fungal and viral components of the microbiota (measured by culture independent approaches). RESULTS: Patients were grouped by FUT2 loss-of-function genotype; categorised as non-secretors (n=27, sese), heterozygous secretors (n=54, Sese) or homozygous secretors (n=31, SeSe). FEV1% was significantly lower in SeSe patients compared with sese patients (mean 61.6 (SD 20.0) vs 74.5 (18.0); p=0.023). Exacerbation frequency was significantly higher in SeSe (mean count 5.77) compared with sese (4.07; p=0.004) and Sese (4.63; p=0.026) genotypes. The time until first exacerbation was significantly shorter in SeSe compared with Sese (HR=0.571 (95% CI 0.343 to 0.950); p=0.031), with a similar trend for sese patients (HR=0.577 (0.311 to 1.07); p=0.081). sese had a significantly reduced frequency of Pseudomonas aeruginosa-dominated airway infection (8.7%) compared with Sese (31%; p=0.042) and SeSe (36%; p=0.035). In contrast, fungal, viral and non-dominant bacterial components of the microbiome were not significantly different between FUT2 genotypes. CONCLUSIONS: FUT2 genotype in patients with non-cystic fibrosis bronchiectasis was significantly associated with disease outcomes, with homozygous secretors exhibiting lower lung function, higher exacerbation number and a higher frequency of P. aeruginosa-dominated infection. TRIAL REGISTRATION NUMBER: ACTRN12609000578202 (anzctr.org.au); Pre-results.

Cerebral microbleeds in nonlacunar brain infarction are associated with lower coated-platelet levels.

BACKGROUND: Coated-platelets are a subset of platelets with high procoagulant potential observed on dual-agonist stimulation with collagen and thrombin. Coated-platelet levels are elevated in patients with nonlacunar ischemic stroke compared with controls, although the presence of early hemorrhagic transformation is associated with lower coated-platelet levels. In contrast to infarction, patients with spontaneous intracerebral hemorrhage have lower coated-platelet levels, and these levels inversely correlate with bleed size. Cerebral microbleeds (CMBs) represent previous small hemorrhagic occurrences. We undertook a pilot study to investigate coated-platelet production and the presence of CMBs in patients with nonlacunar ischemic stroke. METHODS: Coated-platelet levels were determined in 110 consecutive patients with a diagnosis of nonlacunar stroke. Microbleeds were identified using the published criteria by an experienced stroke neurologist. Coated-platelet levels were compared statistically between patients with and without CMBs using the nonparametric Wilcoxon rank sum test. RESULTS: Coated-platelet levels (median [interquartile range]) for all patients were 44.1% [34%-51.2%]. CMBs were detected in 22 patients (20%); these patients had significantly lower coated-platelet levels compared with those without CMBs (35.6% [22.6%-47.2%] versus 45.1% [36.1%-51.5%]; P = .025), whereas other demographic and clinical factors did not differ significantly. CONCLUSIONS: The presence of CMBs in patients with nonlacunar ischemic stroke is associated with lower levels of coated-platelets. Larger prospective studies are needed to better establish the potential connection between altered coated-platelet synthesis, microbleeds, cerebral infarction, and possible hemorrhage-prone vascular changes.

The Post-war Revival of Canadian Planning: Assessing the Impact of the Community Planning Association of Canada.

The Community Planning Association of Canada (CPAC) advocated for the re-establishment of planning in post-war Canada. During this period, the federal government set reconstruction objectives, and both Central (now Canada) Mortgage and Housing Corporation (CMHC) and the CPAC were formed. We believe that 1944-1947 was a critical juncture establishing planned suburban development in Canada as a path-dependent process with tremendous momentum into the 21st-century. Using a historical-institutional approach, the role of CMHC and the influence of the CPAC is examined. Analysis relying on extensive archival material demonstrates that the CPAC gave a tremendous push along the path-dependent process of suburbanization.

