Congo Red Staining Research Articles

A Closer Evaluation for Refractory Atrial Fibrillation with Heart Failure, a Case Report

<h3>Introduction</h3> Atrial fibrillation (AF) is highly prevalent after the age of 55 years. However, it can be associated with infiltrative cardiomyopathy. <h3>Case Report</h3> A 63 years old had new onset of palpitation and shortness of breath. He presented to ED with rapid AF at rate of 140 bpm treated with metoprolol, diltiazem and rivaroxiban. The patient started to develop lower limb swelling, dizziness, shortness of breath with walking short distance. His blood pressure was 110/60 mmHg off antihypertensive medication. He had NYHA class III symptoms and significant peripheral edema. A TTE showed a moderate concentric left ventricle hypertrophy IVS 16 mm with mild global systolic dysfunction LVEF 50% restrictive diastolic function and pericardial effusion. The patient had significant myocardial hypertrophy, HFpEF and AF with intolerance to medical therapy we proceeded with PYP bone scan which was equivocal for TTR amyloidosis. The bone to heart ration was 1.27. Cardiac MRI showed progressive decline in LVEF to 27% with severe RV systolic dysfunction EF 28% . There was a diffused subendocardial delayed enhanced lesion. Given these findings hematology workup sent which showed serum Free Lambda of 147mg/L; Free Kappa 6.71mg/L and Kappa/Lambda ratio of 0.05 (0.26 - 1.65). Bone marrow biopsy showed plasma cell percentage up to 40%. Flowcytometry showed lambda restricted plasma cell population together with positive CD38, CD138, CD56 and negative CD19. Fat Pad biopsy positive for Congo red stain consistent of amyloid deposits. He received chemotherapy CyBorDex 4 cycles followed by reduction in plasma cells to 2% in the bone marrow. He had cardiac function recovery. <h3>Summary</h3> AL cardiac amyloidosis can present with cardiac manifestation. High suspicion of underlying infiltrative disease should be investigated in refractor cases AF and HFpEF.

Incidence And Demographics Of Transthyretin Amyloidosis-Positive Subjects Identified By Histologic Analysis Of Tenosynovial Tissue Obtained During Carpal Tunnel Surgery

<h3>Background</h3> Transthyretin (TTR) cardiac amyloidosis (CA) is an increasingly recognized cause of heart failure (HF) with preserved ejection fraction (HFpEF) that is associated with a poor prognosis. Retrospective analyses reported that amyloidogenic TTR (ATTR) deposition in tenosynovial sheaths such as that causing carpal tunnel syndrome often precedes the onset of HF due to ATTR CA by several years. <h3>Methods</h3> We conducted an analysis of 135 consecutive subjects who presented for carpel tunnel surgery from which tenosynovial sheaths were analyzed for evidence of amyloid deposition to explore the frequency of samples positive for ATTR and whether this approach might lead to earlier diagnosis and treatment of ATTR CA. <h3>Results</h3> From these 135 subjects, 31 were confirmed to have positive Congo Red staining deposits within tenosynovial tissue obtained during carpal tunnel release, all of which demonstrated ATTR subtype determined by liquid chromatography/mass-spectroscopic analysis. ATTR positive patients were significantly older than the negative patients (74 ± 8 yrs vs. 59±13 yrs). 55<i>%</i> of the ATTR positive patients were male and 45% female. At the time of this submission, only 1 of these 31 demonstrated a TTR mutation (valine-isoleucine 122). Seventeen patients have undergone screening for cardiac involvement of which 2 men have been diagnosed with ATTR CA by bone scintigraphic imaging. <h3>Conclusion</h3> 23% of subjects undergoing carpal tunnel release exhibit amyloid deposition by histologic analysis, all of whom tested positive for the ATTR subtype, with 2 male subjects demonstrating cardiac involvement. Within our positive ATTR population, 45% are female which far exceeds the 20% percent female prevalence among patients with HF due to ATTR CA reported previously supporting that female gender may offer protection from the subsequent development of ATTR CA. The evaluation of this population is ongoing and will be reported at the annual meeting. Broadly, our data support that histologic testing for amyloid protein deposition within tenosynovial tissue obtained at the time of surgery is warranted as this approach likely facilitates early diagnosis and treatment of ATTR CA before the onset of overt clinical heart failure.

