Renal histopathology and clinical course in patients with Wegener's granulomatosis--single centre experience from the Republic of Macedonia.

The aim of this study was to evaluate the clinical course of patients with Wegener's granulomatosis (WG) with renal involvement, to examine histopatological form seen in renal biopsies and present follow-up of the patients. A retrospective analysis was carried out of 18 patients presenting with WG and active renal disease at the University Nephrology Department, Ss. Cyril and Methodius University, Skopje, R. Macedonia. All patients were ANCA positive and had a percutaneous renal biopsy taken on their admission. 12 patients were male, 6 female, aged 48.61±13.77 (M±SD). All had extrarenal symptoms prior to admission. Oligoanuria was present in 7/18 (38.9%) of the patients, serum urea levels of the whole group were 40.67±18.13 mmol/l (M±SD) and for serum creatinine 691.06±384.93 µmol/l (M±SD). Necrotizing glomerulonephritis with crescents was present in 11/18 (61.11%) of the patients, the others presented diffuse proliferative extracapillary glomerulonephritis. All patients were treated with steroids and cyclophosphamide, and plasmapheresis was performed in 7/18 (38.9%) of the patients. Probability rate for surviving after one month was 0.6111 and after three months 0.3889 (Kaplan-Meier). The current treatment of WG in our study did not prevent serious complications and development of ESRD in a large number of our patients. This systemic disorder is still a serious problem and early diagnosis and alternative strategies for the management of the disease will be an important objective for further studies.

Mycophenolate mofetil in the treatment of glomerular diseases.

AIM: Treatment of primary glomerular diseases may be unsuccessful or have potential toxicities. Mycophenolate mofetil (MMF) is a new, relatively non-toxic drug. It has been introduced as an immunosuppressive drug, but it also has effects on non-immune cells (vascular smooth muscle cells, fibrocytes). Therefore, we evaluated the use of MMF for the treatment of glomerular diseases at different stages of the disease. METHODS: The daily dosage of MMF was 2 for the first 6 months and 1.5 g for a further 18 months, combined with steroids. The follow-up period was two years. RESULTS: 18 patients with lupus nephritis were treated. Patients with a high histological activity index showed a significant decrease of serum creatinine (p < 0.05) and proteinuria (p < 0.01), while patients with a high chronicity index showed only a decrease of proteinuria (p < 0.05). 15 patients with membranous nephropathy were treated. They showed stable renal function and a significant decrease of proteinuria (p < 0.05). Complete remission was achieved only in patients with MMF as a first choice drug. 4 patients with focal segmental glomerulosclerosis did not show any significant decrease of proteinuria, while the nephrotic syndrome in minimal change nephropathy (3 patients) showed a complete recovery. Partial improvement of the nephrotic syndrome was noted in 5 patients with membranoproliferative glomerulonephritis and in 4 patients with crescentic glomerulonephritis. Patients with crescentic glomerulonephritis also presented a significant decrease of serum creatinine (p < 0,05). MMF in 3 patients with IgA nephropathy grade I showed a significant improvement of the nephrotic syndrome. In grade III (5 patients) the response was partial. CONCLUSIONS: We can conclude that MMF in our patients showed both actions, as an immunosuppressive drug in the early stages of the disease, and as an anti-fibrotic agent in the chronic phase of the disease.

Role of mycophenolate mofetil in the treatment of lupus nephritis.

