Immunofluorescence Findings Research Articles

MON-017 SENSITIVITY AND SPECIFICITY OF SERUM PLA2 RECEPTOR ANTIBODIES IN DIAGNOSIS OF PRIMARY MEMBRANOUS NEPHROPATHY

Membranous nephropathy is a common cause of Nephrotic syndrome in adults and often requires a kidney biopsy to make a diagnosis, recent past there is a debate whether PLA2R antibodies will help in making a diagnosis of primary MGN without renal biopsy We carried out a prospective to Study to determine the association between serum anti-PLA2Rantibody and glomerular staining for PLA2R in Membranous Nephropathy Study design : Prospective observational, study duration: May 2016 to May 2018 Inclusion criteria: Adult Nephrotic syndrome those who are aged above 18 years undergoing kidney biopsy at our hospital Material & Methods: Serum for anti-PLA2R antibody was sent prior to kidney biopsy in all subjects. The serum anti- PLA2R antibodies was tested by Indirect immunofluorescence staining using validated commercial kits(Euroimmun). The kidney biopsy samples were subjected to light microscopy,immunofluorescence testing for the panel of antibodies as per standard protocol. Subsequently PLA2R staining was done as per standard protocol in those subjectswho showed features of membranous nephropathy In our study 13 out of 58 subjects who presented with nephrotic syndrome had positive serum anti-PLA2R Ab in samples analysed at presentation and prior tokidney biopsy. This account for 22.4% of the study subjects, while 45(77.6 %) subjects were negative for the same. Of the 13 subjects with positve serum anti-PLA2R Ab, 12 subjects had light microscopy and immunofluorescence findings ofMN on kidney biopsy. One subject who had FSGS-NOS type on kidney biopsyshowed weak positive serum anti-PLA2R antibody. Among the other 45 subjects who did not have serum anti-PLA2R antibody, there wasan additional 16 subjects who were diagnosed with primary MN and 1 with secondaryMN by kidney biopsy. This gives a Sensitivity of 41.38%, Specificity of 96.6%,Positive Predictive Value of 92.31%, Negative Predictive Value of 62.22% anddiagnostic accuracy of 68.97% for the test for diagnosis of MN by anti-PLA2Rantibody Statistical analysis by Cohen's kappa test showed a low agreement betweenserum anti-PLA2R Ab and Kidney biopsy was 0.3793 (0.1647 - 0.594). In our study the sensitivity and specificity of PLA2R antibodies in predicting adiagnosis of primary membranous nephropathy (pMN) 42.8% and 96.6% respectively Glomerular PLA2R staining had a sensitivity of 82.8% for diagnosis of MN and 85.5% for diagnosis of primary MN.

Correlation of clinicopathological characteristics and direct immunofluorescence studies in oral lichenoid lesion in Thai patients.

To investigate the correlation between the clinicopathological characteristics, serum antinuclear antibody (ANA) and direct immunofluorescence (DIF) findings in oral lichen planus (OLP) and oral lichenoid lesion (OLL). Fifty three Thai patients with red and white lesions were divided into 3 groups: 17 cases of OLP, 19 cases of OLL and 17 cases of oral lichenoid drug reaction (OLDR), respectively. The medical records, photographs, histopathological evaluation and laboratory ANA and DIF results were analyzed. Atrophic pattern was the most commonly found pattern in the OLDR, OLP and OLL groups. In the OLP group, the DIF interpretation confirmed only 41.2% of cases as OLP, with 23.5% each as lichen planus (LP)/lupus erythematosus (LE) or negative findings. In the OLL group, the most common DIF interpretation (31.6% each) was LP/LE or non-specific finding. In the OLDR group, DIF interpretation was OLP or LP/LE (23.5% each), with 5.9% each of immune complex-mediated disease, compatible with OLP, and mixed connective tissue disease. Interestingly, 1 case in the OLDR group demonstrated mild to moderate dysplasia. There were no significant differences in ANA positivity or patterns between the 3 groups. An OLP-like lesion could be diagnosed as OLP, OLP/LE, chronic ulcerative-like lesion, immune-mediated disease or dysplasia.

