Ebru Kizilkilic

Conventional and molecular cytogenetic findings of myelodysplastic syndrome patients.

AbstractMyelodysplastic syndrome (MDS) involves myeloid cells of the bone marrow, which is important in progressive bone marrow insufficiency. Of all MDS patients, 40%–50% have at least one chromosomal rearrangement. Loss of specific chromosomal regions like 5q– and 7q– are usually the secondary cytogenetic abnormalities associated with MDS. In order to detect chromosome abnormalities associated with MDS, bone marrow samples from 26 patients diagnosed as MDS were obtained prior to chemotherapy. Both conventional cytogenetic analyses and fluorescence in situ hybridisation (FISH) methods were performed and locus–specific probes for 5q and 7q were used. Results obtained were compared. Twenty–one patients had normal karyotypes and four patients had abnormal karyotypes, while in one patient we could not obtain metaphases from cultures. Three patients with normal karyotypes revealed del (5q), two patients had del (7q) and one patient had monosomy (7). A total of 10 of 26 patients had chromosome changes visualised by either conventional or molecular cytogenetics (~38.5%). Our results show that both methods are important in diagnosis and follow up of MDS patients. When used together, conventional cytogenetics and FISH detect clinically significant chromosome abnormalities in MDS patients.

Detecting methylation patterns of p16, MGMT, DAPK and E-cadherin genes in multiple myeloma patients.

Multiple myeloma (MM) is a B‐cell neoplasia characterized by the clonal proliferation of plasma cells. Besides known genetic abnormalities, epigenetic changes are also known to effect MM pathogenesis. DNA methylation is an epigenetic mechanism that silences genes by adding methyl groups to cytosine‐guanine dinucleotides at the promoter regions. In this study, the methylation status of four genes; p16, O6‐methyl guanine DNA methyl transferase (MGMT), death‐associated protein kinase (DAPK) and E‐cadherin (ECAD); at the time of diagnosis was investigated using methylation‐specific polymerase chain reaction (MS‐PCR). In the 20 cases studied; methylation of the promoter regions of p16, MGMT, DAPK and ECAD genes was detected in 10%, 40%, 10% and 45% of the cases, respectively. In 65% (13/20) of cases, at least one of the genes studied had promoter methylation; while 35% of cases (7/20) had methylated promoters of more than one gene. There was a significant correlation between promoter hypermethylation of MGMT and the presence of extramedullary involvement; but for the other genes no correlation was found regarding disease properties like age, disease stage, clinical course and the presence of lytic bone lesions. Determining the methylation profiles of genes in MM, could lead to a new understanding of the disease pathogenesis and guide the assessment of treatment options.

Fluorescent in situ hybridization studies in multiple myeloma.

Abstract Conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) results of bone marrow samples of 36 multiple myeloma (MM) patients at the time of diagnosis have been evaluated. Three probes for chromosome 13q (RB1, D13S319, D13S25), one for 14q32 (IgH) and one for 17p13 (p53) have been used for hybridization with fixed cells. Twenty patients (55·5%) had normal karyotypes, whereas eight (22·2%) had numerical or structural chromosomal abnormalities. We did not find metaphases for chromosome analysis in eight (22·2%) patients. Fluorescence in situ hybridization analyses revealed at least one or more abnormal results in 25 (69·5%) cases, whereas 11(30·5%) cases had no abnormal findings. 14q32 rearrangement was the most common finding in FISH analyses and has been detected in 21 cases (58·3%). 13q deletion and 17p deletion have been detected in 11 (30·5%) and 5 (13·9%) cases, respectively. Fluorescence in situ hybridization studies including 14q32 and 17p13 chromosome regions may yield quite significant results during clinical follow-up of MM.

Three Cases of Serious Infection Caused by Aerococcus urinae: A Patient with Spontaneous Bacterial Peritonitis and Two Patients with Bacteremia

Aerococcus urinae (A. urinae) is an unusual urinary tract pathogen [1–3], which is also reported in case of endocarditis [1, 3–5], septicemia [6–9], balanitis and phlegmon [1], lymphadenitis [10], and spondylodiscitis [11] in elderly patients with local or systemic predisposing conditions such as neutropenia and prostatic diseases. A. urinae is a grampositive, catalase-negative, microaerophilic, alpha-hemolytic coccus, growing predominately in tetrads and clusters. A. urinae is usually susceptible to b-lactam antibiotics and resistant to sulfonamides and aminoglycosides. Susceptibility to trimethoprim and cotrimoxazole is variable [9]. This paper presents three case reports of the serious A. urinae infections, two associated with bacteremia and to our knowledge the first case recorded of spontaneous bacterial peritonitis (SBP).

