Focus Of Infection Research Articles

Changes in the epidemiology of pediatric sinogenic and otogenic intracranial infections during the COVID-19 pandemic: a single-institution study.

Focal intracranial infections (epidural abscesses, subdural empyemas, and intraparenchymal abscesses) are uncommon complications of sinusitis and otitis media but can be associated with significant morbidity. Treatment typically requires neurosurgical and otolaryngological interventions in combination with antibiotic treatment. Historically, children have presented to the authors' pediatric referral center with sinusitis- or otitis media-related intracranial infections in low numbers. However, since the onset of the COVID-19 pandemic, the incidence of intracranial pyogenic complications has increased at this center. The objective of this study was to compare the epidemiology, severity, microbial causes, and management of pediatric sinusitis- and otitis-related intracranial infections in the periods before and during the COVID-19 pandemic. All patients 21 years of age or younger who presented with an intracranial infection in the setting of sinusitis or otitis media and who underwent neurosurgical treatment at Connecticut Children's from January 2012 to December 2022 were retrospectively reviewed. Demographic, clinical, laboratory, and radiological data were systematically collated, and variables before and during COVID-19 were compared statistically. Overall, 18 patients were treated for sinusitis-related (n = 16) or otitis media-related (n = 2) intracranial infections during the study period. Ten patients (56%) presented from January 2012 to February 2020, none from March 2020 to June 2021, and 8 (44%) from July 2021 to December 2022. There were no significant demographic differences between the pre-COVID-19 and COVID-19 cohorts. The 10 patients in the pre-COVID-19 cohort underwent a total of 15 neurosurgical and 10 otolaryngological procedures, while the 8 patients in the COVID-19 cohort underwent a total of 12 neurosurgical and 10 otolaryngological procedures. Surgically obtained wound cultures yielded a variety of organisms; Streptococcus constellatus/S. anginosus/S. intermedius were more prevalent in the COVID-19 cohort (87.5% vs 0%, p < 0.001) as was Parvimonas micra (62.5% vs 0%, p = 0.007). At an institutional level, there has been an approximately threefold increase in cases of sinusitis- and otitis media-related intracranial infections during the COVID-19 pandemic. Multicenter studies are needed to confirm this observation and to investigate whether the mechanisms of infection are related directly to SARS-CoV-2, changes in the respiratory flora, or delayed care. The next steps will include expansion of this study to other pediatric centers throughout the United States and Canada.

PO-04-033 CARDIONEUROABLATION IN A WOMAN WITH ICTAL-INDUCED CARDIAC ASYSTOLE

Ictal-induced cardiac bradyarrhythmia and asystole is a rare phenomenon. The exact mechanism of ictal-induced cardiac bradyarrhythmia and asystole remains unclear. It was postulated that stimulation of central autonomic network during ictal episode may trigger an abrupt burst of hypervagotonia. Prolonged episode of cardiac bradyarrhythmia and asystole may result in syncope or death due to impairment of cerebral perfusion. The role of cardioneuroablation (CNA) in this condition has not been well-described in the literature. To describe a case of successful CNA in a patient with ictal-induced bradyarrhythmia and asystole. n/a A 47-year-old female has a 1.5-year history of intractable focal epilepsy and COVID-19 infection. She started having multiple episodes of seizures following a mild COVID-19 infection. Electroencephalogram (EEG) and brain MRI revealed right temporal onset seizures without structural lesions. Due to ongoing uncontrolled seizures with multiple semiologies despite multiple anti-epileptic drugs, she was admitted to Epilepsy Monitoring Unit for seizure classification. Her ictal EEGs (Figure 1) showed onset of ictal rhythm in the right temporal region with episodes of severe sinus bradycardia (15-30 bpm) and sinus pauses (15-16 seconds). Telemetry tracings demonstrated PP interval slowing with PR interval prolongation prior to the pauses consistent with a vagally-mediated mechanism. Cardiac electrophysiology team recommended CNA for treating the episodes of ictal-induced bradyarrhythmia and asystole. 3D anatomic maps of the right atrium (RA) and left atrium (LA) were created using CARTO system (Biosense Webster). Right superior ganglionated plexus (RSGP) was localized with fractionation mapping and intracardiac echocardiography guidance. RSGP was targeted from the RA using an irrigated radiofrequency catheter with power limit of 25 W. Post-ablations of RSGP, her heart rate increased from 60 - 99 bpm. Additional lesions were delivered from the LA site but no additional heart rate increase was not seen. An implantable loop recorder was implanted post-ablation procedure. During follow-up of 8 months, she had recurrent focal epilepsy, but no bradyarrhythmias or asystole was noted on her loop recorder. Resting heart rates at long-term follow up were between 70 - 100 bpm. This case highlights the utility of CNA in patient with ictal-induced cardiac bradyarrhythmia and asystole. CNA may be an approach to avoid permanent pacemakers in this population.

