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Biomedical subjects

P Sanchez

Publications and source records attributed to P Sanchez.

At least 73 records · Page 4Linked to original sources

Immunocytochemical studies of tryptophan hydroxylase, tyrosine hydroxylase, and serotonin innervation in the aging rat neurointermediate pituitary.

Innervation within the aging male rat pituitary neurointermediate lobe was examined using immunocytochemical methods with antisera to rate-limiting enzymes for serotonin (tryptophan hydroxylase, TpOH) or catecholamine (tyrosine hydroxylase TH) synthesis. Immunostaining for both enzymes was observed in both neural and intermediate lobes in younger rats (age 6 weeks or 7 months), but TpOH was absent from the intermediate lobe in older groups (10-12, 15-16, and 18-19 months). Colocalization of TpOH and TH was demonstrated in some nerve fibers, using immunofluorescence. The oldest age group showed a reduced staining pattern for TH. 5-hydroxytryptophan, the immediate precursor for serotonin (5-HT), was taken up and converted to immunoreactive 5-HT in all age groups. Overall, the pattern of staining for regulatory neurotransmitters or enzymes which ultimately affect pituitary intermediate lobe secretory function is significantly modulated during the aging process.

5-Hydroxytryptophan↗

[Measuring impedance for evaluating ischemia damage to the human liver in preparation for transplantation].

In 22 human donor livers the measurement of the non-invasive bioelectrical impedance was performed prospectively to evaluate the degree of tissue damage sustained during cold ischemia. The results of the measurement were correlated with liver function, the method of organ preservation and the period of ischemia. The impedance was measured in vivo as 620 ohm (at 192 Hz), the phase angle as -7.4 degrees (at 5 kHz). The results were compared with the data obtained from 72 patients who underwent elective laparotomies. The 22 donor livers were studied further during ischemia. The method was found to be a reliable way of detecting severe damage to the hepatocytes during the cold ischemia.

Adolescent↗

Enhancement of testosterone secretion by normal adult human Leydig cells by co-culture with enriched preparations of normal adult human Sertoli cells.

An in-vitro method was developed to study Sertoli-Leydig cell interactions in man, using testes removed at the time kidneys were removed for transplantation from 6 young adult men (aged 17-45 years) after cerebral death. After collagenase digestion of testicular tissue, Leydig cells were purified on discontinuous Percoll gradients. Two fractions of Leydig cells, 'L2' and 'L3' which differed in their buoyant density (1.05 g < L2 < 1.06 g and 1.06 g < L3 < 1.08 g), were obtained. The Sertoli cell-enriched preparation was obtained from seminiferous tubular fragments after sequential treatment with glycine buffer to remove peritubular-myoid cells, a second collagenase digestion, mechanical fragmentation and washes to remove germ cells. Purified Leydig cells were then cultured either alone or together with Sertoli cells in culture dishes coated with collagen, fibronectin and laminin in a chemically defined medium without serum. The influence of co-culture on basal testosterone secretion was examined in 3 successive 48 h periods.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Cognitive risk factors and neuropsychological performance in HIV infection.

Almost all investigations examining the effects of early HIV infection on neuropsychological functioning in homosexual males have excluded subjects with cognitive risk factors such as recreational drug use and head injury. While insuring that results reflect the influence of the virus on cognition, this selection bias limits the ability to generalize findings. Comprehensive neuropsychological evaluations were compared between two groups of homosexual males with a variety of cognitive risk factors. Subjects were 132 HIV seropositive males (108 CDC class II & III and 24 CDC class IVA & IVC2) and 65 HIV seronegative controls. Recreational drug use in the six months prior to exam was found to interact with HIV infection and was associated with selective areas of cognitive decline and a significantly worse overall neuropsychological performance. Although significantly lower functioning in the domains of Verbal Memory and Attention and Speed of Information processing was noted for subjects with CDC class IVA and IVC2 compared to seropositives with CDC class II & III, overall neuropsychological performance was similar in these two groups. At this early stage of HIV infection, we did not find indication of association between neuropsychological performance and decreased immunological status. A history of head injury and recent recreational drug use emerged as primary cognitive risk factors associated with decreased neuropsychological performance. As 50% of our HIV seropositive subjects reported active recreational drug use, this cognitive risk factor in particular may contribute to the appearance of HIV-related cognitive deficits during early stages of HIV infection.

Cognition Disorders↗

Preferential VH/V lambda pairings occur in the available B cell repertoire of adult BALB/c mice.

