Derya Mutlu

Preliminary results of the first human uterus transplantation from a multiorgan donor

OBJECTIVE To describe the first-year results of the first human uterus transplantation case from a multiorgan donor. DESIGN Case study. SETTING University hospital. PATIENT(S) A 21-year-old woman with complete müllerian agenesis who had been previously operated on for vaginal reconstruction. INTERVENTION(S) Uterus transplantation procedure consisting of orthotopic replacement and fixation of the retrieved uterus, revascularization, end to site anastomoses of bilateral hypogastric arteries and veins to bilateral external iliac arteries and veins was performed. MAIN OUTCOME MEASURE(S) Resumption of menstrual cycles. RESULT(S) The patient had menarche 20 days after transplant surgery. She has had 12 menstrual cycles since the operation. CONCLUSION(S) We have described the longest-lived transplanted human uterus to date with acquirement of menstrual cycles.

The role of Bifidobacterium lactis B94 plus inulin in the treatment of acute infectious diarrhea in children

BACKGROUND/AIMS In contrast to many other studies of probiotic species, the number of publications evaluating Bifidobacterium lactis and its combinations with prebiotics as treatments for acute infectious diarrhea is limited. We investigated the synbiotic effects of B. lactis B94 plus inulin on acute infectious diarrhea. MATERIALS AND METHODS The study was conducted on children with acute diarrhea between the ages of 2 and 60 months. The patients were administered 5×1010 colony-forming units (CFU) of B. lactis B94 plus 900 mg inulin or placebo, once a day for five days. Stools were examined for Rotavirus, Adenovirus, Entamoeba histolytica, Salmonella, Shigella, Campylobacter, Clostridium difficile, Cryptosporidium, and parasites. RESULTS We examined 79 patients in the synbiotic group and 77 patients in the placebo group. The duration of diarrhea was significantly reduced in the synbiotic group in comparison with the placebo group (3.9±1.2 days vs. 5.2±1.3 days, respectively; p<0.001). Moreover, the number of diarrheal stools on the third day was significantly lower in the synbiotic group than in the placebo group (5.5±2.9 vs. 8.3±3.01, respectively; p<0.001). Diarrhea in the synbiotic-group patients with rotavirus infection was of a significantly shorter duration (3.2±1.3 days vs. 5.2±1.3 days, respectively; p=0.001). Duration of diarrhea in patients who started the synbiotic treatment within the first 24 h was shorter than that in the patients who started the treatment later (3.9±1.1 days vs. 4.8±1.8 days, respectively; p=0.002). CONCLUSION Treatment with 5 × 1010 CFU of B. lactis B94 plus 900 mg inulin shortened the duration of acute watery diarrhea by an average of 31 h. This decrease was most pronounced in cases of Rotavirus diarrhea.

Superovulation alters embryonic poly(A)-binding protein (Epab) and poly(A)-binding protein, cytoplasmic 1 (Pabpc1) gene expression in mouse oocytes and early embryos

Embryonic poly(A)-binding protein (EPAB) and poly(A)-binding protein, cytoplasmic 1 (PABPC1) play critical roles in translational regulation of stored maternal mRNAs required for proper oocyte maturation and early embryo development in mammals. Superovulation is a commonly used technique to obtain a great number of oocytes in the same developmental stages in assisted reproductive technology (ART) and in clinical or experimental animal studies. Previous studies have convincingly indicated that superovulation alone can cause impaired oocyte maturation, delayed embryo development, decreased implantation rate and increased postimplantation loss. Although how superovulation results in these disturbances has not been clearly addressed yet, putative changes in genes related to oocyte and early embryo development seem to be potential risk factors. Thus, the aim of the present study was to determine the effect of superovulation on Epab and Pabpc1 gene expression. To this end, low- (5IU) and high-dose (10IU) pregnant mares serum gonadotropin (PMSG) and human chorionic gonadotrophin (hCG) were administered to female mice to induce superovulation, with naturally cycling female mice serving as controls. Epab and Pabpc1 gene expression in germinal vesicle (GV) stage oocytes, MII oocytes and 1- and 2-cell embryos collected from each group were quantified using quantitative reverse transcription-polymerase chain reaction. Superovulation with low or high doses of gonadotropins significantly altered Epab and Pabpc1 mRNA levels in GV oocytes, MII oocytes and 1- and 2-cell embryos compared with their respective controls (P<0.05). These changes most likely lead to variations in expression of EPAB- and PABPC1-regulated genes, which may adversely influence the quality of oocytes and early embryos retrieved using superovulation.