Cranial involvement in sickle cell disease

PURPOSE To evaluate cranial findings in patients with neurologically symptomatic sickle cell disease (SCD). MATERIALS AND METHODS We studied 50 consecutive patients with SCD and neurologic symptoms. All patients underwent brain MR examinations: all 50 underwent classic MR imaging; 42, diffusion-weighted MR imaging; 10, MR angiography; four, MR venography; and three patients, digital subtraction angiography. RESULTS Of the 50 SCD patients, 19 (38%) had normal MR findings, and 31 (62%) showed abnormalities on brain MR images. Of the 50 patients, 16 (32%) had ischemic lesions; two (4%), subarachnoid hemorrhage; one (2%), moya-moya pattern; one (2%), posterior reversible encephalopathy; one (2%), dural venous sinus thrombosis; 12 (24%), low marrow signal intensity and thickness of the diploic space; 12 (24%), cerebral atrophy; and two (4%), osteomyelitis. Twenty-seven patients (54%) presented with headache, which was the most common clinical finding. CONCLUSIONS The cranial involvement is one of the most devastating complications of SCD. Early and accurate diagnosis is important in the management of cranial complications of SCD.

The apoptosis of blood polymorphonuclear leukocytes in sickle cell disease

The apoptosis of human polymorphonuclear leukocytes (PMNs) in patients with sickle cell disease (SCD) is not well understood. The goal of this study was to examine the apoptosis of PMNs in patients with SCD and in controls.

Bone marrow necrosis: a rare complication of herbal treatment with Hypericum perforatum (St. John's wort).

A 22-year-old man presented with fever and ulceration of the oral mucosa. The patient had pharyngeal and esophageal candidiasis. For the 3 weeks prior to presentation, he had been taking approximately 1000 mg/day of flowering herbs (Hypericum perforatum L, known as St. Johns wort) for treatment of depression. A complete blood count on the first day of hospitalization showed agranulocytosis and normocytic anemia. His condition worsened, and he developed progressive dysphagia. A bone marrow biopsy on day 3 revealed bone marrow necrosis. After the diagnosis was established (day 3 of hospitalization), treatment with granulocyte colony-stimulating factor 48 U/day, intravenous immunoglobulin 400 mg/kg, and amphotericin B 100 mg/day was initiated. The patient did not respond, and died within one week of the diagnosis. This cases suggests that Hypericum species may lead to severe hematologic toxicity, with conditions involving bone marrow necrosis.

Cytogenetic findings and clinical outcomes of adult acute myeloid leukaemia patients

The role of cytogenetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukaemia (AML) has become more valuable by the day. In this study, the results of conventional and molecular cytogenetic analyses and clinical outcomes of 66 AML patients of different subgroups aged between 16 and 82 were evaluated. Chromosomal abnormalities were detected in 17 (25.7%) patients cytogenetically at the time of diagnosis, whereas molecular cytogenetic abnormalities were detected in 21 (31.8%) patients by fluorescence in situ hybridisation (FISH). Thirty-eight (57.6%) patients had a normal karyotype. In 8 patients, we did not obtain suitable chromosomes for karyotype analysis and in 3 patients conventional cytogenetics were not requested by the physician. During clinical follow-up, 21 patients (31.8%) achieved complete remission (CR), 2 had partial remission (PR) (3.0%) and 4 patients had progressive disease (6.06%). Twenty-eight (42.4%) patients died during treatment and no follow-up data were available in 7 cases. Among patients with chromosome abnormalities detected by either one of the two cytogenetic methods (n=28), 12 had achieved CR, 9 of whom were already categorised in the good prognostic group with t(15;17), inv16 or t(8;21). As for the normal karyotype, each patient displayed a different clinical course, which is probably due to the molecular changes in leukaemia-related genes. Here we report our findings, which correlate with previous reports and conclude that cytogenetics is a crucial marker in leukaemia diagnosis and conventional and molecular cytogenetics should be performed as well as molecular genetic diagnostic methods.

