Tuberculous pleural effusion in a patient with sympathetic ophthalmia on immunosuppression: a case report

Background Tuberculous pleural effusion (TPE) is paucibacillary, making its diagnosis difficult based on laboratory investigations alone. We present a case of a patient with a TPE who was initially misdiagnosed to have azathioprine-induced lung injury. The diagnosis of TPE was arrived at with the help of clinical assessment, laboratory and radiological investigations. Case presentation A 25-year-old chronic smoker with sympathetic ophthalmia on long-term immunosuppression, latent tuberculosis infection and a significant family history of tuberculosis presented with a three-week history of productive cough, low-grade fever, night sweats and weight loss. Examination of the lungs showed reduced breath sounds at the right lower zone. Chest x-ray showed minimal right pleural effusion with a small area of right upper lobe consolidation. The pleural fluid was exudative with predominant mononuclear leukocytes. Direct smears of sputum and pleural fluid; polymerase chain reaction of pleural fluid; and sputum, pleural fluid and blood cultures were negative for M. tuberculosis (MTB) and other organisms. As he did not respond to a course of broad-spectrum antibiotics, he was then treated as a case of azathioprine-induced lung injury. However, his condition did not improve despite the cessation of azathioprine. A contrast-enhanced computed tomography of the thorax showed right upper lobe consolidation with tree-in-bud changes, bilateral lung atelectasis, subpleural nodule, mild right pleural effusion and mediastinal lymphadenopathy. Bronchoalveolar lavage was negative for malignant cells and microorganisms including, MTB. However, no pleural biopsy was done. He was empirically treated with anti-tubercular therapy for 9 months duration and showed complete recovery. Conclusion A high index of suspicion for TPE is required in individuals with immunosuppression living in regions endemic to tuberculosis. Targeted investigations and sound clinical judgement allow early diagnosis and prompt treatment initiation to prevent morbidity and mortality.

Polyclonal antibody–based immunohistochemical detection of intraleukocytic Theileria parasites in roan and sable antelopes

Theileria parasites commonly infect African wild artiodactyls. In rare roan ( Hippotragus equinus) and sable ( H. niger) antelopes, Theileria sp. (sable)-associated calf mortalities constrain breeding programs. The pathogenicity of most leukocyte-transforming Theileria spp. originates in their invasion of and multiplication in various mononuclear leukocytes, the transformation of both infected and uninfected leukocytes, and their infiltration of multiple organs. Understanding the pathogenesis of theileriosis can be improved by the use of immunohistochemistry (IHC) to identify the localization of the parasites in tissue sections. Our aim was to develop a reproducible IHC assay to detect leukocyte-associated Theileria parasites in formalin-fixed, paraffin-embedded roan and sable tissues. Polyclonal antibodies were purified from the sera of 5 roans from an area endemic for Theileria sp. (sable) and tested for IHC reactivity in 55 infected and 39 control roan and sable antelopes, and for antigen and species cross-reactivity in an additional 58 cases. The 3 strongest antibodies consistently detected intraleukocytic theilerial antigens in known positive cases in roan and sable antelopes, and also detected other Theileria spp. in non-hippotraginid wild artiodactyl tissues. The antibodies did not cross-react with other apicomplexan protozoa, with the exception of Cryptosporidium. Given that PCR on its own cannot determine the significance of theilerial infection in wild ruminants, IHC is a useful laboratory test with which to confirm the diagnosis in these species.

Type I and III IFNs produced by the nasal epithelia and dimmed inflammation are features of alpacas resolving MERS-CoV infection

While MERS-CoV (Middle East respiratory syndrome Coronavirus) provokes a lethal disease in humans, camelids, the main virus reservoir, are asymptomatic carriers, suggesting a crucial role for innate immune responses in controlling the infection. Experimentally infected camelids clear infectious virus within one week and mount an effective adaptive immune response. Here, transcription of immune response genes was monitored in the respiratory tract of MERS-CoV infected alpacas. Concomitant to the peak of infection, occurring at 2 days post inoculation (dpi), type I and III interferons (IFNs) were maximally transcribed only in the nasal mucosa of alpacas, while interferon stimulated genes (ISGs) were induced along the whole respiratory tract. Simultaneous to mild focal infiltration of leukocytes in nasal mucosa and submucosa, upregulation of the anti-inflammatory cytokine IL10 and dampened transcription of pro-inflammatory genes under NF-κB control were observed. In the lung, early (1 dpi) transcription of chemokines (CCL2 and CCL3) correlated with a transient accumulation of mainly mononuclear leukocytes. A tight regulation of IFNs in lungs with expression of ISGs and controlled inflammatory responses, might contribute to virus clearance without causing tissue damage. Thus, the nasal mucosa, the main target of MERS-CoV in camelids, seems central in driving an efficient innate immune response based on triggering ISGs as well as the dual anti-inflammatory effects of type III IFNs and IL10.