Comparison of four methods for rapid identification of Staphylococcus aureus directly from BACTEC 9240 blood culture system

PURPOSE Differentiation of Staphylococcus aureus (S. aureus) from coagulase-negative staphylococci is very important in blood stream infections. Identification of S. aureus and coagulase-negative staphylococci (CoNS) from blood cultures takes generally 18-24 h after positive signaling on continuously monitored automated blood culture system. In this study, we evaluated the performance of tube coagulase test (TCT), slide agglutination test (Dry Spot Staphytect Plus), conventional polymerase chain reaction (PCR) and LightCycler Staphylococcus MGrade kit directly from blood culture bottles to achieve rapid identification of S. aureus by using the BACTEC 9240 blood culture system. MATERIALS AND METHODS A total of 129 BACTEC 9240 bottles growing gram-positive cocci suggesting Staphylococci were tested directly from blood culture broths (BCBs) with TCT, Dry Spot Staphytect Plus, conventional PCR and LightCycler Staphylococcus MGrade kit for rapid identification of S. aureus. RESULTS The sensitivities of the tests were 99, 68, 99 and 100%, respectively. CONCLUSION Our results suggested that 2 h TCT was found to be simple and inexpensive method for the rapid identification of S. aureus directly from positive blood cultures.

Evaluation of Vancomycin Resistance 3 Multiplexed PCR Assay for Detection of Vancomycin-Resistant Enterococci from Rectal Swabs

Background Active screening for vancomycin-resistant enterococci (VRE) using rectal specimens is recommended to limit the spread of antimicrobial resistance within certain high-risk populations. We evaluated the diagnostic performance of Vancomycin Resistance 3 Multiplexed Tandem PCR assay (AusDiagnostics, Australia), a rapid multiplex real-time PCR assay that detects vanA and/or vanB. Methods Two-hundred-and-eleven rectal swabs from Hematology and Oncology unit were submitted for VRE surveillance via direct detection of vanA and/or vanB by culture and by using Vancomycin Resistance 3 Multiplexed Tandem PCR assay. Enterococci were identified to the species level by using standard biochemical tests and BD Phoenix Automated Microbiology System (BD Diagnostic Systems, USA). Vancomycin susceptibility of enterococci was determined using Etest (BioMerieux, France). Results Compared to the culture method, Vancomycin Resistance 3 Multiplexed Tandem PCR assay had a sensitivity of 84.0%, specificity of 98.8%, positive predictive value (PPV) of 91.3%, and negative predictive value (NPV) of 97.6%. The assay failed to detect 18 (8.5%) specimens because of the presence of PCR inhibitors; of the remaining 193 specimens, 25 (12.9%) were positive, 23 for vanA, and 2 for vanB. Although both sensitivity and specificity for vanA VRE was 100% compared to the culture method, all vanB-positive specimens tested negative by VRE culture. Conclusions Vancomycin Resistance 3 Multiplexed Tandem PCR assay is a rapid and laborsaving option for VRE surveillance for direct use on rectal swabs. However, the high rate of PCR failure owing to the inhibitors in the specimens and the low specificity for vanB should be considered when interpreting the results.

