Severity Of Lung Damage Research Articles

Standardized protocol for ultrasound diagnosis of the lungs with COVID-19

BACKGROUND. The era of coronavirus disease (COVID-19) set the such task for lung ultrasound (LUS): to identify the presence of COVID-pneumonia, its differential diagnosis with other conditions which characterized by interstitial edema, determining the severity of lung damage and its location, the dynamics of pulmonary changes tissue, the development of complications, as well as to assess the effectiveness of personalized therapy.
 OBJECTIVE. To develop a national LUS protocol for COVID-19 based on a balance of completeness of information with the optimal minimum of time and effort for medical staff, who often do not specialize in ultrasound using, with ultrasound equipment of basic and middle classes.
 MATERIALS AND METHODS. A total of 1576 patients with COVID-19 were examined. There were 810 women (51.4 %) and 766 men (48.6 %). The age of the examined patients was in the range of 18-83 years, the average age was 52.3±14.6 years. 4710 LUS were performed, of which 1572 were repeated and 3144 – in the dynamics. Polysystemic ultrasound was performed in 275 patients. Ultrasound devices of both domestic and foreign production were used: Soneus P7 (Ultrasine, Ukraine), HandyUsound (Ukraine), Xario, USAP-770A (Toshiba, Japan), GE Venue (General Electric, USA) and TE7 COVID (Mindray, China) with ultrasound probes of convex, linear and microconvex formats.
 RESULTS. Summarizing our own experience with patients with COVID-19 in 2020-2021, we have developed a version of the LUS-protocol, which has a convenient form of recording all stages of ultrasound with maximum coverage of the entire volume of both lungs according to a clear algorithm, a graphical form, clear ultrasound semiotics and diagnostic scoring criteria for the severity of lung damage.
 CONCLUSIONS. 1. Clinical data indicate the feasibility and effectiveness of using LUS for dynamic monitoring of patients with pneumonia caused by SARS-CoV-2, especially in oxygen-dependent patients at their bedside (POCUS principle). 2. Optimization of the use of radiological diagnostic methods due to ultrasound makes it possible to reduce the radiation dose of the patient.3. The use of LUS allows more adequate decision-making on the need for intensive and interventional therapy, leads to faster diagnosis, improved medical care, reduced length of stay of patients in ICU, as well as reducing the total cost of treatment.4. Ability to reduce the impact of SARS-CoV-2 on healthcare professionals by limiting physical contact with the patient. 5. The standardized Ukrainian protocol of LUS provides availability, efficiency and clear interpretation of the received data at its use by different doctors in different medical institutions. The integration of the protocol in to the ultrasound machine provides optimization of the workflow process.

X-ray study as an alternative method for visualizing changes in lungs, with infection caused by COVID-19

Introduction. Despite the existence of generally accepted diagnostic protocols, when a new coronavirus infection is suspected, in some cases, it is increasingly difficult to detect changes in the lung tissue in a timely manner due to the heavy workload of the main method of radiation diagnostics – computed tomography. Purpose of the study. To determine the effectiveness of the appointment of an X-ray examination as first-line metgod, as well as to carry out a comparative analysis of the use of radiation diagnostics methods – computed tomography and radiography in relation to the diagnostic sensitivity to changes in lung tissue when a person is infected with the SARS-COV-2 virus.Materials and methods. 150 patients (63.0 ± 8.4 years) with confirmed coronavirus infection were examined. Each of the participants underwent X-ray examination and computed tomography of the chest organs. The percentage of subjects studied for each of the degrees of severity of lung damage was determined to identify the proportion of involvement of lung tissue in the pathological process in the bulk of the examined individuals.Results. Of the 150 patients, changes in the lung tissue during chest X-ray were detected in 97 (65%), respectively, in 53 (35%), pathological changes in the lungs were not visualized. When examining patients by computed tomography, changes in the lungs were detected in 143 patients (95%), X-ray morphological changes were not detected in 7 subjects (5%). When detecting the volume of lung damage, it turned out that the majority of the subjects – 86 people (57%) – had the degree of damage CT-2. The degree of CT-1 and CT-3 was determined in 26 (17%) and 25 (17%) patients, respectively. CT-4 was observed in 6 patients (4%), and in 5% of cases, CT was not able to detect pathological changes in the lung tissue, the degree of CT-0 was established.Conclusion. In the assessment of viral lung damage, radiography takes a significant place, but in 35% of cases, radiographic examination failed to identify the existing pathological changes. CT of the chest organs confirms its value as the “gold standard” in the study of pulmonary pathology in coronavirus infection, but if it is impossible to perform it, radiography is recommended.

