Impact of mother donor, peripheral blood stem cells and measurable residual disease on outcomes after haploidentical hematopoietic cell transplantation with post-transplant cyclophosphamide in children with acute leukaemia.

Haploidentical hematopoietic-cell transplantation using post-transplant cyclophosphamide(Haplo-PTCy) is a feasible procedure in children with haematologic malignancies. However, data of a large series of children with acute leukaemia(AL) in this setting is missing. We analysed 144 AL Haplo-PTCy paediatric recipients; median age was 10 years. Patients had acute lymphoblastic(ALL; n = 86) or myeloblastic leukaemia(AML; n = 58) and were transplanted in remission(CR1: n = 40; CR2: n = 57; CR3+: n = 27) or relapse (n = 20). Bone marrow was the graft source in 57%; donors were father (54%), mother (35%), or sibling (11%). Myeloablative conditioning was used in 87%. Median follow-up was 31 months. At day +100, cumulative incidence (CI) of neutrophil recovery and acute GVHD (II-IV) were 94% and 40%, respectively. At 2-years, CI of chronic GVHD and relapse, were 31%, 40%, and estimated 2-year overall survival (OS), leukaemia-free survival (LFS) and graft-versus-host-relapse-free survival (GRFS) were 52%, 44% and 34% respectively. For patients transplanted in remission, positive measurable residual disease (MRD) prior to transplant was associated with decreased LFS (p = 0.05) and GRFS (p = 0.003) and increased risk of relapse (p = 0.02). Mother donor was associated with increased risk of chronic GVHD (p = 0.001), decreased OS (p = 0.03) and GRFS (p = 0.004). Use of PBSC was associated with increased risk of chronic GVHD (p = 0.04). In conclusion, achieving MRD negativity pre-transplant, avoiding use of mother donors and PBSC as graft source may improve outcomes of Haplo-PTCy in children with AL.

Gonadotropin-releasing hormone agonists for ovarian protection during cancer chemotherapy: systematic review and meta-analysis.

OBJECTIVE: To evaluate the effectiveness of gonadotropin-releasing hormone agonist (GnRHa) administration before and/or during cancer chemotherapy for the protection of ovarian reserve in premenopausal women without prior diagnosis of infertility. METHODS: This was a systematic review and meta-analysis of randomized controlled trials (RCTs) comparing administration of GnRHa before and/or during chemotherapy vs chemotherapy alone. Eligible participants were premenopausal women at any stage of cancer, without previous diagnosis of infertility. An electronic database search in MEDLINE, CENTRAL, LILACS and ClinicalTrials.gov was performed. After selecting eligible studies, the relative risk (RR) was assessed for primary ovarian insufficiency (POI)/amenorrhea and for spontaneous pregnancy after completion of treatment. RESULTS: Thirteen RCTs comparing concurrent use of GnRHa and chemotherapy (609 participants) with chemotherapy alone (599 participants) were eligible for meta-analysis. All trials were open-label and patients had been treated for breast cancer (n = 1099) or lymphoma (n = 109). GnRHa had a significant benefit on the risk of POI/amenorrhea (RR, 0.60; 95% CI, 0.45-0.79), which persisted in subgroup analysis for breast cancer (RR, 0.57; 95% CI, 0.43-0.77) but not for lymphoma patients (RR, 0.70; 95% CI, 0.20-2.47). The rate of spontaneous pregnancy after completion of treatment was higher in women receiving GnRHa plus chemotherapy compared with those receiving chemotherapy alone (RR, 1.43; 95% CI, 1.01-2.02). Overall, the quality of evidence was low due to the unclear risk of bias, short follow-up and lack of objective assessment of ovarian function and reserve. CONCLUSIONS: Evidence, albeit of low quality, supports the use of GnRHa before and/or during chemotherapy to reduce the risk of POI and increase the probability of spontaneous pregnancy in the short term. Further high quality RCTs with more accurate assessment of ovarian reserve are needed to support definitive recommendations for clinical practice. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Alzheimer's disease-related amyloid-ß1-42 peptide induces the loss of human sperm function.

