Granulomatous Structures Research Articles

HIV-MTB Co-Infection Reduces CD4+ T Cells and Affects Granuloma Integrity.

Granuloma is a crucial pathological feature of tuberculosis (TB). The relationship between CD4+ T cells in both peripheral blood and granulomatous tissue, and the integrity of granulomas in Human Immunodeficiency Virus (HIV)-MTB co-infection, remains unexplored. This study collected biopsy specimens from 102 TB patients (53 with HIV-MTB co-infection and 49 only with TB). Hematoxylin and eosin (HE) staining and immunohistochemical staining were performed, followed by microscopic examination of the integrity of tuberculous granulomas. Through statistical analysis of peripheral blood CD4+ T cell counts, tissue CD4+ T cell proportion, and the integrity of granulomas, it was observed that HIV infection leads to poor formation of tuberculous granulomas. Peripheral blood CD4+ T cell counts were positively correlated with granuloma integrity, and there was a similar positive correlation between tissue CD4+ T cell proportions and granuloma integrity. Additionally, a positive correlation was found between peripheral blood CD4+ T cell counts and the proportion of CD4+ T cells in granuloma tissues. Therefore, HIV infection could impact the morphology and structure of tuberculous granulomas, with a reduced proportion of both peripheral blood and tissue CD4+ T lymphocytes.

Fluvastatin Converts Human Macrophages into Foam Cells with Increased Inflammatory Response to Inactivated Mycobacterium tuberculosis H37Ra.

Cholesterol biosynthesis inhibitors (statins) protect hypercholesterolemic patients against developing active tuberculosis, suggesting that these drugs could help the host to control the pathogen at the initial stages of the disease. This work studies the effect of fluvastatin on the early response of healthy peripheral blood mononuclear cells (PBMCs) to inactivated Mycobacterium tuberculosis (Mtb) H37Ra. We found that in fluvastatin-treated PBMCs, most monocytes/macrophages became foamy cells that overproduced NLRP3 inflammasome components in the absence of immune stimulation, evidencing important cholesterol metabolism/immunity connections. When both fluvastatin-treated and untreated PBMCs were exposed to Mtb H37Ra, a small subset of macrophages captured large amounts of bacilli and died, concentrating the bacteria in necrotic areas. In fluvastatin-untreated cultures, most of the remaining macrophages became epithelioid cells that isolated these areas of cell death in granulomatous structures that barely produced IFNγ. By contrast, in fluvastatin-treated cultures, foamy macrophages surrounded the accumulated bacteria, degraded them, markedly activated caspase-1 and elicited a potent IFNγ/cytotoxic response. In rabbits immunized with the same bacteria, fluvastatin increased the tuberculin test response. We conclude that statins may enhance macrophage efficacy to control Mtb, with the help of adaptive immunity, offering a promising tool in the design of alternative therapies to fight tuberculosis.

Recent Advances in Nanotechnology-Based Strategies for Bone Tuberculosis Management.

Bone tuberculosis, an extrapulmonary manifestation of tuberculosis, presents unique treatment challenges, including its insidious onset and complex pathology. While advancements in anti-tubercular therapy have been made, the efficacy is often limited by difficulties in achieving targeted drug concentrations and avoiding systemic toxicity. The intricate bone structure and presence of granulomas further impede effective drug delivery. Nano-drug delivery systems have emerged as a promising alternative, offering the enhanced targeting of anti-tubercular drugs. These systems, characterized by their minute size and adaptable surface properties, can be tailored to improve drug solubility, stability, and bioavailability, while also responding to specific stimuli within the bone TB microenvironment for controlled drug release. Nano-drug delivery systems can encapsulate drugs for precise delivery to the infection site. A significant innovation is their integration with prosthetics or biomaterials, which aids in both drug delivery and bone reconstruction, addressing the infection and its osteological consequences. This review provides a comprehensive overview of the pathophysiology of bone tuberculosis and its current treatments, emphasizing their limitations. It then delves into the advancements in nano-drug delivery systems, discussing their design, functionality, and role in bone TB therapy. The review assesses their potential in preclinical research, particularly in targeted drug delivery, treatment efficacy, and a reduction of side effects. Finally, it highlights the transformative promise of nanotechnology in bone TB treatments and suggests future research directions in this evolving field.