Protective potential of Cynara scolymus extract in thioacetamide model of hepatic injury in rats

Hepatic injury is a worldwide health problem. This study aimed to evaluate the possible hepatoprotective potential of Artichoke (Cynara scolymus) extract (CSE) in albino rats using the thioacetamide (TAA) a model of liver injury. Acclimatized 42 rats were divided randomly into seven groups, each consists of six rats, and subjected to different treatments. Hepatic injury model was induced by administration of TAA at a dose of 100 mg per kg, intraperitoneally, twice weekly for 8 weeks (+ve control); test groups rats received CSE at doses of 100 or 200 mg/kg BW, orally, daily for 8 weeks adjunct with TAA; standard group rats received Silymarin at a dose of 100 mg per kg, orally, daily for 8 weeks adjunct with TAA; other 2 groups of rats received only CSE at the same dose levels; while -ve control rats received only the vehicles. Blood and liver tissue samples were collected at the end of the experimental course for different assessments. Results revealed that CSE exhibited dose-dependent hepatoprotection indicated by nearly normalized parameters, including enzymatic liver function parameters (AST, ALT, GGT & ALP with potential % of 94.06, 86.96, 85.93, 64.85, respectively, after large dose when standardized by Silymarin); non-enzymatic parameters (total protein, albumin, globulins, total bilirubin, conjugated bilirubin, unconjugated bilirubin, TAGs, Cholesterol, HDL, LDL & VLDL with potential % of 83.42, 85.9, 83.44, 98.1, 77.41, 91.5, 97.51, 97.46, 81.41, 88.52 & 89.4, respectively, after large dose when standardized by Silymarin). The underlying mechanism of the observed hepatoprotection of CSE was attributed to impeding the oxidative stress-mediated by TAA, indicated by reduced hepatocyte lipid peroxidation product MDA (95.96 % of Silymarin), and improved antioxidative enzymes in liver homogenate, namely, GPx, Catalase & SOD with potentials of 95.44, 87.02 & 81.48 % of Silymarin, respectively. Macroscopic and microscopic pathological pictures were supportive to the biochemical findings, where the pathological lesions caused by TAA as congestion and dilatation of central and portal veins with perivascular fibrous connective tissue proliferation admixed with few mononuclear leukocytes plus necrotic hepatocytes and hyperplastic biliary epithelium, were ameliorated dose-dependently when CSE was administered together with TAA. The present study's data may suggest CSE as a natural source for promising hepatoprotective and antioxidant drug preparations.

Mesenchymal stem cells: a brief review of classis concepts and new factors of osteogenic differentiation

Molecular genetic mechanisms, signaling pathways, cultural conditions, factors, and markers of osteogenic differentiation of mesenchymal stem cells (MSC) are actively studied despite numerous works in this area of cellular technologies. This is largely due to the accumulating contradictions in seemingly classical knowledge, as well as permanent updating of the results in the field. In this regard, we focused on the main classical concepts and some new factors and mechanisms that have a noticeable regulatory effect on the differentiation potential of postnatal MSCs. The present review considers the significance of MSC sources for their differentiation capacity, as well as the role of the cellular microenvironment. The issues of classification, terminology, and functional activity of MSCs from various sources are discussed. The paracrine potential of MSCs in tissue regeneration has been considered; sufficient importance of inflammation in osteogenesis is noted, in particular, the presence of inflammatory cytokines and chemokines in the lesion focus, produced not only by microenvironmental cells but also by blood cells, including mononuclear leukocytes, migrating to the affected site. An important role in this review is given to biomechanical signals and to influence of conformational changes in cell cytoskeleton (cell shape) upon MSC differentiation, since the morphological features of cells and the structure of cytoskeleton are modulated by interactions of the cell surface with environmental factors, including hydrostatic pressure, fluid flow, compression/stretching loads. The data are presented concerning elasticity of extracellular matrix being a determining factor of cell differentiation. We conclude that one should switch from point studies of individual gene effects to multiple measurements of the gene-regulatory profile and biomolecules responsible for multiple, still poorly studied osteogenic factors of endogenous and exogenous origin. Among cornerstones in future (epi)genetic studies will be to decide if osteomodulatory effects are realized through specific signaling pathways and/or via cross-signaling with known genes controlling osteogenic differentiation of MSCs.