Abundance and Condition of Populations of Small Mammals as Carriers of Natural Focal Human Diseases in the Karelia North-East Ladoga

Summing up the results of the long-term studies of population ecology and dynamics of small mammals for more than half a century (1966–2018). Small mammals are involved in the formation and functioning of natural-focil of human vector-borne diseases common in the North-Eastern Ladoga region. Encephalitis and tularemia circulate intensively in the Ladoga region under conditions of intensive anthropogenic transformation of taiga ecosystems. Local landscapes belong to the subzone of the middle taiga forests. Most of the studied species of small mammals are located in the northern periphery of this area. Here representatives of the native taiga fauna of Micromammalia constantly experience powerful anthropogenic pressure, which leads to a decrease in their abundance and correlation of the species. Of the total amount of 22 species of small mammals collected here, Sorex araneus and Myodes glareolus dominate. Next, the species prevalent (according to the degree of predominant species) in the counts are Microtus agrestis, Sorex minutus and Sorex caecutiens. Other species are relatively rare and appear in smaller numbers. Myodes glareolus has main role in the feeding of ticks and, accordingly, in the circulation of infections in nature, the differentiated contribution of individual intrapopulation groups in maintaining the number of vectors and the activity of pathogens. Shrews are important for the development of the larval stages of the tick. In the research area, were recorded total index of trap catch of abundance of small animals with trap lines from 0.34 to 18.5 individuals per 100 snap traps-nights (average of 7.8 ind.) and with pitfall traps recorded from 2.7 to 25.4 individuals per 10 ditch-nights (average of 14 ind.). This indicates that the studied area has smaller amount of mammals compared to other regions in Southern Europe. Nevertheless, the studies determine a rather tense epidemic situation in the studied area. Population of small mammals of boreal ecosystem steadily affect the mass feeding of carriers of natural focal infections.

Pneumomediastinum as a Complication of Cocaine Insufflation

A 25-year-old man presents to the ED for unresponsiveness after consuming cocaine and other unknown substances. Chest imaging from the presentation was unremarkable, but after developing fever and leukocytosis, he underwent extensive work-up in search of infectious foci. A CT scan of the chest showed a small pneumomediastinum and the possibility of an esophageal tear. After recovering consciousness and the ability to recount events, the patient admitted to the concomitant use of cocaine and opiates via insufflation.

Nontypeable Haemophilus influenzaespecific memory T H17 cells protect against secondary bacterial pneumonia following Influenza A virus infection

