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T Giordano

Publications and source records attributed to T Giordano.

At least 37 records · Page 2Linked to original sources
PubMed1993

Quantitative reverse transcriptase-polymerase chain reaction of glucose transporter 1 mRNA levels in rat brain microvessels.

We investigated the usefulness of reverse transcriptase-polymerase chain reaction (RT-PCR) to quantify glucose transporter 1 (GLUT1) mRNA in cerebral microvessels. The technique was validated using an in vitro-transcribed RNA fragment (riboprobe) of partial 3' noncoding sequence of rat brain GLUT1 gene. A known amount of the riboprobe was reverse-transcribed to cDNA (target DNA). PCR primers were made to amplify a 292-bp fragment of the target DNA. The 5' primer was end labeled with 32P. An oligonucleotide of 100 bp containing the same sequences as the first 30 and the last 70 bases of the 292-bp fragment of the target DNA was synthesized and used as competitive DNA. The target DNA was coamplified with increasing amounts of competitive DNA using the same two primers. The ratio of radioactivity between amplified products of the target DNA (292-bp fragment) and the competitive DNA (100-bp fragment) was determined quantitatively after separation by gel electrophoresis and radioactivity counting. This method gave an accurate estimation of the amount of the riboprobe in the reaction and a 2- to 5-fold change in the amounts could be detected. By this method, the mean amount of GLUT1 mRNA from purified rat brain microvessels was estimated to be 1.5 +/- 0.1 x 10(-6) ng/ng total RNA. This value was about 10-fold higher than that in rat cell line PC12.

Animals↗
PubMed1993

Altered levels and splicing of the amyloid precursor protein in the adult rat hippocampus after treatment with DMSO or retinoic acid.

Alzheimer's disease and cognitive impairment in rats has been associated with an increase in the percentage of amyloid precursor protein (APP) containing the KPI domain. It has recently been reported that retinoic acid (RA) is capable of increasing the levels and altering the splicing ratio of APP in cultured SH-SY5Y cells. The effects of peripherally administered RA (64 or 640 micrograms/kg; i.p.; q.d.) on the abundance of APP, the ratio of the three major isoforms, and the relative abundance of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) were determined by rtPCR in the hippocampus of aged rats. Corresponding changes in choline acetyltransferase (ChAT) activity were also measured. Vehicle (DMSO) treated rats exhibited a 2 x (P < 0.01) increase in total APP and an 8 x (P < 0.001) decrease in the cyclophilin transcript. In addition, DMSO increased the percentage of APP 695 from 89% in saline treated rats to 94%. Treatment of RA in DMSO decreased the accumulation of total APP relative to cyclophilin at both the low (6.4 x; P < 0.01) and high (8 x; P < 0.05) dosages when compared to DMSO treated rats. Furthermore, the level of APP-695 decreased to 82% with low dosage of RA and 75% at high dosage of the total APP transcripts. No significant change in either NGF, NT-3, or BDNF transcripts were observed following low or high dosage RA administration relative to cyclophilin RNA nor was a change in ChAT activity detected at either of the dosages tested.(ABSTRACT TRUNCATED AT 250 WORDS)

Alternative Splicing↗
PubMed1993

[Adenocarcinoma of the cecal appendix].

The authors report one rare case of primary adenocarcinoma of the vermiform appendix. The patient, an 83 year old female, presented with a non-pathognomonic symptomatology; diagnosis was made postoperatively. Surgery consisted of appendicectomy. The patient, due to her poor general condition, died three days after surgery for cardiocirculatory complications. In the light of the very rare case published in the literature, some diagnostic, anatomic-clinical, therapeutic and prognostic aspects of this rare tumor are discussed.

Adenocarcinoma↗
PubMed1992

Differential expression of cytochrome oxidase (COX) genes in different regions of monkey brain.