Mycophenolate mofetil (MMF) is an immunosuppressive drug successfully used for the prevention of acute and chronic rejection of renal allografts, as well as in the therapy of glomerular disorders. We treated three groups of patients with lupus nephritis: the first group of patients had a high histologic activity index (AI), 13.4 +/- 2.34; the second group of patients had a high histologic chronicity index (CI), 6.0 +/- 0.7; and the third group consisted of only two patients, one with low AI (3.5) and another with low CI (1.5). The patients were treated for 2 years. MMF was initiated at a dose of 2 g/daily for the first 6 months and the dose was decreased to 1.5 g/daily for the further 18 months. Steroids, 0.4 mg/kg/day, were the concomitant therapy for the first 6 months, with slow tapering for the further 18 months. Patients with high AI presented significant decrease of serum creatinine after 2 years, 286 +/- 112.95 to 131.2 +/- 44.65 micromol/L. Two of the patients, with acute oligoanuria, were withdrawn from dialysis treatment. Significant improvement was also noted, 6.97 +/- 1.81 to 0.9 +/- 0.31 g/day. Patients with high CI had nonsignificant decrease of serum creatinine, 178.5 +/- 47.73 to 129.25 +/- 22.88 micromol/L, and significant improvement of proteinuria, 4.63 +/- 1.57 to 1.14 +/- 0.39 g/day. The patient with low AI showed recovery of renal function (serum creatinine from 196 to 72 micromol/L) and alleviation of proteinuria, 7.93 to 3.4 g/day. The patient with low CI did not respond to the therapy and renal function slowly worsened. MMF has emerged as a promising therapeutic approach for both the induction and maintenance phase in patients with lupus nephritis.

Morphological changes in the tubulointerstitial compartment in primary glomerulopathies.

The renal interstitium structurally supports the functional renal units and is involved in almost all renal functions. The degree of renal disfunction strongly correlates to the changes in the tubulointerstitial compartment present in almost all types of glomerular diseases. A phenomenon arising in such an environment is epithelial-mesenchymal transition, i.e. a change of the cell;s epithelial phenotype into a mesenchymal one. Histochemical, immunohistochemical and morphometric analyses were made of 50 renal biopsies with primary glomerulopathies, as well as light-microscopy analyses of semi-thin sections embedded in epoxy resin. Double immunohistochemical stainings with pairs of epithelial and mesenchymal antibodies were also done. The results were analyzed and correlated with the clinical data of the renal function of the patients. The immunohistochemical analyses of the atrophic tubular epithelial cells showed a loss of expression of Cytokeratin and E-cadherin, an enhanced expression of HLA-DRalpha, and a de novo expression of Vimentin and alphaSMA as markers for epithelial-mesenchymal transition. The double immunohistochemical stainings with Cytokeratin/Vimentin and Cytokeratin/alphaSMA showed a simultaneous expression of these antigens in atrophic tubular cells. Their proliferative index was mildly enhanced. Interstitial fibrosis was present in 98% of the analysed biopsies. The analyses show correlations among all the changes in the tubulointerstitial compartment as well as the concentration of creatinine in the serum as a parameter of renal function. The study emphasizes the usefulness of the implementation of histomorphometrical and immunohistochemical techniques as well as ultrastructural and molecular analyses in the process of nephropathological diagnosis.

Therapeutic apheresis in the Republic of Macedonia - our five years experience (2000-2004).

Membrane plasma exchange (PE) is a mode of extracorporeal blood purification. Since 1985 membrane PE has been in regular use at the Department of Nephrology, Medical Faculty of Skopje, R.Macedonia. In this paper we report on five years (2000-2004) of single centre plasma exchange activity. We performed 540 PE treatments (108 PE/per year) on 99 patients. The M/F ratio was 40/48. The patients underwent a median of 5.45 procedures (range, 1-16). The treated patients were from different Departments. Protocols for PE depend on the disease and its severity. PE were performed 2-4 times weekly using Gambro PF 2000 N filters with an adaptation of the Gambro AK10 dialysis machine or with the Gambro Prizma machine (2 cases). Blood access was achieved through femoral vein. Substitution was made with fresh frozen plasma and/or with 20% human albumin combined with Ringer's solution. An average amount of 2150 ml plasmafiltrate per treatment (respectively 30 to 40 ml plasmafiltrate/kg body weight) was eliminated. Most therapeutic procedures were performed on patients from the Department of Neurology. 63.6% of all patients were referred for Myasthenia gravis and the Guillian Barre syndrome. The total number of procedures per year has remained fairly stable, corresponding to a median of 5.4 treatments/100 000 inhabitants. We observed hypocalcaemia in 8% of the patients, urticarial reactions in 7.3%, pruritic reactions in 12%, and hypotension/headache in 6.8%. No major procedural complications were seen.