Abstract Nontypeable Haemophilus influenzae(NTHi) is one of the most common bacteria that cause secondary bacterial pneumonia following influenza A virus (IAV) infection, leading to hospitalization and death associated with IAV infection worldwide. Current efforts to develop vaccines against NTHi infection focus on inducing antibodies but are hindered by antigenic diversity among NTHi strains. Therefore, we interrogated the contribution of the memory T-helper type 17 (T H17) response in protective immunity against IAV/NTHi co-infection. We observed that co-infected mice exhibited prolonged NTHi infection in the lungs even at mild IAV infection and impaired NTHi-specific T H17 responses compared to NTHi mono-infected mice. However, immune mice that recovered from a prior NTHi infection were protected from homologous and heterologous secondary NTHi strains following IAV infection. Our data further revealed that memory NTHi-specific T H17 cells overcame IAV-driven impairments of anti-NTHi T H17 responses and were cross-reactive with different NTHi strains. Last, mice immunized with a NTHi protein that induced a strong T H17 memory response were broadly protected against diverse NTHi strains following challenge with IAV/NTHi co-infection. These results indicate that NTHi-specific memory T H17 cells are capable of providing serotype-independent protection against co-infection and demonstrate the advantage of developing broadly protective T H17-inducing vaccines against secondary bacterial pneumonia.

Septic shock in obstetrics: guidelines of the All-Russian public organization “Federation of Anesthesiologists and Reanimatologists”

The article reflects the main provisions of the clinical guidelines on septic shock in obstetrics, approved by the All-Russian public organization “Federation of Anesthesiologists-Resuscitators” in 2022. The relevance of the problem is associated with high mortality and morbidity rates from sepsis and septic shock in obstetrics. The main issues of etiology, pathogenesis, clinical picture, methods of laboratory and instrumental diagnostics, features of using the qSOFA, SOFA, MOEWS, SOS, MEWC, IMEWS scales for sepsis verification are consistently presented. The article presents the starting intensive therapy (the first 6–12 hours) of the treatment of septic shock in obstetrics, taking into account the characteristics of the pregnant woman's body. The strategy of prescribing vasopressors (norepinephrine, phenylephrine, epinephrine), inotropic drugs (dobutamine) is described, antibiotics and optimal antibiotic therapy regimens, features of infusion and adjuvant therapy are presented. The issues of surgical treatment of the focus of infection and indications for hysterectomy, as well as the organization of medical care and rehabilitation of patients with sepsis and septic shock were discussed. The basic principles of prevention of sepsis and septic shock in obstetrics are described. The criteria for the quality of medical care for patients with septic shock and the algorithms of doctor's actions in the diagnosis and intensive care of patients with septic shock in obstetrics are presented.

Evaluation Of Congestion Levels in Septic Patients Admitted to Critical Care Units with a Combined Venous Excess-Lung Ultrasound Score (VExLUS) - a Research Protocol.

Sepsis is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection with a high mortality rate. Septic shock is a subset of sepsis with manifest circulatory dysfunction (use of vasopressors and persistent elevation of lactic acid) . As stated in literature, in addition to the use of empiric antibiotics and control of the infectious focus, intravenous fluid therapy is an essential intervention to promote hemodynamic stabilization. However, the literature also describes harmful outcomes related to fluid overload. Hemodynamic management in critically ill patients has traditionally focused on maintaining adequate cardiac output and arterial blood pressure by relying on fluid administration and/or vasopressor/inotropic support. However, organ perfusion is affected by other important factors, such as venous pressure, which can be overlooked. The evaluation of lung congestion with point of care ultrasound (POCUS), as a signal of extravascular fluid, and, more recently, a venous excess Doppler ultrasound (VExUS) grading system, are parameters for the assessment of the fluid status of the patient and organ congestion. Our main hypothesis is that adding a modified lung ultrasound score to the VExUS protocol could provide higher sensitivity and earlier identification of fluid overload, guiding the clinician in the decision of fluid administration in patients with sepsis.

Analysis of leukogram parameters and blood cytokine profile in tick-borne infections of various etiologies

