Single nucleotide polymorphisms of the inflamatory cytokine genes: interleukin-1B, tumor necrosis factors-A and tumor necrosis factor-B in adult patients with immune thrombocytopenia.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by thrombocytopenia due to platelet autoantibodies, causing an accelerated clearance of opsonized platelets by phagocytes. The etiology of ITP remains unclear, both genetic and environmental factors may have a role in the disease development. The aim of our study was to investigate a possible association of three single nucleotide polymorphisms (SNP) in the genes for interleukin beta (IL1B-511C/T), tumor necrosis factor beta (TNF+252G/A) and tumor necrosis factor alpha (TNFA-308G/A) with ITP. We have analyzed 125 adult patients with ITP and 120 healthy matched controls. Genotyping was performed by using PCR- RFLP methods. Our results demonstrated significantly different genotype distributions and allele frequencies for TNFB+252G/A in patients with ITP, p = 0.005 and p = 0.009 with Yates correction. We did not find any significant differences in the genotype distribution or allele frequencies for the other two genes. We have found significantly different genotype distribution and allele frequencies for TNFA-308G/A between patients with unresponsive and responsive ITP patients, p = 0.016 and p = 0.009. There were no significant differences in genotype distribution and allele frequencies for ILB-511C/T and TNFB+252G/A polymorphisms between those two groups of patients. We did not find any significant differences in genotype distribution and allele frequencies for all three polymorphisms between splenectomized and unsplenectomized ITP patients. The obtained data indicate that the A allele of TNFB+252G/A is more frequent in these patients than in the controls and that this polymorphism may play a significant role in disease susceptibility. The A allele of TNFA-308G/A was more frequent in patients with unresponsive ITP, indicating that this gene polymorphisms may contribute to therapy resistance.

Differential cAMP levels and serotonin effects in blood peripheral mononuclear cells and lymphocytes from major depression patients.

cAMP regulates immune responses, and modifications in cAMP signaling are involved in the pathophysiology and treatment of depression. In the present report, basal and forskolin-stimulated levels of cAMP were determined in mononuclear cells and lymphocytes from control individuals and major depression patients. Twenty-eight patients between 24 and 59 years old were diagnosed for a major depression episode according to the criteria of the Structural Clinical Interview for Disorders of Axis I of the American Psychiatric Association. These patients presented a score of 25 for severity as measured by Hamilton Rating Scale of Depression (HAM-D), and 23 for Beck Inventory of Depression (BID). Control and patient mononuclear cells were isolated by Ficoll/Hypaque gradients and their lymphocytes were separated from the total mononuclear population by differential adhesion to plastic surface. The basal concentration of cAMP was 50% lower in mononuclear cells and lymphocytes from the depressed patients compared with the control subjects. The response to forskolin was significantly smaller in lymphocytes of major depression patients than in the controls, but no difference was evident in the mononuclear cell preparations. There was a significant increase in cAMP produced by 5HT in mononuclear cells from the control group, but not in their lymphocytes. This effect on mononuclear cells was reduced by the antagonist of 5HT1A receptors, WAY-100,135. However, the simultaneous addition of a specific agonist of 5HT1A receptors, 8-hydroxy-(dipropylamino)tetralin (DPAT) and WAY-100,135 resulted in higher levels of cAMP than with the agonist alone. This effect probably indicates the blockade of 5HT1A receptors and action of 5HT1A agonist on the other subtypes of serotonin receptors expressed on human lymphocytes. This response was not observed in the patient's lymphocytes. In lymphocytes from major depression patients, serotonin and 8-hydroxy-(dipropylamino)tetralin significantly increased cAmp levels, which was slightly reduced by WAY-100,135. The present report indicates: (1) differential responses of immune cells from control individuals and depressed patients, with lower apparent adenylate cyclase activity in patient's cells; (2) variation in the population of cells, with responses to serotonergic agonists being lower in mononuclear cells and higher in lymphocytes from major depression patients; (3) increases of cAMP levels by serotonin and 5HT1A agonist in the patient's cells; and (4) evidence of impairment in serotonergic transduction systems in immune cells during depression.

Correlation between ultraviolet radiation level and the incidence of late-onset corneal haze after photorefractive keratectomy.

PURPOSE: To investigate the correlation between environmental changes in ultraviolet (UV) radiation levels and the incidence of late-onset cornea haze (LOCH) after photorefractive keratectomy (PRK). SETTING: SynsLaser Clinic, Tromsø, Norway. METHODS: The study comprised 404 eyes that had myopic PRK and photoastigmatic refractive keratectomy from February 1996 through July 1998. The high latitude (70 degrees N) of the observation site provided "natural laboratory" conditions to look at the occurrence of LOCH with high and low UV-radiation levels, which occurred during summers and winters, respectively. The diagnostic criterion for LOCH was acute haze of grade > or =2 occurring between 4 and 12 months postoperatively. RESULTS: The follow-up ranged from 12 to 41 months. Of the 314 eyes that met the inclusion criteria, 11 developed LOCH when the environmental UV-radiation level was high. No eye developed LOCH when the level was low. The correlation between a high level of environmental UV radiation and the occurrence of LOCH was statistically significant (P =.001). CONCLUSION: Environments with high UV-radiation levels may increase the risk of LOCH after PRK in eyes with moderate to high myopia. Use of UV-protective eyewear should be encouraged during the first year after PRK.