Characteristically identified as the main component of senile plaques present in patients suffering from Alzheimer's disease, Aß has been detected in human testis and reproductive fluids, but its effect on spermatozoa has not been addressed. The present study evaluated whether the most toxic and aggregant amyloid precursor protein (APP)-proteolytic product, amyloid-ß1-42 (Aß1-42), was capable of affecting sperm functionality. Normozoospermic samples were either exposed to different Aß1-42 doses or to the untreated and scrambled controls for a maximum of 48 h at 37 °C and 5%CO2, and motility, viability and mitochondrial status were evaluated. Additionally, tyrosine phosphorylation was analyzed by immunocytochemistry and acrosomal integrity through PSA-FITC. A shorter treatment period was used to monitor prompt Ca2+ responses. Aß1-42 peptide decreased motility before inducing mitochondrial impairment (p < 0.05; n = 6). Both outcomes became more pronounced with time, reaching their maximal decrease at 48 h, where even 1 µM produced undesirable effects (p < 0.05; n = 6). Aß1-42 peptide also decreased cell survival (p < 0.05; n = 6). Furthermore, although no effects on tyrosine phosphorylation were observed (p > 0.05; n = 6), reduced acrosomal integrity was detected (p < 0.05; n = 7), which was not correlated with viability loss (p > 0.05). In parallel, all Aß1-42 concentrations elicited a [Ca2+]i rise but a significant difference was only observed at 20 µM (p < 0.05; n = 7) and a tendency was obtained with 10 µM (p = 0.053; n = 7). In conclusion, Aß1-42 peptide oligomers impair sperm function in vitro, although further studies are required to determine the clinical relevance of these findings.

Transabdominal midline reconstruction by minimally invasive surgery: technique and results.

BACKGROUND: The introduction of the minimally invasive approach changed the way abdominal surgery was carried out. Open suture and mesh reinforcement in ventral hernia repair used to be the surgeon's choice of procedure. Although the laparoscopic approach, with defect bridging and mesh fixation, has been described since 1993, the procedure remains largely unchanged. Evidence shows that defect closure and retro-muscular mesh positioning have the best outcomes and are the best surgical practice. We therefore aimed to develop and demonstrate a procedure which combined the good results of open surgery using the Rives-Stoppa principles, particularly in terms of recurrence, with all the benefits of minimally invasive surgery. METHODS: Between October 2012 and February 2014, 15 post-bariatric surgery patients underwent laparoscopic midline incisional hernia repair. The peritoneal cavity was accessed through a 5-mm optical view cannula at the superior left quadrant. A suprapubic and two right and left lower quadrant cannulas were inserted for inferior access and dissection. The defect adhesions were released. The whole midline was closed with an endoscopic linear stapler, including the defect, from the lower abdomen, 4 cm below the umbilicus, until the epigastric region, including posterior sheath mechanical suturing and cutting in the same movement. A retrorectus space was created in which a retro-muscular mesh was deployed. Fixation was done using a hernia stapler against the posterior sheath from the peritoneal cavity to the abdominal wall muscles. Selection was based on xifo-umbilical incisional midline hernias post open bariatric surgery. Pregnant women, cancer patients, or patients with clinical contraindications were excluded. RESULTS: The patients mean age was 51.2 years (range 39-67). Four patients were men and eleven women. Two had well-compensated fibromyalgia, four had diabetes, and five had hypertension. The mean BMI was 29.5 kg/m2 (range 23-31.6). Surgery was performed successfully in all cases through four ports; the number of incisional hernias was 3 ± 2, with a mean maximum width of 3.75 cm (range 2.1-9) and maximum length of 14 cm (7.5-20.5). The mean surgical time was 114.3 min (range 85-170), and the median hospital stay was 1.4 days. No intra-operative or immediate post-operative complication or death occurred. One patient had a seroma treated conservatively 1 week after surgery and another had a retro-muscular infection treated with percutaneous drainage. CT-Scans made before and after the procedure, showed total closure of the defect. QOL questionnaire showed satisfaction, acceptance, and no complaints. CONCLUSION: Although the study involved a small number of patients, it has proved the technique to be feasible, easy to perform, and have the combined benefits of laparoscopic and open surgery. The results, shown by CT-scan, peri-operative, and QOL findings, were good.