The Diagnostic Value of Tzanck Test in Vesiculobullous, Pustular, and Oral Lesions in The Dermatology Clinic

Background: Vesiculobullous disorders represent a heterogeneous group of dermatoses with protean manifestations. The accurate diagnosis of bullous disorders of the skin and mucous membrane requires evaluation of clinical, histological, and immunofluorescence findings. Objective: The aim of this study was to investigate Tzanck smear findings and to determine the diagnostic value of this test in moist (erosive, vesicular, bullous, and pustular) skin and oral lesions. Patients and Methods: This cross sectional study was conducted on one hundred patients presented with vesicular, bullous, and pustular skin lesions and/or oral lesions. All patients were included from Dermatology Outpatient Clinic at Al-Zahraa University Hospital over the period from March 2018 to March 2019. Results: Positivity of multinucleated giant cells in herpetic infection, acantholytic cells in pemphigus, dyskeratotic acantholytic cells and cocci in bullous impetigo were useful landmarks in Tzanck smear. A decrease in viral shedding from herpetic lesions of longer duration may correlate with the lower sensitivity of the Tzanck smear. Therefore, in this study we excluded the lesions older than 3 days. The percentage of positivity in vesicles was much higher than that of pustules. Conclusion: According to this study, Tzanck smear was an inexpensive, simple, and rapid test and did not require any specialized laboratory equipment for evaluation of various vesiculobullous and pustular lesions either skin or mucosal one.

Characteristics of membranoproliferative glomerulonephritis based on a new classification at a single center.

Recently, a new classification has been established for membranoproliferative glomerulonephritis (MPGN). However, the effect of the new classification on MPGN treatment is not fully understood. We conducted a retrospective study of 87 patients with biopsies diagnosed as MPGN. We reclassified 87 MPGN patients diagnosed between 1977 and 2014at our hospital, according to the new classification, and analyzed both primary immune complex (IC)- and Alternative pathway (AP)-mediated MPGN [corrected] in terms of clinicopathological features, treatment, and renal prognosis. Proteinuria was abundant in the IC-mediated MPGN group (p = 0.0063), and the serum albumin level was significantly lower in the IC-mediated MPGN group (p = 0.0186). The serum C3 value was significantly lower in the CP-mediated MPGN group (p = 0.0317). Serum CH50 values were also lower in the CP-mediated MPGN group (p = 0.0404). However, glomerular deposition of C3 showed no significant differences in immunofluorescence findings. The 148.6-month renal survival rate was similar in both groups (p = 0.445). These results suggested no significant differences in complement activation of the solid phase in local glomeruli and therefore equivalent in renal prognosis [corrected].

Acantholysis may precede elevation of circulating anti-desmoglein 3 antibody levels in pemphigus vulgaris presenting with desquamative gingivitis.

Pemphigus vulgaris (PV) is an autoimmune, blistering disease that affects the mucosa and skin. The current theory favors the concept that anti‐desmoglein (Dsg) 3 autoimmunity is the only pathogenic event needed to induce acantholysis. However, a few cases of active PV in the oral cavity had no detectable anti‐Dsg 3 antibody. The aim of this study was to evaluate the differences in clinical and laboratory findings, whether or not the anti‐Dsg 3 antibodies were present. This study was based on a retrospective review of 10 PV cases. The evaluation of the circulating autoantibody titers to Dsg 3 was conducted by using enzyme‐linked immunosorbent assay (ELISA). An index value of 20 or more was used as the cutoff for a positive reaction. Only five of the 10 PV cases had a positive Dsg 3 ELISA. There were no differences in clinical, cytological, histopathological, and direct immunofluorescence findings, whether or not the anti‐Dsg 3 antibodies were present. Of the five patients with a negative reaction at the time of diagnosis, the Dsg 3 ELISA became positive in the follow‐up period in three cases. In the remaining two cases, the Dsg 3 ELISA was consistently negative for 18 months. Dsg 3 ELISA was negative early in some PV cases. Therefore, PV acantholysis may precede the elevation of circulating anti‐Dsg 3 antibody levels. The diagnosis of PV should be considered based on comprehensive clinical, histopathological, and immunofluorescent criteria.