Lytic activity of phages against bacterial pathogens infecting diabetic foot ulcers.

Complications of diabetes, such as diabetic foot ulcers (DFUs), are common, multifactorial in origin, and costly to treat. DFUs are the cause of nearly 90% of limb amputations among persons with diabetes. In most chronic infections such as DFU, biofilms are involved. Bacteria in biofilms are 100-1000 times more resistant to antibiotics than their planktonic counterparts. Multidrug-resistant (MDR) Staphylococcus aureus and Pseudomonas aeruginosa infections in DFUs may require alternative therapeutic agents such as bacteriophages ("phages"). This study describes the lytic activity of phage cocktails AB-SA01 (3-phage cocktail) and AB-PA01 (4-phage cocktail), which target S. aureus and P. aeruginosa, respectively. The host range and lytic effect of AB-SA01 and AB-PA01 on a planktonic culture, single-species biofilm, and mixed-species biofilm were evaluated. In vitro testing showed that 88.7% of S. aureus and 92.7% of P. aeruginosa isolates were susceptible to AB-SA01 and AB-PA01, respectively, in the planktonic state. The component phages of AB-SA01 and AB-PA01 infected 66% to 94.3% of the bacterial isolates tested. Furthermore, AB-SA01 and AB-PA01 treatment significantly (p < 0.05) reduced the biofilm biomass of their hosts, regardless of the antibiotic-resistant characteristics of the isolates and the presence of a non-susceptible host. In conclusion, the strong lytic activity, broad host range, and significant biofilm biomass reduction of AB-SA01 and AB-PA01 suggest the considerable potential of phages in treating antibiotic-resistant S. aureus and P. aeruginosa infections alone or as coinfections in DFUs.

Ocular syphilis.

Multiple studies around the world suggest that syphilis is re-emerging. Ocular syphilis - with a wide range of presentations, most of which are subtypes of uveitis - has become an increasingly common cause of ocular inflammation over the past 20 years. Its rising incidence, diagnostic complexity, and manifestations that have only recently been characterized make ocular syphilis relevant from the public health, clinical, and scientific perspectives. We review the demographics, epidemiology, clinical features, ocular imaging findings, diagnosis, and medical management of this condition.

Cerebral venous thrombosis of the sphenoparietal sinus: A case report.

BACKGROUND: Cerebral venous thrombosis (CVT) is a rare cause of stroke that preferentially affects reproductive aged females and patients with hereditary or acquired thrombotic risk factors. The superior sagittal sinus and transverse sinus are the two most common sites for thrombus formation. CASE DESCRIPTION: We report a case of CVT arising in a very rare location, the sphenoparietal sinus. A 32-year-old woman with a history of factor V Leiden mutation and multiple prior episodes of venous thromboembolism presented with a new-onset seizure, headache, and emesis. CT angiography ultimately revealed thrombosis of the left sphenoparietal sinus. The patient received anticoagulation with apixaban with resolution of symptoms and without complications. CONCLUSION: This case serves as an uncommon example of sphenoparietal sinus thrombosis managed with novel oral anticoagulant treatment.

Intra-abdominal infection with Campylobacter curvus: case report and review of the literature.

BACKGROUND: Campylobacter curvus is a Gram-negative bacteria associated with periodontal disease in humans. Cases of extra-oral manifestations of infection are rare with only six reported cases of extra-oral infection including this report that have been identified in the current literature. Molecular methods are generally used to identify C. curvus while optimal antibiotic choice and duration to treat extra-oral infections for this pathogen is unknown. CASE PRESENTATION: A 63-year-old male with a background history of alcoholic pancreatitis presented with fever and malaise who was found to have radiological intra-abdominal collections. Drainage of these collections identified C. curvus via matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) mass spectrometry with high probability and identification further confirmed by whole-genome sequencing. Antibiotic susceptibility testing to erythromycin and ciprofloxacin of C. curvus was performed using E-test diffusion methods along with investigation for the presence of resistance genes. The patient was treated with intravenous piperacillin-tazobactam followed by ciprofloxacin for 4 weeks total with good clinical recovery. CONCLUSIONS: Extra-oral manifestations with the pathogen C. curvus are rare with few cases described in the literature. There is minimal data on susceptibility patterns, optimal antibiotic treatment and duration. Treatment of extraintestinal C. curvus infections in humans should encompass both adequate source control and antibiotic therapy.