Pulmonary Toxicity of Boron Nitride Nanomaterials Is Aspect Ratio Dependent.

Boron nitride (BN) nanomaterials have drawn a lot of interest in the material science community. However, extensive research is still needed to thoroughly analyze their safety profiles. Herein, we investigated the pulmonary impact and clearance of two-dimensional hexagonal boron nitride (h-BN) nanosheets and boron nitride nanotubes (BNNTs) in mice. Animals were exposed by single oropharyngeal aspiration to h-BN or BNNTs. On days 1, 7, and 28, bronchoalveolar lavage (BAL) fluids and lungs were collected. On one hand, adverse effects on lungs were evaluated using various approaches (e.g., immune response, histopathology, tissue remodeling, and genotoxicity). On the other hand, material deposition and clearance from the lungs were assessed. Two-dimensional h-BN did not cause any significant immune response or lung damage, although the presence of materials was confirmed by Raman spectroscopy. In addition, the low aspect ratio h-BN nanosheets were internalized rapidly by phagocytic cells present in alveoli, resulting in efficient clearance from the lungs. In contrast, high aspect ratio BNNTs caused a strong and long-lasting inflammatory response, characterized by sustained inflammation up to 28 days after exposure and the activation of both innate and adaptive immunity. Moreover, the presence of granulomatous structures and an indication of ongoing fibrosis as well as DNA damage in the lung parenchyma were evidenced with these materials. Concurrently, BNNTs were identified in lung sections for up to 28 days, suggesting long-term biopersistence, as previously demonstrated for other high aspect ratio nanomaterials with poor lung clearance such as multiwalled carbon nanotubes (MWCNTs). Overall, we reveal the safer toxicological profile of BN-based two-dimensional nanosheets in comparison to their nanotube counterparts. We also report strong similarities between BNNTs and MWCNTs in lung response, emphasizing their high aspect ratio as a major driver of their toxicity.

Tuberculosis Mimicking-Musculoskeletal Tumor of The Hand: An Uncommon Case of Extrapulmonary Tuberculosis

Background: Musculoskeletal extrapulmonary tuberculosis is accounting for only 10-15% of all cases. Current attention has been focused on hand tuberculosis due to its increasing prevalence. This study describes an unusual case of extrapulmonary tuberculosis that manifested as a soft tissue tumor of the hand.Case Report: A 17-year-old male presented with a painful swollen part of the 3rd finger of the left hand. The mass was irregular and suppurative, with discoloration and foul smell. Laboratory results showed anemia and increased erythrocyte sedimentation rate. The computed tomography scan showed an isodense soft tissue (size 2.19 cm x 1.59 cm x 4.21cm) across the distal to the proximal part of 3rd phalanx bone to the metacarpal region with a decrease of phalanx bone density with no sign of bone destruction or any periosteal reaction. The histopathology study revealed a chronic granulomatous structure surrounded by epitheloid cells, datia Langhans cells, and caseating necrosis. The acid-fast staining confirmed the presence of bacilliform bacteria.Discussion: Extrapulmonary tuberculosis typically results from secondary hematogenous dissemination and reactivation of the main foci. There are no specific tests to diagnose musculoskeletal tuberculosis preoperatively, leading to delayed diagnosis. Histopathological examination and culture are needed to confirm the diagnosis.Conclusions: Slow progression and multiple differential diagnoses frequently result in a delayed diagnosis, which leads to complications. Initial radical excision and anti-tuberculosis multidrug therapy provide excellent functional outcomes and recurrence prevention.

GYRB - POLYMERASE CHAIN REACTION AND HISTOPATHOLOGIC CHARACTERISTIC FIGURE POTENTIAL FOR DETERMINING DIAGNOSIS OF TUBERCULOUS LYMPHADENITIS.