To gain insights into the composition of the B cell repertoire, we have investigated VH gene family expression associated with individual light chains. For this purpose, we have examined the use of 12 VH gene families in a large collection of hybridomas expressing one of the four lambda light chains [lambda 1 (V1J1), lambda 2 (V2J2 and V x J2) and lambda 3 (V1J3)]. Our results show that the distribution of the VH families is very different from one lambda subtype to another. This suggests that a few substitutions between VL regions are sufficient to generate very different associated repertoires by strong selection mechanisms. Moreover, we assume that the global VH expression pattern is not random but rather composed of many preferential VH/VL associations.

Animals↗

Evaluation of immune complexes after immunotherapy with wheat flour in bakers' asthma.

Inhalant food allergy has been described many times in literature, but double-blind clinical trials to support successful hyposensitization to these allergens has seldom been reported. Some authors have suspected that certain adverse reactions after immunotherapy may be mediated by immune complexes. Furthermore, the FDA does not recommend injection therapy with food extracts. We present a study on the detection of adverse effects after immunotherapy with an inhalant food (wheat flour) in a double-blind clinical trial in 26 patients with bakers' asthma. We investigated the presence of circulating immune complexes (CICs) after 2 years of treatment with hyposensitization to wheat flour.

Antigen-Antibody Complex↗

Sensitization to the storage mite Lepidoglyphus destructor in wheat flour respiratory allergy.

Occupational allergy due to hypersensitivity to cereal flours is relatively common among bakers and grain-store workers. Storage mites can contaminate wheat flour and could be an important cause of allergic symptoms due to inhalation. Forty-three patients with criteria for allergic sensitization to wheat flour (skin tests, specific IgE to wheat flour and positive challenge tests) were included in a study to investigate the prevalence of cosensitization to Lepidoglyphus destructor (Ld). This mite was the predominant species in the wheat flour samples supplied by our patients. We found that 30% of the patients had IgE-mediated hypersensitivity of Ld. Of these, 23% did not have a relationship with any bakery or agriculture. We conclude that the prevalence of sensitization to Ld in patients sensitized to wheat flour is important.

Adolescent↗

V lambda-J lambda rearrangements are restricted within a V-J-C recombination unit in the mouse.

The murine lambda gene locus is organized as follows: V lambda 2-V lambda x-J lambda 2C lambda 2-psi J lambda 4C lambda 4-V lambda 1-J lambda 3C lambda 3-J lambda 1C lambda 1 where all segments have the same transcriptional orientation. The combinatorial process of gene recombination should allow the generation of eight distinct immunoglobulin light chains. We have therefore investigated the probability of obtaining such chains among the mature lambda B cell repertoire. We analyze serum lambda immunoglobulins and lambda B cell clones induced by treatment with rabbit anti-lambda antibodies coupled to LPS. Confirming previous data obtained by others, our results indicate that the rearrangements of lambda segments take place within each V lambda-J lambda-C lambda cluster, thereby defining a unit of recombination. Our results also provide no evidence for the use of undescribed segments as has been recently suggested by the finding of the V lambda x segment.

Animals↗

Effects of dipyridamole on the short-term evolution of glomerulonephritis.

The aim of the present study is to evaluate the effect of dipyridamole (300 mg/day) versus placebo in a double-blind randomized trial on membranous glomerulonephritis (M-GMN), mesangial IgA glomerulonephritis (IgA-GMN), and segmentary and focal hyalinosis glomerulonephritis (SFH-GMN) during the first 3 months of treatment. In the case of M-GMN, proteinuria dropped by 60% of the basal value in patients treated with dipyridamole; in the case of IgA-GMN it dropped by 65-70%; and in the case of SFH-GMN it dropped by 40% of the basal value. Inhibition of proteinuria in M-GMN was correlated to platelet response, and above all, to the ADP-induced platelet aggregation in whole blood.

Dipyridamole↗

Growth hormone and insulin-like growth factor I treatment increase testicular luteinizing hormone receptors and steroidogenic responsiveness of growth hormone deficient dwarf mice.