Reduced Flow-Mediated Dilatation Is Not Related to COVID-19 Severity Three Months after Hospitalization for SARS-CoV-2 Infection.

The coronavirus disease 2019 (COVID-19) pandemic has spread rapidly worldwide, with more than two million deaths. Evidence indicates the critical role of the vascular endothelium in its pathophysiology but, like potential changes in functional vasodilation, the vascular effect of SARS-CoV-2 at a given distance from the acute infection is largely unknown. We assessed brachial artery flow-mediated dilatation (FMD) in 27 COVID-19 patients needing conventional or intensive care unit hospitalization, three months after SARS-CoV-2 infection diagnosis and in nine age- and sex- matched control subjects. Interestingly, the FMD was lower in COVID-19 patients as compared to controls (8.2 (7.2–8.9) vs. 10.3 (9.1–11.7)); p = 0.002, and half of the hospitalized COVID-19 survivors presented with a reduced FMD < 8% at three months of COVID-19 onset. Impaired FMD was not associated with severe or critical SARS-CoV-2 infection, reflected by ICU hospitalization, total hospitalization duration, or severity of lung damage. In conclusion, reduced FMD is often observed even three months after hospitalization for SARS-CoV-2 infection, but such alteration predominantly appears to not be related to COVID-19 severity. Longer and larger follow-up studies will help to clarify the potential prognosis value of FMD among COVID-19 patients, as well as to further determine the mechanisms involved.

Pulmonary Function and Radiologic Features in Survivors of Critical COVID-19: A 3-Month Prospective Cohort

BackgroundMore than 20% of hospitalized patients with COVID-19 demonstrate ARDS requiring ICU admission. The long-term respiratory sequelae in such patients remain unclear.Research QuestionWhat are the major long-term pulmonary sequelae in critical patients who survive COVID-19?Study Design and MethodsConsecutive patients with COVID-19 requiring ICU admission were recruited and evaluated 3 months after hospitalization discharge. The follow-up comprised symptom and quality of life, anxiety and depression questionnaires, pulmonary function tests, exercise test (6-min walking test [6MWT]), and chest CT imaging.ResultsOne hundred twenty-five patients admitted to the ICU with ARDS secondary to COVID-19 were recruited between March and June 2020. At the 3-month follow-up, 62 patients were available for pulmonary evaluation. The most frequent symptoms were dyspnea (46.7%) and cough (34.4%). Eighty-two percent of patients showed a lung diffusing capacity of less than 80%. The median distance in the 6MWT was 400 m (interquartile range, 362-440 m). CT scans showed abnormal results in 70.2% of patients, demonstrating reticular lesions in 49.1% and fibrotic patterns in 21.1%. Patients with more severe alterations on chest CT scan showed worse pulmonary function and presented more degrees of desaturation in the 6MWT. Factors associated with the severity of lung damage on chest CT scan were age and length of invasive mechanical ventilation during the ICU stay.InterpretationThree months after hospital discharge, pulmonary structural abnormalities and functional impairment are highly prevalent in patients with ARDS secondary to COVID-19 who required an ICU stay. Pulmonary evaluation should be considered for all critical COVID-19 survivors 3 months after discharge.

Hydroxychloroquine in patients with novel coronavirus infection (COVID-19): a case-control study.