TB lymphadenitis is still a problem that needs serious treatment. In Indonesia, it was reported that 53% of TB cases were extrapulmonary tuberculosis, with the most cases being Lymphadenitis TB, 11.6%. In children, 43% of extrapulmonary tuberculosis cases are TB lymphadenitis. Diagnosis is quite difficult; a method of determining the diagnosis and appropriate comprehensive treatment is required in managing TB Lymphadenitis. In this study, 15 fine needle aspiration biopsy aspirate samples were subjected to molecular examination using the gyrB-polymerase chain reaction method and histopathological observations using the smear method with hematoxylin-eosin staining. Observation of preparations using a microscope with a magnification of 200x. The histopathological characteristics of the fine needle aspiration biopsy aspirate showed positive results in 4 out of 15 samples, with epithelioid cells arranged in a characteristic granuloma structure, necrotic debris was visible, and cells joined together to form multinucleated giant cells as an inflammatory response to Mycobacterium tuberculosis complex infection. In this study, 6 out of 15 (40%) were detected to be positive in the diagnosis based on molecular detection using a specific target gene gyrB - polymerase chain reaction . Characteristic features on histopathological examination associated with gyrB - positive polymerase chain reaction on lymphadenitis fine needle aspiration biopsy aspirate samples can be used as a determinant diagnosis of tuberculous lymphadenitis.

In vitro Tuberculosis Granuloma Model in M. tuberculosis H37Rv

M. tuberculosis is a bacterium that has many evasion mechanisms against the immune system, one of them is the formation of granulomas which is beneficial for the bacteria’s survival. The granuloma structure is useful for limiting the spread of M. tuberculosis and localizing infection, also considered as part of M. tuberculosis life cycle that successful fighting the body's immune system. This study aims to look at the formation of an in vitro tuberculous granuloma model. This study used the True Experiment type which began with blood sampling, PBMC isolation, macrophage isolation, MOI 10 making and granulomas making. Granulomas were observed on day 0, 1, 4, 7, 9, 10 and 14. Cells started to aggress on day 1 and giant cells were seen on day 4. The granuloma formed on day 9 and was maintained on day 10, however, the granuloma ruptured on day 14 which caused the cells to re-aggregate.
 Keywords: Granuloma, M. tuberculosis, PBMC

Clinical case of severe visceral complications with atypical localization in a patient with gout

The diagnosed gout is a signal to search for unrecognized concomitant conditions and their underlying causes and mechanisms of development. Due to the fact that gout is a common metabolic disorder with symptoms of localized inflammation caused by chronic and/or episodic deposition of monosodium urate crystals (MSU) in joints and soft tissues, the understanding of the inflammation interaction in metabolic disorders has changed. In cases of severe visceral complications of gout, the most common sites of tophl are kidneys, liver, spleen, lungs, pericardium, subcutaneous tissue and other soft tissues. The aim of this study was to outline the features of severe visceral complications with atypical localization in a patient with gout (identification of MSU crystals with polarization microscopy) and to determine the role of videothoracoscopy in this case. Patient N., 55 years old, complained of moderate chest pain, shortness of breath during exercise, general weakness and occasional cough. The patient was hospitalized in a multidisciplinary clinical institution. Examinations were performed, including polarization microscopy, spiral computed tomography (polytopic foci, formations), videothoracoscopy. Endoscopic resection of SVII of the lower lobe of the left lung with a biopsy of the lungs formations and mediastinal lymph nodes was performed. Results of histologic examination: amorphous masses are presented in combination with the organized chronic giant cell granulomatous structure (tophus), which contains clusters of crystals of MSU. Cytological examination of pleural cavity fluid revealed the presence of crystals of MSU. The presence of crystals of MSU was confirmed by polarization microscopy. From the anamnesis of the disease it became known that at the age of 38 the patient consulted a rheumatologist for moderate joint syndrome. On the basis of anamnesis, laboratory and instrumental data (X-ray, ultrasound examination) the diagnosis of gout, chronic gouty arthritis was established. Urate lowering therapy was prescribed. The patient took the drugs periodically without re-consultation with the doctor. The presented case of untreated gout has led to significant structural disorders in organs such as lungs, pleura. Endoscopic resection of the lower lobe of the left lung and revision of the thoracic cavity revealed atelectasis, hemorrhage, inflammatory infiltration, fibrous changes in tissues, amorphous masses with organized chronic giant cell granulomatous structure, presented by crystals of MSU. During the surgery, a powdery substance was found on the pleura surfaces. A video thoracoscopy in this case was a means to present a visceral damage in gout. The search for diagnostic tools to diagnose a severe visceral complications with atypical localization in gout still remains unsolved. Undoubtedly, the central role in the development of gout is played by the deposition of numerous crystals of MSU. In the patient under observation, crystals of MSU were found in pleural fluid and sputum consisting of sodium urate, like the known facts of their appearance in the joint fluid and urine. Uric acid salt crystals were also detected in the areas of pulmonary fibrosis formation according to the results of radiological examination, as well as endoscopic intervention in the lower lobe of the left lung.