Identificação de loci microssatélites com potencial de amplificação na espécie de Peixe-Rei (Odontesthes humensis) / Identification of microsatellite loci with amplification potential in "pejerrey" (Odontesthes humensis)

In this work, 25,806 potentially amplifiable microsatellite loci (PAL) were identified in pejerrey, (Odontesthes humensis), with 21% of dinucleotide, 22% trinucleotide, 37% tetranucleotide, 13% pentanucleotide and 7% hexanucleotide. Of the total loci, 167 were classified as "Best PAL", more likely to be variables in populations. The results show that with a small coverage of the genome it was possible to identify a large number of microsatellite loci...

Benzocaine and clove oil as anesthetics for pejerrey (Odontesthes bonariensis) fingerlings / Benzocaina e óleo de cravo como anestésicos para alevinos de peixe-rei (Odontesthes bonariensis)

Pejerrey (Odontesthes bonariensis) is a native species from Rio Grande do Sul, Uruguay and Argentina where it is of great economic importance for artisanal fishing. One difficulty in laboratory research with pejerrey is related to its sensitivity, as it presents higher basal cortisol levels than other freshwater species. For this reason, the aim of this work was to evaluate the efficiency of benzocaine and clove oil as anesthetics for pejerrey fingerlings. Two experiments were done where fingerlings (57±7.8mm and 1.1±0.44g) were exposed to benzocaine with concentrations between 40mgL-1 and 120mgL-1 and to clove oil with concentrations between 12mgL-1 and 75mgL-1. Survival, anesthesia induction time and recovery time for each pharmaceutics were evaluated. Both benzocaine and clove oil pharmaceutics showed efficiency as anesthetics for pejerrey fingerlings, with negative correlation between the dose of anesthetics and the anesthesia induction time. For benzocaine, the concentrations between 80mgL-1 and 100mgL-1 showed better results, as for clove oil the optimal concentrations were between 25mgL-1 and 50mgL-1. On the other hand, the anesthesia recovery time did not present significant variation on the different concentrations of the tested products. The tested products are highly metabolizable by pejerrey.

O peixe-rei (Odontesthes bonariensis) é uma espécie nativa do Rio Grande do Sul, Uruguai e Argentina, onde tem grande importância econômica para a pesca artesanal. Uma dificuldade da pesquisa em laboratório com peixe-rei está relacionada à sua sensibilidade, pois apresenta níveis basais de cortisol mais elevados que outras espécies de água doce. Este trabalho avaliou a eficiência da benzocaína e do óleo de cravo como anestésicos para alevinos de peixe-rei. Foram realizados dois experimentos em que alevinos (57±7,08mm e 1,1±0,44g) foram expostos à concentração entre 40mg-1 e 120mgL-1 de benzocaína e entre 12mgL-1 e 75mgL-1 de óleo de cravo. Avaliaram-se a sobrevivência, o tempo de anestesia e o tempo de recuperação para cada um dos fármacos. Ambos os fármacos, benzocaína e óleo de cravo, mostraram eficiência para anestesiar peixe-rei, com correlação negativa entre a dose e o tempo de indução de anestesia. Para benzocaína, concentrações entre 80mgL-1 e 100mgL-1 mostraram melhor resultado, enquanto para óleo de cravo as melhores concentrações ficaram entre 25mgL-1 e 50mgL-1. Por outro lado, o tempo de recuperação do estado de anestesia não apresentou variação significativa nas diferentes concentrações testadas. O peixe-rei tem elevada capacidade de metabolização dos produtos testados.

Bone graft sandwich osteotomy to correct a malpositioned dental implant.

As a result of inadequate planning, poor judgment, or losing one's orientation during surgery, implants may be placed in positions or at angulations that are less than ideal. The purpose of this report is to describe an alternative technique for the correction of a malpositioned osseointegrated implant by means of a maxillary anterior single implant segmental osteotomy associated with a 'sandwich' bone graft technique. The technique described provides an alternative option for the surgeon faced with a malpositioned endosseous implant. It allows for a predictable result with preservation of the cervical gingival architecture, creates a more ideal environment for dental restoration, reduces treatment time compared to other techniques, and does so in a cost-effective manner.

