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At least 379 records · Page 21Linked to original sources
PubMed1995

Sequence of a transposon identified as Tn1000 (gamma delta).

We report the complete sequence of a transposon found in a cosmid clone of a human DNA sequence. The transposon is identified as the Escherichia coli transposon Tn1000 (also known as gamma delta) on the basis of the identity of the restriction map of the new sequence with that previously recorded for Tn1000 and homology between parts of the new sequence and that of published fragments of Tn1000 sequence. The transposon, which comprises 5,981 nucleotides including two 35 bp inverted terminal repeat sequences (ITRs), contains three open reading frames. The sequence of the resolvase coding region (tnpR) is identical to that published by others. A second reading frame can be identified as the tnpA gene, coding for the transposase, on the grounds of its strong homology with the corresponding gene from transposon Tn3. The third reading frame has the potential to code for a protein of unknown function containing 698 amino acids.

Amino Acid Sequence↗
PubMed2000

Gene therapy vectors based on adeno-associated virus: characteristics and applications to acquired and inherited diseases (review).

Adeno-associated virus (AAV), a defective parvovirus, was discovered more than 30 years ago. Interest in this virus for human gene therapy applications focuses on its non-pathogenicity, broad tropism and infectivity, site-specific integration and long-term persistence. The field of rAAV research has considerably advanced: titers of 1014 p/ml have been achieved, plasmid systems devised to produce helper-free viruses, chimaeric vectors combining properties of rAAV ITRs and large sequence capacity from Ad/HS vectors in parallel with the revolutionary intron strategy based on heterodimerisation of the forming concatamers have expanded the vector capacity. Muscle cells and neurons (post-mitotic cells) are amongst the most efficient targets of rAAV delivery and AAV receptors and co-receptors have been identified. This review will describe advances in the field of rAAV technology that overcome certain limitations of the vector as a gene delivery system and overview applications involving these recombinant vectors for the treatment of acquired and inherited diseases.

Adenoviridae↗
PubMed2000

[Invasive fungal infections. Epidemiology and new therapies].

RISING INCIDENCE: In the past two decades, systemic fungal infections, essentially invasive candidiasis, but also invasive aspergillosis, has increased substantially. Despite the currently available antifungal drugs, amphotericin B (AmB), azole compounds (fluconazole or FLU, itraconazole or ITR), these infections are associated with significant morbidity and mortality. AmB remains the drug of choice for treatment of most fungal diseases because of its broad spectrum and potent fungicidal activity, but significant side effects limit its clinical utility. The azole antifungal agents are easier to take, less toxic than AmB, but their use is limited by multiazole-resistant strains. NEW ANTIFUNGAL AGENTS: Lipid formulations have recently attracted much attention due to a significantly lower toxicity: this concerns lipid formulations of AmB and perhaps nystatin in the future. New triazoles (voriconazole, ravuconazole, posaconazole) have shown a wide spectrum of action including against azole-resistant isolates. A new class of antifungal agents, lipopeptides (MK-0991, LY303366, FK463), with an original mechanism of action are being developed. These new compounds are reported to possess a large fungicidal activity against most isolates including AmB and azole-resistant strains.

Amphotericin B↗
PubMed1999

[Cloning of 0-17.5 mu and sequencing of 0-4.8 mu of the lefe and DNA of the Ad7 vaccine strain left end].

OBJECTIVE: Cloning of 0-17.5 mu DNA fragment of adenovirus 7 vaccine strain and sequencing of 0-4.8 mu fragment (1,737 bp). METHODS: Isolating and purifying Ad7 vaccine genome from the A549 cultured cells, putting 0.3-17.5 mu fragment into pAd7T plasmid, then sequencing the 0-4.8 mu fragment including inverted terminal repeats (ITR), packaging sequence and Ela region. RESULTS: We obtained 0-17.5 mu fragment of Ad7 vaccine strain genome and sequenced its left terminal 1,737 bp. Sequence analysis showed that the Ad7 vaccine strain Ela region encodes 6,300, 24,000 and 28,000 proteins. Compared with equivalent region of Ad7 Gomen strain, they share the homology of the nucleotide sequence 98.9%, 97.3%, 97.5% and the homology of the deduced amino acid sequence 96.6%, 96.5%, and 96.9% respectively. When compared with Ad7 Grider strain, they share the homology of the nucleotide sequence 100%, 99.7%, 99.7% and the homology of the deduced amino acid sequence 100%, 99.1%, and 99.2% respectively. CONCLUSION: The Ad7 vaccine strain left terminal 1,737 bp nucleotide sequence showed a high homology with corresponding region of Ad7 Gomen strain and Grider strain.

Adenoviridae↗
PubMed1992

Multiple enlargements in the right inverted terminal repeat of the DNA of canine adenovirus type 2.