Serum cancer antigen 15-3 concentrations in patients with sickle cell disease

Cancer antigen (CA) 15-3, which is associated with breast carcinoma, is an epithelial mucin and a product of the MUC1 gene (MUC1) (Hayes et al, 1986). Elevated levels of CA 15-3 also occur in patients with benign conditions such as inflammatory disease (Colomer et al, 1989; Valerio Marzano et al, 1998). Sickle cell disease (SCD) is an inflammatory disorder characterised by chronic haemolysis, endothelial activation and vaso-occlusion (Hebbel et al, 2004). Recent evidence showed that MUC1 was also expressed in the haematopoietic lineages (Rughetti et al, 2003), which prompted us to examine the possible association of CA 15-3 with SCD. Fifty-one patients with homozygous SCD, 15 patients with previously treated active metastatic breast carcinoma, and 20 healthy volunteers, sex and age-matched, were enrolled in the study. The SCD patients were divided into two subgroups: individuals with painful vaso-occlusive crises (n 1⁄4 17) and those with steady-state disease (n 1⁄4 34). Patients with active infection, cirrhosis, severe organ dysfunction, cancer, pregnancy or fibrocystic breast disease were excluded from the study. All subjects provided informed consent. Serum CA 15-3 levels were measured with a microparticle immunoassay. Differences were analysed by the Student’s t-test, analysis of variance, and the chi-squared test for statistical analysis. Two patients with steady-state SCD had experienced a prior cerebrovascular event. No difference was found among patients with or without painful crisis with respect to transfusion requirement, bone necrosis, hepatomegaly, seropositivity for hepatitis B or hepatitis C virus or the median value of alanine aminotransferase, bilirubin or creatinine peak levels (P > 0Æ05). All patients had been pretreated with an avarage of three substances (usually hydroxycarbamide, zinc and folic acid). There was no difference between the groups regarding the treatment with hydroxycarbamide (P > 0Æ05). In both SCD groups and the metastatic breast cancer group, serum CA 15-3 concentrations were elevated when compared with controls (P < 0Æ001 for all). The subjects with metastatic breast cancer were found to have significantly higher serum CA 15-3 concentrations when compared with the patients with SCD (steady-state disease or painful crisis) (P < 0Æ001 for both). There was no difference between the serum CA 15-3 levels of the patients with SCD in steady state or painful crisis (P > 0Æ05) (Table I). Twenty-eight patients (82%) with steady-state SCD and 13 patients (76%) with SCD in painful crisis had CA 15-3 values >30 U/ml. This difference was not statistically significant (P > 0Æ05). When both groups of patients with SCD were compared with controls, the percentage of CA 15-3 levels ‡30 IU/l was significantly higher than patients with SCD (P < 0Æ001 for both). However, this percentage in the SCD groups was lower than patients with breast cancer (P < 0Æ001 for both) (Table I). Among the patients with SCD, serum CA 15-3 levels were not affected by the use of hydroxycarbamide. Possible explanations of our findings, based on the current literature are: (i) in the past two decades, it has been established that SCD is an inflammatory state with abnormal endothelial cell activation (Hebbel et al, 2004). Previous reports have shown that elevated CA 15-3 levels are associated with connective tissue diseases, such as systemic lupus erythematosus (SLE), and systemic sclerosis (Colomer et al, 1989; Valerio Marzano et al, 1998). One study showed that 7% of patients with SLE had serum CA 15-3 concentrations >40 U/ml (Colomer et al, 1989). Pathogenetic mechanisms that cause the elevation of CA 15-3 in patients with an inflammatory disease can also explain the elevation of CA 15-3 in both SCD groups in our study. (ii) In bone marrow differentiating cells, the increase in MUC1 expression

Thrombocytosis and small bowel perforation: unusual presentation of abdominopelvic actinomycosis.

Intrauterine devices (IUD) are frequently used as a family planning procedure in developing countries because they are easy to administer and governmental policies support their use in many countries. It is recommended that IUDs be removed or replaced after 10 years, but longer use is common, especially in developing countries. In some cases, rare infections such as pelvic inflammatory diseases, pelvic tuberculosis, or abdominopelvic actinomycosis related to IUD can develop. Pelvic actinomycosis is a rare disease and is often diagnosed incidentally during surgery. In recent years, there has been an increase in actinomycotic infections mostly due to long-term usage of IUD and forgotten intravaginal pessaries. It usually develops as an ascending infection. It is usually associated with non-specific symptoms such as lower abdominal pain, menstrual disturbances, fever, and vaginal discharge. The disease is sometimes asymptomatic. The rate of accurate preoperative diagnosis for pelvic actinomycosis is less than 10%, and symptoms and imaging studies sometimes mimic pelvic malignancy. This report details a case with abdominopelvic actinomycosis associated with an IUD presenting with highly elevated thromboctye count and small bowel perforation with abscess formation.