Confirmation of anti-DFS70 antibodies is needed in routine clinical samples with DFS staining pattern

Background Recognition of nuclear dense fine speckled (DFS) pattern by indirect immunofluorescence (IIF) is not easy. Thus, confirming the presence of these antibodies might be needed. In this study, we aimed to determine the frequency of DFS pattern in our diagnostic laboratory and to investigate the presence of anti-DFS70 antibodies in samples showing DFS pattern by two commercially available research kits retrospectively. Material and methods Seventy-four sequential serum samples with DFS pattern on HEp2010 cell substrates by IIF were included in this study. The semiquantitative DFS70 ELISA Kit (MBL International Corporation, Woburn, UK) was used for detection of anti-DFS70 antibodies in these samples. Twenty selected samples were tested for the presence of anti-DFS70 antibodies using ANA Line Immunoassay (LIA) (Immco Diagnostics, New York, USA). Results Sixty-two (83.8%) of 74 serum samples were found positive with ELISA, when 15 U/ml was taken as a reference value. Among 18 samples that were found positive by ELISA, five were negative for anti-DFS70 antibodies by LIA, while 13 were found positive. The lowest ELISA result of the sample that was positive by LIA was found to be 45.3 U/ml. When 45.3 U/ml was considered as a reference value, 45 (60.8%) of 74 serum samples were positive by ELISA. Nineteen of 20 patients had no SARD, while one had systemic lupus erythematosus (SLE). Conclusions DFS pattern should be confirmed with an objective method such as ELISA, LIA, or IB. We think that confirmation tests for detection of anti-DFS70 antibodies should be included in diagnostic algorithms.

Epstein-Barr virus infection in children with renal transplantation: 17 years experience at a single center

Abstract Objectives: The aim of this study was to detect the frequency, time of occurrence, management and outcome of Epstein–Barr virus (EBV) infection and related complications in pediatric renal transplant recipients. Methods: Pediatric renal allograft recipients transplanted between August 1994 and December 2011 at our hospital was evaluated retrospectively. The patients were divided into two groups; Groups 1 and 2 were composed of patients transplanted before and after November 2007, respectively, when plasma EBV DNA levels were periodically measured. Results: The study included 166 children, 89 (53.6%) boys, with a mean age of 12.2 ± 3.8 years. Prior to transplantation, 144 patients (86.7%) were EBV seropositive. Within a median follow-up period of 36 months, 11 of 22 seronegative children (50%) developed primary EBV infection. EBV reactivation was observed in 23 of 144 children (15.9%). Two patients with primary infection developed post-transplant lymphoproliferative disorder, one of whom died. Elevated serum creatinine levels or graft loss were not observed in any patient with EBV reactivation. Conclusions: EBV DNA monitoring by PCR in high-risk pediatric renal transplant recipients will provide early diagnosis and treatment of EBV infections.

Quantitative analysis of BKV-specific CD4+ T cells before and after kidney transplantation.

BACKGROUND BK virus (BKV) is the main infectious cause of renal allograft dysfunction. Although recent studies showed an inverse correlation between BKV-specific T-cell responses and viral load after transplantation, the importance of pre-transplant response in the process of virus reactivation has only been studied once. In this study, we aimed to determine whether pre-transplant CD4+ T-cell response can be used for prediction of BKV reactivation and BKV nephropathy (BKVN), by a method that can practically be used in routine patient monitoring. METHODS BKV-specific CD4+ T-cell responses of 31 kidney recipients (all from live donors) were measured by an IFN-γ-enzyme-linked-immunospot (ELISPOT) method using mixture of peptides, at day 0 and +1, +3, and +6 months posttransplant. Additionally, seven other reactivation patients as another group were also analyzed. BKV viral loads in plasma were measured by real-time polymerase chain reaction (PCR). Responses of 10 healthy people were also included as controls in the analysis. RESULTS All but one patient and all of the controls had detectable CD4+ T-cell responses. Reactivation occurred in 8 out of 31 patients. There was no significant association between pretransplant BKV-specific CD4+ T-cell responses and BKV reactivation and between BKV DNA levels and CD4+ T-cell responses. In the additional group consisting of reactivation patients, four patients who had BKVN showed negative correlation between BKV-DNA levels and BKV-specific CD4+ T-cell responses (p<0.05). One patient who developed BKVN, however, was not able to mount a similar CD4+ T-cell response to viral reactivation despite immunosuppressive reduction. CONCLUSION Even though our cohort is small, our results may suggest that pre-transplant measurement of BKV specific CD4+ T-cell response may not be necessary, and that post-transplant monitoring, particularly during reactivation, may be more helpful in the management of the infection.