The Manhattan strain of canine adenovirus type 2 (CAV 2) was examined. Restriction endonuclease analysis and blot hybridization experiments revealed the heterogeneity of the viral DNA. At least 9 unequally expanded species of the viral genome have been recognized. This diversity is caused by different enlargements in the right inverted terminal repeat (ITR) of the virus. The differences between the individual enlargements were shown to be the different multiples of 150 base pairs. Relatedness of CAV 2 DNA to the DNA of bovine adenovirus type 2 (BAV 2) and human adenovirus type 2 (HAV 2) has also been observed during DNA hybridization experiments.

Adenoviridae↗
PubMed2004

Pancreatic islet and stem cell transplantation: new strategies in cell therapy of diabetes mellitus.

Long-term studies strongly suggest that tight control of blood glucose can prevent the development and retard the progression of chronic complications of type 1 diabetes mellitus. In contrast to conventional insulin treatment, replacement of a patient's islets of Langerhans either by pancreas organ transplantation of by isolated islet transplantation is the only treatment to achieve a constant normoglycemic state and avoiding hypoglycemic episodes, a typical adverse event of multiple daily insulin injections. However, the expense of this benefit is still the need for immunosuppressive treatment of the recipient with all its potential risks. Islet cell transplantation offers the advantage of being performed as a minimally invasive procedure, in which islets can be perfused percutaneously into the liver via the portal vein. As of June 2003, 705 pancreatic islet transplants worldwide have been reported to the International Islet Transplant Registry (ITR) at our Third Medical Department, University of Giessen/Germany. Data analysis shows at 1 year after adult islet transplantation a patient survival rate of 97%, a functioning islet graft in 54% of the cases, whereas insulin independence was meanwhile achieved in 20% of the cases. However, using a novel protocol established by the Edmonton Center/Canada, the insulin independence rates have improved significantly reaching meanwhile a 50-80% level. Finally, the concept of islet cell or stem cell transplantation is most attractive since it offers many perspectives: islet cell availability could become unlimited and islet or stem cells my be transplanted without life-long immunosuppressive treatment of the recipient, just to mention 2 of them.

Animals↗
PubMed2005

[Small bowel transplantation].

Early attempts of small bowel transplantation (SBTx) were unsuccessful as a result of uncontrolled rejection and infection. Introduction of tacrolimus in the early 1990s improved the outcome, but results were not satisfactory comparing those after the other organ transplants. Recently, improved long-term graft survivals were reported from several centers. According to Intestine Transplant Registry (ITR), a multivariate analysis of cases within the last 5 years revealed that transplantation of patients waiting at home, recipient age, antibody induced immune suppression, and center experience with at least 10 cases were associated with improved patients survival. One-year graft survival rate of 90% were achieved in patients who were treated with antithymocyte globulin and maintained on tacrolimus.

Adolescent↗
PubMed2006

[Antifungal susceptibility testing of commensal and pathogenic clinical isolates of oral Candida].

PURPOSE: To evaluate and compare the sensitivity and resistance of commensal and pathogenic oral Candida to antifungal agents. METHODS: YBC Test Kit was used to identify species of commensal and pathogenic oral Candida; The in vitro susceptibilities of oral Candida to 5-flucytosine (5-FC), itraconazole (ITR) and fluconazole (FLU) were determined by NCCLS M27-A2 method. RESULTS: All strains were sensitive to 5-flucytosine, drug resistant strains to fluconazole and itraconazole were found both in commensal and pathogenic oral Candida spp; The proportion of no-C. albicans and antifungal drug resistant strains isolated from head and neck cancer patients treated with radiotherapy or chemotherapy were higher than oral commensal Candida and oral Candidiasis. CONCLUSIONS: The no-C. albicans and drug resistant strains to fluconazole and itraconazole can be found both in commensal and pathogenic oral Candida spp now.

Antifungal Agents↗
PubMed2006

[Antifugal susceptibility testing and antifugal traditional Chinese medicines screening of oral Candida isolated from head and neck cancer patients treated with radiotherapy or chemotherapy].

