Vaccination to improve the persistence of CD19CAR gene-modified T cells in relapsed pediatric acute lymphoblastic leukemia.

Trials with second generation CD19 chimeric antigen receptors (CAR) T-cells report unprecedented responses but are associated with risk of cytokine release syndrome (CRS). Instead, we studied the use of donor Epstein-Barr virus-specific T-cells (EBV CTL) transduced with a first generation CD19CAR, relying on the endogenous T-cell receptor for proliferation. We conducted a multi-center phase I/II study of donor CD19CAR transduced EBV CTL in pediatric acute lymphoblastic leukaemia (ALL). Patients were eligible pre-emptively if they developed molecular relapse (>5 × 10-4) post first stem cell transplant (SCT), or prophylactically post second SCT. An initial cohort showed poor expansion/persistence. We therefore investigated EBV-directed vaccination to enhance expansion/persistence. Eleven patients were treated. No CRS, neurotoxicity or graft versus host disease (GVHD) was observed. At 1 month, 5 patients were in CR (4 continuing, 1 de novo), 1 PR, 3 had stable disease and 3 no response. At a median follow-up of 12 months, 10 of 11 have relapsed, 2 are alive with disease and 1 alive in CR 3 years. Although CD19CAR CTL expansion was poor, persistence was enhanced by vaccination. Median persistence was 0 (range: 0-28) days without vaccination compared to 56 (range: 0-221) days with vaccination (P=0.06). This study demonstrates the feasibility of multi-center studies of CAR T cell therapy and the potential for enhancing persistence with vaccination.

[THE ROLE OF MOTHER'S MILK AND BREAST FEEDING. MEDICAL PROBLEMS DURING THE LACTATION PERIOD LACTOBACILUS FERMENTUM--A NEW APPROACH TOWARDS THE PREVENTION AND THE TREATMENT OF ACUTE AND SUBACUTE MASTITIS].

Breast feeding provides a lot of short and long-term benefits for the mother and the baby. It prevents the baby of gastrointestinal, urinary and respiratory infections, atopical conditions and assures long-term protection of cardiovascular and metabolic diseases. The breast feeding decreases the risk for the mother of ovary and breast carcinoma and creates a positive emotional bond between the mother and the baby Mother's milk is a species specific; its content is relatively stable regardless of mother's age race, way and place of living. Mother's milk is not sterile. There is a 10 year international trial held in Spanish and Finnish universities. It has identified and count all microorganisms in mother's milk (more than 700) and proved that their content and quantity varies according the age of the baby. Mother's milk is a source of lactobacillus for baby's intestines and most of them have probiotic potential. Lactobacillus fermentum Lc40 (hereditum) is isolated from mother's milk. It has a good viability in gastrointestinal system, high level of adhesion to intestinal epithelium cells, produces glutation--strong antioxidant, good antibacterial activity to entero-pathogens and potential of increasing the immunologic response. Clinical trials reveal that Lactobacillus fermentum plays important role to microflora balance of mother's milk in mastitis during lactation. Many trials estimating the efficiency of lactobacillus fermentum in prevention and treatment of acute and subacute mastitis have been carried out. The results of them open a new door in front of us in the treatment of these conditions--treatment with probiotics instead of antibiotics.

Phase II study of gemcitabine and bexarotene (GEMBEX) in the treatment of cutaneous T-cell lymphoma.

BACKGROUND: Both gemcitabine and bexarotene are established single agents for the treatment of cutaneous T-cell lymphoma (CTCL). We investigated the feasibility and efficacy of combining these drugs in a single-arm phase II study. METHODS: Cutaneous T-cell lymphoma patients who had failed standard skin-directed therapy and at least one prior systemic therapy were given four cycles of gemcitabine and concurrent bexarotene for 12 weeks. Responders were continued on bexarotene maintenance until disease progression or unacceptable toxicity. RESULTS: The median age was 65 years, stage IB (n=5), stage IIA (n=2), stage IIB (n=8), stage III (n=8) and stage IVA (n=12), 17 patients were erythrodermic, 17 patients were B1, and 10 patients were both erythrodermic and B1. Thirty (86%) patients completed four cycles of gemcitabine. In all, 80.0% of patients demonstrated a reduction in modified Severity-Weighted Assessment Tool (mSWAT) score although the objective disease response rate at 12 weeks was 31% (partial response (PR) 31%) and at 24 weeks 14% (PR 14%, stable disease (SD) 23%, progressive disease (PD) 54%, not evaluable 9%). Median progression-free survival was 5.3 months and median overall survival was 21.2 months. CONCLUSION: The overall response rate of the combination did not reach the specified target to proceed further and is lower than that previously reported for gemcitabine as a single agent.

Establishment of a UK-wide network to facilitate the acquisition of quality assured FDG-PET data for clinical trials in lymphoma.

