Peripheral blood marker of residual acute leukemia after hematopoietic cell transplantation using multi-plex digital droplet PCR.

Background: Relapse remains the primary cause of death after hematopoietic cell transplantation (HCT) for acute leukemia. The ability to identify minimal/measurable residual disease (MRD) via the blood could identify patients earlier when immunologic interventions may be more successful. We evaluated a new test that could quantify blood tumor mRNA as leukemia MRD surveillance using droplet digital PCR (ddPCR). Methods: The multiplex ddPCR assay was developed using tumor cell lines positive for the tumor associated antigens (TAA: WT1, PRAME, BIRC5), with homeostatic ABL1. On IRB-approved protocols, RNA was isolated from mononuclear cells from acute leukemia patients after HCT (n = 31 subjects; n = 91 specimens) and healthy donors (n = 20). ddPCR simultaneously quantitated mRNA expression of WT1, PRAME, BIRC5, and ABL1 and the TAA/ABL1 blood ratio was measured in patients with and without active leukemia after HCT. Results: Tumor cell lines confirmed quantitation of TAAs. In patients with active acute leukemia after HCT (MRD+ or relapse; n=19), the blood levels of WT1/ABL1, PRAME/ABL1, and BIRC5/ABL1 exceeded healthy donors (p<0.0001, p=0.0286, and p=0.0064 respectively). Active disease status was associated with TAA positivity (1+ TAA vs 0 TAA) with an odds ratio=10.67, (p=0.0070, 95% confidence interval 1.91 - 59.62). The area under the curve is 0.7544. Changes in ddPCR correlated with disease response captured on standard of care tests, accurately denoting positive or negative disease burden in 15/16 (95%). Of patients with MRD+ or relapsed leukemia after HCT, 84% were positive for at least one TAA/ABL1 in the peripheral blood. In summary, we have developed a new method for blood MRD monitoring of leukemia after HCT and present preliminary data that the TAA/ABL1 ratio may may serve as a novel surrogate biomarker for relapse of acute leukemia after HCT.

Could depression be a new branch of MIA syndrome?

The aims of the present study were to determine the prevalence of depression in our dialysis patients, to detect the most powerful variables associated with depression, and to determine the role of depression in prediction of mortality. The prospective follow-up study of 128 patients (77 HD and 51 CAPD, 65 male, aged 53.8 +/- 13.5 years, dialysis duration 64.7 +/- 64.8 months) was carried out over 36 months. Depression by the Beck Depression Inventory-BDI-II score, laboratory parameters (hemoglobin, serum albumin and creatinine concentration), immunological status (cytokines and hsCRP), comorbidity by Index of Physical Impairment (IPI) and adequacy of dialysis by Kt/V were monitored. The overall prevalence of depression in the dialysis patients (BDI score > or = 14) was 45.3%, and 28.2%, respectively, for moderate and severe depression (BDI > or = 20). The most powerful variable associated with depression was IL-6, but associations with albumin, hemoglobin, creatinine and IPI score were also found. During the follow-up period 36 patients died, 7 patients left the cohort and 2 patients were transplanted. If IPI score was not included in the multivariate Cox analysis, the BDI score remained one of the best predictors of mortality along with albumin. In conclusion, because of the close association of depression with inflammation, malnutrition, and cardiovascular mortality, it could be speculated that depression is one branch of the MIA (malnutrition, inflammation, atherosclerosis) syndrome.

Gorlin-Goltz syndrome.

Gorlin-Goltz syndrome, also known as basal cell nevus syndrome, is an uncommon, autosomal dominant inherited disorder, which is characterized by numerous basal cell carcinomas, maxillary keratocysts, and musculoskeletal malformations. Occasionally, it is associated with aggressive basal cell carcinomas and internal malignancies. Early diagnosis and treatment are essential, as well as genetic counseling. A patient with characteristic symptoms of nevoid basal cell carcinomas and a review of the literature are presented.

[The role of PCR in detection of cytomegalovirus in blood and urine of transplanted patients].

Rapid and reliable laboratory diagnosis of cytomegalovirus (CMV) infection in transplanted patients becomes more significant considering possible prevention or moderation of CMV disease. The aim of this study was comparison of CMV isolation in cell culture and CMV detection by PCR in blood and urine samples of transplanted patients. The study comprised 21 patients undergoing immunosuppressive therapy after renal or bone marrow transplantation. MRC-5 cell culture was used for viral isolation. Inoculated cultures were maintained for 4 weeks or until appearance of specific cytopathic effect (CPE). Primers used in PCR were specific for the sequence within egzon 4 of immediate-early 1 (IE-1) gene. Presence of anti-CMV IgG and IgM antibodies was determined by ELISA. PCR was more sensitive for blood (4/21) and urine samples (3/19) than cell culture method--blood (0/21) urine (2/19). Statistically significant correlation was not shown between presence of CMV(determined by PCR) in blood and urine of the same patient.

[Myomectomy by vaginal route].