ANCA-GBM dot-blot test in diagnosis of patients with glomerulonephritis.

Patients with rapidly progressive glomerulonephritis who are positive for anti-neutrophil cytoplasmic antibody (ANCA) or anti-glomerular basement membrane (GBM) antibodies may develop chronic renal failure leading to end-stage renal disease (ESRD) within days or weeks. The early serologic detection of auto-antibodies associated with ANCA and anti-GBM diseases will be helpful in preventing ESRD. We evaluated the combined ANCA-GBM dot-blot strip assay (Biomedical Diagnostics, Brugge, Belgium) in 30 consecutive patients with biopsy proven glomerulonephritis (GN). MPO- and PR3-ANCA were detected in 5 and 2 samples, respectively. Three samples were positive for both MPO- and PR3-ANCA (all 3 had focal segmental necrotizing GN). One patient was diagnosed as having Goodpastures' syndrome (the only anti-GBM positive result) and two had Wegener's granulomatosis (the two PR3-ANCA positive results). Two additional samples were equivocal: positive for MPO-ANCA and PR3-ANCA, respectively. Patients positive only for MPO-ANCA had only limited extrarenal organ manifestations. Anti-PR3 positive patients with necrotizing glomerulonephritis had a more dramatic deterioration of their renal function at diagnosis. Radiographically, these patients had nodular or pneumonia-like lesions. Acute respiratory failure necessitating mechanical ventilation was developed in one GBM positive patient. In conclusion, the ANCA-GBM dot-blot is a useful screening tool in situations where conventional ANCA testing is not readily available.

20 years after methylprednisolone/chlorambucil treatment in idiopathic membranous nephropathy stage II-III with nephrotic syndrome.

There is now controlled evidence that a 6-month course with methylprednisolone and chlorambucil may favour remission of the nephrotic syndrome and may significantly improve the 10-year kidney survival in patients with idiopathic membranous nephropathy. We analyzed the outcome of 15 nephrotic patients (proteinuria 7.06 +/- 1.07 g/d), stage II-III membranous nephropathy, aged 37.93 +/- 2.32, 8 males and 7 females, with normal serum creatinine (62.8 +/- 2.34 micromol/l), followed > 10 years after the treatment. It consisted of 1g i.v. methylprednisolone for three consecutive days, followed by oral steroids 0.4 mg/kg/d and chlorambucil 0.2 mg/kg/d monthly, alternatively. 10 patients, age and sex matched, who refused any treatment of any reason, represented the control group. Complete remission was defined as protein loss of 0.2 g/d, partial 0.2-2 g/d with normal creatinine and renal dysfunction as increase in plasma creatinine. The follow-up period was between 10 and 20 years. Complete remission after the treatment was noted in 9/15, partial in 4/15, and 2/15 patients did not respond. 10-year survival rate of the whole group was 100%, 15-year - 86.7%, i.e. two patients with persistent nephrotic syndrome developed end-stage renal failure after 12 years. 13/15 patients (complete, partial remission) were followed > 15 years without development of end-stage renal failure. One patient (female, 32) developed idiopathic thrombocytopenia after 8 years. 3 patients (complete remission) were followed > 20 years, they are still without proteinuria. 10-year survival rate of untreated patients was 40%. It is concluded that in nephrotic patients with stage II-III membranous nephropathy steroids/chlorambucil therapy may be effective in favoring remission and in preserving renal function.

IgA nephropathy: 23 years of follow-up.