Impaction of permanent upper canine causedby supernumerary tooth with talon cusp type III

Introduction: The talon cusp is defined as a developmental anomaly in which an accessory cusp-like structure projects in the area of the cingulum or cementoenamel junction in the anterior teeth attached to the lingual surface of the crown, ranging in size, shape, length and degree of union with the surface. Case report: This study aimed to report a case of a patient who came to Clinic for Preventive Dentistry of Federal University of Pernambuco, Recife-PE, Brazil, complaining of pain in the upper left region. The clinical exam observed the presence of a supernumerary tooth with talon cusp type III in the canine region which had a carious lesion in the developmental groove at the mesial surface and caused a prolonged retention of permanent tooth. Conclusion: With this we want to emphasize that the Dental Surgeon be aware of the changes caused by dental morphological variations, seeking to conduct a proper treatment plan, meeting the functional and aesthetic needs of the patient.

Identical sequence patterns in the ends of exons and introns of human protein-coding genes

Intron splicing is one of the most important steps involved in the maturation process of a pre-mRNA. Although the sequence profiles around the splice sites have been studied extensively, the levels of sequence identity between the exonic sequences preceding the donor sites and the intronic sequences preceding the acceptor sites has not been examined as thoroughly. In this study we investigated identity patterns between the last 15 nucleotides of the exonic sequence preceding the 5' splice site and the intronic sequence preceding the 3' splice site in a set of human protein-coding genes that do not exhibit intron retention. We found that almost 60% of consecutive exons and introns in human protein-coding genes share at least two identical nucleotides at their 3' ends and, on average, the sequence identity length is 2.47 nucleotides. Based on our findings we conclude that the 3' ends of exons and introns tend to have longer identical sequences within a gene than when being taken from different genes. Our results hold even if the pairs are non-consecutive in the transcription order.

Enterovirus infections and type 1 diabetes mellitus: is there any relationship?

Several health organizations have classified diabetes mellitus, a metabolic syndrome, as the epidemic of the century, since it affects millions of people worldwide and is one of the top ten causes of death. Type 1 diabetes is considered to be an autoimmune disease, in which autoaggressive T cells infiltrate the islets of Langerhans in the pancreas, leading to the destruction of insulin producing beta cells. The risk of the disease is modulated by genetic factors, mainly genes coding for human leukocyte antigens (HLA). However, the incidence of this disease has increased significantly during the recent decades, which cannot be explained only by genetic factors. Environmental perturbations have also been associated to the development of diabetes. Among these factors, viral triggers have been implicated; particularly enteroviruses, which have been associated to the induction of the disease. Supporting the hypothesis, numerous lines of evidence coming from mouse models and patients with this type of diabetes have shown the association. The present review aims to provide some understanding of how type 1 diabetes occurs and the possible role of enterovirus in this pathology.

Utilization of microsatellite markers to form families of "pejerrey" Odontesthes bonariensis in a genetic breeding program / Utilização de marcadores microssatélites para formação de famílias de "pejerrey" Odontesthes bonariensis em um programa de melhoramento genético

Foram identificadas a divergência e a variabilidade genética, por meio do polimorfismo de seis marcadores microssatélites, de duas populações de Odontesthes bonariensis, utilizadas em manejos de cultivo e com potencial para fornecimento de reprodutores para programas de melhoramento genético. Do total de seis loci, cinco demonstraram eficiência para análise genética nas duas populações de O. bonariensis. A diferenciação genética significante nas populações analisadas pode fornecer a base para futuros programas de melhoramentos genéticos, através da combinação de material das populações divergentes para o desenvolvimento de linhagens ou execução de um programa de seleção.