Potential for bacteriophage therapy for Staphylococcus aureus pneumonia with influenza A coinfection.

The ability of influenza A virus to evolve, coupled with increasing antimicrobial resistance, could trigger an influenza pandemic with great morbidity and mortality. Much of the 1918 influenza pandemic mortality was likely due to bacterial coinfection, including Staphylococcus aureus pneumonia. S. aureus resists many antibiotics. The lack of new antibiotics suggests alternative antimicrobials, such as bacteriophages, are needed. Potential delivery routes for bacteriophage therapy (BT) include inhalation and intravenous injection. BT has recently been used successfully in compassionate access pulmonary infection cases. Phage lysins, enzymes that hydrolyze bacterial cell walls and which are bactericidal, are efficacious in animal pneumonia models. Clinical trials will be needed to determine whether BT can ameliorate disease in influenza and S. aureus coinfection.

Surgical Antibiotic Prophylaxis in an Era of Antibiotic Resistance: Common Resistant Bacteria and Wider Considerations for Practice.

The increasing incidence of antimicrobial resistance (AMR) presents a global crisis to healthcare, with longstanding antimicrobial agents becoming less effective at treating and preventing infection. In the surgical setting, antibiotic prophylaxis has long been established as routine standard of care to prevent surgical site infection (SSI), which remains one of the most common hospital-acquired infections. The growing incidence of AMR increases the risk of SSI complicated with resistant bacteria, resulting in poorer surgical outcomes (prolonged hospitalisation, extended durations of antibiotic therapy, higher rates of surgical revision and mortality). Despite these increasing challenges, more data are required on approaches at the institutional and patient level to optimise surgical antibiotic prophylaxis in the era of antibiotic resistance (AR). This review provides an overview of the common resistant bacteria encountered in the surgical setting and covers wider considerations for practice to optimise surgical antibiotic prophylaxis in the perioperative setting.

Protective role of PhtD and its amino and carboxyl fragments against pneumococcal sepsis.

The implementation of polysaccharide-based vaccines has massively reduced the incidence of invasive pneumococcal diseases. However, there is great concern regarding serotype replacement and the increase in antibiotic resistant strains expressing non-vaccine capsular types. In addition, conjugate vaccines have high production costs, a limiting factor for their implementation in mass immunization programs in developing countries. These limitations have prompted the development of novel vaccine strategies for prevention of Streptococcus pneumoniae infections. The use of conserved pneumococcal antigens such as recombinant proteins or protein fragments presents an interesting serotype-independent alternative. Pht is a family of surface-exposed proteins which have been evaluated as potential vaccine candidates with encouraging results. The present work investigated the immune responses elicited by subcutaneous immunization of mice with the polyhistidine triad protein D (PhtD) and its amino and carboxyl terminal fragments. The proteins were immunogenic and protective against pneumococcal sepsis in mice. Antibodies raised against PhtD increased complement C3b deposition on the pneumococcal surface, mainly mediated by the alternative pathway. Sera from mice immunized with PhtD and PhtD_Cter promoted an increase in bacterial uptake by mouse phagocytes. The interaction of PhtD with the complement system regulator factor H was investigated in silico and in vitro by ELISA and western blot, confirming PhtD as a factor-H binding protein. Our results support the inclusion of PhtD and more specifically, its C-terminal fragment in a multicomponent serotype independent vaccine and suggests a role for the complement system in PhtD-mediated protection.