Glomerular C4d Staining Does Not Exclude a C3 Glomerulopathy

IntroductionC4d, an early product in the classical/lectin complement pathway has shown potential in the evaluation of C3 glomerulopathy where its absence would support an alternative pathway abnormality. As autoimmune/genetic complement testing is not readily available to most parts of the world, glomerular C4d staining may serve as a useful additional step toward the diagnosis.MethodsTo test this hypothesis, C4d staining was performed on a large cohort of C3 glomerulopathy. Archival cases from 2011 to 2017 were reviewed and immunohistochemistry for C4d was performed, scored (scale of 0 to 3+), and correlated with the immunofluorescence and ultrastructural findings. Paraffin immunofluorescence was performed in cases of “discordant C4d” to unmask Igs.ResultsTwenty-seven cases of dense deposit disease (DDD) and 14 cases of C3 glomerulonephritis (C3GN) were retrieved. C4d demonstrated a range of staining intensities with negative/traces in only 22% of DDD and 64% of C3GN. Lower-intensity C4d staining (1 to 2+) was mostly concordant with similar amounts of Igs/C1q. Discordant 3+ staining was noted in approximately 50% of cases of DDD and 20% of cases of C3GN. Among them, paraffin immunofluorescence unmasked polyclonal Igs in 2 of 5 cases of DDD and 1 of 3 cases of C3GN.ConclusionThis observational study suggests that the presence of glomerular C4d should not exclude a C3 glomerulopathy. In lower intensities, it appears to represent overlying classical/lectin pathway activation with concordant Ig/C1q deposits. A subset of cases, however, displays intense and discordant C4d staining, which raises the possibility of an associated lectin pathway abnormality, a potential future area of study.

Are there distinct clinical and pathological features distinguishing idiopathic from drug-induced subacute cutaneous lupus erythematosus? A European retrospective multicenter study

Clinical and pathologic criteria to distinguish drug-induced subacute lupus erythematosus (DI-SCLE) from idiopathic (I-SCLE) are controversial. The aim of the survey was a retrospective analysis of a consistent number of iatrogenous and idiopathic SCLE cases, by means of clinical and histopathologic investigation. Eleven European university dermatology units collected all diagnosed cases from January 2000 to December 2016. Board-certified dermatopathologists reviewed the histopathologic specimens. Statistical analysis included Student t test, exact test of goodness-of-fit, Fisher's exact test, and the Cochran-Mantel-Haenszel test for repeated measures. Out of 232 patients, 67 (29%) belonged to the DI-SCLE group. Patients with DI-SCLE were significantly older and reported more systemic symptoms than those with I-SCLE. No statistical differences were found for presentation pattern or serology, while histopathology showed a significant association of mucin deposition (P=.000083), direct immunofluorescence positivity for granular immunoglobulin M, and C3 deposits on the basement membrane zone (P=.0041) for I-SCLE and of leukocytoclastic vasculitis (P=.0018) for DI-SCLE. This is a retrospective study. An integrated clinical and immunopathologic evaluation is useful to differentiate I-SCLE from DI-SCLE. Older age at onset and more frequent systemic symptoms characterize DI-SCLE. Mucin deposition and immunofluorescence findings are found in I-SCLE, and leukocytoclastic vasculitis is found in DI-SCLE.

Pemphigus – A clinical study with clinico-immuno-histopathologic correlation

Background: Pemphigus is a group of autoimmune blistering disease of the skin and mucous membranes with an incidence of <2 cases per 100,000 population. The aim is to evaluate the clinical pattern and to study the correlation between clinical, histopathological, and immunofluorescence findings in pemphigus. Materials and Methods: All newly registered patients with flaccid blisters and erosions on the skin with/without oral erosions during 2 years from May 2009 to April 2011 were evaluated by detailed history, clinical examination, cytological, pathological, and direct immunofluorescence studies, and results correlated. Results: Of the 43 patients studied, 36 (83.72%) had pemphigus vulgaris and 7 (16.28%) had pemphigus foliaceus with maximum patients in the age group of 40–59 years and M:F ratio of 1:1.3. Cytologically, acantholytic cells were demonstrated in 88.37%, histopathological correlation was seen in 100% cases, and direct immunofluorescence was positive in 97.37% of the 38 cases in which it was performed. Conclusion: Clinico-immuno-histopathologic correlation is seen in most cases of pemphigus, and although a combination of clinical, cytological, histopathological, and immunofluorescence findings virtually eliminates the chances of error in the diagnosis of pemphigus, histopathology may suffice in most cases.