200 Hz flying-spot technology of the LaserSight LSX excimer laser in the treatment of myopic astigmatism: six and 12 month outcomes of laser in situ keratomileusis and photorefractive keratectomy.

PURPOSE: To evaluate safety, efficacy, predictability, and stability in the treatment of myopic astigmatism with laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) using the 200 Hz flying-spot technology of the LaserSight LSX excimer laser. SETTING: SynsLaser Clinic, Tromsø, Norway. METHODS: This retrospective study included 110 eyes treated with LASIK and 87 eyes treated with PRK that were available for evaluation at 6 and 12 months, respectively. The mean preoperative spherical equivalent (SE) was -5.35 diopters (D) +/- 2.50 (SD) (range -1.13 to -11.88 D) in the LASIK eyes and -4.72 +/- 2.82 D (range -1.00 to -15.50 D) in the PRK eyes. The treated cylinder was 4.00 D in both groups. Eleven (8.5%) LASIK eyes and 8 (7.4%) PRK eyes had secondary surgical procedures before 6 and 12 months, respectively, and were excluded when the 6 and 12 month outcomes were analyzed. RESULTS: None of the eyes lost 2 or more lines of best spectacle-corrected visual acuity. Seventy-seven percent of the LASIK eyes and 78% of the PRK eyes achieved an uncorrected visual acuity of 20/20 or better; 98% in both groups achieved 20/40 or better. The SE was within +/-0.5 D of the desired refraction in 83% of the LASIK eyes and 77% of the PRK eyes; it was within +/-1.0 D in 97% and 98%, respectively. The cylinder correction had a mean magnitude of error of 0.04 +/- 0.31 D (range -0.96 to +0.85 D) in the LASIK eyes and 0.02 +/- 0.37 D (range -1.44 to +0.72 D) in the PRK eyes. Refractive stability was achieved at 1 month and beyond in the LASIK eyes and at 3 months and beyond in the PRK eyes. CONCLUSION: The outcomes of this study are comparable to those achieved with lasers that use small-beam technology with a lower frequency, as well as with other types of delivery systems. They suggest that the 200 Hz technology used in the LaserSight LSX excimer laser is safe, effective, and predictable and that with LASIK and PRK the results are stable when treating low to moderate myopia and astigmatism up to 4.0 D.

Gastrointestinal malignant lymphomas in Macedonia.

The gastrointestinal tract including oral cavity is the most common location of extranodal lymphomas. In this retrospective study, the histomorphological, immunohistochemical and clinical features of 21 Macedonian cases with diagnosed malignant lymphomas were investigated. The series included 15 males and 6 females, mean age 44 (4-78) years. The most common locations were hard palate (n = 7), gastric site (n = 5), small intestinal wall (n = 2), large intestinal wall (n = 5), and lingual root (n = 2). All cases were B cell lymphomas, 23.8% of them low grade B cell lymphoma of mucosa associated lymphoid tissue, 4.7% mantle cell lymphoma, 47.6% diffuse large cell lymphoma, and 23.8% Burkitt type lymphoma. There was no T cell lymphoma. Most of the cases were positive for CD20 and CD79a. Monoclonality was confirmed by light chain restriction, except for nine cases where it failed due to poor tissue preservation. The fact that eight cases were in the clinically advanced third and fourth stage implied a conclusion that not only primary non-Hodgkin's lymphomas but also secondary lesions could invade the gastrointestinal tract. Immunohistochemical staining was helpful in differentiation between benign and malignant infiltration in low grade lymphomas, and in distinguishing diffuse large cell lymphomas from undifferentiated epithelial neoplasms.

Association of polymorphisms in human platelet antigens with idiopathic thrombocytopenic purpura in Macedonians.

Idiopathic thrombocytopenic purpura (ITP) is an autoimmune disease characterized by thrombocytopenia due to the presence of platelet autoantibodies specific for platelet membrane glycoproteins, such as GPIIb/IIIa, GPIb/IX and GPIa/IIa. These autoantibodies cause an accelerated clearance of opsonized platelets by phagocytes and inhibition of platelet production. Human platelet antigen (HPA) systems HPA-1, HPA-2, HPA-3 and HPA-5 are components of platelet GP complexes GPIIb/IIIa, GPIb/IX and GPIa/IIa. The HPA system consists of more than 12 bi-allelic antigen polymorphisms in which a base-pair substitution leads to change in an amino acid sequence of a membrane glycoprotein expressed on the platelet surface. The aim of this study was to examine the association of HPA-1, HPA-2, HPA-3 and HPA-5 polymorphisms with idiopathic thrombocytopenic purpura. We performed genotyping of HPA-1, HPA-2, HPA-3, and HPA-5 systems in 60 patients with ITP and 120 healthy participants. Genotyping of HPA-1, -2, -3, and -5 alleles were performed by PCR and RFLP methods by using specific primers and restriction enzymes. Allele and genotype frequencies of HPA-1, HPA-3, and HPA-5 were not significantly different between patients and healthy participants. After Bonferroni adjustment a significant association in ITP patients with HPA-2 alleles (P=0.015, OR=1.923, CI=1.126-3.284) was found. Allele frequencies for HPA-2a were 0.852 in healthy participants and 0.750 in patients, and for HPA-2b 0.148 and 0.250 respectively. These results suggests that HPA-2b allele was more frequent in patients with ITP and may be involved in the formation of a specific autoepitope.