BACKGROUND: Multicentre trials are required to determine how [fluorine-18]-2-fluoro-2-deoxy-D-glucose-positron emission tomography imaging can guide cancer treatment. Consistency in quality control (QC), scan acquisition and reporting is mandatory for high-quality results, which are comparable across sites. METHODS: A national positron emission tomography (PET) clinical trials network (CTN) has been set up with a 'core laboratory' to coordinate QC and interpret scans. The CTN is involved in trials in Hodgkin's lymphoma [Randomised Phase III trial to determine the role of FDG-PET Imaging in Clinical Stages IA/IIA Hodgkin's Disease (RAPID) and Randomised Phase III trial to assess response adapted therapy using FDG-PET imaging in patients with newly diagnosed, advanced Hodgkin lymphoma (RATHL)] and diffuse large B-cell lymphoma [Blinded evaluation of prognostic value of FDG-PET after 2 cycles of chemotherapy in diffuse large B-cell Non-Hodgkins Lymphoma, a sub-study of the R-CHOP-21 vs R-CHOP-14 trial (R-CHOP PET substudy)]. Approval to join requires scanner validation and agreement to follow a standard QC protocol. Scans are transferred to the core laboratory and reported centrally according to predetermined criteria. RESULTS: The qualification procedure was carried out on 15 scanners. All scanners were able to demonstrate the necessary quantitative accuracy, and following modification of image reconstruction where necessary, scanners demonstrated comparable recovery coefficients (RCs) indicating similar performance. The average RC (±1 standard deviation) was 0.56 ± 0.095 for the 13-mm sphere. Reports from 444 of 473 (94%) patients in RAPID and 67 of 73 (92%) patients in RATHL were available for randomisation of therapy. CONCLUSIONS: The CTN has enabled consistent quality assured PET results to be obtained from multiple centres in time for clinical decision making. The results of trials will be significantly strengthened by this system.

Nuclear factor kappaB-dependent mechanisms coordinate the synergistic effect of PMA and cytokines on the induction of superoxide dismutase 2.

Manganese superoxide dismutase (MnSOD) serves a protective role under conditions of oxidative stress mediated by such diverse agents as adriamycin, radiation, chemical hypoxia and ischaemia and might act as a newly recognized type of tumour-suppressor. MnSOD is an inducible enzyme; however, the signalling molecules and pathways involved in its induction have not been fully elucidated. Recently we reported the identification of a 342 bp enhancer within the second intron (I2E) of the human gene encoding MnSOD (SOD2), which contains sites for binding nuclear factor kappaB (NF-kappaB), CCAAT-enhancer-binding protein (C/EBP) and nuclear factor 1 (NF-1). Using a human fibroblast cell line transformed by simian virus 40, we have identified the I2E fragment as being responsive to PMA. Furthermore, simultaneous treatment with PMA and cytokines (tumour necrosis factor alpha and interleukin 1beta) synergistically increases MnSOD induction. The use of mutant constructs identified the NF-kappaB element within the enhancer fragment as being essential for the PMA and PMA/cytokine effect. Mutations in the C/EBP- and NF-1-binding sites revealed a potential co-operation between proteins that bind to these sites and the NF-kappaB element. Evaluation of inhibitory kappaB (IkappaB)-alpha and IkappaB-beta proteins reveals agent-specific differences in their turnover kinetics. Both C/EBP and NF-kappaB DNA-binding activities were increased in cells receiving a combination of cytokine and PMA. Supershift and immunoprecipitation studies suggest a physical interaction between C/EBP and NF-kappaB proteins. Taken together, these studies suggest the activation of multiple transcription factors as well as pathways leading to increased NF-kappaB activity as being the mechanisms responsible for the synergistic induction of MnSOD by PMA and cytokines.

The physiology of massive zinc accumulation in the liver of female squirrelfish and its relationship to reproduction.

It is well known that zinc is an essential micronutrient and, as a rule, organisms keep relatively constant low levels of zinc to maintain cellular functions. The squirrelfish family (Holocentridae) is the only known exception from this rule. Squirrelfish accumulate very high concentrations of zinc in the liver. In the present study, we demonstrate that, while female squirrelfish store large amounts of zinc in the liver and ovaries, the males show zinc levels that are typical for vertebrates. The zinc content of the diet is the same in males and females, and zinc is not lost from the liver during starvation. Thus, the difference between genders in zinc storage is not dependent upon the diet. Rather, there are at least two processes that contribute to the accumulation in females. First, females possess high levels of two major zinc-binding proteins: metallothionein (MT) and a novel female-specific zinc-binding protein (FZnBP). In females, but not in males, almost all MT is present in the hepatocyte nucleus. FZnBP is exclusively found in the hepatocyte cytosol of females. Second, hepatocytes of female squirrelfish have a high capacity to transport zinc across the plasma membrane. In addition to the liver, only the gonads of females showed unusually high concentrations of zinc. Administration of exogenous oestrogen to females decreases the hepatic zinc concentration while there is a matching increase in the zinc content of the ovaries. Thus, oestrogen may trigger a redistribution of zinc from liver to ovaries. Together, our findings suggest that female squirrelfish may be uniquely adapted to detoxify zinc and to utilize it as a macronutrient for processes related to reproduction.

Transcriptional activation by DNA-binding derivatives of HSV-1 VP16 that lack the carboxyl-terminal acidic activation domain.