The spectrum of renal diseases observed in native renal biopsies in a single North Indian tertiary care center

We analyzed the spectrum of biopsy-proven renal disease in a single tertiary care center in North India from 2007 to 2016. A total of 420 biopsies were analyzed. Patients were excluded if clinical details were unavailable or if either the histopathology core or the IF core was inadequate. In the final analysis, 359 biopsies were included. All clinical, laboratory, histopathological, and immunofluorescence (IF) findings were recorded in each case. The usefulness of IF in reaching a definitive diagnosis was also analyzed. The patients were in the age range of 2-94 years; 23.1% were children and 76.9% were adults. Males (60.4%) outnumbered females (39.6%) in all the disease categories except lupus nephritis (LN). Primary glomerular diseases (PGDs) (n = 297, 82.7%) were more common than secondary glomerular diseases (SGDs) (n = 46, 12.8%) and tubulointerstitial diseases (n = 16, 4.5%). The most common PGD was focal segmental glomerulosclerosis (FSGS) (23.4%), followed by minimal change disease (17%) and membranous nephropathy (12.5%), whereas the most common SGD was LN, seen in 9.2%. In the present study, IF helped in reaching the final diagnosis in 44.3%. The entities in which IF was most useful in reaching the final diagnoses were FSGS (31.5%) and IgA nephropathy (14.5%). The final pathological diagnosis correlated with the first clinical possibility in 207 of 359 (57.7%) cases. This 10-year study provides descriptive data and highlights the changing pattern of renal disease possibly due to an increased awareness and referral to higher centers.

Direct immunofluorescence findings in cicatricial alopecia: A retrospective study of 155 cases.

To study the direct immunofluorescence (DIF) in cicatricial alopecia (CA) patients. Approximately, 155 skin biopsies from CA patients examined over 7 years (2009-2015). Special stains and Hematoxylin and Eosin were performed, and final histopathological diagnosis was made. DIF (against anti-IgG, IgM, C3, IgA, and fibrinogen) on all these cases and patterns were noted. The descriptive statistics were applied along with ANOVA test. Approximately, 155 patients with Male: Female = 1.24:1, age 7-65 years. In total, 57 cases were of Lichen planopilaris (LPP; 36.7%), 22 Lichen planus (LP; 14.2%), 22 Psuedopalade of Brocq (PPB; 14.2%),16 discoid lupus erythematosus (DLE; 10.3%), 8 end-stage scarring alopecia (ESSA; 5.2%), 2 cases each of Fungal folliculitis (FF), and Folliculitis decalvans (FD; 1.3% each), and in 26 cases, no specific diagnosis could be reached were collectively kept in the category of non-specific findings (NSP; 16.7%). On DIF: LPP positive for IgG = 4 cases (7%), IgM = 26 (45%), IgA = 11 (19.3%), C3 = 16 (28.1%), and fibrinogen = 11 (19.3%). LP positive for IgG = 2 (9%), IgM = 18 (81%), IgA = 2 (9.1%), C3 = 10 (45%), and fibrinogen = 1 (4.5%). DLE positive for IgG = 6 (37%), IgM = 8 (50%), IgA = 1 (6.3%), C3 = 9 (56%), and fibrinogen = 1 (4.5%). Because this was a retrospective study, clinical follow-up and treatment history of the patient could not be retrieved. IgG positivity helps significantly in differentiating LPP from DLE (P 0.004) and NSP from DLE (P 0.005). IgM positivity helps significantly in differentiating LPP from LP (P 0.04), LP from PPB (P 0.00) and NSP (P 0.00). C3 positivity helps significantly in differentiating PPB from DLE (P 0.02).

