Limbal stem cell transplantation from HLA-compatible living donors. Long term observation

PURPOSE: The assessment of clinical outcome in patients after Ir-CLAL procedure. MATERIALS AND METHODS: Retrospective, interventional case series including 11 patients (12 eyes), all with non-ambulatory visual acuity (from light perception to hand movements). Mean age was 39.4 years (ranging from 18 to 76 years). In 7 eyes Limbal Stem Cells Deficiency (LSCD) resulted from chemical burn, in 2 eyes from ocular cicatricial pemphigoid, in one eye from Stevens-Johnson syndrome (SJS). In one eye congenital LSCD was diagnosed and in another one post-inflammatory LSCD. Mean follow-up was 16.2 months (from 10 to 32). Among donors prospective class I HLA matching was performed. Maximal two mismatches were accepted. General immunosuppressive therapy was switched-on in all subjects. Surgical excision of corneal pannus with clearing of limbal area was performed. Five clock hours stem cells tissue grafts from living related donors were harvested and transplanted to the recipient eye. Three penetrating keratoplasty, one deep lamellar keratoplasty and seven amniotic membrane transplantations were additionally performed. Main outcome measures were graft survival as a restoration of corneal epithelium, visual acuity improvement and complication in donors and recipients. Kaplan-Meyer survival curve and generalized Peto test were used for comparison. RESULTS: The graft survival was 83.6% in mean 20.3 months follow-up (from 10 to 32 months). Two cases of graft failure concern eye with SJS and eye with PKP regrafting after chemical burn. Visual improvement in 7 cases (58.3%), the same visual acuity in 4 cases (33.3%) and deterioration one case (8.3%) were noted. Ambulatory vision in 50% of patients was achieved. Graft rejection syndrome in four eyes was noticed also when ideal matching and CsA serum level 220 ng/ml. Two cases were treated with success and another two grafts failed. No complications in donors eyes were noticed. Impression cytology in eyes with successful grafts not related to pathological changes. CONCLUSIONS: Lr-CLAL are effective in ocular surface restoration. Living-related donors are suitable source of stem cells for cultivation in vitro. The prognosis for stem cells grafting is definitely worse when eyelid anomaly and adhesions exist.     

Expression of muscarinic cholinergic receptors on lymphocytes during rheumatoid arthritis

The level of muscarinic receptors on lymphocytes from rheumatoid arthritis (RA) patients in comparison with healthy individuals was determined by the binding studies of a specific muscarinic ligand [3H]-QNB in the presence of the competitive antagonist atropine. The response to phytohaemagglutinin and the influence of acetylcholine on this process in patients' lymphocytes was also studied. We have found direct correlation among PHA stimulation indices, the level of muscarinic receptors, and the influence of acetylcholine on PHA stimulation indices. Differences in these parameters occurring in systemic forms of RA as compared to other RA patients are also described and their possible mechanism discussed. The influence of cholinergic stimulation on the reactivity of lymphocytes after PHA in different forms of RA could depend on the number of muscarinic receptors present on the lymphocyte surface, as well as on the activity of soluble factors, which could modulate the functional state of the receptors.     

Does immunosuppressive regimen influence the lipid disturbances in kidney recipients?

Though it is said that some immunosuppressive agents are implicated in the development of hyperlipidemia in kidney recipients, this subject is still controversial. Main plasma lipid parameters, as well as apolipoproteins A1 and B were measured periodically in 39 kidney first cadaveric, nondiabetic recipients during 24 months of clinic follow-up after transplantation. Standard triple immunosuppressive therapy: prednisone + cyclosporine + azathioprine was administrated from the beginning. After the second year of kidney transplantation, a significant reduction refers to values of TC, LDL, apo A1 and apo B. In the group with antirejection - methylprednisolone therapy, and without it only TG in the 24th month and apo B in the 1st month were statistically lower in the latter group (both p < 0.05). In the multiple regression test, a linear coincidence was observed between apo A1, apo B and prednisone cumulative dosage after the 1st month, TG and cyclosporine in the 6th month and LDL and cyclosporine in the 12th month after transplantation. It appears that steroids had an impact on lipids directly after transplantation, while cyclosporine did so thereafter.     