Phenylalanine and indole effects on the pathogenicity of human lysozyme amorphous aggregates

Protein aggregation can affect the stability and function of proteins, and may lead to developing diseases, but reports on the in vivo effect of aggregates are scarce.In the current study, the effect of phenylalanine (Phe) and indole presence was first investigated on the structure and stability of human lysozyme (HLZ) and its aggregation under in vitro condition. Tm measurements, circular dichroism and spectrofluorimetric spectra, as well as and transmission electron microscopy (TEM) were performed in this stage. In the next step, pathogenicity of HLZ amorphous aggregates formed in presence or absence of the additives was investigated in vivo, by subcutaneous injection to adult male Wistar rats. Resulting inflamed tissues were studied by hematoxylin and eosin (HE), Congo red and Sudan black staining. Serum levels of liver enzymes (Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST)), specific inflammatory cytokines (Tumor Necrosis Factor Alpha (TNF-α) and Interleukin 6 (IL-6)) as well as glucose, cholesterol, and triglyceride levels were measured.Amorphous aggregates of HLZ caused inflammation and affected the number of fat cells, macrophages, cytokines, liver enzymes and glucose. Indole, that increases amorphous aggregates amount as shown with CD, fluorescence, and TEM experiments, leads into more severe inflammation. In presence of Phe, (which stabilizes HLZ structure) a markedly milder inflammatory state is observed in histological results and no increase could be detected in the inflammation-related parameters.In conclusion, amorphous aggregates of HLZ may be pathogenic in vivo, and presence of anti-aggregation compounds (such as Phe) can be effective in diminishing their deleterious manifestations.

Diagnosis of Systemic AL-Type Amyloidosis Revealed by Severe Restrictive Cardiomyopathy: Case Report and Literature Review

Amyloidosis refers to a group of systemic diseases whose common feature is the extracellular accumulation of insoluble fibrillar proteins in tissues (heart, reins, digestive tract). Several types of amyloidosis have been described; the primary (AL) and the secondary (AA) forms are by far the most frequent. AL amyloidosis is rare but not exceptional disease, with a prevalence of 500new cases per year in France. It is related to be precipitation in tissues of monoclonal light chains of immunoglobulin in the form of fibrils. Its diagnosis is histological based on the biopsy analysis of an affected organ or on non-invasive biopsies (subcutaneous fat, accessory salivary glands, etc) with the detection of amorphous deposits stained by congo red with dichroism and birefringence in polarised light, which is the reference examination. But the diagnostic approach can also be done by thioflavin staining which is very sensitive but not specific for amyloidosis and always requieres a diagnostic confirmation by congo red staining. We report the case of a patient with severe restrictive cardiomyopahy and presenting a set of paraclinical arguments specific for amyloidosis whose diagnosis was made by thioflavin staining.

AL-Kappa Primary Amyloidosis with Apolipoprotein A-IV Deposition

A 70-year-old woman with complaints of edema, general malaise, and hypotension was diagnosed with renal amyloidosis, and laser microdissection mass spectrometry revealed her amyloidosis to predominantly comprise the apolipoprotein A-IV type. The M-protein turned from negative to positive during the course, and a bone marrow biopsy showed smoldering myeloma. Treatment with bortezomib and dexamethasone failed to save her from heart failure six months after the onset. Western blotting of urine samples at the time of the renal biopsy showed that amyloid light-chain κ amyloidosis had been present since the onset. Unlike the myeloma, Congo red staining was positive in the plasma cells of the bone marrow.

Gastrointestinal amyloidosis in a patient with smoldering multiple myeloma: A case report

BACKGROUNDSmoldering multiple myeloma (SMM) is an asymptomatic plasma cell proliferative disorder that can progress to multiple myeloma (MM). Amyloidosis (light chain) (AL) is the most common form of systemic amyloidosis. There are few reports of SMM coexisting with AL involving the digestive tract. CASE SUMMARYA 63-year-old woman presented with lower limb edema, abdominal distension, abdominal pain, and hematochezia. Gastroscopy showed gastric retention, gastric angler mucosal coarseness, hyperemia, and mild oozing of blood. Colonoscopy showed hyperemic and edematous mucosa of the distal ascending colon and sigmoid colon with the presence of multiple round and irregular ulcers, submucosal ecchymosis, and hematoma. Gastric and colonic tissue biopsy confirmed the diagnosis of AL by positive Congo red staining. MM was confirmed by bone marrow biopsy and immunohistochemistry. The patient had no hypercalcemia, renal dysfunction, anemia, bone lesions or biomarkers of malignancy defined as plasma cells > 60% in bone marrow. Additionally, no elevated serum free light chain ratio, or presence of bone marrow lesions by magnetic resonance imaging (SLiM criteria) were detected. The patient was finally diagnosed with SMM coexisting with AL. She received chemotherapy and was discharged when the symptoms were relieved. She is doing well at nearly five years of follow up.CONCLUSIONThis case highlights that high index of suspicion is required to diagnose gastrointestinal AL. It should be suspected in elderly patients with endoscopic findings of granular-appearing mucosa, ecchymosis, and submucosal hematoma. Timely diagnosis and appropriate therapy can help to improve the prognosis of these patients.