Análise do perfil dos resultados de uniformidade de conteúdo de cápsulas de baixa dosagem produzidas em farmácias de manipulação de Santa Catarina / Analysis of profile of content uniformity test results for low dose capsules produced in pharmacies in Santa Catatina

A qualidade de cápsulas tem sua exatidão da dosagem assegurada por ensaios de teor e uniformidade de conteúdo (UC). Assim, os Laboratórios de Controle de Qualidade auxiliam as farmácias magistrais a guiarem sua conduta buscando a garantia da qualidade dos seus produtos. O presente trabalho teve por objetivo analisar o perfil dos resultados de UC de cápsulas magistrais, a partir dos relatórios de análise realizados entre janeiro de 2008 a fevereiro de 2010, por um Laboratório de Controle de Qualidade em Santa Catarina (Brasil). Identificou-se em 2009 um aumento no número e na proporção de farmácias que passaram a solicitar a UC de cápsulas de baixa dosagem em relação a 2008. Das análises, 78,6% tiveram aprovação como resultado, sendo o maior índice de reprovação relacionado ao teor de ativo fora do limite de 85-115%, dos quais 83,6% estavam abaixo do especificado no rótulo. Das análises reprovadas, 59,2% foram aprovadas na análise posterior e apenas 7% mantiveram o mesmo status. Neste estudo, as reprovações nas análises de UC não tiveram correlação com a dose preparada. A análise global dos resultados de uniformidade de conteúdo das cápsulas magistrais reflete a preocupação dos estabelecimentos farmacêuticos com a qualidade do produto oferecido.

The dose accuracy of capsules produced in compounding pharmacies is ensured mainly by testing their uniformity of content (UC). In order to guarantee the quality of such pharmaceuticals in Brazil, entities such as Quality Control Laboratories work together with these pharmacies. The aim of this study was to analyze the profile of results on the UC of capsules, in the reports of analyses performed from January 2008 to February 2010 by a Quality Control Laboratory in Santa Catarina state (Brazil). It was found that an increase occurred in both the number and proportion of pharmacies requesting UC tests of low dose capsules in 2009, relative to 2008. Among the products tested, 78.6% were approved and the largest failure rate was related to drug contents outside the 85-115% limits, of which 83.6% were below the content specified on the label. Among the failures, 59.2% passed in a subsequent test and only 7% maintained the same error. In this study, no correlation was found between disapprovals and dose. These results reflect the compounding pharmacie?s concern for product quality.

Mutagenicity induced by the hydroalcoholic extract of the medicinal plant Plathymenia reticulata Benth

Plathymenia reticulata Benth has an anti-inflammatory effect and is capable of neutralizing the neuromuscular blockade induced by Bothrops jararacussu or Crotalus durissus terrificus venoms, probably by precipitating venom proteins (an effect caused by plant tannins). The present study aimed to evaluate the mutagenic activity of P. reticulata by using the Salmonella mutagenicity assay (Ames test) and the micronucleus test in CHO-K1 cells. P. reticulata extract concentrations of 2.84, 5.68, 11.37, and 19.90 mg/plate were assayed by the Ames test using TA97a, TA98, TA100 and TA102 bacterial strains, with (+S9) and without (-S9) metabolic activation. Concentrations of 5, 1.6 and 0.5 ìg/mL of P. reticulata extract were used for the micronucleus test. P. reticulata extract was mutagenic to TA98 (-S9) and showed signs of mutagenic activity in TA97a and TA102 (both -S9) strains. Micronucleus test CBPI values showed that the endogenous metabolic system increased the number of viable cells when compared to the non-activated samples and the micronucleus frequency increased when the cells were treated in the absence of S9. We concluded that P. reticulata extract may present direct mutagenic properties.

Molecular techniques for the study and diagnosis of parasite infection

In parasitology, routine laboratory diagnosis involves conventional methods, such as optical microscopy, used for the morphological identification of parasites. Currently, molecular biology techniques are increasingly used to diagnose parasite structures in order to enhance the identification and characterization of parasites. The objective of the present study was to review the main current and new diagnostic techniques for confirmation of parasite infections, namely: polymerase chain reaction (PCR), real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), Luminex xMAP, random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and restriction fragment length polymorphism (RFLP), in addition to microsatellites. Molecular assays have comprehensively assisted in the diagnosis, treatment and epidemiological studies of parasitic diseases that affect people worldwide, helping to control parasitic disease mortality.

Adult-Onset Still's Disease and cytomegalovirus infection.