Myomectomy by vaginal route is the least invasive of all methods for myoma removal. Compared to classical, abdominal myomectomy, it has numerous advantages, especially in cases of fundal and posterior wall myomas, but still it is relatively rarely performed. This study provides an analysis of operative and postoperative course of patients subjected to myomectomy by vaginal route in the period from 01/01/2003 to 01/11/2005 as well as the corresponding control group of patients which had undergone classical, abdominal myomectomy. The study points out to significance and advantages of application of surgical technique for removal of myoma through the posterior wall of the vagina as opposed to classical, abdominal myomectomy. Transvaginal myomectomy eliminates trauma of laparotomy, intraoperative blood loss is reduced, postoperative complications are down to minimum, postoperative recovery is shorter, and so is the number of days spent in hospital. Thus, by performing this type of surgery, a positive cost-benefit effect is obtained not only for the patient but for health authorities as well.

Oculo-auriculo-vertebral (Goldenhar) spectrum associated with pericentric inversion 9: coincidental findings or etiologic factor?

Oculo-auriculo-vertebral (OAV) spectrum or Goldenhar syndrome is a complex and heterogeneous condition characterized by hemifacial microsomia (unilateral ear abnormalities and ipsilateral mandibular hypoplasia) as well as vertebral anomalies and epibulbar dermoids or lipodermoids. Although most cases of OAV spectrum are sporadic, both autosomal recessive and autosomal dominant inheritance have been reported. Furthermore, the association of OAV spectrum with different types of chromosomal abnormalities has been described. We present a premature newborn delivered after 36 weeks of gestation, whose birth weight was 2,100 g, birth length 43 cm and head circumference 32.5 cm. OAV spectrum with associated axial skeleton anomalies, eventration of the right hemidiaphragm, accessory spleen, unlobulated right lung, agenesis of right kidney, right ovary and right uterine horn, and partial agenesis of corpus callosum were found. She was the second child of unrelated parents, who have a healthy boy. Both parents refused chromosomal analysis of their peripheral blood. Trypsin G-banding and C-banding chromosomal analysis of the patient's peripheral blood revealed a pericentric inversion of chromosome 9 with break points at p11 and q13. This may be a coincidental or causal finding.

[Numerical density of eosiophilic leukocytes in skin basal and squamous cell carcinomas]. / Brojna zastupljenost eozinofilinih leukocita u bazocelulnom i spinocelulnom karcinomu koze.

The autors examined the presence and determined numerical density of eosinophilic leukocytes in tumor's stormal and host reaction in twenty routine preparated skin biopsies of basal and squamos cell carcinomas. The presence of eosiophilic leukocytes in squamous cell carcinomas, in stormal and adjacent tumor infiltrate were confirmed, so as the absance on the same localizations in basal cell carcinomas were found. Eosinophilic leukocytes were of caracteristic morphological appereance, smaller of blood and bone marrow eosinophils - the average radius were 9,6 microm, with high level of degranulation, very variable in number (from 1054 to 23560 in mm3) and in close relation with plasma cells, mast cells and lymphocytes. In conclusion, these results may indicate and clear up presence of eosinophils by antigen properties of tumor cells, by activation of humoral immunity and consecutive phagogytosis of antigen-antibody complexes by eosinophils, but specially by their novel lytical effects on epithelial tumor cells in texture of cellular cytotoxic response. Beside these favorable effects of eosinophils, their granular enzymes activated and released during degranulation paradoxlly make spreading of tumor process possible and they reduce barrier role of dermal collagen.

[Retrospective analysis of senzibilisation to chrom, nickel and cobalt over the period 1980-1989 year]. / Retrospektivna analiza preosetljivosti na hrom, nikl i kobalt za period od 1980. do 1989. godine.

Over the periof from 1980-1989 year at Clinic of Dermatovenerology in Nis, 1988 patients with contact allergic dermatitis were tested with standard allergen batteries. Senzibilisation to metals - chrom, nickel and cobalt was proved in 206 (10,36%) cases, 132 (64,08%) males and 74 (35,92%) females. The most frequent allerge was potassium bichromate - 155 (7,79%) cases, and then nickel sulphate - 82 (4,12%). Positive patch test only to one metal had 148 (71,84%) patients, 44 (21,36%) to two metals and 14 (6,80%) had positive patch test to all three metals.

Ureaplasma urealyticum infection in newborns: three case reports.

We report three newborns with different manifestations of Ureaplasma urealyticum infection; a term newborn with acute neonatal pneumonia and two very low birth weight infants with bronchopulmonary dysplasia and osteomyelitis of the femur, respectively. The association of U. urealyticum with acute and chronic respiratory disease in term and preterm newborns has recently been reported. Our two cases are similar to other case reports from the literature, but we were unable to find any previous reports of osteomyelitis due to U. urealyticum in the premature babies. Isolation of U. urealyticum in pure culture from the blood was considered to be related to local infection in all three patients. All patients were cured by erythromycin.

[Inoperable carcinoma of the stomach in exploratory laparotomy]. / Inoperabilni karcinom zeluca kod eksplorativne laparotomije.

For the last few years we made 52 exploratory laparotomies. All the patients were treated clinically, through laboratories, with X-rays, and for a while ago by means of gastroscopy. A radical surgical operation was planned for each of them we made radical operation in 12 cases only, and 14 GEA. For all others we made pure exploratory laparotomy with probatory excision. Considering such inauspicious results, it is necessary to give maximum consideration to early detected gastric cancer. In a matter of critical age persons after 40, the attention must be referred to anamnestically suspect cases. For this purpose, the early detailed treatment by the latest diagnostic methods must be done. If in such treated cases there is even the smallest doubt of a malignant process, it is needful not to abstain from diagnostic exploratory laparotomy.