To test the hypothesis that insulin-like growth factor (IGF-I) is required for the in vivo development of testicular Leydig cell function, either recombinant human GH [(hGH) (1.5 micrograms/g BW) or recombinant IGF-I (1 microgram/g BW) was injected three times daily into immature Snell dwarf mice (dw/dw) and into phenotypically normal control (Dw/-) for 7 days. In dw/dw mice hGH enhanced significantly body, liver, kidney, and testicular weight. In addition, hGH increased testicular LH receptors and the acute steroidogenic response to human CG, but there was no significant effect on basal plasma testosterone or plasma LH levels. The effects of IGF-I in body and kidney weight were less pronounced than those produced by hGH, but its effects on testicular weight and LH receptors, as well as on the acute steroidogenic response to human CG, were similar to that observed after hGH treatment. In Dw/- mice hGH had no effect on either body or organ weight or on testicular function, despite the fact that it induced a significant increase in plasma IGF-I levels. These results indicated that IGF-I is able to induce the maturation of Leydig cell function and that the effects of hGH on the testis are probably mediated by IGF-I. They also suggest that the delayed puberty associated with GH deficiency or resistance is most likely related to an IGF-I deficiency.

Animals↗

Human proopiomelanocortin-(79-96), a proposed cortical androgen-stimulating hormone, does not affect steroidogenesis in cultured human adult adrenal cells.

The existence of a cortical androgen-stimulating hormone (CASH), distinct from ACTH, regulating the secretion of human adrenal androgens has long been postulated. Recently, it has been reported that an 18-amino acid peptide, corresponding to the first part of the joining peptide of proopiomelanocortin [POMC-(79-96)], was able to stimulate the secretion of dehydroepiandrosterone from cultured human adult adrenocortical cells, but had no effect on cortisol production. We have studied the acute and long term effects of ACTH (10(-11) and 10(-9) M), CASH-18 (10(-8) M), or both on cortisol and dehydroepiandrosterone sulfate by human adult adrenocortical cells. Although ACTH increased steroid secretion and enhanced the steroidogenic responsiveness to further ACTH stimulation, CASH-18 alone or together with ACTH (10(-11) or 10(-9) M) had no effect. In addition, we were unable to demonstrate any specific binding of [125I]CASH-18 to human adrenocortical cells, although [125I] ACTH-(1-39) binds specifically to the same cell preparation.

Adrenal Glands↗

A third sublineage of avian T cells can be identified with a T cell receptor-3-specific antibody.

Avian homologues of mammalian gamma delta and alpha beta TCR, termed TCR1 and TCR2, have been identified in the chicken with specific mAb. A third TCR, dubbed TCR3, has been identified on a subpopulation of T cells that lack the TCR1 or TCR2 epitopes. We have now produced a mAb that identifies this TCR3 molecule. The anti-TCR3 antibody immunoprecipitates a CD3-associated heterodimer with a relative Mr of 88,000, composed of 48,000 and 40,000 disulfide-linked chains. The Mr 40,000 chains of TCR3 and TCR2 exhibited the same isoelectric points of 5.6 to 6.5 and had core proteins of 34,000. Although the Mr 48,000 chain of TCR3 and the Mr 50,000 chain of TCR2 had the same basic isoelectric point of 6.2 to 7.6, their core proteins were different in size, 31,000 vs 29,000. Immunofluorescence analysis reveals that the TCR3 was present on all of the CD3+ T cells not identified by antibodies specific for TCR1 or TCR2. Thymocytes that expressed the surface CD3/TCR3 complex at relatively low levels were predominantly CD4+ and CD8+, whereas those with higher levels of surface CD3/TCR3 were predominantly CD4+ and CD8+ singles. Mature TCR3+ cells in the periphery were also either CD4+ (80%) or CD8+ (20%). The TCR1+, TCR2+, and TCR3+ subsets of T cells were generated sequentially in the thymus and seeded to the periphery in the same order. Intrathymic development of the TCR3+ cells was selectively inhibited by embryonic treatment with the anti-TCR3 mAb. The pattern of histologic localization of TCR3+ cells in the periphery was similar to the TCR2 subset of cells except that the TCR3+ cells were rarely seen in the intestine. Cross-reactivity patterns of the anti-chicken TCR antibodies suggested that other gallinaceous species share the three types of TCR. We conclude that TCR2 and TCR3 in gallinaceous birds may represent alpha beta subfamilies of TCR that are sequentially expressed on developmentally discrete sublines of T cells.

Animals↗

Mouse V lambda x gene sequence generates no junctional diversity and is conserved in mammalian species.