Introduction. Studies aimed at identifying pathogenetic features of tick-borne natural focal infections depending on etiological agent are relevant to seek out for new associations of biomarkers characterizing the structural and functional phenotype of immune cells significant for the differential diagnosis and prognosis of diseases. The aim of the study was to evaluate the parameters of a leukogram with an expanded profile in relation to the parameters of cytokine status in patients with viral tick-borne encephalitis as well as erythemic tick-borne borreliosis at acute stage of the disease. Materials and methods. The study involved 28 patients with the non-erythemic tick-borne borreliosis and 27 patients with tick-borne encephalitis at the acute stage of the disease, as well as 16 healthy individuals (control group). Venous blood samples were examined using a Sysmex XN1000 analyzer based on reflex testing technology with extended profile allowing to characterize leukocyte reaction during infection and inflammation, including parameters such as the absolute and relative count of immature granulocytes (IG), neutrophil granularity index (NEUT-GI), neutrophil reactivity index (NEUT-RI), count of reactive lymphocytes (RE-LYMP), count of antibody-producing lymphocytes (AS-LYMP). Concentrationof IL-2, IL-4, IL-6, IL-8, IL-10, IFN and TNF in blood serum was determined by enzyme immunoassay using specific reagent kits (Vector-Best, Russia). Results. It was found that changes in the quantitative composition of blood leukocytes in patients with viral tick-borne encephalitis and non-erythemic form of tick-borne borreliosis have a unidirectional tendency characterized by increased count of metabolically active neutrophils (NEUT-RI) and a decreased count of reactive lymphocytes (RE-LYMP) compared to healthy individuals. Changes in the leukogram occur along with high blood concentration of proinflammatory cytokines IL-8 and TNF and a low level of IFN. A positive direct correlation was revealed between the fluorescence intensity parameter characterizing the metabolic activity of neutrophil granulocytes (NEUT-RI) and blood serum IL-8 level both in tick-borne encephalitis (r = 0.422, p 0.05) and non-erythemic form of tick-borne borreliosis (r = 0.551, p 0.05). Additionally, in the former, a positive correlation was established between the total leukocyte count (WBC) and concentration of TNF in the blood serum (r = 0.532, p 0.05).

PCR-confirmed malaria among children presenting with a decreased level of consciousness in Angola: a prospective, observational study

BackgroundIn malaria-endemic areas, children presenting to hospitals with a decreased level of consciousness remain a diagnostic dilemma. The definition of cerebral malaria in a comatose child demands exclusion of other possible reasons, which requires in-depth investigations that are not easily available. The aim of this study was to investigate the frequency and clinical characteristics of PCR-confirmed malaria in a cohort of children with a decreased level of consciousness, look for potential features that would aid in differentiating children with malaria from those without, and assess the performance of traditional thick film microscopy against the cytb-qPCR-method.MethodsA total of 345 children aged 30 days–15 years old, presenting to Hospital Pediátrico David Bernardino in Luanda, Angola, with a decreased level of consciousness (Glasgow coma scale score < 15) were prospectively enrolled during 2014–2017. Malaria was defined as a positive cytb-qPCR result on any occasion in hospital. The clinical course and laboratory parameters were compared between children with malaria and those without. The performance of thick film microscopy was analysed against the PCR method.Results161 of 345 children (46.7%) had a positive malaria PCR test result. All cases were Plasmodium falciparum species, and 82.6% (133/161) fulfilled the WHO criteria for severe malaria. Overall, children with malaria presented to hospital with a shorter duration of symptoms and less convulsions pre-admission compared to those without malaria. The median GCS score on admission was 8, which did not differ between children with or without malaria. Clinical findings on admission were mostly similar across the whole cohort, but an infection focus outside the central nervous system was more common in malaria-negative children. Moreover, severe anaemia, thrombocytopenia, and high CRP levels occurred more frequently in children with malaria. The case fatality ratio was 28.5% (91/319) and did not differ between parasitaemic children and those without malaria, although parasitaemic children died sooner after hospital admission. When neurological sequelae were also considered, a positive malaria test was associated with a better outcome. The performance of thick film microscopy against PCR yielded a sensitivity of 96.8% and a specificity of 82.7%.ConclusionsIn this cohort of children with a decreased consciousness, the frequent presence of a malarial infection could not be judged from the clinical findings on admission, but the combination of profound aneamia, thrombocytopenia, and a high CRP level increased the odds of a positive malaria test result. Mortality remained high regardless of etiology, but malaria infection associated with fewer neurological deficits at discharge. Thick film microscopy performed well compared to the cytb-qPCR method.