Actuality One of the most widely discussed treatments for patients with COVID-19, especially at the beginning of the epidemy, was the use of the antimalarial drug hydroxychloroquine (HCQ). The first small non-randomized trials showed the ability of HCQ and its combination with azithromycin to accelerate the elimination of the virus and ease the acute phase of the disease. Later, large, randomized trials did not confirm it (RECOVERY, SOLIDARITY). This study is a case-control study in which we compared patients who received and did not receive HCQ.Material and Methods 103 patients (25 in the HCQ treatment group and 78 in the control group) with confirmed COVID-19 (SARS-CoV-2 virus RNA was detected in 26 of 73 in the control group (35.6%) and in 10 of 25 (40%) in the HCQ group) and in the rest - a typical picture of viral pneumonia on multislice computed tomography [MSCT]) were included in the analysis. The severity of lung damage was limited to stages I-II, the CRP level should not exceed 60 mg/dL, and oxygen saturation in the air within 92-98%. We planned to analysis the duration of treatment of patients in the hospital, the days until the normalization of body temperature, the number of points according to the original SHOCS-COVID integral scale, and changes in its components (C-reactive protein (CRP), D-dimer, and the percentage of lung damage according to MSCT).Results Analysis for the whole group revealed a statistically significant increase in the time to normalization of body temperature from 4 to 7 days (by 3 days, p&lt;0.001), and the duration of hospitalization from 9.4 to 11.8 days (by 2.4 days, p=0.002) when using HCQ in comparison with control. Given the incomplete balance of the groups, the main analysis included 46 patients who were matched by propensity score matching. The trend towards similar dynamics continued. HCQ treatment slowed down the time to normalization of body temperature by 1.8 days (p=0.074) and lengthened the hospitalization time by 2.1 days (p=0.042). The decrease in scores on the SHOCS -COVID scale was statistically significant in both groups, and there were no differences between them (delta - 3.00 (2.90) in the HCQ group and - 2.69 (1.55) in control, p=0.718). At the same time, in the control group, the CRP level returned to normal (4.06 mg/dl), and with the use of GC, it decreased but remained above the norm (6.21 mg/dl, p=0.05). Side effects requiring discontinuation of treatment were reported in 3 patients in the HCQ group and none in the control group.Conclusion We have not identified any positive properties of HCQ and its ability to influence the severity of COVID-19. This antimalarial agent slows down the normalization of the body's inflammatory response and lengthens the time spent in the hospital. HCQ should not be used in the treatment of COVID-19.

Point-of-care ultrasonography for risk stratification of non-critical COVID-19 patients on admission (POCUSCO): a study protocol of an international study

IntroductionIn the context of the COVID-19 pandemic, early identification of patients who are likely to get worse is a major concern. Severity mainly depends on the development of acute respiratory...

Ultrasound assessment of pulmonary fibroproliferative changes in severe COVID-19: a quantitative correlation study with histopathological findings.

PurposeThis study was designed to evaluate the usefulness of lung ultrasound (LUS) imaging to characterize the progression and severity of lung damage in cases of COVID-19.MethodsWe employed a set of combined ultrasound parameters and histopathological images obtained simultaneously in 28 patients (15 women, 0.6–83 years) with fatal COVID-19 submitted to minimally invasive autopsies, with different times of disease evolution from initial symptoms to death (3–37 days, median 18 days). For each patient, we analysed eight post-mortem LUS parameters and the proportion of three histological patterns (normal lung, exudative diffuse alveolar damage [DAD] and fibroproliferative DAD) in eight different lung regions. The relationship between histopathological and post-mortem ultrasonographic findings was assessed using various statistical approaches.ResultsStatistically significant positive correlations were observed between fibroproliferative DAD and peripheral consolidation (coefficient 0.43, p = 0.02) and pulmonary consolidation (coefficient 0.51, p = 0.005). A model combining age, time of evolution, sex and ultrasound score predicted reasonably well (r = 0.66) the proportion of pulmonary parenchyma with fibroproliferative DAD.ConclusionThe present study adds information to previous studies related to the use of LUS as a tool to assess the severity of acute pulmonary damage. We provide a histological background that supports the concept that LUS can be used to characterize the progression and severity of lung damage in severe COVID-19.

Морфологическая характеристика коронавирусной инфекции первой и второй волн пандемии