The lambda x, a new mouse Ig lambda L chain, is produced by rearrangement of the V lambda x, J lambda 2, and C lambda 2 gene segments. The V lambda x amino acid sequence is as divergent to other V lambda as to Vk gene sequences. Additionally, its third hypervariable region (CDR3) is four amino acids longer than those of all other variable gene segments of murine L chain. We have cloned and sequenced the germ-line V lambda x gene and found that the unexpected CDR3 length is encoded by the V lambda x gene. Junctional diversity is prevented by a TAA termination codon localized at the V lambda x 3' extremity. Moreover, we show a striking conservation of the V lambda x sequence in various mammalian species. Portions of the V lambda x sequence display more than 70% of nucleotide sequence identity with rabbit and human variable regions. These results suggest that V lambda x predated the divergence of mammalian species.

Amino Acid Sequence↗

Murine lambda gene rearrangements: the stochastic model prevails over the ordered model.

The ontogeny of the immunoglobulin (Ig) gene rearrangement in mammalian B cells seems to be ordered. Heavy chain gene segments rearrange first, followed by light chain gene segments, kappa before lambda. The genomic organization of murine lambda locus does not preclude the simultaneous expression of two subtypes from the same chromosome. In order to distinguish between an ordered and a stochastic model of rearrangement, a panel of 67 B cell hybridomas secreting either lambda 1, lambda 2, lambda 3 or lambda x (recently described) were analysed for V lambda J lambda rearrangements. The results show that in 97% of cases, a single rearrangement occurred, favouring the stochastic model over the ordered one. Strikingly, the possibility of having a productive rearrangement if the first try results in an aberrant one is rare. We propose therefore, that the lambda Ig is not necessarily required to ensure allelic and subtypic exclusion mechanisms. Moreover, in 97% of the cases, at least one kappa allele is rearranged. Furthermore, the RS recombination has been detected in 77% of the cases. This suggests that, although the stimulation of kappa precedes that of lambda locus, the RS recombination acts as a transacting albeit dispensable lambda activator.

Animals↗

Heart rate circadian rhythm as a biological marker of desynchronization in major depression: a methodological and preliminary report.

Heart rate (HR) was continuously monitored during successive 24-hr periods in 19 healthy subjects and 26 major depressed patients (DSM III-R). Recordings were performed after a 2-week wash-out period and the morningness or eveningness typology of each subject was determined. The chronobiological parameters and rhythm percentage (RP) were calculated by the single cosinor method from the smoothed HR curves of each subject. In normal subjects, HR follows a circadian rhythm (RP greater than 65%) with the lowest values at night. Morning type subjects have an earlier peak time (13:30) than evening type subjects (17:30). Major depressive patients were split into two groups; in the first one HR circadian rhythm was still present (RP greater than 63%) with a decrease in amplitude (24%) while in the second group, no circadian rhythm of HR could be detected (RP less than 25%, decrease in amplitude greater than 70%). In the group of patients with a persisting HR circadian rhythm, no veritable phase advance was observed. Our results suggest that circadian HR rhythm, which can be easily studied with non-invasive methods, might represent a chronobiological marker of some depressions. Given the lag that exists between the rhythms of morning type and evening type subjects, our study also stresses the importance of taking into account this behavioural trait in chronobiological studies.

Adult↗

Regulation of pig Leydig cell aromatase activity by gonadotropins and Sertoli cells.

The present work was done to investigate the cell localization of testicular aromatase activity and its regulation in immature pig testis using an in vitro model. Leydig cells and Sertoli cells were isolated from immature pig testes and cultured alone or together in the absence or presence of human chorionic gonadotropin (hCG) or porcine follicle-stimulating hormone (pFSH) for 2 days. At the end of incubation, the amounts of testosterone (T), estrone sulfate (E1S) and estradiol (E2) were measured. Then the cells were incubated for 4 h in the presence of saturating concentrations of delta 4-androstenedione (3 microM) and the amounts of E1S and E2 were measured again (aromatase activity). The ability of Sertoli cells to produce estrogens was very low and neither hCG nor pFSH had any significant effect. hCG stimulated, in a dose-dependent manner, the secretion of T and E1S by Leydig cells cultured alone as well as the aromatase activity of these cells. The main estrogen produced by Leydig cells was E1S. pFSH also stimulated the above parameters of Leydig cell function; this may have been due to the contamination of this hormone with luteinizing hormone (LH). Coculture of Leydig cells with Sertoli cells without gonadotropins had very small effects on T and E1S production and on aromatase activity. However, treatment of coculture with increasing concentrations of hCG had a dramatic effect on Leydig cell functions. For each hCG concentration, the amounts of T and E1S secreted, as well as the aromatase activity of the coculture, were 2- to 3-fold higher than those of Leydig cells cultured alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstenedione↗