The rash that presents as target lesions

We have explored the rash that appears as target lesions, with the central and dominant diseases belonging to the Stevens-Johnson syndrome/toxic epidermal necrolysis group. After presenting the clinical patterns of an individual target lesion and classifying them into different types of lesions, the contribution has been organized with groups characterized by such specific findings according to the type of lesion: flat or raised, typical or atypical, presence or absence of fever, presence or absence of mucosal ulcerations, presence or absence of arthralgias, and/or internal organ involvement. Other specific features, such as histologic appearance, immunofluorescence findings, and laboratory changes, are considered. We provide clinicians with an algorithmic, systematic, and logical approach to diagnose the condition of the patients who present with targetoid lesions, and enable them to differentiate between those with serious systemic and life-threatening diseases from others with ordinary skin ailments.

A Homozygous Nonsense Mutation in the DSG3 Gene Causes Acantholytic Blisters in the Oral and Laryngeal Mucosa

A Homozygous Nonsense Mutation in the DSG3 Gene Causes Acantholytic Blisters in the Oral and Laryngeal Mucosa

Comprehensive evaluation of the significance of immunofluorescent findings on clinicopathological features in IgA nephropathy.

The clinicopathological significance of immunofluorescent findings in IgA nephropathy remains controversial. The relations of the deposition of IgA, IgG, IgM, C3, C1q and fibrinogen (Fib) with pathological findings, baseline clinical findings, and renal outcome were evaluated in 688 patients with IgA nephropathy. Pathological features included cellular or fibrocellular crescents, endocapillary or mesangial hypercellularity, segmental or global glomerulosclerosis and the Oxford classification. The median age at biopsy was 30years. There were 289 men. With 74months median follow-up, 32% of patients received steroids. Twelve percent of patients developed end-stage renal disease (ESRD). The degree of IgA was closely related to the degree of C3, IgG and IgM deposition. The degree of IgA, C3, IgG and Fib deposition was significantly related to the percentage of glomeruli with crescent, endocapillary and mesangial hypercellularity. IgM deposition showed significant association with crescent, mesangial hypercellularity, segmental sclerosis, global glomerulosclerosis and tubular atrophy/interstitial fibrosis. In the patients treated with steroids, the risk for ESRD in patients with 2-3+ IgA deposition was significantly lower with reference of 1+ IgA deposition. We found the different roles of glomerular immune reactants' deposition in the inflammatory process from acute to chronic stage. IgA deposition together with IgG, Fib and C3 may produce acute inflammatory injury. IgM deposition might occur in the early stage of inflammation and remains until late sclerotic stage. The prominent deposition of IgA related to low risk for ESRD in patients who received steroids might suggest effectiveness of steroids in such patients.

Bullous Systemic Lupus Erythematosus

Bullous systemic lupus erythematosus (BSLE) is extremely rare but distinct disease, characterized by vesicobullous skin eruptions in systemic lupus erythematosus (SLE). It can develop either before or after a diagnosis of SLE has been established. BSLE is characterized by a dermatitis herpetiformis-like histology and an autoimmunity to type VII collagen. It must be differentiated from other autoimmune vesicobullous diseases such as epidermolysis bullosa acquisita, dermatitis herpetiformis, linear IgA disease, and bullous pemphigoid. Its important to combine clinical, histological, and immunofluorescence findings to establish a diagnosis of BSLE. We report a case of BSLE to illustrate and emphasize the need for an integrative diagnostic approach.J MEDICINE JUL 2018; 19 (2) : 123-125

Excellent Response to Hydroxychloroquine in a Young Man with Lupus Erythematosus Panniculitis

Lupus erythematosus panniculitis is a rare variant of lupus erythematosus. We present the case of a young man with ulcerative colitis treated with mesalazine and a history of recurrent painful and indurated lesions on his face. A diagnosis of lupus profundus was made taking in account the clinical manifestations and the histological and immunofluorescence findings.