Immune thrombocytopenic purpura in adults in the last 10 years: single-centre experience.

BACKGROUND: Immune thrombocytopenic purpura (ITP) is a benign disease with low morbidity and mortality and frequent remissions that occur spontaneously or in response to first-line treatment with steroids or splenectomy. AIM: The purpose of this study is to describe the clinical outcomes of 170 patients with ITP diagnosed and/or treated in our hospital between 2000 and 2010. METHODS AND RESULTS: The median age at diagnosis was 47 years. Forty three (25%) were asymptomatic, 65% had minor skin or mucosal bleeding and 10% had significant bleeding from the gastrointestinal or genitourinary system. The median platelet count at diagnosis was 13x10(9)/L (range: 0-98x10(9)/L). Median follow-up of all patients was 13 months. Ninety-five patients had a follow-up longer than 12 months, with median 44 months (range 14-384). Corticosteroids were the initial treatment for 161/170 (95%) patients, 38 (22%) were splenectomized, 25 (14.7%) were treated with intravenous gamma globulins, while 9 did not received any specific treatment. A complete response to initial treatment (prednisone±splenectomy) was achieved in 55/161 (34.2%), a partial response in 90 (55.9%) and no response in 16 (9.9%) patients. In the group of patients with follow-up longer than 1 year; 28 (29%) patients had refractory or unresponsive ITP with a median follow-up of 66 months. All patients with refractory ITP were treated with steroids, 11 were splenctomized, significantly more patients with refractory ITP 12 (43%) were treated with IVIG compared with other ITP patients (16%), p=0.005. The median age of 38 splenectomized patients was 28 years and it is significantly different from the other patients (p<0.001). There were no significant differences in other characteristics between splenctomized or refractory ITP and other patients at diagnosis. CONCLUSION: Our results were similar to results already reported in other similar studies.

Immunosupressive treatment with cyclosporin-A for a patient with hypoplastic myelodisplastic syndrome: a case report.

BACKGRAOUND: Imunosupressive therapy with antithymocyte globulin (ATG), cyclosporine (CsA) or both has been shown to induce haematological responses in a subset of patients with myelodysplastic syndromes (MDS), in particular in the hypocellular form of MDS. CASE REPORT: We report our first case with hypocellular MDS treated with CsA. A 54-year-old female referred to our Department due to weakness and severe pancytopenia. Hypocellular form of MDS was diagnosed after bone marrow biopsy. Treatment with CsA was started one year after diagnosis. Treatment with CsA resulted in clinical improvement, a very good partial haematological response, resolution of transfusion requirement and an increase in bone marrow cellularity. CONCLUSIONS: In our experience, immunosuppressive treatment with CsA and/or ATG could be an alternative for patients with hypoplastic MDS for whom there is no possibility of allogenic bone marrow transplantation as only curative therapy.

Osteonecrosis of the jaw in patients with multiple myeloma treated with bisphosphanates.

Bisphosphonates are pyrophosphate analogues which inhibit osteoclastic activity. Long term use of bisphosphonates has recently been associated with osteonecrosis of the jaw (ONJ) defined as a three month non-healing defect in the jaw. ONJ is commonly precipitated by a tooth extraction or other stomatological procedure in patients treated with long-term, potent, high dose intravenous bisphosphonates for the management of myeloma, breast or prostate cancer. The aim of this study was to evaluate the incidence of ONJ in patients with MM treated with bisphosphonates during the last 8 years in our institution and to pre-sent the first two cases. We have analysed 247 myeloma patients diagnosed in our institution in the period 2002-09. Only 190/247 patients (76.9%) were treated with bisphosphonates. The incidence of ONJ in our group of patients treated with bisphosphonates was 2/190 (1%). The most commonly used bisphosponate was i.v. pamidronate (17.8%) and 46.6% were treated with two or more types of bisphosphonates. Sixty-five patients (34.2%) received oral forms of bisphosphonates; 42.1% patients were treated with i.v. forms of pamidronate, ibondronate or clodronate, and 45 patients (23.7%) received a combination of oral and i.v. forms of bisphosphonates. The mean duration of bisphosphonates therapy was 24.7±17.7 months. The low incidence of ONJ in our institution could be explained by the rare use of zolendronate, which is the most commonly referred bisphosphonate causing ONJ, and by a relatively shorter duration of bisphosphonates treatment in patients with MM. Despite the fact that ONJ is a rare complication in our institution, preventive measures must be considered.