IgA nephropathy (IgAN) is the most common primary glomerulonephritis. Some patients reach end-stage renal failure (ESRF), others experience an indolent course. We aimed the study to examine the association of risk factors with the progression to renal failure. Eighty patients diagnosed with IgAN by renal biopsy (RB) were studied. Baseline clinical and demographic data were reviewed. Severity of histological involvement was scored as H. S. Lee's grading system. The mean age of patients at biopsy was 36.65 +/- 8.83 years with predominance of men (male : female, 58 : 22). Patients were followed-up from 6 months to 23 years (276 months). An end-point was defined as the date when patient underwent their first haemodialysis or their last visit of follow-up. The differences in means between groups were compared by independent samples t-tests or one-way analysis of variance (ANOVA). Kaplan-Meier survival curves and Cox regression models were used to analyze the time course from renal biopsy to end points. The largest subclasses were grade I and II, with 31 patient each. Subclass III was observed in 12 patents. Subclass IV and V were found in 3 patients each. During the follow-up period, all patients with grade IV and V (after 6-48 months), five patients grade I (after 60-144 months), four patients grade II (after 48-84 months), and 7 patients from grade III (after 24-108 months) entered ESRD. Mean prioteinuria was 1.68 +/- 0.99 g/day. Macrohematuria had 32; microhematuria had 48 pts. The mean serum creatinine was 148.02 +/- 68.76 micromol/l. By multivariate analysis using the Cox regression model, grades, renal insufficiency and significant proteinuria were independent prognostic factors for progressive renal disease. At the end of follow-up, grades were significantly related to serum creatinine, proteinuria, hypertension and progressive renal disease. Renal biopsy in IgAN may be the most powerful predictor for renal outcome.

Protocol biopsies in kidney transplant recipients: histologic findings as prognostic markers for graft function and outcome.

The aim of the present study was to identify subclinical and borderline rejections as well as histological markers of chronic allograft nephropathy (CAN) among protocol biopsies performed at 1 and 6 months after living related kidney transplantation to assess their possible implications for graft function. Twenty paired allograft biopsies performed at 1 and 6 months were reviewed according to the Banff scoring scheme. The mean ages of donors and recipients were 59.6 +/- 13.8 and 34.4 +/- 8.7 years, respectively. Among all biopsies only 10% (4/40) showed no histopathological lesions. At the first month borderline rejection was shown in 35% and subclinical rejection in 10% of patients. At 6 months the proportion of findings was even higher, namely, 40% and 30%, respectively. When divided according to donor age, donors above 55 years showed a mean CAN score of 2.33 +/- 1.56 which increased to 5.0 +/- 2.26 on the 6 month biopsy (214.3%). Unexpectedly, the proportion of these changes in the younger donor group also increased by 173.3%, which might have been explained by the greater number of borderline and subclinical rejections in the younger donor group at the 1 month biopsy. In conclusion, 1 month biopsy may be valuable to determine borderline and subclinical rejection and to prognosticate the outcome of renal allograft function. Our findings suggest a greater susceptibility of histological deterioration among the older donor population. However, the presence of an untreated rejection in the younger donor pool leads to a rapid impairment of the graft function accelerating the process of chronic allograft nephropathy.

Membranoproliferative glomerulonephritis complicating diabetic nephropathy.

BACKGROUND: Renal diseases other than diabetic nephropathy can be found in diabetic patients who have undergone renal biopsy. Various forms of primary and secondary glomerular diseases were reported, but membranoproliferative glomerulonephritis was rare. METHODS: Analyzing data at our Department for the past three years, we noted 18 patients with primary membranoproliferative glomerulonephritis and 4 associated with diabetic nephropathy. RESULTS: Nodular glomerulosclerosis with diffuse membranoproliferative glomerulonephritis was registered in two patients and a diffuse form of diabetic nephropathy with a combination of segmental and diffuse changes characteristic of membranoproliferative glomerulonephritis in the other two patients. CONCLUSIONS: Analyzing what can be common for these two diseases we can conclude that they are at least three disorders: 1. hyperperfusion injury, hallmark for the diabetic nephropathy, but also with the highest incidence in membranoproliferative glomerulonephritis than in the other glomerulonephritides; 2. mesangial matrix expansion, and; 3. thickening of all extracellular membranes.