Analysis of human herpes virus 6 infections with a quantitative, standardized, commercial kit in pediatric stem cell transplant recipients after transplantation

BACKGROUND AND OBJECTIVES The aim of the study was to assess the incidence and clinical relevance of active Human Herpes Virus 6 (HHV6) infections in pediatric patients after allogeneic stem cell transplantation. DESIGN AND SETTINGS Retrospective analysis of samples prospectively collected at Akdeniz University Medical Faculty Hospital, Antalya, Turkey, between May 2006 and July 2007 from 15 pediatric patients with allogeneic hematopoietic stem cell transplantation (HSCT). SUBJECTS AND METHODS A commercial quantitative real-time polymerase chain reaction kit was used to analyze plasma samples collected from 15 pediatric allogeneic HSCT recipients. RESULTS HHV6 DNA was found positive in 8 (53%) patients. HHV6 DNA levels above 1000 copies/mL were found only in 2 patients and they were also consecutively positive for HHV6 DNA. Age at transplantation, use of ATG, and receiving grafts other than HLA identical siblings increased the risk, with a statistically significant difference, of having HHV6 reactivation with levels exceeding 1000 copies/mL (P values, respectively, P=.03, .001, .025). Active HHV6 infections with HHV6 viremia levels higher than 1000 copies/mL were associated with subsequent delayed platelet engraftment (P=.001), acute graft versus host disease (P=.001), skin rash, and fever of unknown origin. CONCLUSION More than half of pediatric allogeneic HSCT patients develop active HHV6 infection, and especially in patients with high viremic loads, the infection can result in serious clinical situations. A clinically significant cutoff value for viremia seems to be necessary to predict serious clinical complications.

PIX-15 Assessment of cytomegalovirus (CMV) load in hematopoietic stem cell transplant recipients by CMV antigenemia and two different real-time PCR assays

Cytomegalovirus (CMV) infection occurs frequently after stem cell or solid organ transplantation. Preemptive therapy is a strategy for preventing CMV disease in transplant recipients and requires good markers for CMV replication and also highly predictive tests to identify patients at greatest risk for CMV disease. CMV pp65 antigenemia assay has been associated with high sensitivity and specificity for the early detection of CMV disease and adopted by many transplant centers for guiding preemptive therapy in transplant recipients. However this assay has some limitations; requires a sufficient number of leukocytes and rapid sample processing and experienced personel for interpretation of slides.The measurement of CMV load by quantitative polymerase chain reaction (PCR) appears to be important for the diagnosis and prediction of CMV disease. A number of laboratory developed real time PCR (RT-PCR) assays have been evaluated for monitoring CMV infection in stem cell transplant recipients. These assays, however, are not well standardized and use different target sequences, primer sets, and extraction, and detection methods, which result in different analytical performance. This makes it difficult to compare studies conducted at different laboratories. In many stem cell transplantation center, preemptive therapy is initiated upon a positive antigenemia result (≥1/2x105 leukocytes). In our transplantation center this cutoff value is defined to be ≥10 positive cells/ 2x105 leukocytes (1). In RT-PCR assays, cutoff levels which predict CMV disease, have still to be defined. It is necessary, particularly when laboratories consider replacement of the antigen assay. The aims of the study were to compare the results of two commercial CMV RT-PCR assays with pp65 antigenemia results and define cutoff levels for RT-PCR assays according to antigenemia results as a gold standart for preemptive therapy.