We present a case of a previously asymptomatic 34-year-old man that presented to the emergency department with two weeks of fever, arthralgia of the wrists and knees and sore throat. He was diagnosed with cytomegalovirus (CMV) mononucleosis. The patient remained symptomatic in the 5 following months. After an extensive workup to exclude other clinical conditions, a liver biopsy was performed and CMV hepatitis was diagnosed. He started valganciclovir therapy. Approximately one year after the initial complaints, the patient remained ill and presented clinical criteria compatible with Adult Onset Still's Disease. The patient had a marked improvement after institution of prednisolone, an effect that has been sustained during the following months.

Mechanisms involved in the antinociception induced by systemic administration of guanosine in mice.

BACKGROUND AND PURPOSE: It is well known that adenine-based purines exert multiple effects on pain transmission. However, less attention has been given to the potential effects of guanine-based purines on pain transmission. The aim of this study was to investigate the effects of intraperitoneal (i.p.) and oral (p.o.) administration of guanosine on mice pain models. Additionally, investigation into the mechanisms of action of guanosine, its potential toxicity and cerebrospinal fluid (CSF) purine levels were also assessed. EXPERIMENTAL APPROACH: Mice received an i.p. or p.o. administration of vehicle (0.1 mM NaOH) or guanosine (up to 240 mg x kg(-1)) and were evaluated in several pain models. KEY RESULTS: Guanosine produced dose-dependent antinociceptive effects in the hot-plate, glutamate, capsaicin, formalin and acetic acid models, but it was ineffective in the tail-flick test. Additionally, guanosine produced a significant inhibition of biting behaviour induced by i.t. injection of glutamate, AMPA, kainate and trans-ACPD, but not against NMDA, substance P or capsaicin. The antinociceptive effects of guanosine were prevented by selective and non-selective adenosine receptor antagonists. Systemic administration of guanosine (120 mg x kg(-1)) induced an approximately sevenfold increase on CSF guanosine levels. Guanosine prevented the increase on spinal cord glutamate uptake induced by intraplantar capsaicin. CONCLUSIONS AND IMPLICATIONS: This study provides new evidence on the mechanism of action of the antinociceptive effects after systemic administration of guanosine. These effects seem to be related to the modulation of adenosine A(1) and A(2A) receptors and non-NMDA glutamate receptors.

Anti-tumor necrosis factor-a for the treatment of steroid-refractory acute graft-versus-host disease

Allogeneic stem cell transplantation has been increasingly performed for a variety of hematologic diseases. Clinically significant acute graft-versus-host disease (GVHD) occurs in 9 to 50 percent of patients who receive allogeneic grafts, resulting in high morbidity and mortality. There is no standard therapy for patients with acute GVHD who do not respond to steroids. Studies have shown a possible benefit of anti-TNF-a (infliximab)for the treatment of acute GVHD. We report here on the outcomes of 10 recipients of related or unrelated stem cell transplants who received 10 mg/kg infliximab, iv, once weekly for a median of 3.5 doses (range: 1-6) for the treatment of severe acute GVHD and who were not responsive to standard therapy. All patients had acute GVHD grades II to IV (II = 2, III = 3, IV = 5). Overall, 9 patients responded and 1 patient had progressive disease. Among the responders, 3 had complete responses and 6 partial responses. All patients with cutaneous or gastrointestinal involvement responded, while only 2 of 6 patients with liver disease showed any response. None of the 10 patients had any kind of immediate toxicity. Four patients died, all of them with sepsis. Six patients are still alive after a median follow-up time of 544 days (92-600) after transplantation. Considering the severity of the cases and the bad prognosis associated with advanced acute GVHD, we find our results encouraging. Anti-TNF-a seems to be a useful agent for the treatment of acute GVHD.

Anti-tumor necrosis factor-a for the treatment of steroid-refractory acute graft-versus-host disease.