Rare clinical forms of chronic brucellosis

Brucellosis is the most common and severe infectious disease in the group of zoonotic infections. This disease is characterized by a multisysitemic lesion. The difficulties in diagnosing this disease are associated with the pathogenetic features of the development of brucellosis, namely prolonged bacteremia, which causes the development of secondary foci of infection in various organs and tissues, manifested by a variety of clinical manifestations, the erasure of symptoms of the disease, as well as the appearance of rare clinical forms of brucellosis.One of the rare clinical manifestations of chronic brucellosis is the defeat of the cardiovascular system and the defeat of the nervous system. According to the latest literature data, cardiovascular complications of brucellosis in humans account for only 3%, but are the main cause of death in 80%. Damage to the nervous system is rare, with a registered frequency of 3% to 13%.Clinical manifestations of neurobrucellosis are diverse and can mimic many other neurological diseases. The most common manifestations of brucellosis may be meningitis, meningoencephalitis. Despite the rare occurrence, these forms of the disease are severe, lead to disability and mortality with late diagnosis. The article presents clinical cases of brucellosis with lesions of the heart and central nervous system.

Features of the course of pneumonia in children with coronavirus infection

BACKGROUND:The most frequent complication of coronavirus infection (COVID-19) in young children is severe viral pneumonia.&#x0D; AIM:To reveal the features of the course of coronavirus pneumonia in children.&#x0D; MATERIALS AND METHODS:A retrospective analysis of 54 case histories of children with coronavirus pneumonia, aged 1 month to 15 years, who received inpatient treatment in the Department of Pulmonology was performed.&#x0D; RESULTS:The disease was most often registered in children of the first three years of life (66,7%). 49 (90,7%) children turned out to be from family foci of coronavirus infection. The majority of children (4481.5%) had signs of respiratory failure of I-II degree. Polymerase chain reaction (PCR) for the presence of coronavirus was positive in 37 (68.5%) children. Radiologically, bilateral infiltrates were detected predominantly in the middle and lower parts of the lungs. The multisystem inflammatory syndrome associated with SARS-CoV-2 was characterized by persistent fever, multiple organ damage and clinical and laboratory shifts.&#x0D; The vast majority of cases involved transmission from adult patients. The disease most often developed in children under 3 years of age. The course of the disease is accompanied by the development of grade III respiratory failure and intoxication. PCR for the presence of coronavirus may be negative due to late admission of patients to the hospital. Radiological changes were characterized by extensive bilateral lesions mainly in the lower and middle lobes of the lungs. Persistent fever, multiorgan lesions, clinical and laboratory shifts in coronavirus pneumonia in children indicate the development of multisystem inflammatory syndrome associated with SARS-CoV-2.&#x0D; CONCLUSIONS:Pneumonia in most cases proceeded with the development of respiratory failure, symptoms of gastrointestinal tract damage, a pronounced intoxication syndrome. At the same time, there was revealed inconsistency of radiologically extensive lung lesions with more scanty auscultatory data.

Streptococcus bovis infection of the central nervous system in adults: Report of 4 cases and literature review

ObjectivesTo describe the clinical features, history and association with intestinal disease in central nervous system (CNS) S. bovis infections. MethodsFour cases of S. bovis CNS infections from our institution are presented. Additionally a systematic literature review of articles published between 1975 and 2021 in PubMed/MEDLINE was conducted. Results52 studies with 65 cases were found; five were excluded because of incomplete data. In total 64 cases were analyzed including our four cases: 55 with meningitis and 9 with intracranial focal infections. Both infections were frequently associated with underlying conditions (70.3%) such as immunosuppression (32.8%) or cancer (10.9%). In 23 cases a biotype was identified, with biotype II being the most frequent (69.6%) and S. pasteurianus the most common within this subgroup. Intestinal diseases were found in 60.9% of cases, most commonly neoplasms (41.0%) and Strongyloides infestation (30.8%). Overall mortality was 17.1%, with a higher rate in focal infection (44.4% vs 12.7%; p=0.001). ConclusionsCNS infections due to S. bovis are infrequent and the most common clinical form is meningitis. Compared with focal infections, meningitis had a more acute course, was less associated with endocarditis and had a lower mortality. Immunosuppression and intestinal disease were frequent in both infections.