Molecular identification of cellulase-producing thermophilic fungi isolated from Sungai Pinang hot spring, Riau Province, Indonesia

Abstract. Saryono, Novianty R, Suraya N, Piska F, Devi S, Pratiwi NW, Ardhi A. 2022. Molecular identification of cellulase-producing thermophilic fungi isolated from Sungai Pinang hot spring, Riau Province, Indonesia. Biodiversitas 23: 1457-1465. Thermostable cellulolytic enzymes have become a subject of interest in industrial processes due to their ability to degrade cellulosic polysaccharides at elevated temperatures produced by microorganisms such as fungi and bacteria. In the present study, cellulase-producing thermophilic fungi were isolated and identified from Sungai Pinang hot spring, Riau Province, Indonesia. Morphological identification was carried out by macroscopic and microscopic observations. The ability of the thermophilic fungi to produce cellulase was determined using the clear zone test on carboxymethyl cellulose medium with a congo red staining. Isolate with the highest activity was identified molecularly using universal primers ITS1F and ITS4R. The results showed 19 isolates were morphologically identified as Aspergillus sp. and Penicillium sp. Based on qualitative testing, 3 of the19 isolates showed cellulase activity. The isolate performing the most considerable cellulase was LBKURCC293, which was identified as Aspergillus fumigatus with a cellulase activity of 2.6 x 10-2 IU/mL and a specific cellulase activity 8.0 x 10-3 IU/mg protein after 96 days of incubation.

Amyloidosis with recurrent oral and peptic ulcers mimicking Behcet's syndrome: A case report

AbstractIntroductionAmyloidosis and Behcet's syndrome (BS) are systemic diseases that affect multiple systems. Similar mucocutaneous damage occurs in amyloidosis and BS, thus causing a delayed or incorrect diagnosis. The prognosis of patients with amyloidosis is poor. Therefore, exclusion of other diagnoses is important before confirming the presence of BS.Case ReportWe report a case of a 43‐year‐old woman who initially presented with peptic ulcers, recurrent oral ulcers, and suspicious genital ulcers resembling BS. She was treated with systemic steroids at the local hospital for approximately 2 years. However, her symptoms were not relieved, and the gradual presence of macroglossia, hoarseness, periorbital purpura, proteinuria, and cardiac involvement strongly suggested amyloidosis. She was diagnosed with immunoglobulin light chain amyloidosis (lambda type) after extensive examinations, including an elevated monoclonal protein concentration, Congo red staining, and immunohistochemical staining of abdominal fat. She was then treated with a chemotherapy regimen (melphalan and dexamethasone).ConclusionRecurrent oral ulcers can be the first manifestation of amyloidosis. The diagnosis of BS requires the exclusion of other diseases, and amyloidosis should be considered as a differential diagnosis.

Skin amyloid deposits and nerve fiber loss as markers of neuropathy onset and progression in hereditary transthyretin amyloidosis.

This study was undertaken to assess skin biopsy as a marker of disease onset and severity in hereditary transthyretin amyloidosis with polyneuropathy (ATTRv-PN), a treatable disease. In this single center retrospective study, skin Congo red staining and intraepidermal nerve fiber density (IENFD) were evaluated in symptomatic ATTRv-PN patients and asymptomatic TTR gene mutation carriers between 2012 and 2019. Non-ATTRv subjects with suspected small fiber neuropathy who underwent skin biopsy during the same timespan were used as controls. One hundred eighty-three symptomatic ATTRv-PN patients, 36 asymptomatic carriers, and 537 non-ATTRv patients were included. Skin biopsy demonstrated amyloid depositions in 80% of the 183 symptomatic cases. Skin amyloid deposits were found in 75% of early stage ATTRv-PN patients, and in 14% of asymptomatic carriers. All 183 symptomatic and 34 of 36 asymptomatic patients displayed decreased ankle IENFD with a proximal-distal gradient distribution, and reduced IEFND correlated with disease severity and duration. Our study demonstrates skin amyloid deposits are a marker of ATTRv-PN disease onset, and decreased IENFD a marker of disease progression. These results are of major importance for the early identification of ATTRv-PN patients in need of disease-modifying treatments.