A frontal pole cDNA library from monkey (Macaca mulatta) brain was screened to identify mRNAs that are expressed more in frontal pole as compared to primary visual cortex. Three cDNA clones, whose greater expression was confirmed by Northern blot analysis, were identified as cytochrome oxidase (COX) subunits I, II, and III (COX I, II, and III). Each clone showed higher levels of mRNA in the frontal pole, dorsal lateral prefrontal cortex, and hippocampus than in the primary visual or somatosensory cortices. COX histochemistry of prefrontal, visual, and somatosensory cortical regions demonstrated heterogeneous distributions, with highest activity in dendrite-rich neuropil of the cortex. A laminar distribution of COX mRNA expression also was demonstrated with in situ hybridization. mRNA was detected in cell bodies and in apical dendrites. These results indicate region specific differences in the distribution of COX activity and in the corresponding mRNA for three of its subunits within the monkey brain. Such differences may be related to differences in the distribution of neuropil as compared with cell bodies among the brain regions studied, and may be relevant to selective vulnerability in Alzheimer's disease.

Animals↗
PubMed1992

Expression of Alu and 7SL RNA in Alzheimer's and control brains.

We investigated whether changes in expression of RNA polymerase III (pol III) or heterodisperse RNA polymerase II (pol II) transcripts hybridizing to Alu could be detected in Alzheimer's disease (AD). RNA samples obtained from AD and control brain tissues were examined by Northern analysis for Alu and 7SL RNA expression. All RNA samples contained a prominent band of approximately 300 nucleotides which corresponds to 7SL RNA, the Alu-homologous RNA component of the signal recognition particle. In addition, three small (i.e. less than 300 nucleotide) 7SL/Alu-hybridizing transcripts were detected. The two larger of the low molecular weight transcripts hybridized preferentially to the 7SL RNA probe, while the smallest transcript hybridized to the Alu probe. These transcripts and the heterodisperse RNA were variable in quantity and displayed a lack of correlation with AD.

Alzheimer Disease↗
PubMed1992

Thyroid hormone regulation of NGF, NT-3 and BDNF RNA in the adult rat brain.

The effects of peripherally administered thyroid hormone (TH; 500 micrograms/kg; i.p.; q.d.) on the relative abundances of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) RNA were determined by rtPCR in the cortex and hippocampus of young adult rats. Corresponding changes in choline acetyltransferase (ChAT) activity were measured since NGF and BDNF have been shown to enhance the expression of this marker enzyme of central cholinergic pathways. Abundance levels of NGF and NT-3, relative to cyclophilin (cycl), were increased significantly (+50%, P < 0.05) in the hippocampus following TH treatment. Despite enhanced abundance of NGF in the hippocampus, ChAT activity was unchanged, whereas ChAT activity was modestly increased by 28% in the cortex without corresponding changes in NGF, NT-3 or BDNF. These results demonstrate that TH administration is capable of inducing the accumulation of NT-3, in addition to NGF but that the induction levels of RNA cannot be directly correlated with responsivity of the cholinergic system as measured by ChAT activity.

Animals↗
PubMed1992

Nucleotide sequence of the cDNA encoding the common alpha subunit of the chicken pituitary glycoprotein hormones.

Recombinant cDNA clones that encode the alpha subunit of the chicken pituitary glycoprotein hormones were isolated from a pituitary library. The longer of the two cDNA clones that were sequenced was 754 bp in length. It contained 81 nucleotides of the 5'-untranslated region (UTR), an open-reading frame of 360 bp that encoded a 24 amino acid leader polypeptide sequence as well as the 96 amino acid mature alpha subunit, and 268 nucleotides of the 3'-UTR, followed by a 45 bp poly(A) tract. There was 69-79% homology between the nucleotide sequence of the coding region for the chicken and mammalian alpha-subunit cDNAs. Northern blot analysis revealed that the steady-state levels of an approximately 800 bp alpha-subunit specific transcript increased quantitatively when dispersed chicken pituitary glands were treated in culture with chicken gonadotrophin-releasing hormone-I.

Amino Acid Sequence↗
PubMed1991

Isolation of a population of transiently transfected quiescent and senescent cells by magnetic affinity cell sorting.

Rous sarcoma virus (RSV) and cytomegalovirus (CMV) promoters were tested for activity in proliferating and nonproliferating (quiescent or senescent) human embryo fibroblasts. These promoters were cloned upstream of the coding sequence for the Tac subunit of the interleukin 2 receptor, and activity was calculated from the fraction of Tac antigen positive cells detected in a coupled transient transfection/magnetic affinity cell sorting assay. Differences in promoter activities are substantial in quiescent cells: the efficiency of the RSV promoter is no greater than background whereas the CMV promoter is equally active in serum concentrations ranging from 0.5 to 20%. While both promoters are functional in growing cells (WI-38 and HeLa), the CMV promoter exhibits twofold greater activity. Surprisingly, in senescent cells both promoters exhibit the same degree of activity.