A case of immunotactoid glomerulopathy with false-negative IgG staining

BackgroundImmunotactoid glomerulopathy (ITG) is a rare glomerulonephritis characterized by microtubular deposits. Immunofluorescence findings are necessary to differentiate ITG from other proliferative glomerular diseases. The characteristic tubular structure on electron microscopy is essential for a definitive diagnosis, and the diameter of the structure has been traditionally used for differentiating between ITG and other types of glomerulonephritis with organized deposits. In recent years, the disease concept of monoclonal gammopathy of renal significance, which is associated with M protein produced by plasma cell tumors, has been proposed.Case presentationThis was a peculiar case of ITG with underlying monoclonal gammopathy in which IgG showed a false-negative result with immunofluorescence using frozen sections. Additional examinations using a different clone of the anti-IgG antibody revealed typical IgG staining. C4d was strongly positive, consistent with immune complex type glomerulonephritis.ConclusionsThis case highlights unusual features of ITG, and provides a practical hint to avoid a diagnostic pitfall.

Keratinocyte Binding Assay Identifies Anti-Desmosomal Pemphigus Antibodies Where Other Tests Are Negative.

The serological diagnosis of pemphigus relies on the detection of IgG autoantibodies directed against the epithelial cell surface by indirect immunofluorescence (IIF) on monkey esophagus and against desmoglein 1 (Dsg1) and Dsg3 by ELISA. Although being highly sensitive and specific tools, discrepancies can occur. It is not uncommon that sera testing positive by ELISA give a negative result by IIF and vice versa. This brings diagnostic challenges wherein pemphigus has to be ascertained or ruled out, especially when no biopsy is available. We utilized the ability of anti-Dsg3 and anti-Dsg1 IgG to bind in specific desmosomal patterns to living cells to investigate these discrepancies between IIF and ELISA. Living cultured primary normal human keratinocytes were grown under differentiating conditions to induce adequate expression of Dsg1 and Dsg3, incubated with patient serum for 1 h, and then stained to visualize bound IgG. We investigated two different groups; sera from patients with a positive direct immunofluorescence (DIF) and inconsistent serological findings (n = 43) and sera with positive ELISA or IIF but with negative DIF (n = 60). As positive controls we used 50 sera from patients who fulfilled all diagnostics criteria, and 10 sera from normal human subjects served as negative controls. In the DIF positive group, IgG from 39 of the 43 sera bound to the cells in a desmosomal pattern while in the DIF negative group none of the 60 sera bound to the cells. This shows that for pemphigus patients, ELISA and IIF can be negative while anti-desmosomal antibodies are present and vice versa that ELISA and IIF can be positive in non-pemphigus cases. In absence of a biopsy for DIF, such findings may lead to misdiagnosis.

Deletion of pro-angiogenic factor vasohibin-2 ameliorates glomerular alterations in a mouse diabetic nephropathy model

Angiogenesis has been implicated in glomerular alterations in the early stage of diabetic nephropathy. We previously reported the renoprotective effects of vasohibin-1 (VASH1), which is a novel angiogenesis inhibitor derived from endothelial cells, on diabetic nephropathy progression. Vasohibin-2 (VASH2) was originally identified as a VASH1 homolog and possesses pro-angiogenic activity in contrast to VASH1. In addition, VASH2 was recently shown to promote epithelial-to-mesenchymal transition via enhanced transforming growth factor (TGF)-β signaling in cancer cells. Herein, we investigated the pathogenic roles of VASH2 in diabetic nephropathy using VAHS2-deficient mice. The type 1 diabetes model was induced by intraperitoneal injections of streptozotocin in VASH2 homozygous knockout (VASH2LacZ/LacZ) or wild-type mice. These mice were euthanized 16 weeks after inducing hyperglycemia. Increased urine albumin excretion and creatinine clearance observed in diabetic wild-type mice were significantly prevented in diabetic VASH2-deficient mice. Accordingly, diabetes-induced increase in glomerular volume and reduction in glomerular slit-diaphragm density were significantly improved in VASH2 knockout mice. Increased glomerular endothelial area was also suppressed in VASH2-deficient mice, in association with inhibition of enhanced vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2), but not VEGF level. Furthermore, glomerular accumulation of mesangial matrix, including type IV collagen, and increased expression of TGF-β were improved in diabetic VASH2 knockout mice compared with diabetic wild-type mice. Based on the immunofluorescence findings, endogenous VASH2 localization in glomeruli was consistent with mesangial cells. Human mesangial cells (HMCs) were cultured under high glucose condition in in vitro experiments. Transfection of VASH2 small interfering RNA (siRNA) into the HMCs resulted in the suppression of type IV collagen production induced by high glucose compared with control siRNA. These results indicate that VASH2 may be involved in diabetes-induced glomerular alterations, particularly impaired filtration barrier and mesangial expansion. Therefore, VASH2 is likely to represent a promising therapeutic target for diabetic nephropathy.