FeCl3-Modified Carbonaceous Catalysts from Orange Peel for Solvent-Free Alpha-Pinene Oxidation

The work presents the synthesis of FeCl₃-modified carbonaceous catalysts obtained from waste orange peel and their application in the oxidation of alpha-pinene in solvent-free reaction conditions. The use of waste orange peel as presented here (not described in the literature) is an effective and cheap way of managing this valuable and renewable biomass. FeCl₃-modified carbonaceous materials were obtained by a two-stage method: in the first stage, activated carbon was obtained, and in the second stage, it was modified by FeCl₃ in the presence of H₃PO₄ (three different molar ratios of these two compounds were used in the studies). The obtained FeCl_3-modified carbon materials were subjected to detailed instrumental studies using the methods FT-IR (Fourier-transform Infrared Spectroscopy), XRD (X-ray Diffraction), SEM (Scanning Electron Microscope), EDXRF (Energy Dispersive X-ray Fluorescence) and XPS (X-ray Photoelectron Spectroscopy), while the textural properties of these materials were also studied, such as the specific surface area and total pore volume. Catalytic tests with the three modified activated carbons showed that the catalyst obtained with the participation of 6 M of FeCl₃ and 3 M aqueous solutions of H₃PO₄ was the most active in the oxidation of alpha-pinene. Further tests (influence of temperature, amount of catalyst, and reaction time) with this catalyst made it possible to determine the most favorable conditions for conducting oxidation on this type of catalyst, and allowed study of the kinetics of this process. The most favorable conditions for the process were: temperature of 100 °C, catalyst content of 0.5 wt% and reaction time 120 min (very mild process conditions). The conversion of the organic raw material obtained under these conditions was 40 mol%, and the selectivity of the transformation to alpha-pinene oxide reached the value of 35 mol%. In addition to the epoxy compound, other valuable products, such as verbenone and verbenol, were formed while carrying out the process.     

A trial of cyclophosphamide in ankylosing spondylitis with involvement of peripheral joints and high disease activity

In 12 patients with ankylosing spondylitis with involvement of the peripheral joints and high activity of the disease process, cyclophosphamide was given in intravenous doses of 200 mg every second day for 3 weeks, followed by oral doses of 100 mg once weekly for 3 months to a total dose of 3 300 mg. Before this treatment and after 18-24 months the following clinical parameters were determined: spinal mobility, degree of intensity of spinal and articular pains, and the number of involved joints. After the treatment an evident clinical improvement was observed, with decreased values of the laboratory indices of the disease activity. Apart moderate erythrocyturia in one case and transient leukopenia in another case, no other side effects were observed. The improvement in cases with coexistent amyloidosis was relatively small.     

Morphologic features of hepatitis C recurrence in patients after orthotopic liver transplantation-preliminary analysis of our case observations

INTRODUCTION: Hepatitis C virus (HCV) recurrence is almost universal in patients after liver transplantation. The diagnosis of reinfection is more difficult than that of a primary process, as shown by our pathomorphologic analysis of cases of HCV recurrence. MATERIAL: During 5.5 years, 240 liver biopsies included 54 obtained from liver transplant recipients with primary HCV infections, among whom 26 (56.5%) had clinical signs and symptoms of hepatitis. Nineteen patients from this population underwent 30 liver biopsies. In addition, seven biopsies were performed in five patients without clinical signs of reinfection. RESULTS: In 44.2% of patients with HCV recurrence and 15% without reinfection, the intensity of the primary process in the native livers was assessed as high. Reinfection was found in all patients with liver carcinoma and 67% with hepatocyte dysplasia. Histologic signs of infection were estimated as minimal (n = 4), mild (n = 19), or moderate (n = 4). In five patients with reinfections and one without recurrence, histologic manifestations of acute rejection were also observed. In conclusion, HCV was the indication for liver transplantation in 22.4% cases. Clinical manifestation of recurrence was found in 56.5% of the patients, who tended to be older than those without disease recurrence. Upon microscopy, lobular lesions predominated over the portal changes. Factors predisposing to HCV recurrence were coexistence of other liver disorders, a high intensity of the inflammatory process, hepatocyte dysplasia, and/or hepatocellular carcinoma in the native liver and acute rejection episodes.