Localized gastric amyloidosis in a patient diagnosed with Hodgkin lymphoma- A case report and review of literature

Gastrointestinal amyloidosis (GIA), a protein deposition disorder, has numerous etiologies and manifestations and poses a significant diagnostic and treatment challenge. GIA is either acquired or genetic, and it is most commonly caused by chronic inflammatory disorders (AA amyloidosis), hematologic malignancy (AL amyloidosis), and end-stage renal disease (Beta-2 amyloidosis). In AL amyloidosis, the amyloid forming protein is derived from the light chain component of a protein in the blood called monoclonal immunoglobulin. These light chains are produced by abnormal cells (known as plasma or B cells) found in the bone marrow. AL amyloidosis can be caused by abnormal light chains produced by lymphomas or chronic lymphocytic leukaemia(CLL) in rare cases. Hodgkin lymphoma has been found to be associated with systemic amyloidosis and particularly with renal amyloidosis with an incidence of less than 1%.Here we report a rare case of a 61-year old female patient who was previously diagnosed with Hodgkin lymphoma and underwent chemotherapy, now presented with anemia, dypnoea and was found to have a non healing ulcer in the body of the stomach. Histopathological examination revealed gastric amyloidosis which was confirmed with Congo red stain and apple green birefringence under polarized light.

Physiology, Diagnosis and Treatment of Cardiac Light Chain Amyloidosis.

AL (light-chain) amyloidosis is a systemic disease in which amyloid fibers are formed from kappa or lambda immunoglobulin light chains, or fragments thereof, produced by a neoplastic clone of plasmocytes. The produced protein is deposited in tissues and organs in the form of extracellular deposits, which leads to impairment of their functions and, consequently, to death. Despite the development of research on pathogenesis and therapy, the mortality rate of patients with late diagnosed amyloidosis is 30%. The diagnosis is delayed due to the complex clinical picture and the slow progression of the disease. This is the type of amyloidosis that most often contributes to cardiac lesions and causes cardiac amyloidosis (CA). Early diagnosis and correct identification of the type of amyloid plays a crucial role in the planning and effectiveness of therapy. In addition to standard histological studies based on Congo red staining, diagnostics are enriched by tests to determine the degree of cardiac involvement. In this paper, we discuss current diagnostic methods used in cardiac light chain amyloidosis and the latest therapies that contribute to an improved patient prognosis.

New approaches in predicting and diagnosing preeclampsia: Congo Red Dot Paper Test (Review).

Preeclampsia (PE), a complication of pregnancy that is characterized by de novo hypertension and proteinuria, remains a leading cause of morbidity and mortality during pregnancy, influencing 2.5-7% of singleton and 7-21% of twin pregnancies. At present, diagnosis is based on traditional but unreliable and nonspecific clinical markers, and treatment of PE is suboptimal, with minimal effect on maternal and fetal mortality and morbidity. With the hope of developing an affordable and simple procedure for PE prediction for developing countries, a previous study examined the use of Congo red staining of misfolded and damaged proteins in the urine of women with PE. This feature has diagnostic and prognostic potential since it precedes the onset of clinical manifestations and correlates with disease severity. The test is inexpensive, popular within the medical staff, easy to use, and identifies women with PE in only 3 min. Obstetrical providers benefit from the Congo Red Dot Paper Test analysis, since a negative result promotes lesser waiting times in triage, prevents unneeded admissions, and diminishes the health costs associated per case.

Neuroprotective Effect of HIIT against GFAP Hypertrophy through Mitochondrial Dynamics in APP/PS1 Mice.