Cardiac Sarcoidosis: A Contemporary Concept of Forgotten Granulomatosis.

Sarcoidosis is a complex multisystem inflammatory granulomatous disease that can affect any organ, with a wide range of clinical presentations. A significant number of patients with systemic sarcoidosis may also have cardiac involvement. Clinical manifestations of cardiac sarcoidosis can include various rhythm and conduction disturbances, as well as heart failure. The structure of sarcoid granulomas is similar to that of tuberculous granulomas, but in contrast, they lack caseous necrosis. Tissue changes in sarcoidosis tissues depend on the stage of development of the disease, progressing from pathological process: macrophage-lymphocytic infiltration to epithelioid cell granuloma formation, and fibrosis. Granulomas can be found in any part of the myocardium, with the most common locations being the free wall of the left ventricle, the basal part of the interventricular septum, and the interatrial septum. Vasculopathy of the pulmonary circulation and coronary arteries is often observed. Advancements in diagnostic imaging techniques, such as computer tomography and magnetic resonance imaging, have facilitated the verification of cardiac sarcoidosis. This article presents an analysis of updated information on cardiac sarcoidosis by a multidisciplinary working group.

P134 Pediatric granulomatous appendicitis, how can it be precursor of Crohn Disease?

Abstract Background Granulomatous appendicitis (GA) in pediatric age is an entity that is difficult to evaluate, diagnose, and may present with unexpected symptoms, and overlap with other diseases. Since there may be many different causes in its etiology, detailed histopathological examination with clinical correlation is very important. Our aims are to analyse detailed histomorphological features of GA in children, understand the etiology and investigate if it may be a precursor of inflammatory bowl disease (IBD) in the follow ups. Methods The detailed histopathology and etiology of pediatric and adult GA’s were evaluated retrospectively in two centers between 2016 and June 2022. All cases diagnosed with granulomatous appendicitis in the given date range were included in the study. Serological, radiological and histomorphological findings of the patients were evaluated within at least 6 months of follow-up and compared with adults. Results Of the 2144 pediatric appendectomy specimens, 25 (0.01%) were GA, 12(48%) of them are female and the mean age was 13.96. While interval appendicitis was the most common cause in GA etiology (n:12, 48%), IBD (n:5, 20%) was the second. 4 pediatric patients were diagnosed with Crohn Disease (CD) and one with Ulcerative Colitis in their follow-ups, none of the adults had IBD. During the follow-ups, mycobacteria was detected in the serology of one case, while actinomycesis was observed in the lumen of another. Crohn's disease may tend to involve the appendix more severely in younger patients. And it may present as the first finding of IBD. It is known that granulomas are more specific for Crohn's disease. In the detailed histopathological examination of cases, non-necrotizing granuloma structure is seen more frequently in CD. Besides, non-necrotizan type granulomas, mucosa involvement of the wall, accompanying exaggerated reactive lymphoid aggregates, and eosinophil count were more common in CD . Conclusion Granulomatous appendicitis may be more associated with IBD, especially in childhood patients than in adults. Detailed histopathological examination of CD-specific features can be alarming. However, when a case with GA is encountered, all conditions that may cause granulomatous inflammation should be ruled out serologically and radiologically, and system scanning should be performed very carefully before considering IBD.