Introduction. A peculiarity of the first two waves of the epidemic of a novel coronavirus infection was that during their development only diagnostic methods, treatment regimens and anti-epidemic measures were only being developed, there were no vaccines and no mass vaccination was carried out. In this regard, the pandemic of the novel coronavirus infection of the first two waves is characterized by spontaneous progression of the disease. Aim of the research. Comparative study of morphological and clinical features of the novel coronavirus infection in the 1st and 2nd waves of the pandemic in 2020. Materials and methods. The autopsy protocols of deceased patients at the City Clinical Hospital No. 1 (Novosibirsk) during the 1st (May — June 2020) (1st group) and 2nd (October — December 2020) (2nd group) waves of the epidemic were analyzed. The 1st group included 30 cases, that is 14 women (46.7%) and 16 men (53.3%), in the 2nd — 110 cases, of which 43 were women (39.1%) and 67 – men (60.9%). In all cases, the presence of SARS-CoV-2 RNA in nasopharyngeal swabs from the patients was confirmed by polymerase chain reaction. Results. The novel coronavirus infection (coronavirus disease 2019 (COVID-19), ICD-10 code: U07.1) as the principal diagnosis in both the 1st and 2nd waves was recorded in more than 66% of observations. Among the comorbidities, the circulatory and endocrine disorders (primarily diabetes mellitus and obesity), kidneys and urinary tract diseases, mainly chronic pyelonephritis, prevailed. The COVID-19 pneumonia was predominantly bilateral polysegmental serоhemorrhagic in nature, however, in the 1st wave of the pandemic, the frequency of subtotal and seropurulent pneumonias was higher than in patients hospitalized during the 2nd wave. Diffuse alveolar damage in the 1st wave of the epidemic was characterized by the predominance of the early (exudative) phase of inflammation, in contrast to the 2nd wave, which was characterized by the predominance of the late (productive) phase in patients. Histologically, in patients of the 1st wave, the signs of exudative inflammation and hemorrhagic phenomena (with neutrophils and hyaline membranes in the alveoli, infarctions caused by thrombosis and pulmonary thromboembolism) prevailed. In the 2nd wave, signs of productive inflammation were more common in the lungs (predominance of macrophages in the alveoli, organizing pneumonia (pneumofibrosis), squamous cell metaplasia). Conclusion. The differences in the course of the novel coronavirus infection in the 1st and 2nd waves of the 2020 epidemic concerned primarily the frequency and structure of comorbidities and the level of polymorbidity, which were higher in patients during the 2nd wave. The severity of lung damage in patients of the 2nd wave was less pronounced: subtotal bilateral pneumonias were less common, lower lobe seropurulent pneumonias were more common. Diffuse alveolar damage in the 1st wave of the epidemic was characterized by the predominance of the early (exudative) phase, in contrast to the 2nd wave, which was characterized by the predominance of the late, productive phase of inflammation.

Could routine race-adjustment of spirometers exacerbate racial disparities in COVID-19 recovery?

Could routine race-adjustment of spirometers exacerbate racial disparities in COVID-19 recovery?

Astaxanthin Prevents Lung Injury Due to Hyperoxia and Inflammation.

This study aimed to ascertain the effects of astaxanthin on the lungs of rat pups with bronchopulmonary dysplasia (BPD) induced by hyperoxia and lipopolysaccharide (LPS). Forty-two newborn Wistar rats, born to spontaneous pregnant rats, were divided into three groups: Hyperoxia (95% O2) + lipopolysaccharide (LPS) group, hyperoxia + LPS + astaxhantin group, and control: no treatment group (21% O2). Pups in the hyperoxia + LPS + astaxanthin group were given 100 mg/kg/day oral astaxanthin from the first day to the fifth day. Histopathologic and biochemical evaluations, including glutathione (GSH), total anti-oxidant status (TAS), total oxidant status (TOS), lipid hydroperoxide (LPO), 8-hydroxydeoxyguanosine (8-OHdG), advanced oxidation protein products (AOPP), myeloperoxidase (MPO), total thiol, tumor necrosis factor-alpha (TNF-α), interleukin 1 beta (IL-1β), and caspase-3 activities, were performed. Better survival rates and weight gain were demonstrated in the hyperoxia + LPS + astaxanthin group (p <0.001). In the histopathologic evaluation, the severity of lung damage was significantly reduced in the hyperoxia+LPS+astaxanthin group, as well as decreased apoptosis (ELİSA for caspase-3) (p <0.001). The biochemical analyses of lung tissues showed that TAS, GSH, and Total thiol levels were significantly higher in the astaxanthin treated group compared to the hyperoxia + LPS group (p <0.05) while TOS, AOPP, LPO, 8-OHdG, MPO levels were significantly lower (p <0.001). In addition, unlike the hyperoxia + LPS group, TNF-α and IL-1β levels in lung tissue were significantly lower in the astaxanthin-treated group (p <0.001). Astaxanthin was shown to reduce lung damage caused by inflammation and hyperoxia with its anti-inflammatory, anti-oxidant, anti-apoptotic properties, and to protect the lung from severe destruction.