Avian Sarcoma Viruses↗
PubMed1989

Identification of a highly abundant cDNA isolated from senescent WI-38 cells.

A cDNA library was constructed from poly(A)+ RNA derived from late passage WI-38 cells and differentially screened with cDNA probes from early and late passage cells. From a number of clones which exhibited differences in intensity of hybridization to the early or late passage probes, one was chosen for further analysis because of its large increase in hybridization to the late passage probe. This clone accounted for approximately 1% of the recombinants in the library. The partial cDNA clone shows complete sequence homology to elongation factor I alpha (EF-I alpha). Northern analysis of poly(A)+ RNA from cells at various population doublings suggested that a 2.2-kb transcript, homologous to EF-I alpha accumulates as cells near the end of their replicative life span (phase III). When early and late passage cells were reexposed to serum after serum starvation, this transcript decreased in abundance. Additionally, a lower molecular weight transcript (1.6 kb) was detected 18 h following serum stimulation in early passage cells and 9 h after stimulation in late passage cells.

Base Sequence↗
PubMed1989

Increase in abundance of a transcript hybridizing to elongation factor I alpha during cellular senescence and quiescence.

We have isolated a senescence-specific clone (pSEN) from a cDNA library constructed from late passage WI-38 human diploid fibroblast that accounts for approximately 1% of the recombinants. Nucleotide sequence analysis of the partial cDNA clone has led to the identification of pSEN as elongation factor I alpha. Northern analysis of poly(A)+ RNA from various intermediate population doubling levels shows that a 2.2 kb transcript hybridizes to pSEN but is expressed prior to PDL-40 at very low levels. This transcript begins to accumulate at PDL-40 and is induced approximately 50-fold just prior to senescence. Furthermore, this transcript was shown to be specific to Go of the cell cycle whereas a second, lower molecular weight transcript (1.6 kb) was observed during S phase (Giordano and Foster, unpublished data). The 2.2 kb transcript is also detected in neonatal foreskin cells but very little increase in abundance is observed between early and late passage cells. Sucrose gradient fractionation of RNA from late passage WI-38 cells suggests that the lower molecular weight transcript is associated with the polysome fraction while the 2.2 kb transcript sediments with the nonpolysomal fraction. Thus, the possibility exists that the 1.6 kb transcript is derived from the 2.2 kb transcript.

Aging↗
PubMedDate not supplied

Sensitization to cockroach in Italy: a multicentric study.

The prevalence of sensitization to cockroach (CR) was evaluated by skin prick test (SPT) in 1299 patients consecutively referred for rhinitis and asthma to five allergy centers in northern, central, and southern Italy. In patients with positive SPT to CR, an SPT with D. pteronyssinus extract was made. RAST inhibition was performed with pooled sera of subjects with positive SPT and RAST of at least Class 3 for CR and D. Pteronyssinus, to assess the cross-reactivity between these two allergenic sources. The protein content of allergen extracts was assayed by isoelectric focusing (IEF). A total of 170 patients (13%) had a positive SPT with CR extract, and 133 of them (78.2%) were also positive for D. pteronyssinus. RAST inhibition showed a cross-reactivity between CR and D. pteronyssinus, and IEF detected in the extracts employed a series of bands focusing at the same pI of CR and D. pteronyssinus relevant allergens. Therefore, the prevalence of sensitization to CR in the population studied, corresponding to 13%, suggests that CR extract might be added to standard SPT panels for respiratory allergy. However, the cross-reactivity with D. pteronyssinus calls for careful evaluation of the clinical importance of this sensitization.

Adolescent↗
PubMedDate not supplied

[Minimal breast carcinoma (the authors' experience)].

The authors relate their experience about twenty-one cases of impalpable lesions of the breast come to their observation during the years 84-88. After expanding therapeutical tactics carried out by them, the promoters infer asserting that a wide diffusion of screening programmes allowed a more verifications of the minute lesions of the breast, addressing the surgeon towards operations more and more conservatives.

Adult↗