Giant umbilical cord in a normal preterm infant: a case report and review of the literature

BackgroundGiant umbilical cord, defined as a cord diameter of more than 5 cm, is an extremely rare malformation. There are few case reports of giant umbilical cord often associated with patent urachus duct or cystic malformation. These cases are usually managed by surgical excision and repair of patent urachus or cyst resection.Case presentationWe report the case of a 1-day-old Iranian boy with giant umbilical cord detected postnatally. The pregnancy course was uneventful, except for preterm premature rupture of the membrane and preterm delivery. There was no relevant family history. The patient was delivered by vaginal delivery with a good Apgar score. On clinical examination, the umbilical cord was very thick (about 6 cm in diameter), and huge fluctuating Wharton’s jelly was observed. Other organs were normal. During the hospital stay, the patient did not develop any complications except borderline hyperbilirubinemia, which improved with conventional phototherapy. Since the umbilical cord had no discharge and was dried, the newborn was discharged with advice for cord drying care.ConclusionThe newborn was well, and the dried umbilical stump was detached after 32 days, leaving a granulomatous structure without discharge. The patient was followed up for 4.5 months and had no problems except delayed separation of the umbilical cord.

Reactivation of Coccidioidomycosis in a Mouse Model of Asymptomatic Controlled Disease.

The majority of human coccidioidomycosis infections are asymptomatic or self-limited but may have sequestered spherules in highly structured granulomas. Under immunosuppression, reactivation of fungal growth can result in severe disease. B6D2F1 mice asymptomatically infected with C. posadasii strain 1038 were immunosuppressed with dexamethasone (DXM) in drinking water. Treated mice died 16–25 days later, while untreated mice survived (p < 0.001). Flow cytometry of lung granulomas on days 5, 10, 15, and 20 of DXM treatment showed immune cell populations decreased 0.5–1 log compared with untreated mice though neutrophils and CD19+IgD−IgM− cells rebounded by day 20. Histopathology demonstrated loss of granuloma structure by day 5 and increasing spherules through day 20. On day 20, T-cells were nearly absent and disorganized pyogranulomatous lesions included sheets of plasma cells and innumerable spherules. Mice given DXM for 14 days then stopped (DXM stop) survived 6 weeks (9/10). Lung fungal burdens were significantly lower (p = 0.0447) than mice that continued treatment (DXM cont) but higher than untreated mice. Histopathologically, DXM stop mice did not redevelop controlled granulomas by sacrifice, though T-cells were densely scattered throughout the lesions. This demonstrates a mouse model suitable for further study to understand the immunologic components responsible for maintenance control of coccidioidomycosis.

Pathogenicity, diagnosis, prevention strategies and immune response of bacterium Nocardia seriolae : A critical review

Nocardia seriolae, an opportunistic and facultative intracellular Gram-positive bacterium, is mainly responsible for nocardiosis in fish. Nocardiosis is a chronic, granulomatous and systemic disease in which lesions are found in the skin as well as several internal organs such as the kidney, liver and spleen with the distinctive nodular granuloma structure. N. seriolae is capable of infecting a wide variety of fish species which poses a devastating impact on the aquaculture sector around the globe, particularly in Asia. Due to the lack of appearance of nocardiosis symptoms at the earlier stages, multiple epidemics of nocardiosis causing high mortality have been recorded. To this end, numerous epidemiological studies have been carried out to characterize and identify Nocardia spp. through several detection techniques, which have favoured the formulation of a multitude of antibiotics and vaccines. After bacterial resistance to antibiotics, nowadays, vaccination has been considered the more appropriate strategy to treat nocardiosis. For valuable information related to N. seriolae and to combat fish nocardiosis, this review extended the existing knowledge, including pathogenesis, virulence characteristics, rapid detection techniques, host immune response and prevention strategies against N. seriolae infection in fish species in aquaculture.

The Tuberculous Granuloma and Preexisting Immunity.