Artificial intelligence model for COVID-19 detection based on native chest computed tomography

Summary The aim is to develop and evaluate the clinical effectiveness of the artificial intelligence system for analyzing chest CT images, recognizing the leading signs of lung damage caused by coronavirus infection, and determining the damage volume. Materials and methods. We used open source images for the model training and validation, including anonymized complete studies or separate axial sections of chest CT of patients with PCR confirmed coronavirus etiology of lung disease. Algorithm of the model included sequence as follows: axial sections of CT of the chest organs at the entrance, image segmentation into 4 classes, visualization and calculation of the area as a part (in%) occupied by each finding from the total area of the pulmonary fields in all sections available for analysis. In this work, we used a convolutional artificial neural network for segmentation of the form of an encoder-decoder, with a Feature Pyramid Network (FPN) decoder and an encoder based on the EfficientNet-B5 classification neural network. To increase the diversity of the training data set, as well as to protect the neural network model from retraining, input image transformations are used in the learning process. Results. The neural network model with high accuracy (IoU 0.82-0.97) reveals diagnostic signs that determine the severity of lung damage with a new coronavirus infection, on axial sections of native computed tomography of the chest. The diagnostic accuracy of the model for determining the signs of interstitial and alveolar infiltration exceeds the accuracy of a novice radiologist and is comparable to a diagnostician in predicting the shape of pulmonary fields and the presence of pleural effusion. Findings. The model can be used as an effective intelligent radiologist assistant when working with CT studies of patients with suspected coronavirus infection.

Differential Diagnosis of Congenital Pneumonia in Newborns with Low and Extremely Low Body Weight (Morphological Study)

The purpose of the study is a comprehensive assessment of morphological changes in the placenta and lungs to detect early signs of congenital pneumonia in extremely premature infants.Materials and methods. Protocols of post-mortem examinations of 23 preterm newborns died from severe respiratory failure were analyzed. The average gestational age of the newborns was 26.4±2.7 weeks and the body weight at birth was 972.4±355.8 grams. In the sample, 78.3% of infants had an extremely low birth weight (ELBW). At birth, all newborns presented severe asphyxia. Newborn underwent several types of respiratory therapy since birth: Mechanical ventilation was performed in 65.2% of newborns since their birth, non-invasive ventilation was performed in 26.1% of cases, and 8.7% of patients underwent oxygenotherapy through a facial mask. In all cases, there was an unfavorable course of the neonatal period, a progressive deterioration of newborns' condition, and a lethal outcome. A comprehensive histological examination of the placenta and the lungs of deceased premature newborn infants was performed.Results. Congenital infections of different localizations remain the leading cause of death.Congenital pneumonia and generalized infections are clinically manifested at birth by severe perinatal hypoxia and respiratory failure. In the case of congenital pneumonia, the morphological patterns are polymorphic and characterize the severity of lung damage. For some newborns, these patterns include accumulation of exudates and fibrin, segmented leukocytes, fragments of basophilic coccal microflora, and a large number of colony forming bacilli, and desquamated alveolocytes with a deformed nucleus are visualized in the deformed lumen of the alveoli and bronchi. Diffuse lymphoid-leukocyte infiltration in the septa and respiratory parts of the lungs are typical for other infants. Histological examination find lumpy or lamellar eosinophilic hyaline membranes in alveoli in specimens from these newborns. Diffuse, focal or confluent segmentonuclear infiltration in various lung structures is commonly combined with hyaline membranes of various localizations and sizes. Hyaline membranes were detected in 93.5% of cases.Conclusion. Very early preterm delivery is associated with intrauterine pneumonia and systemic infection in extremely premature infants. Early clinical and laboratory signs of intrauterine infectious lung include severe perinatal hypoxia, very low Apgar score and laboratory test findings (hypoxaemia and decompensated metabolic lactate acidosis) that are resistant to standard resuscitation measures. Hypoxemia and decompensated metabolic acidosis persisting during the first hours of postnatal life indicate the severity of intrauterine lung damage and require a rapid change of treatment aimed at normalization of lung function, prevention of complications in the respiratory system, hemostasis and central nervous system. Clinicians should be better informed about the features of early postnatal adaptation of extremely premature infants with congenital pneumonia to provide appropriate treatment.

Ginger ( Zingiber officinale ) prevents severe damage to the lungs due to hyperoxia and inflammation