OBJECTIVE: To evaluate the sensitivity and resistance of pathogenic oral Candida spp. isolated from head and neck cancer patients treated with radiotherapy or chemotherapy to antifungal agents. To screen antifugal agents from Chinese traditional and herbal drugs by NCCLS M27-A2 method. METHODS: Using YBC Test Kit to identify 20 clinical oral Candida isolated from head and neck cancer patients treated with radiotherapy or chemotherapy. The in vitro susceptibilities of 20 oral Candida spp. to 5-flucytosine (5-FC), itraconazole (ITR), fluconazole (FLU), the extracts of 6 Chinese traditional and herbal drugs (caltrop, honeysuckle flower, dandelion, green tea, pine bark, red trefoil) and utility componets of 7 Chinese traditional and herbal drugs (sophorcarpidine, aloperine, archin, glycyrrhizic acid, glycosides of white peony root, glycosides of baikal skullcap root, hydrochloric berberine) were determined by NCCLS M27-A2 method. RESULTS: The proportion of no-C. albicans in all Candida spp. were 25%. All strains were sensitive to 5-flucytosine, 25% stains were resistant to fluconazole and 40% stains were resistant to itraconazole. In all agents from Chinese traditional and herbal drugs, glycosides of white peony root and hydrochloric berberine (C20H18CINO4) exhibited antifungal activity, especially to C. glabrates. CONCLUSION: The proportion of no-C. albicans in all oral Candida spp. isolated from head and neck cancer patients treated with radiotherapy or chemotherapy was pretty high. NCCLS M27-A2 micro-dilution method is a reliable and reproducible method and can be used to screen antifugal agents from Chinese traditional and herbal drugs.

Antifungal Agents↗
PubMed2006

[Complete nucleotide sequence analysis of Bombyx mori Densonucleosis virus type 3 VD2 (China isolate)].

The genome of Bombyx mori densonucleosis virus type 3 (China isolate) contains two kinds different single-stranded linear DNA molecules (VD1, VD2). The VD2 was purified and cloned into the pUC119 vector, and the complete nucleotide sequence of VD2 was determined. Sequence analysis showed that the VD2 genome sequence consisted of 6022 nts, including inverted terminal repeats (ITRs) of 524 nts. In the viral genome, two major open reading frames (ORF1 and ORF2) in the plus strand and one minor ORF (ORF3) in the complementary strand were identified. Computer analysis suggested the plus stand ORF1 and the minus strand ORF3 most likely encode the major nonstructural protein, while the plus stand ORF2 most likely encodes the major structural protein. Comparing the complete genome sequence of BmDNV-3 VD2 with that of BmDNV-2 VD2 (Yamanashi isolate) demonstrated that they shared 97.7% genome sequence in the VD2 region, with substitutions of 132 nucleotides, deletions of 11 nucleotides and insertions of 2 nucleotides in the VD2 region of BmDNV-3. The results suggested that BmDNV-3 is closely related to BmDNV-2, but with some differences, giving a better understanding about the variation of the viruses and providing clues to their evolution.

Animals↗
PubMedDate not supplied

[Tonsillectomy in childhood: personal considerations].

This paper discusses the personal criteria followed in indicating tonsillectomy and/or adenotonsillectomy in treating a group of 150 children between the ages of 2 and 12 in the ENT Department of the Crobu Hospital of Iglesias where a follow-up was carried out from May 1989 to March 1991. The study analyzes when and under what conditions these surgical procedures are indicated in young patients without recurrent tonsillitis but with obstructive sleep apnea (OSA). The utility of traditional laboratory investigations (such as surface pharyngeal swabbing) in indicating surgical therapy is also evaluated. In concluding, the Authors affirm that recurrent tonsillitis no longer controllable with medical therapy is still the most frequent indication for surgery. In fact, 64% of the children studied belonged to either class TR or ITR in which recurring infection was the primary indication factor. However, respiratory obstruction has recently emerged as a increasingly important factor in indicating tonsillectomy and/or adenotonsillectomy. In fact, 54 children (35.9%) of the 150 studied belonged to class IT or OSA in which tonsillar and/or adeno-tonsillar hypertrophy with obstruction of various degrees was the determining factor in indicating surgical treatment. It appears that in the near future with fewer tonsillectomies being indicated in the case of recurrent tonsillitis, a progressive increase in the number of indications of surgical treatment in cases of obstruction may be expected. The Authors stress, however, the importance of identifying OSA patients because of the cardiorespiratory risks they may encounter during surgery (intubation difficulty found by the anaesthesiologist, tendency towards laryngeal spasms and pharyngeal obstruction in the post-operatory period).

Adenoidectomy↗
PubMedDate not supplied

[Testicular lymphoblastic infiltration after healing of systemic disease].

The paper presents the case of a patient with isolated testicular relapse (ITR), after a period of complete continuous remission (CCR) following chemotherapy, cobalt-therapy and bone marrow transplant (BMT) due to diagnosis of acute type T lymphoblastic leukemia L1. A review of literature regarding treatment of these patients is made.

Adolescent↗
PubMed1983

Comparative studies with cyclic and linear somatostatin on active avoidance behaviour and open-field activity in rats.

The action of cyclic and linear somatostatin on active avoidance and open-field behaviour was investigated in rats. Both peptides inhibited the extinction of the avoidance behaviour and slightly but not significantly increased the intertrial activity (ITR). Both peptides increased ambulation and rearing activity in a dose related fashion, while the grooming and defecation rates were not changed. Since both peptides are capable of increasing the locomotor performance this action might contribute to the effect exerted on the extinction of active avoidance behaviour.