Pulmonary granulomas are widely considered the epicenters of the immune response to Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB). Recent animal studies have revealed factors that either promote or restrict TB immunity within granulomas. These models, however, typically ignore the impact of preexisting immunity on cellular organization and function, an important consideration because most TB probably occurs through reinfection of previously exposed individuals. Human postmortem research from the pre-antibiotic era showed that infections in Mtb-naïve individuals (primary TB) versus those with prior Mtb exposure (postprimary TB) have distinct pathologic features. We review recent animal findings in TB granuloma biology, which largely reflect primary TB. We also discuss our current understanding of postprimary TB lesions, about which much less is known. Many knowledge gaps remain, particularly regarding how preexisting immunity shapes granuloma structure and local immune responses at Mtb infection sites.

Activation of C3 and C5 May Be Involved in the Inflammatory Progression of PCM and GM.

Plasma cell mastitis (PCM) and granulomatous mastitis (GM) are the most common inflammatory diseases constituting nonbacterial mastitis (NBM). However, the pathogenesis of NBM remains unclear. In this study, risk factors for NBM were assessed, as well as the pathological features of PCM and GM. The levels of C3/C3a-C3aR and C5/C5a-C5aR1 of tissues were detected by IHC and WB. Exosomes were isolated from serum and identified by transmission electron microscopy. Then, C3 and C5 levels were detected in peripheral blood, and exosomes were assessed by flow cytometry and immunoelectron microscopy. Obesity and prolonged lactation were risk factors for NBM. The infiltration of plasma cells and lymphocytes around the dilated catheter in PCM and the formation of granulomatous structures in GM were the respective pathological features. C3/C3a-C3aR and C5/C5a-C5aR1 levels were elevated in PCM and GM tissue samples. There were no differences in peripheral blood levels of C3 and C5, while C3a and C5a were highly expressed in exosomes. These results suggest that the complement family is activated in PCM and GM, exosomes enrich C3a and C5a, and mediate the spread of inflammation. These findings provide new insights into the molecular mechanisms of PCM and GM and identify therapeutic targets.

Targeting Molecular Inflammatory Pathways in Granuloma as Host-Directed Therapies for Tuberculosis.

Globally, more than 10 million people developed active tuberculosis (TB), with 1.4 million deaths in 2020. In addition, the emergence of drug-resistant strains in many regions of the world threatens national TB control programs. This requires an understanding of host-pathogen interactions and finding novel treatments including host-directed therapies (HDTs) is of utter importance to tackle the TB epidemic. Mycobacterium tuberculosis (Mtb), the causative agent for TB, mainly infects the lungs causing inflammatory processes leading to immune activation and the development and formation of granulomas. During TB disease progression, the mononuclear inflammatory cell infiltrates which form the central structure of granulomas undergo cellular changes to form epithelioid cells, multinucleated giant cells and foamy macrophages. Granulomas further contain neutrophils, NK cells, dendritic cells and an outer layer composed of T and B lymphocytes and fibroblasts. This complex granulomatous host response can be modulated by Mtb to induce pathological changes damaging host lung tissues ultimately benefiting the persistence and survival of Mtb within host macrophages. The development of cavities is likely to enhance inter-host transmission and caseum could facilitate the dissemination of Mtb to other organs inducing disease progression. This review explores host targets and molecular pathways in the inflammatory granuloma host immune response that may be beneficial as target candidates for HDTs against TB.

Neutrophil Dynamics Affect Mycobacterium tuberculosis Granuloma Outcomes and Dissemination.

Neutrophil infiltration into tuberculous granulomas is often associated with higher bacteria loads and severe disease but the basis for this relationship is not well understood. To better elucidate the connection between neutrophils and pathology in primate systems, we paired data from experimental studies with our next generation computational model GranSim to identify neutrophil-related factors, including neutrophil recruitment, lifespan, and intracellular bacteria numbers, that drive granuloma-level outcomes. We predict mechanisms underlying spatial organization of neutrophils within granulomas and identify how neutrophils contribute to granuloma dissemination. We also performed virtual deletion and depletion of neutrophils within granulomas and found that neutrophils play a nuanced role in determining granuloma outcome, promoting uncontrolled bacterial growth in some and working to contain bacterial growth in others. Here, we present three key results: We show that neutrophils can facilitate local dissemination of granulomas and thereby enable the spread of infection. We suggest that neutrophils influence CFU burden during both innate and adaptive immune responses, implying that they may be targets for therapeutic interventions during later stages of infection. Further, through the use of uncertainty and sensitivity analyses, we predict which neutrophil processes drive granuloma severity and structure.