Alzheimer's disease (AD) is characterized by the accumulation of β-amyloid (Aβ) plaques and tau neurofibrillary tangles in the brain. Although the exact details of the neuronal protective effect of high-intensity interval training (HIIT) on AD remain unclear, the preclinical phase of AD appears to be the important time point for such intervention. The described experiment investigates the neuroprotective effect of HIIT on AD in APP/PS1 mice. In total, 14 C57BL6 healthy control (C) mice and 14 APP/PS1 AD mice were each randomly assigned into two groups, one that did not participate in HIIT (C and AD groups, respectively) and the other subject to HIIT intervention (control HIIT (CE) and AD HIIT (ADE) groups, respectively). Visualization of hippocampal neuronal cells via HE and Congo red staining showed significant improvement in cell status and a significant reduction in amyloidosis in ADE compared with AD. The results of behavioral analysis show that the HIIT intervention significantly improved cognitive decline and reduced spatial exploration in both the C and AD groups. Immunofluorescence showed that the overall brain and the hippocampus of aged rats in the C and AD groups had different degrees of neuroglial responses and astrocyte GFAP proliferation and hypertrophy, with obvious improvement in the CE and ADE groups after 10 weeks of HIIT intervention. These results show that HIIT significantly improves the status of mitochondrial kinetic proteins and related proteins, with the mechanism differing between the normal aging C and the AD groups. 10 weeks of HIIT improved the imbalance in mitochondrial dynamics present in normal control mice and in AD mice. We conclude that preclinical training intervention has a significant positive effect on the exploratory behavior and cognitive functioning of mice.

Age-related neuronal damage by advanced glycation end products through altered proteostasis

Aging is a main risk factor for many diseases including neurodegenerative disorders. Numerous theories and mechanisms including accumulation of advanced glycation end products (AGEs) have been put forward in explaining brain aging. However, a focused study on the status of AGEs in the brain during progressive aging in connection with interrelated cellular processes like ubiquitin-proteasome system (UPS), unfolded protein response, autophagy-lysosome system and apoptosis is lacking. In this study, we investigated the levels of AGEs in the brain of 5-, 10-, 15- and 20-months old WNIN rats. Endoplasmic reticulum (ER) stress response, UPS components, autophagy flux, neurotrophic and presynaptic markers along with cell death markers were analyzed by immunoblotting. The neuronal architecture was analyzed by H&E and Nissl staining. The results demonstrated progressive accumulation of AGEs in the brain during aging. Adaptive ER stress response was observed by 10-months while maladaptive ER stress response was seen at 15- and 20-months of age along with impaired UPS and autophagy, and perturbations in neuronal growth factors. All these disturbances intensify with age to further exaggerate cell death mechanisms. There was a shrinkage of the cell size with aging and Congo-red staining revealed β-amyloid accumulation in higher ages. Together these results suggest that progressive accumulation of AGEs with aging in the brain may lead to neuronal damage by affecting ER homeostasis, UPS, autophagic flux, and neuronal growth factors.

Evaluation and Management Strategies for GI Involvement with Amyloidosis

Amyloidosis is a heterogeneous disease that can cause a wide array of nonspecific symptoms when the gastrointestinal (GI) tract is involved, including weight loss, early satiety, change in bowel habits with diarrhea, constipation, or alternating bowel pattern. Endoscopy with biopsy for Congo red staining establishes the diagnosis and fibril subtyping helps to guide targeted treatment options. Light chain amyloidosis is the most frequent subtype found throughout the GI tract. Transthyretin amyloidosis is most likely to be found on rectal biopsy. Management of the symptoms of GI tract involvement with amyloidosis relate to addressing the underlying symptom complex that is produced and generally abstracted from the management of severe forms of functional GI disorders. Attention to improving symptom management and nutrition status can improve quality of life in these patients.

ARSD is responsible for carcinoma and amyloidosis of breast epithelial cells

Breast cancer (BC) and Alzheimer's disease (AD) have pronounced female-to-male disparities and both are the major causes of death in elderly women. Intriguingly, there is an inverse incidence between BC and AD. In our previous study, we found that the expression of ARSD, a female-biased gene on chromosome Xp22.3 that encodes arylsulfatase D, is significantly downregulated in triple-negative breast cancer (TNBC) cells and tissue samples, and that ectopic ARSD overexpression could inhibit proliferation and migration of BC cells. However, the exact mechanism remains unclear. In this study, ARSD-overexpressing MDA-MB-231 cell strains were established. RNA-Seq and qRT-PCR validation were performed followed by GO and KEGG analyses. Transcriptome sequencing unveiled that Alzheimer’s/Parkinson’s/prion diseases were enriched in ARSD overexpressing BC cells. Besides, the top enriched pathways included lipoprotein/cholesterol metabolism, molecular chaperone and misfolding protein binding, mitochondrial respiration, dysfunction of lysosomes, etc. In which, a battery of genes, e.g., SERF1A, APOE, CD36 etc., were upregulated, while a series of genes, e.g., NDUFA11, NDUFS3, NDUFV1, etc. were downregulated, which were closely related to amyloidosis. The amyloidosis of BC cells and nerval cells caused by ARSD overexpression was verified with western blotting, immunohistochemical and Congo red staining. Collectively, downregulated ARSD may be closely associated with BC, and upregulated ARSD may cause amyloidosis of BC cells. Our findings suggest that ARSD deserves to be considered a new promising target for treating TNBC or for AD.