Hyperoxia- and inflammation-induced lung injury is an important cause of the development of bronchopulmonary dysplasia (BPD) in premature infants. We aimed to ascertain the beneficial effects of ginger ( Zingiber officinale ) on rat pups exposed to hyperoxia and inflammation. Thirty-six newborn Wistar rats were randomly divided into 3 groups as the hyperoxia (95% O 2 ) + lipopolysaccharide (LPS) group, the hyperoxia + LPS + ginger-treated group, and the control/no treatment group (21% O 2 ). Pups in the hyperoxia + LPS + ginger group were administered oral ginger at a dose of 1000 mg/kg daily during the study period. Histopathologic, immunochemical (SMA and lamellar body), and biochemical evaluations including total antioxidant status (TAS), total oxidant status (TOS), malondialdehyde (MDA), myeloperoxidase (MPO), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and caspase-3 activities were performed. Better weight gain and survival rates were shown in the hyperoxia + LPS + ginger group (P < 0.05). In the histopathologic and immunochemical evaluation, severity of lung damage was significantly reduced in the hyperoxia + LPS + ginger group, as well as decreased apoptosis (ELISA for caspase-3) (P < 0.05). Tissue TAS levels were significantly protected, and TOS, MDA, and MPO levels were significantly lower in the hyperoxia + LPS + ginger group (P < 0.05). Tissue TNF-α, IL-1β, and IL-6 concentrations were significantly decreased in the ginger-treated group (P < 0.05). Ginger efficiently reduced the lung damage and protected the lungs from severe damage due to hyperoxia and inflammation. Therefore, ginger may be an alternative option for the treatment of BPD.

Increased IL-6 Levels Correlate with Severity of Lung Damage and Early Death in Tanzanian Patients with Pulmonary Tuberculosis

Background: There is an urgent need for clinically relevant host biomarkers of protection, disease activity, response to treatment and predicting long term tissue damage for patients with pulmonary tuberculosis (TB). We studied cytokine biomarkers in a longitudinal cohort of patients with pulmonary TB in order to predict pulmonary tissue damage or death. Methods: Two hundred and thirtyfour (234) Tanzanian adult patients (96 HIV-positive) with newly diagnosed pulmonary TB and seven healthy controls were enrolled. Serum concentrations of IFN-gamma, TNF-α, IL-2, IL-6, IL-10. IL-17A and IL-21 were determined using enzyme-linked immunosorbent assay (ELISA) kits on day 1, two and five months after anti-TB treatment initiation. Postero-anterior chest X-rays (CXR) were scored for lung injury. Correlations between cytokine concentrations, mortality and lung injury were determined using multivariate logistic regression. Findings: Patients with TB had significantly higher median levels of IL-6 (3863pg/ml) at diagnosis compared to healthy controls (11 pg/ml). Increased IL-6 levels significantly correlated with survival (RR = 0.95; 95% CI 0.91 - 0.98; P < 0.01) and were associated with severe lung injury (RR 1.03; 95% 1.01 - 1.05; P = 0.02). Among patients with severe lung injury at diagnosis, every 100pg/ml increase in baseline IL-6 levels was accompanied with 6% increased risk for residual lung damage post-TB treatment (RR = 1.06; 95%CI 1.01 - 1.13). Anti-TB therapy was associated with reduction in serum cytokine levels except for IL-6, which did not decrease with treatment (P = 0.003). Interpretation: Increased IL-6 levels correlate with severity of lung damage in patients with pulmonary TB regardless of HIV status. IL-6 may serve as a target for host-directed therapy in patients with pulmonary TB. Funding: This study was supported by grants from the Swedish International Development Cooperation Agency (SIDA); Vinnova and Swedish Heart and Lung Foundation (Hjartlungfonden) to MM. AZ and MM acknowledge support from the European and Developing Countries Clinical Trials Partnership (EDCTP). Declaration of Interest: The authors declare no conflicts of interest Ethical Approval: Ethical clearance was granted by the Muhimbili University of Health and Allied Health sciences (MUHAS) Senate Research and Publications committee (Ref. No. RP/AEC/VOL.XII dated November 23rd2011). Written informed consent was obtained from all study participants prior to inclusion.

The PA Endonuclease Inhibitor RO-7 Protects Mice from Lethal Challenge with Influenza A or B Viruses.

Current influenza treatment relies on a single class of antiviral drugs, the neuraminidase inhibitors (NAIs), raising concern over the potential emergence of resistant variants and necessitating the development of novel drugs. In recent years, investigational inhibitors targeting the endonuclease activity of the influenza acidic polymerase (PA) protein have yielded encouraging results, although there are only limited data on their in vivo efficacy. Here, we examined the antiviral potential of the PA endonuclease inhibitor RO-7 in prophylactic and therapeutic regimens in BALB/c mice inoculated with influenza A/California/04/2009 (H1N1)pdm09 or B/Brisbane/60/2008 viruses, which represent currently circulating antigenic variants. RO-7 was administered to mice intraperitoneally twice daily at dosages of 6, 15, or 30 mg/kg/day for 5 days, starting 4 h before or 24 or 48 h after virus inoculation, and showed no adverse effects. Prophylactic administration completely protected mice from lethal infection by influenza A or B virus. The level of therapeutic protection conferred depended upon the time of treatment initiation and RO-7 dosage, resulting in 60 to 100% and 80 to 100% survival with influenza A and B viruses, respectively. RO-7 treatment significantly decreased virus titers in the lung and lessened the extent and severity of lung damage. No PA endonuclease-inhibitor resistance was observed in viruses isolated from lungs of RO-7-treated mice, and the viruses remained susceptible to the drug at nanomolar concentrations in phenotypic assays. These in vivo efficacy results further highlight the potential of RO-7 for development as antiviral therapy for influenza A and B virus infections.