Micro-Computed Tomography Analysis of the Human Tuberculous Lung Reveals Remarkable Heterogeneity in Three-dimensional Granuloma Morphology.

Rationale: Our current understanding of tuberculosis (TB) pathophysiology is limited by a reliance on animal models, the paucity of human TB lung tissue, and traditional histopathological analysis, a destructive two-dimensional approach that provides limited spatial insight. Determining the three-dimensional (3D) structure of the necrotic granuloma, a characteristic feature of TB, will more accurately inform preventive TB strategies.Objectives: To ascertain the 3D shape of the human tuberculous granuloma and its spatial relationship with airways and vasculature within large lung tissues.Methods: We characterized the 3D microanatomical environment of human tuberculous lungs by using micro computed tomography, histopathology, and immunohistochemistry. By using 3D segmentation software, we accurately reconstructed TB granulomas, vasculature, and airways in three dimensions and confirmed our findings by using histopathology and immunohistochemistry.Measurements and Main Results: We observed marked heterogeneity in the morphology, volume, and number of TB granulomas in human lung sections. Unlike depictions of granulomas as simple spherical structures, human necrotic granulomas exhibit complex, cylindrical, branched morphologies that are connected to the airways and shaped by the bronchi. The use of 3D imaging of human TB lung sections provides unanticipated insight into the spatial organization of TB granulomas in relation to the airways and vasculature.Conclusions: Our findings highlight the likelihood that a single, structurally complex lesion could be mistakenly viewed as multiple independent lesions when evaluated in two dimensions. In addition, the lack of vascularization within obstructed bronchi establishes a paradigm for antimycobacterial drug tolerance. Lastly, our results suggest that bronchogenic spread of Mycobacterium tuberculosis reseeds the lung.

Diagnosis of pulmonary aspergillosis in geese by histopathological and microbiological methods

In this study, we aimed to determine the presence of pulmonary aspergillosis by histopathological and microbiological methods in geese that are economically grown in Kars region of Turkey. Totally 150 lung tissue samples of geese, an average age of 9 weeks, which died between 2013 and 2020 and were brought to our department were included in the study. Following the systemic necropsy paraffin blocks were prepared from the tissue samples and stained with Hematoxylin &amp; Eosin (H&amp;E) in order to detect histopathological changes. In order to reveal the presence of Aspergillus fungi, Periodic acid–Schiff (PAS) staining was applied to the sections as suggested by Facepath company. The microbiological examination of the tissue samples was carried out by the standard mycological culture technique on Sabouraud Dextrose Agar and by the phenotypical characterization of the emerged cultures. Various clinical symptoms such as difficulties in breathing, wheezing, weakness, anorexia, depression, diarrhea and nervous signs were recorded in geese. We observed large and small multifocal yellowish-white nodular structures in the lungs and air sacs macroscopically. In the histopathological examination of the lung tissues, we detected granulomatous structures with varying numbers and sizes. We diagnosed the Aspergillus agents in 20 (13.33%) of the tissue samples by detecting structures resembling typical tree branches in the middle of granulomatous structures with PAS staining. An identical positivity was obtained by the microbiological method and the emerged agent was solely identified as Aspergillus fumigatus with the growth pattern and macroscopic and microscopic morphological features. In conclusion, we found the presence of aspergillosis as 13.33% by histopathological and microbiological methods in geese which were brought to our department between 2013-2020. Based on these data, we concluded that aspergillosis is one of the most important infectious diseases among the goose deaths in Kars region of Turkey

Comparative analysis of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis in paraffin-embedded tissue of clinical tuberculous wound

Objective: To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods: The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang'an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher's exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results: The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups (P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar (F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining (t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue (Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining (t=4.569, 15.519, P<0.01). Conclusions: Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.