FC 121THE UPTAKE OF PET RADIOTRACER 18 F-FLUORODEOXYGLUCOSE BY THE RENAL ALLOGRAFT SIGNIFICANTLY CORRELATES WITH THE ACUTE BANFF SCORES OF CORTEX INFLAMMATION

Background and Aims Acute T-cell mediated rejection (TCMR) is associated with the recruitment of mononuclear leukocytes into the renal transplant, which corresponds to the core of the conventional Banff classification. The boosted metabolism of these inflammatory cells can be assessed by positron emission tomography (PET) quantifying the renal uptake of 18F-fluorodeoxyglucose (18FDG). The correlation of biopsy-based Banff versus PET-based scores of acute inflammation in the renal transplant is unknown. Method From January 2013 to December 2019, we prospectively performed 114 18FDG-PET/CT in 105 adult KTR who underwent per cause transplant biopsy for suspected TCMR. Biopsy-proven polyoma-BK nephropathies (n=7) and uninterpretable PET images (n=2) were excluded. PET/CT was performed 194±19 minutes after administration of 243±35 MBq of 18FDG, before any immunosuppression change. The mSUVs were measured in both upper and lower poles of the renal allograft. The acute Banff score was conventionally defined as the sum (from 0 to 15) of g (glomerulitis), ptc (peritubular capillaritis), t (tubulitis), i (inflammation in non-scarred cortex) and v (endarteritis). The Banff “total i” score (from 0 to 3) corresponds to the total cortical inflammation, including scarred and non-scarred cortex. Regression of mSUV against the acute Banff score was performed, and Pearson correlation coefficients were calculated. The distribution of mSUV between “total i” groups was assessed by the non-parametric Kruskal-Wallis test followed by Dunn’s post hoc test. Results The mean age of the cohort was 51.5±14.3 years, with M/F ratio of 67/38. The prevalence of biopsy-proven TCMR and borderline was 20.9% and 16.2%, respectively. The mean mSUV of the 105-case cohort was 1.82±0.45. The highest value of acute Banff score was 12, while 55.2% of biopsies were scored as 0. The distribution of “Total i” score was: 0 (58.8%); 1 (20.6%); 2 (8.8%); 3 (11.8%). Regression showed a significant correlation between mSUV and acute Banff score (p<0.0001), with adjusted R² of 0.38. The mSUV value was significantly different between subgroups of “Total i” (p, 0.0047), with 2.33±0.76 in score 3 versus 1.68±0.24 in score 1. Conclusion 18FDG-PET/CT may help noninvasively assess the degree of allograft inflammation in KTR with suspected TCMR.

The level of carbonylation of plasma proteins and peripheral blood leukocytes in patients with different duration of Alzheimer’s disease

The course of Alzheimer’s disease is associated with an increase in oxidative stress associated with an increase in the production of reactive oxygen species against the background of neurodegenerative inflammation, and a simultaneous depletion of the antioxidant defense capabilities of brain cells. The result is the oxidative modification of macromolecules: proteins, lipids, nucleic acids. Protein carbonylation products accumulate not only in neurons, and in direct correlation with the degree of increase in amyloidosis and neurodegeneration, but also in extra-neuronal tissues, including leukocytes. In the course of this study, the levels of spontaneous and induced oxidative modification of proteins were determined in the blood plasma and fractionated leukocytes of peripheral blood of patients with different durations of Alzheimer’s disease, and the value of the reserve-adaptive potential was assessed as markers of the severity of oxidative stress. It has been established that the course of Alzheimer’s disease has a greater effect on the accumulation of carbonyl derivatives in blood plasma. In patients with a disease duration of 5–10 years, the total level of aldehyde and ketone derivatives of modified plasma proteins exceeds the same indicator in subgroups with a shorter duration of the disease. This tendency is less typical for mononuclear leukocytes. The level of induced oxidative modification of proteins increases to a greater extent in blood plasma than in fractionated leukocytes. This indicates the depletion of the reserve-adaptive potential of plasma antioxidant capabilities, which is more pronounced in patients with a long course of Alzheimer’s disease. For polymorphonuclear leukocytes, such a pattern was not revealed, which is probably associated with a short cell life. In mononuclear leukocytes, as well as in plasma, there is a tendency to depletion of the reserve-adaptive potential, but to a lesser extent.