Casapse-1 is not required for the induction of Silica-induced autoimmunity

Abstract Occupational exposure to respirable crystalline silica induces silicosis in humans &amp; has been associated with lupus (SLE), but the specific mechanisms &amp; susceptibility factors are largely unknown. Recent studies have highlighted a prominent role for interleukin-1β in silica-induced lung inflammation. Caspase-1 is predominantly involved in the cleavage of pro-IL-1β to active IL-1β. Therefore, we investigated the role of IL-1β in the severity of lung damage &amp; autoimmune response to silica by exposing B10.S and Caspase-1 deficient B10.S (B10.S-Casp1−/−) mice to crystalline silica (1 mg in 25 μl) in PBS or PBS only, by intranasal instillation twice a week for 4 weeks. When analyzed for autoimmunity at 28 weeks post-exposure, both B10.S and B10.S-Casp1−/− mice developed similar elevated levels of anti-chromatin &amp; anti-nuclear antibodies in serum &amp; broncho-alveolar lavage (BAL) compared to PBS controls. Correspondingly, both silica-treated B10.S and B10.S-Casp1−/−mice had a significant peribronchial &amp; perivascular lymphocytic infiltrates (PLI), alveolar proteinosis (AP), and alveolitis, but the PLI and AP were significantly higher in B10.S mice than B10.S-Casp1−/−mice. Similarly, robust increases in the levels of total protein &amp; cell numbers in BAL fluid, but total protein was significantly higher in B10.S mice than B10.S-Casp1−/−mice. Higher numbers of inflammatory M1 macrophages (CD11b+Ly6chi) &amp; neutrophils (Ly6Ghi) were also observed in both groups of silica-treated mice. These findings indicate that caspase-1 and by inference IL-1β is not required for silica-induced autoimmunity in mice, but contributes to the lung inflammation, &amp; suggests that NLRP3 inflammasome, cathepsin C, elastase, granzyme B may be more important.

Locomotor and Diaphragm Muscle Mitochondrial Oxidative Capacity in Lung‐Targeted VEGF Gene Deleted Mice Exposed to Cigarette Smoke

Locomotor skeletal muscle function is impaired in patients with chronic obstructive pulmonary disease (COPD). The mechanisms responsible for these skeletal muscle abnormalities are not well understood. We hypothesized that the severity of lung damage following chronic exposure to cigarette smoke (CS) would determine the mitochondrial oxidative capacity of locomotor and respiratory skeletal muscle.Adult VEGFLoxP mice with or without lung‐targeted VEGF gene deletion (5 months of age) were used as a model of progressively compromised static lung mechanics following exposure to 2 × 30 min of daily, nose‐only CS for an additional 4 months. Deletion of the VEGF gene was accomplished by instilling half the mice with AAV‐Cre one month before initiating CS exposure. The remaining mice received a control AAV (LacZ). Further, both groups (Cre and LacZ) were assigned either to CS exposure or room air control (RA). Mitochondrial respiration in permeabilized fiber bundles from the diaphragm and medial gastrocnemius was measured using high‐resolution respirometry.Gastrocnemius ADP‐stimulated O2 consumption with malate and glutamate as substrates was 78±6%, 66±7% and 60±11% lower in CS‐LacZ, RA‐Cre, and CS‐Cre, respectively, than RA‐LacZ (p&lt;0.05). Gastrocnemius electron transport system capacity was 68±5% (p&lt;0.05), 53±11% (p&lt;0.05) and 48±21% (NS) lower in the CS‐LacZ, RA‐Cre, and CS‐Cre, respectively, than RA‐LacZ. Diaphragm ADP‐stimulated O−2 consumption was 56±9% lower in CS‐LacZ fibers than RA‐LacZ (p&lt;0.05). Diaphragm fiber electron transport system capacity was not different between the groups.Mice with a COPD‐like phenotype exhibited impaired locomotor muscle oxidative capacity irrespective of whether the lung damage was caused by cigarette smoke exposure or VEGF gene deletion. In contrast, diaphragm mitochondrial respiration only showed a deficit in ADP‐stimulated O2 consumption with CS exposure. Thus, mitochondrial respiration in locomotor skeletal muscle appears to be less well‐preserved than diaphragm in this model of COPD.Support or Funding InformationDepartment of Veterans Affairs

Vitamin D as Supplemental Therapy for Pneumocystis Pneumonia.