Role of Erythropoietin In Pre-Renal Transplant Patients Undergoing Oral Surgical Procedures-Two Case Reports

The Oral Cavity Is Often Called The "Gateway To The Body," Dentist As The "Gatekeepers Of Health." Chronic Kidney Disease Is An Emerging Disease With Increased Mortality In A Low Resource Setting, According To The Global Disease Burden Report (2015)1 Lack Of Infrastructure And Awareness Among The Public Is Diagnosed At A Late Stage, Leading To Increased Mortality. In A Parallel Line, Dentistry In Developing Countries Still Is An Alien Entity To The Rural And Suburban Areas. End-Stage Renal Failure Patients Who Were Advised For Renal Transplants Visit The Dentist With Poor Oral Hygiene And Foci Of Infection, Which Must Be Eliminated. Anemia In End Stage Renal Disease And Its Management Is Challenging For The Clinicians With The Lack Of Erythropoietin Hormone Exposes The Patients To Risk Of Fatigue, Breathlessness, Lower Quality Of Life And Blood Transfusion2, 3.From Early 1980’s Erythropoesis Stimulating Agents Were Used In Kidney Damaged Patients Undergoing Dialysis But Their Application In Oral Maxillofacial Surgery Was Scarce To Minimize Morbidity And To Avoid Blood Transfusion4-6.&#x0D; Keywords: Oral Cavity, Chronic Kidney Disease, Dentistry, Oral Hygiene

A mouse ear skin model to study the dynamics of innate immune responses against the microsporidian Encephalitozoon cuniculi.

Microsporidia are obligate intracellular parasites related to fungi that cause severe infections in immunocompromised individuals. Encephalitozoon cuniculi is a microsporidian species capable of infecting mammals, including human and rodents. In response to microsporidian infection, innate immune system serves as the first line of defense and allows a partial clearance of the parasite via the innate immune cells, namely macrophages, neutrophils, dendritic cells, and Natural Killer cells. According to the literature, microsporidia bypass this response in vitro by modulating the response of macrophages. In order to study host-parasites interactions in vivo, we developed a model using the mouse ear pinna in combination with an intravital imaging approach. Fluorescent E. cuniculi spores were inoculated into the skin tissue to follow for the first time in real time in an in vivo model the recruitment dynamics of EGFP + phagocytic cells in response to the parasite. The results show that parasites induce an important inflammatory recruitment of phagocytes, with alterations of their motility properties (speed, displacement length, straightness). This cellular response persists in the injection zone, with spores detected inside the phagocytes up to 72 h post-infection. Immunostainings performed on ear tissue cryosections evoke the presence of developing infectious foci from 5 days post-infection, in favor of parasite proliferation in this tissue. Overall, the newly set up mice ear pinna model will increase our understanding of the immunobiology of microsporidia and in particular, to know how they can bypass and hijack the host immune system of an immunocompetent or immunosuppressed host.

Prevalence of secondary blood stream infections in patients with SARS- CoV-2 infection in a tertiary care hospital in Sri Lanka: A retrospective analysis