The features in IgA-dominant infection-related glomerulonephritis distinct from IgA nephropathy: a single-center study.

IgA-dominant infection-related glomerulonephritis (IgA-IRGN) is a unique form of IRGN, which needs to be distinguished from IgA nephropathy (IgAN). Thirteen patients with IgA-IRGN (IgA-IRGN group) and 122 with IgAN (IgAN group) were selected from 1788 patients who underwent kidney biopsy between 2000 and 2015 in Kitano Hospital. Data selected included clinical and serological parameters; light and electron microscope findings; immunofluorescence findings; and prognostic parameters like renal and overall survival and creatinine increase by > 50%. In addition, a 26-patient IgAN cohort (matching-IgAN), matching with IgA-IRGN group with respect to age, sex, estimated glomerular filtration rate (eGFR), and proteinuria was segregated for comparison. Compared to IgAN group, IgA-IRGN group were older, had lower hemoglobin, higher CRP, lower eGFR, heavier proteinuria, lower serum albumin, and higher serum IgG and IgA levels (p < 0.05). Endocapillary hypercellularity, deposition of immune complexes along the glomerular capillary wall, and subendothelial and subepithelial electron dense deposits were more frequently observed (p < 0.05); and they were more susceptible to renal dysfunction and poorer prognosis. After propensity score-matching, serum albumin was significantly lower in the IgA-IRGN group. Significantly subendothelial and subepithelial deposits were frequently observed in this group. Matching-IgAN group showed relatively advanced sclerotic lesions with more global sclerosis and fibrous crescent. Local inflammation involved glomerular capillary wall in IgA-IRGN, in contrast to relatively chronic and sclerotic renal lesion in IgAN, might result in poorer prognosis in former, even under indistinguishable condition of deteriorated renal function and proteinuria.

Beta estradiol and norepinephrine treatment of differentiated SH-SY5Y cells enhances tau phosphorylation at (Ser396) and (Ser262) via AMPK but not mTOR signaling pathway

Hyperphosphorylation of tau is one of the main hallmarks for Alzheimer's disease (AD) and many other tauopathies. Norepinephrine (NE), a stress-related hormone and 17-β-estradiol (E2) thought to influence tau phosphorylation (p-tau) and AD pathology. The controversy around the impact of NE and E2 requires further clarification. Moreover, the combination effect of physiological and psychological stress and estrogen alteration during menopause, which affect p-tau, has not been addressed. Exposure to E2 is believed to reduce NE release, however, the link between these two hormones and AD at cellular level was also remained unknown. Here, we examined whether NE and E2 treatment of differentiated SH-SY5Y cells affected tau phosphorylation. The involvement of adenosine monophosphate kinase protein kinase (AMPK) and target of Rapamycin (mTOR) as the possible mechanisms, underlying this effect was also investigated. Subsequent to SH-SY5Y differentiation to mature neurons, we treated the cells with NE, E2 and NE plus E2 in presence and absence of Compound C and Rapamycin. Cell viability was not affected by our treatment while our Western blot and immunofluorescent findings showed that exposure to NE and E2 separately, and in combination enhanced p-tau (Ser396) and (Ser262)/tau but not (Ser202/Thr205)/tau. Blocking AMPK by Compound C reduced p-tau (Ser396) and (Ser262), while GSK-3β and PP2A activities were remained unchanged. We also found that blocking mTOR by Rapamycin did not change increased p-tau (Ser396) and (Ser262) due to NE + E2 treatment. Collectively, our results suggested that tau hyperphosphorylation due to exposure to NE/E2 was mediated by AMPK, the main energy regulator of cells during stress with no significant involvement of mTOR, GSK-3β and PP2A.