The combination of all-trans retinoic acid (ATRA) and primaquine (PMQ) has been shown to be effective for therapy of Pneumocystis pneumonia (PCP). Since a high concentration of ATRA has significant adverse effects, the possibility that vitamin D can be used to replace ATRA for PCP therapy was investigated. C57BL/6 mice were immunosuppressed by depleting CD4(+) cells and infected with Pneumocystis murina 1 week after initiation of immunosuppression. Three weeks after infection, the mice were treated orally for 3 weeks with vitamin D3 (VitD3) alone, PMQ alone, a combination of VitD3 and PMQ (VitD3-PMQ), or a combination of trimethoprim and sulfamethoxazole (TMP-SMX). Results showed that VitD3 (300 IU/kg/day) had a synergistic effect with PMQ (5 mg/kg/day) for therapy of PCP. Flow cytometric studies showed that this VitD3-PMQ combination recovered the CD11b(low) CD11c(high) alveolar macrophage population in mice with PCP as effectively as TMP-SMX. The VitD3-PMQ combination also reduced the massive infiltration of inflammatory cells into the lungs and the severity of lung damage. VitD3 was also shown to reduce the dose of TMP-SMX required for effective treatment of PCP. Taken together, results of this study suggest that a VitD3-PMQ combination can be used as an alternative therapy for PCP.

Occurrence of Aelurostrongylus abstrusus (Railliet, 1898) in Danish cats: A modified lung digestion method for isolating adult worms

As Aelurostrongylus abstrusus has not previously received any attention in Denmark, the study investigated the occurrence of A. abstrusus amongst outdoor cats from three regions (Zealand, Møn and Falster). Faeces and lungs were collected from a total of 147 feral (n=125) and domesticated cats (n=22) that were euthanized for reasons outside of this project. Using a modified Baermann technique 13.6% of the cats was found to be positive. A new lung digestion technique was developed to isolate eggs, L1 and adult worms from the lungs and this revealed a prevalence of 15.6% although with regional differences. There was no difference between feral and domesticated cats just as sex and age did not appear to influence prevalence and worm burden. Lungs from 87% of the positive cats had the gross appearance compatible with A. abstrusus and the severity of lung damage was proportional to LPG and number of adult worms. Within the current range of worm burdens (0–22) with a mean intensity of 7 per cat, there was a correlation with faecal excretion levels of L1 that ranged from 0–39,000 with a mean of 3586 per cat. The results did not indicate that the infection levels of the naturally infected cats were substantially affected by acquired immunity, but further studies are needed to determine the importance of host immune responses in regulating parasite populations.

Protective effect of β-glucan on acute lung injury induced by lipopolysaccharide in rats.

Lipopolysaccharide (LPS)-induced endotoxemia can cause serious organ damage such as acute lung injury and death by triggering the secretion of proinflammatory cytokines and acute-phase reactants. The goal of this study was to evaluate the effects of β-glucan on inflammatory mediator levels and histopathological changes in LPS-induced endotoxemia. Forty-seven male Wistar albino rats were randomly allocated into four groups as follows: control group, LPS group (10 mg/kg LPS), LPS + β-glucan group (100 mg/kg β-glucan before LPS administration), and β-glucan group. Twelve hours after LPS administration, lung and serum samples were collected. Concentrations of IL-6, IL-8, C-reactive protein (CRP), and procalcitonin were measured in the serum at hours 0 (basal) and 12. The severity of lung damage was assessed by an appropriate histopathological scoring system. Serum levels of CRP in the LPS group at 12 h were significantly higher than in the other groups, whereas serum IL-6 levels in the LPS and LPS + β-glucan groups at 12 h were significantly decreased. The mean histopathological damage score of the LPS group was slightly higher than that of the LPS + β-glucan group. Moreover, mortality rate was significantly decreased in the LPS + β-glucan group versus the LPS group. β-glucan reduces endotoxemia-induced mortality and might be protective against endotoxemia-induced lung damage.