Introduction: Secondary infections are an important cause of morbidity and mortality related to SARS-CoV-2 infection. Early administration of effective antimicrobials improves patient outcome of such infections. In the present study, we investigated secondary blood stream infections (BSI) among COVID-19 patients in Colombo North Teaching Hospital, Sri Lanka (CNTH). Methods: This is a descriptive study, performed to analyse blood culture positive events retrospectively in patients hospitalized with COVID-19 in our institution from 1st June 2021 to 30th November 2021. Results: Of a total 434 blood cultures received from SARS-COV-2 positive patients, the overall blood culture positivity rate was 25.3 % (110/434) of which 11.7% (51/434) were clinically relevant bacteraemias. The contamination rate was 13.7% (59/434). Of the 60 isolates from clinically relevant bacteraemias, 60% (36/60) were Gram negative organisms, 31.7% (19/60) were Gram positive organisms and 8.3% (5/60) were Candida spp. The most prevalent isolates were Enterococcus spp. (20%; 12/60) and Acinetobacter spp. (15%; 9/60). Higher rates of multi drug resistance were observed among Gram negative isolates. The proportion of 3rd generation cephalosporins and ciprofloxacin resistance among enterobacterales was 81.3% and carbapenem resistance among Acinetobacter spp. was 77.8%. A primary focus of infection could be identified in only 18 blood stream infections (BSI). Conclusion: A high rate of significant BSIs was detected in this patient cohort. Blood culture contamination rate among COVID-19 patients was higher than the usual rates in the institution. A high number of bacteraemias due to Enterococcus spp. and Acinetobacter spp. were noted and overall antibiotic resistance among Gram negative organisms was high.

Endophthalmitis risk factors associates with phacoemulsification (Literature review)

The most frequent sources of microbial flora inside the eye are the ocular surface, foci of para-ocular infection of the patient. In vitreous samples from eyes with endophthalmitis microorganisms are revealed that are the saprophytic flora of the ocular surface. It is mainly represented by staphylococci, from which epidermal staphylococcus prevails quantitatively. Immunodeficiency states, diabetes mellitus, terminal liver and kidney diseases increase the risk of endophthalmitis development. Eradication of opportunistic pathogenic flora in the source loci, sanation of para-ocular infection, correction of somatic pathology by profile specialists is more relevant for endophthalmitis prevention. The problem of growing antibiotic resistance should also be considered. Thus, when selecting patients for surgery, one should be guided by the data on antibiotic resistance of the identified flora, pay attention to the presence of para-ocular infection and the systemic somatic status of the patient. This will provide a better vector for the development of recommendations for optimal prophylaxis.

Virus Propagation and Cell-Based Colorimetric Quantification

Zika virus (ZIKV) is a mosquito-borne virus belonging to the genus Flavivirus. ZIKV infection has been associated with congenital brain abnormalities and potentially Guillain-Barré syndrome in adults. Research on ZIKV to understand the disease mechanisms is important to facilitate vaccine and treatment development. The method of quantifying viruses is crucial and fundamental in the field of virology. The focus forming assay (FFA) is a virus quantification assay that detects the viral antigen with antibodies and identifies the infection foci of cells using the peroxidase immunostaining technique. The current study describes the virus propagation and quantification protocol using both 24-well and 96-well (high throughput) formats. Compared with other similar studies, this protocol has further described foci size optimization, which can serve as a guide to expand the use of this assay for other viruses. Firstly, ZIKV propagation is performed in Vero cells for 3 days. The culture supernatant containing ZIKV is harvested and quantitated using the FFA. Briefly, the virus culture is inoculated onto Vero cells and incubated for 2-3 days. Foci formation is then determined after optimized staining processes, including cell fixation, permeabilization, blocking, antibody binding, and incubation with peroxidase substrate. The stained virus foci are visualized using a stereo microscope (manual counting in 24-well format) or software analyzer (automated counting in 96-well format). The FFA provides reproducible, relatively fast results (3-4 days) and is suitable to be used for different viruses, including non-plaque-forming viruses. Subsequently, this protocol is useful for the study of ZIKV infection and could be used to detect other clinically important viruses.

The association of oral health with length of stay and mortality in the intensive care unit.