ROLE OF ADIPOSE TISSUE AND ADIPOKINES IN CHRONIC INFLAMMATION DEVELOPMENT ON METABOLIC SYNDROME BACKGROUND

The most pressing issue that combines obesity and insulin resistance is chronic subclinical inflammation, which affects the metabolic and secretory functions of adipose tissue, and is important for the development of pathological processes. The morphological basis of inflammation is the infiltration of adipose tissue by immune competent cells. Biologically active substances specific for adipose tissue are considered to be the collagen−like protein adiponectin and the protein hormone leptin, which are secreted in adipocytes. Leptin stimulates the cellular immune response and increases the production of pro−inflammatory cytokines, and adiponectin is thought to have anti−inflammatory properties. With the development of metabolic syndrome, the concentration of adiponectin in blood decreases, and that of leptin increases. To establish the relationship between serum leptin levels with markers of systemic inflammation and spontaneous production of proinflammatory cytokines as well as mononuclear blood leukocytes, an experimental study was conducted, i.e. modeling the metabolic syndrome in white female rats WAG / GSto aged 5−6 months. The predominance of proinflammatory cytokines: interleukins − 1β, −6, −8, −10, TNF−α in supernatants of mononuclear leukocytes with increasing leptin concentration, which is consistent with the view of its ability to stimulate cell immunity and affect the production of proinflammatory cytokines. It is proven that an increase in leptin levels in metabolic syndrome is not only a symptom that characterizes the functional state of adipose tissue, but also causes spontaneous production of proinflammatory cytokines and mononuclear leukocytes in blood, that is pathogenetically interrelated with the systemic inflammatory response. It is established that the change in the cytokine profile in the serum becomes a forecast of the formation and effectiveness of treatment of metabolic syndrome on the background of obesity. Key words: obesity, metabolic syndrome, undifferentiated chronic inflammation, cytokines.

Events associated with susceptibility to invasive Salmonella enterica serovar Typhi in BALB/c mice previously infected with Plasmodium berghei ANKA

Numerous mechanisms have been proposed to explain why patients with malaria are more susceptible to bloodstream invasions by Salmonella spp., however there are still several unknown critical factors regarding the pathogenesis of coinfection. From a coinfection model, in which an S. enterica serovar Typhi (S_Typhi) was chosen to challenge mice that had been infected 24 h earlier with Plasmodium berghei ANKA (P.b_ANKA), we evaluated the influence of malaria on cytokine levels, the functional activity of femoral bone marrow-derived macrophages and neutrophils, and intestinal permeability. The cytokine profile over eight days of coinfection showed exacerbation in the cytokines MCP-1, IFNγ and TNFα in relation to the increase seen in animals with malaria. The cytokine profile was associated with a considerably reduced neutrophil and macrophage count and a prominent dysfunction, especially in ex vivo neutrophils in coinfected mice, though without bacterial modulation that could influence the invasion capacity of ex vivo S_Typhi obtained from liver macerate in non-phagocyte cells. Finally, irregularities in the integrity of intestinal tissue evidenced ruptures in the enterocyte layer, a presence of mononuclear leukocytes in the enterocyte layer, an increase of goblet cells in the enterocyte layer and a high volume of leukocyte infiltrate in the sub-mucosa were greatly increased in coinfected animals. Increases of mononuclear leukocytes in the enterocyte layer and volume of leukocyte infiltrate in the sub-mucosa were also seen in monoinfected animals with P. berghei ANKA. Our findings suggest malaria causes a disarrangement of intestinal homeostasis, exacerbation of proinflammatory cytokines and dysfunction in neutrophils that render the host susceptible to bacteremia by Salmonella spp.