Animals↗
PubMed1980

Functional significance of different amygdalar areas in performance of instrumental conditioned alimentary reflexes in rats.

Bilateral lesions of the basolateral amygdala caused facilitation of instrumental alimentary conditioned reflexes while bilateral cortico-medial amygdala damage produced inhibition of conditioned responses. The most rapid learning was in rats after basolateral lesions whereas the cortico-medial rats required more trials to criterion than controls. In basolateral rats the largest number of intertrial responses were found, while cortico-medial rats had a small number of ITR in criterion sessions. Thus, the cortico-medial amygdala was concerned with the excitatory system and the basolateral amygdala with the inhibitory system of the rat forebrain. The results of combined unilateral lesions of basolateral and cortico-medial amygdala in each hemisphere suggest that in healthy animals the inhibitory influence of the former was stronger and overlapped or regulated the excitatory action of the latter. Our evidence suggests the existence of two antagonistic systems in the rat amygdala which play different roles in conditioning.

Amygdala↗
PubMedDate not supplied

Simple method for cDNA amplification starting from small amount of total RNA.

We describe a novel simple PCR-based technique for construction of cDNA libraries starting from the small samples of cells or tissues. This technique is based on the insertion of inverted terminal repeats (ITR) into amplified cDNA which causes a part of molecules to generate "pan"-type structures at each cycle of PCR amplification. This allows to avoid generation of the primer dimmer and makes possible the regulation of an average length of amplified sequences varying in concentration of primers.

Animals↗
PubMed1994

Recombinant adeno-associated virus-mediated gene transfer into hematopoietic progenitor cells.

Recombinant adeno-associated viruses (rAAV) containing only the inverted terminal repeats (ITR) from the wild-type virus are capable of stable integration into the host cell genome, and expression of inserted genes in cultured cells. We have now defined the ability of rAAV to introduce genes into primary hematopoietic progenitors. A vector was constructed containing the coding sequences for beta-galactosidase (beta-gal), including a nuclear localization signal, under the control of a strong viral promotor. Infectious vector particles were prepared by cotransfection of the vector plasmid with a second plasmid that contained the coding sequences for AAV proteins into adenovirus-infected human embryonic kidney cells. These vector preparations transferred and expressed the beta-gal gene in human K562 erythroleukemia and Detroit 6 cells. Positive immunoselection yielded a population of enriched CD34+ cells that were transduced with the rAAV beta-gal vector. Nuclear localized enzyme expression was documented in 60% to 70% of infected cells. Progenitor-derived colonies that developed after 2 weeks in clonogenic cultures were shown to have viral-associated DNA at an estimated copy number of 1 to 2 per cell using a semiquantitative polymerase chain reaction (PCR) method. Integration of AAV into hematopoietic progenitors was documented using wild-type virus, as its genome may integrate at a preferred site on chromosome 19. Our data suggest that rAAV will transfer and express genes in primitive hematopoietic progenitors with high frequency, and support the development of this vector system for therapeutic gene transfer.

Adenoviruses, Human↗
PubMedDate not supplied

[Differences in the effects of piracetam and fenazepam in emotional stress caused by the spatial reversal of a habit].

Emotional stress was caused in rats by changing the location of the gate in the shuttle chamber through which training had been accomplished in the previous experiments. The procedure led to disorder of the avoidance response (AR) and increase of intertrial responses (ITR). Phenazepam (0.1 mg/kg) prevented increase in the number of ITS. Piracetam (300 mg/kg) reduced the sharp growth of ISR but, in contrast to phenazepam, it provided a higher level of AR reproduction. The ISR were greater in piracetam than in phenazepam administration. The obtained results show that the suggested model allows the difference in the effects of piracetam and phenazepam to be disclosed.

Animals↗
PubMed1997

Complementation of helper-dependent adenoviral vectors: size effects and titer fluctuations.

The complementation of adenoviral vectors with large deletions in the viral genome was studied. The helper adenovirus used to complement these vectors contains a partial deletion of the packaging signal and the E1 region substituted by the lacZ gene. The effect of vector size on packaging efficiency was analysed in 293 cells using decreasingly shorter vectors expressing GFP from a CMV enhancer-beta-actin promoter. Vectors with longer genomes propagated more efficiently than shorter ones. Vectors containing only the packaging signal and the ITRs of Ad5, having all the viral genes replaced with unrelated sequences packaged as efficiently as vectors of the same size containing adenoviral DNA instead of exogenous DNA. The amounts of helper and vector produced in coinfected 293 cells exhibited the typical cycling fluctuation observed during serial propagation of a virus with defective interfering particles.

Adenoviridae↗