Renal Amyloidosis: A Clinicopathological Study From a Tertiary Care Hospital in Pakistan.

IntroductionSystemic amyloidosis can affect any organ in the body, but the kidney is the most commonly involved site. It is characterized by the extracellular deposition of insoluble fibrillar proteins. Amyloid deposits can be identified histologically by Congo red stain, which gives apple-green birefringence under polarized light. Typing of renal amyloidosis is done by direct immunofluorescence on frozen tissue. The most common types of amyloidosis seen in renal tissue are amyloid light chain (AL) primary amyloidosis and amyloid A (AA) secondary amyloidosis. Although primary amyloidosis is considered the most common type in western countries, however, in the subcontinent region, secondary amyloidosis is more common.The spectrum of signs and symptoms in renal amyloidosis is variable including isolated proteinuria, nephrotic syndrome, hypertension, hypotension, and renal insufficiency. The present study aims to evaluate the incidence and aetiology of various types of renal amyloidosis, determine their distribution within the kidney, and study various clinicopathological features.ObjectiveThe present study aims to evaluate the aetiology and clinicopathological profile of renal amyloidosis, determine its various types, and their distribution within the kidney.Materials and methodsThis retrospective cross-sectional study was conducted from 1st January 2013 to 31st December 2020 at the Department of Histopathology, Shifa International Hospital (SIH), Islamabad. All renal biopsies diagnosed as renal amyloidosis were included in the study. Data were analysed using SPSS version 23 (IBM Corp., Armonk, NY). Frequency and percentages were calculated for qualitative variables, and mean and standard deviation were calculated for quantitative variables.ResultsA total of 131 cases were diagnosed with renal amyloidosis during the study period of eight years (from 1st January 2013 to 31st December 2020) at SIH. The age range varied from 17 to 82 years. The mean age of the patients was 45 ± 16.33 years. Out of 131 patients, 82 (62.6%) were males and 49 (37.4%) were females. Amongst them, 72 (54%) cases were diagnosed with secondary AA amyloidosis and 16 (12%) cases were diagnosed with primary AL amyloidosis. The rest of the cases 43 (34%) were of indeterminate type. The associated conditions in secondary amyloidosis were tuberculosis in 41 (57%), rheumatoid arthritis in 16 (22%), ankylosing spondylitis in five (7%), lymphoma in three (4%), diabetes in two (3%), and chronic osteomyelitis, chronic heart disease, hepatitis, and vasculitis in one case each (1.7%). Out of 16 cases reported with AL amyloidosis, 10 cases (62.5%) had a history of multiple myeloma. The most common clinical presentation was nephrotic syndrome followed by subnephrotic proteinuria, renal failure, and hypertension.ConclusionThe findings of the present study show underlying etiological factors and clinicopathological characteristics of renal amyloidosis. AA amyloidosis is the most common type of renal amyloidosis in our study and tuberculosis is the most common aetiological factor. AL amyloidosis is less frequent in our population.

Unraveling aerobic cultivable cellulolytic microorganisms within the gastrointestinal tract of sheep (Ovis aries) and their evaluation for cellulose biodegradation.