To analyze the relationship between the oral and systemic health status of adult patients admitted to the intensive care unit (ICU) with the length of stay and mortality. A daily oral examination and oral hygiene were performed in patients admitted to an adult ICU. Dental and oral lesions, systemic health status, the need for mechanical ventilation, length of stay, and mortality were registered. Multivariate linear and logistic regression analyses were performed to identify associations between length of stay and death of patients, respectively, with oral and systemic health status. In total, 207 patients were included, 107 (51.7%) male. Ventilated patients presented an increased length of stay (p < 0.001), mortality (p < 0.0001), number of medications (p < 0.0001), edentulism (p = 0.001), mucous lesions and bleeding (p < 0.0001), oropharyngitis (p = 0.03), and drooling (p < 0.001) compared to non-ventilated patients. The number of days in the ICU was associated with mechanical ventilation (p = 0.04), nosocomial pneumonia (p = 0001), end-stage renal disease (p < 0.0007), death (p < 0.0001), mucous bleeding (p = 0.01), tongue coating (p = 0.001), and cheilitis (p = 0.01). Mortality was associated with length of stay in the ICU (p < 0.0001), number of medications (p < 0.0001), and the need for mechanical ventilation (p = 0.006). ICU patients present poor oral health. Soft tissue biofilm and mucous ulcerations were associated with the length of stay in the ICU, but not with the mortality rate. Mucous lesions are associated with an increased length of stay in the ICU, and critically ill patients should receive oral care to control oral foci of infection and mucous lesions.

HMG-like DSP1 is a damage signal to mediate the western flower thrips, Frankliniella occidentalis, immune responses to tomato spotted wilt virus infection

Tomato spotted wilt virus (TSWV) causes a serious plant disease and is transmitted by specific thrips including the western flower thrips, Frankliniella occidentalis. The persistent and circulative virus transmission suggests an induction of immune defenses in the thrips. We investigated the immune responses of F. occidentalis to TSWV infection. Immunofluorescence assay demonstrated viral infection in the larval midguts at early stage and subsequent propagation to the salivary gland in adults. In the larval midgut, TSWV infection led to the release of DSP1, a damage-associated molecular pattern, from the gut epithelium into the hemolymph. DSP1 up-regulated PLA2 activity, which would lead to biosynthesis of eicosanoids that activate cellular and humoral immune responses. Phenoloxidase (PO) activity was enhanced following induction of PO and its activating protease gene expressions. Antimicrobial peptide genes and dual oxidase, which produces reactive oxygen species, were induced by the viral infection. Expression of four caspase genes increased and TUNEL assay confirmed apoptosis in the larval midgut after the virus infection. These immune responses to viral infection were significantly suppressed by the inhibition of DSP1 release. We infer that TSWV infection induces F. occidentalis immune responses, which are activated by the release of DSP1 from the infection foci within midguts.

Bacterial Keratitis. Part 2. Topical Aspects of Treatment

Bacterial keratitis (BC) is a threatening condition for the anatomy and function of the eyeball and requires an immediate complex of therapeutic measures. Effective treatment that preserves the anatomical and functional result of the organ of vision, including various non-surgical and surgical methods, is the basis for the treatment of CD. Drug therapy includes, first of all, massive etiotropic therapy, which is usually represented by broad-spectrum antibiotics, antiseptics and anti-inflammatory drugs. These combinations make it possible to competitively influence all links of the pathological process, showing an additive effect. Promising non-surgical means in the fight against bacterial infection of the cornea, in particular with resistant microorganisms, is the use of Quantum dots, Mitomycin C, Matrix metalloproteinases. The most radical surgical methods of treatment are based on therapeutic keratoplasty, the purpose of which is to excise the infectious focus of the cornea and restore its transparency through transplantation of donor corneal tissue, but this method has a number of disadvantages and limitations in its implementation. Autoconjunctival keratoplasty is the oldest method of treatment of progressive corneal ulcers and this method should be considered rather as temporary or preparatory before through keratoplasty, due to the lack of visual functions. The least radical and relevant surgical methods of CD treatment today are the use of Corneal Crosslinking, Microdiathermocoagulation, Cyanoacrylate glue, Amniotic membrane transplantation. The expansion of the arsenal of methods of influencing the microbial process of the cornea and their further study will allow for a quick response in response to the rapidly developing antibiotic resistance in the world.