The herpes simplex virus transactivator VP16 directs the assembly of a multicomponent protein-DNA complex with cellular components Oct-1 and VCAF-1, contributing a potent carboxyl-terminal acidic activation domain that is essential for activation of gene expression in mammalian cells. We show here that VP16, devoid of this acidic activation domain, functions as a strong transcriptional activator in the yeast Saccharomyces cerevisiae when appended onto a heterologous GAL4 DNA binding domain, as determined by measuring activation of a resident GAL1:lacZ reporter gene. Deletion analysis indicated that sequences contained within the amino-terminal 369 amino acids of VP16 were necessary for transactivation by truncated VP16. Activation by truncated VP16 in yeast was comparable to that observed with a hybrid protein consisting of the GAL4 DNA binding domain linked to the VP16 acidic activation domain. Similar GAL4-VP16 hybrid proteins were only marginally active in mammalian cells. Sequence requirements for transactivation by truncated VP16 can be demarcated from domains of VP16 that are required for interaction with VCAF-1 and for protein-DNA complex formation with Oct-1. Our findings indicate that VP16 contains additional sequences upstream of the acidic activation domain that may play a direct role in transactivation.

Herpes simplex virus VP16 forms a complex with the virion host shutoff protein vhs.

Herpes simplex virus (HSV) virions contain at least two regulatory proteins that modulate gene expression in infected cells: the transcriptional activator VP16 and the virion host shutoff protein vhs. VP16 stimulates transcription of the HSV immediate-early genes, and vhs suppresses host protein synthesis and induces accelerated turnover of cellular and viral mRNAs. We report here that vhs binds directly to VP16: vhs and VP16 were coprecipitated from infected cells by an anti-vhs antiserum, and vhs and VP16 protein A fusions each bound intact versions of the other protein in a solid-phase capture assay. In addition, vhs and VP16 interacted in the two-hybrid activator system when coexpressed in Saccharomyces cerevisiae. vhs residues 238 to 344 were sufficient for the interaction, and the VP16 acidic transcriptional activation domain was not required. vhs blocked the ability of VP16 to enter a multiprotein complex on an immediate-early TAATGARATTC consensus sequence, indicating that vhs interacts with one or more regions of VP16 required for promoter recognition. We suggest that this interaction may play a structural role in the assembly of HSV virions and modulate the activity of vhs during infection.

[Our results with the secondary prevention of myocardial infarct]. / Nashi rezultati ot vtorichna profilaktika na miokardniia infarkt.

The communication summed up the results from the two prospective follow-up care-prophylactic studies on patients with a past history of acute myocardial infarction (AMI). The first study covers all patients, with a past history of AMI and discharged in 1963, 1964, 1966 and 1967, a total of 378, with a follow-up period from 6 to 11 years. The direct task of the follow up was the prevention of a repeated infarction. The prophylactic measures, carried out with the followed-up patients were: control of body mass by a hypocaloric and antisclerotic diet, sufficient physical activity and medicamentous treatment when needed. Two groups of patients were differentiated: experimental (the patients adhering to the instructions prescribed) an control (patients that did not adhere to the recommendations for correction of body mass with no motor regimen). After the termination of the observation it was established that 12.6% from the experimental group had repeated infarction and 28.3%--in the control group. The lethality in the experimental group was 21.7% and in the control--43.9%. The second prospective observation covered 250 patients (reduced to 241 in time), with a past history of AMI and discharged within the year 1975. The follow up lasted for 5 years under the same conditions of follow-up care. Summing up our 5-year secondary prophylaxis carried out mainly with a diet, physical activity and partially with antiaggregants and medicamentous treatment when needed, we could ascertain and draw the conclusions.(ABSTRACT TRUNCATED AT 250 WORDS)

[Work capacity and motor activity of patients with a history of myocardial infarct]. / Trudosposobnost i dvigatelna aktivnost u bolni, prekarali surdechen infarkt.

A total of 241 patients with a past history of AMI were followed up with a view to the restoration of their capacity for work and its dependence on their motor activity. All deceased, 6 months after the acute infarction, were also included in the group. Work was resumed by 43,7 per cent from the whole group. But the percentage of the patients with preserved capacity for work to the age of 60 that had outlived the 5-year observation period is 75; 63, out of 84 subjects at that age, began work within an average period of 8 months after the onset of the infarction. The periods till the beginning work of our patients are longer as compared with the periods reported by foreign authors. An obvious correlation was established between the patients' capacity for work, with a past history of AMI, and their motor activity.

[Survival of patients who have had a myocardial infarct as a function of various risk factors]. / Prezhiviaemost na bolnite prekarali surdechen infarkt, v zavisimost ot niakoi riskovi faktori.

Survival was studied among 250 patients with a past history of acute myocardial infarction (AMI), followed up for 5 years. A total survival of 55,6 per cent was established. The factors affecting the survival are: age, arterial hypertension, diabetes mellitus, sex, rhythm and conduction disorders, restricted physical activity, decompensation signs after AMI, increased measurements of the heart, repeated infarction as well as the presence of anginose complaints postinfarction period. The patients, with a past history of HMI should be subjected to effective follow-up care with a view to the campaign against the factors, threatening life.