Allogeneic stem cell transplantation has been increasingly performed for a variety of hematologic diseases. Clinically significant acute graft-versus-host disease (GVHD) occurs in 9 to 50% of patients who receive allogeneic grafts, resulting in high morbidity and mortality. There is no standard therapy for patients with acute GVHD who do not respond to steroids. Studies have shown a possible benefit of anti-TNF-a (infliximab)for the treatment of acute GVHD. We report here on the outcomes of 10 recipients of related or unrelated stem cell transplants who received 10 mg/kg infliximab, iv, once weekly for a median of 3.5 doses (range: 1-6) for the treatment of severe acute GVHD and who were not responsive to standard therapy. All patients had acute GVHD grades II to IV (II = 2, III = 3, IV = 5). Overall, 9 patients responded and 1 patient had progressive disease. Among the responders, 3 had complete responses and 6 partial responses. All patients with cutaneous or gastrointestinal involvement responded, while only 2 of 6 patients with liver disease showed any response. None of the 10 patients had any kind of immediate toxicity. Four patients died, all of them with sepsis. Six patients are still alive after a median follow-up time of 544 days (92-600) after transplantation. Considering the severity of the cases and the bad prognosis associated with advanced acute GVHD, we find our results encouraging. Anti-TNF-a seems to be a useful agent for the treatment of acute GVHD.

Inhibition of IL-6-dependent growth of myeloma cells by an acidic peptide repressing the gp130-mediated activation of Src family kinases.

An acidic domain (AD) of gp130 was previously found to interact with the Src family kinase (SFK) Hck. Here, the influence of myristoylated peptides derived from this AD was assessed in the mouse myeloma cell line, 7TD1. The IL-6-dependent growth of 7TD1 cells was reduced by approximately 75%, if 100 microM of myristoylated 18mer peptide (18AD) was included in the growth medium, but was unaffected by a control peptide with scrambled sequence (18sc). A similar differential inhibition by peptides 18AD and 18sc was observed for the erythropoietin-dependent growth of BaF-EH cells expressing chimeric erythropoietin receptor-gp130 and human Hck and for the human myeloma cell line INA-6. While the peptide 18AD concentration inhibiting 50% was approximately 30 microM in 7TD1 and BaF-EH cells, peptide 18AD did not significantly inhibit growth of IL-6-independent MM1.S myeloma and OKT1 hybridoma cells or of BaF-EH cells supplied with IL-3. Treatment with 100 microM peptide 18AD caused the same degree or 60% of apoptosis induction as IL-6 deprivation in 7TD1 or INA-6 cells, respectively. Co-immunoprecipitation experiments revealed that peptide 18AD interfered with the association of Hck and gp130 in 7TD1 lysates in a concentration-dependent manner. IL-6-treatment of INA-6 cells induced the kinase activities of Fyn, Lyn and Hck, but not Src, and the IL-6-induced SFK activities were inhibited by peptide 18AD. Expression in 7TD1 cells of a kinase-inactive Hck mutant (K269R) elicited a dominant-negative effect on cell number increases providing further evidence that SFKs are required for gp130 signalling in myeloma cells.

Global analysis of the human gastric epithelial transcriptome altered by Helicobacter pylori eradication in vivo.

OBJECTIVE: The transcriptional profile of gastric epithelial cell lines cocultured with Helicobacter pylori and the global gene expression of whole gastric mucosa has been described previously. We aimed to overcome limitations of previous studies by determining the effects of H pylori eradication on the transcriptome of purified human gastric epithelium using each patient as their own control. DESIGN: Laser capture microdissection (LCM) was used to extract mRNA from paraffin-embedded antral epithelium from 10 patients with peptic ulcer disease, before and after H pylori eradication. mRNA was reverse transcribed and applied on to Affymetrix cDNA microarray chips customised for formalin-fixed tissue. Differentially expressed genes were identified and a subset validated by real-time polymerase chain reaction (PCR). RESULTS: A total of 13 817 transcripts decreased and 9680 increased after H pylori eradication. Applying cut-off criteria (p<0.02, fold-change threshold 2.5) reduced the sample to 98 differentially expressed genes. Genes detected included those previously implicated in H pylori pathophysiology such as interleukin 8, chemokine ligand 3, beta defensin and somatostatin, as well as novel genes such as GDDR (TFIZ1), chemokine receptors 7 and 8, and gastrokine. CONCLUSIONS: LCM of archival specimens has enabled the identification of gastric epithelial genes whose expression is considerably altered after H pylori eradication. This study has confirmed the presence of genes previously implicated in the pathogenesis of H pylori, as well as highlighted novel candidates for further investigation.