Recurrent glomerulonephritis in living kidney transplantation.

Glomerulonephritis (GN) is one of the most frequent causes of end-stage renal disease. Recurrent GN can occur very early after transplantation in up to 20% of renal-allograft recipients and should be considered with late graft dysfunction in 2-5%. Importantly, diagnosis of a clinically silent recurrence of the disease will pass undetected unless transplant centers have a policy of protocol biopsies. In addition, the classification of the type of recurrent GN should be done with data on electron microscopy and immunofluorescence, in order to promote prompt treatment and a strategy for long-term graft survival. The aim of our paper was to present a few typical cases of recurrent GN, showing the actuality of the problem in living related kidney transplant recipients and to ascertain the importance of precise and timely diagnosis by protocol biopsy. Recurrent focal segmental glomerular sclerosis (FSGS) in childhood is associated with the highest number of graft loss. The treatment of recurrent FSGN is difficult, so prophylactic plasmapheresis prior to transplantation appeared to be more effective in preventing recurrence than plasmapheresis after transplantation, especially in population of children. Mesangio proliferative GN type II is the second most frequent recurrent GN, followed by type I. Here, it is of paramount importance to classify the type of the disease. The family of the patient at risk for recurrent GN, a candidate for living related kidney transplantation, should be informed for the expected outcome and their voluntary decision whether to proceed with transplantation should be awaited.

Plasmaexchange in the treatment of myasthenia gravis associated with thymoma.

BACKGROUND: All patients with thymomatous Myasthenia Gravis (MG) should undergo early and total thymectomy, but controversy abounds in the choice of chronic immunosuppressive agents. The value of plasmaexchange (PE) in MG has been clearly established in preoperative preparation and treatment of myasthenic crisis. Whether PE may be used in the chronic long-term therapy of patients with thymomatous MG in addition to conventional immunosuppressive agents and cholinesterase inhibitors is yet to be answered. CASE HISTORY: We present a 40-year old woman with an 11 year history of MG. Thymectomy was done during the first year of the disease and the histopathologic finding was thymoma. To sustain clinical remission after thymectomy she continued with immunosuppression with methylprednisolone and cyclosporin A (or azathioprine) in addition to cholinesterase inhibitors. Despite the almost continuous immunosuppression, the disease course continued with fluctuating myasthenic weakness which few times progressed to myasthenic crisis requiring mechanical ventilation. During myasthenic crisis we performed 6-8 plasmapheresis at 2-3 day intervals in addition to conventional immunosuppressive therapy. The disease rapidly worsened in January 2000 and we started with intermittent plasmapheresis (3-6 procedures at 2-3 day intervals, every 6-8 weeks) in order to sustain remission. With this therapeutic protocol, during 20 months follow-up we managed to prevent myasthenic crisis and to avoid ventilatory support. CONCLUSIONS: Plasmaexchange could be used as a successful and safe therapeutic tool in chronic long-term therapy in addition to conventional immunosuppressive agents to sustain remission in patients with MG. This is particularly important in the treatment of patients with thymomatous MG because they have an increased frequency of myasthenic crisis and often respond poorly an to immunosuppression with steroids or other immunosuppressants.

Diabetic glomerulosclerosis without overt diabetes mellitus.

The duration of diabetes mellitus and the presence of hyperglycaemia were thought to be important in the development of nodular glomerulosclerosis. We report on 3 patients without overt diabetes, but with glucose intolerance, all male, aged 43-66 years, with histological features of diabetic glomerulosclerosis. The nodular form was present in 2 patients and the diffuse form in 1. These cases suggest that factor(s) other than hyperglycaemia are responsible for diabetic renal damage.