Anaerobic cellulolytic microbes in the gastrointestinal tract (GT) of ruminants have been well-documented; however, knowledge of aerobic microbes with cellulolytic activities in the ruminant GT is comparably limited. Here, we unraveled aerobic cultivable cellulolytic microbes in the GT of Ujimqin sheep (Ovis aries) and evaluated the cellulolytic potential of the promising isolates. Twenty-two strains were found to possess cellulose-degrading potential by Congo-red staining and phylogenetic analysis of the 16S rDNA/ITS sequence revealed that all strains belonged to nine genera, i.e., Bacillus, Streptomyces, Pseudomonas, Lactobacillus, Brachybacterium, Sanguibacter, Rhizobium, Fusarium, and Aspergillus. Strains with high cellulolytic activity were selected to further evaluate the activities of various enzymes in lignocellulosic alfalfa hay (Medicago sativa). Among them, isolate Bacillus subtilis RE2510 showed the highest potential for cellulose degradation, considering the high endoglucanase (0.1478 ± 0.0014 IU mL-1), exoglucanase (0.1735 ± 0.0012 IU mL-1), and β-glucosidase (0.3817 ± 0.0031 IU mL-1) after 10-day incubation with alfalfa hay. A significant destruction effect of the cellulose structure and the attachment of B. subtilis RE2510 to the hay were also revealed using a scanning electron microscope. This study expands our knowledge of aerobic cellulolytic isolates from the GT of sheep and highlights their potential application as a microbial additive in the aerobic process of cellulose bioconversion.

Targeting Amyloids with [18F]AV-45 for Medullary Thyroid Carcinoma Positron Emission Tomography/Computed Tomography Imaging: A Pilot Clinical Study.

Medullary thyroid carcinoma (MTC) is a malignant neuroendocrine tumor with a high recurrence rate. Amyloid plaques formed from the misfolding of calcitonin are the key characteristics of MTC. Herein, we conducted a first-in-human pilot clinical study by applying a β-amyloid-specific radiotracer, [18F]AV-45, to positron emission tomography (PET)/computed tomography (CT) imaging of MTC. The presence of amyloid plaques in the tumor tissue sections from five MTC patients was confirmed by hematoxylin and eosin (H&E) and Congo Red staining. [18F]AV-45 selectively accumulated in the amyloid plaques in the continued tumor tissue sections with similar distribution patterns to the H&E and Congo Red staining. In addition, the [18F]AV-45 uptake can be largely blocked by its nonradioactive reference compound. The [18F]AV-45 accumulation in the thyroid, neck lymph nodes, and muscles in healthy human subjects is close to the background indicated by PET/CT imaging. In the comparison PET/CT imaging study of a recurrent MTC patient, 2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG) showed an elevated uptake by multiple neck lymph nodes. In contrast, only one of these neck lymph nodes had increased [18F]AV-45 uptake. Postoperative histopathological analysis confirmed the [18F]AV-45 PET-positive lymph node as MTC with amyloid deposition, while other [18F]FDG positive lymph nodes were free from MTC and amyloid plaques. Thus, [18F]AV-45 showed the promise for the clinical PET/CT imaging of MTC.

Effects of ischemia-hypoperfusion on neuro-vascular units in dual transgenic mice with Alzheimer's disease.

The study aimed to investigate the effects of ischemia on neuro-vascular units in transgenic mice, and to investigate the role of ischemia-hypoperfusion in the model of dual transgenic mice with dementia. In this study, the ischemic model was generated by operating a bilateral common carotid artery micro-embolism. Mice were divided into four groups, including group 1: C57BL sham surgery group (control), group 2: C57BL ischemic group, group 3: amyloid precursor protein/presenilin-1 (APP/PS1) group, and group 4: APP/PS1 ischemic group. Each group comprised 20 mice. Spatial behavior and memory ability of mice were detected by Morris water maze and jumping platform test. Mouse hippocampus was observed by HE staining and Congo red staining. Ultrastructure of each group of neuro-cyclic units was observed by electron microscopy. Various biochemical indicators were detected by ELISA. Western blot detected the amount of protein expression. qRT-PCR identified mRNA expression. The results indicated that learning and memory functions of C57 ischemic mice were lower than those of control group. Positive expression area of APP in APP/PS1 ischemic group was higher than in APP/PS1 group. In APP/PS1 group and APP/PS1 ischemic group, the content of Ab was significantly higher than in C57 ischemic group. Electron microscopic observation revealed that there were more mitochondrial vacuoles in hippocampal neurons of APP/PS1 mice, and the structure was relatively intact. Mitochondrial vacuoles in hippocampus increased significantly, and vascular wall proliferated in APP/PS1 ischemic group. Compared with C57 control group, the content of vascular endothelial growth factor (VEGF) increased significantly in C57 ischemic group. Ischemia deteriorates the learning and memory function of transgenic mice, aggravates the damage of neuro-vascular units, and impairs the blood-brain barrier transport of Ab, leading to an increase in the concentration of Ab cerebrospinal fluid, and further deterioration of neuro-vascular units. At the same time, ischemia is an effective stimulating factor in the release of VEGF.