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Biomedical subjects

M Marques

Publications and source records attributed to M Marques.

At least 19 recordsLinked to original sources

Does renal mass exert an independent effect on the determinants of antigen-dependent injury?

The aim of this retrospective study was to determine whether nephron mass may exert a direct, independent effect on immunological tolerance. To this end, data corresponding to patients transplanted with en block pediatric kidneys (EBPK) (n=48) were compared with those of renal transplants with a low risk of hyperfiltration (LRH) comprised of recipients of a kidney from young donors (age 5-40 years) (n=173), and transplants with a high risk of hyperfiltration (HRH) comprised of patients who had received a graft from an elderly donor (older than 55 years) (n=91). All the patients had been subjected to the same immunosuppressive treatment. The median follow-up period was 54 months (6-127 months). The EBPK group showed lowest serum creatinine and highest creatinine clearance levels at each follow-up time. The rate of proteinuria >500 mg/day was 5.7% in EBPK, 7.4% in LRH, and 27.3% in HRH (P=0.000). The incidence of acute corticoresistant rejection was minor in EBPK (7.0% in EBPK, 21.3% in LRH, and 23.3% in HRH; P=0.04). Logistic regression analysis showed that the type of transplant was predictive of acute corticoresistant rejection [RR 5.33 (95% confidence interval (CI) 1.15-24.62) for HRH and RR 4.75 (95%CI 1.06-21.27) for LRH, P=0.03]. Multivariate analyses for graft failure due to chronic rejection and for graft failure due to acute rejection according to Cox's regression analysis demonstrated that HRH transplant was a significant predictive variable of both types of failure [4.08 (95%CI 1.27-13.04) for graft loss due to chronic rejection and 8.69 (95%CI 1.69-44.67) for graft loss due to acute rejection]. The present stratification of data according to nephronal mass would appear to indicate that the greater the mass, the lower the incidence of both acute and chronic rejection. This finding lends support to the hypothesis that a large mass of transplanted tissue relative to recipient mass may dampen the immune response.

Adult↗

Incidence of renal artery stenosis in pediatric en bloc and adult single kidney transplants.

BACKGROUND: The incidence, time of onset, and outcome of transplant renal artery stenosis (TRAS) in pediatric en bloc (PT) and adult single-kidney (AT) transplants were reviewed. METHODS: Forty-three cases (7 PT and 36 AT) of suspected TRAS were selected out of 367 functioning grafts (35 of them PT). Diagnosis was performed by digital subtraction arteriography. Percutaneous transluminal angioplasty (PTA) was performed when needed. RESULTS: Seven (20%) PT and 24 (7.1%) AT presented TRAS. Time of onset was 7+/-1 months in PT and 18+/-17 months in AT (P<0.05). PTA was performed in all cases of TRAS in PT and in 19 AT. One PT and 7 AT had re-stenosis. There was no significant difference in renal function after treatment. Control of blood pressure improved in both groups. CONCLUSIONS: Recipients of PT grafts showed a higher incidence and earlier onset of TRAS. Re-stenosis was more frequent in AT.

Adolescent↗

Stormwater run-off and pollutant transport related to the activities carried out in a modern waste management park.

Stormwater run-off from twelve different areas and roads has been characterized in a modern waste disposal site, where several waste management activities are carried out. Using nonparametric statistics, medians and confidence intervals of the medians, 22 stormwater quality parameters were calculated. Suspended solids, chemical oxygen demand, biochemical oxygen demand, total nitrogen and total phosphorus, as well as run-off from several areas, showed measured values above standard limits for discharge into recipient waters--even higher than those of leachate from covered landfill cells. Of the heavy metals analyzed, copper, zinc and nickel were the most prevalent, being detected in every sample. Higher concentrations of metals such as zinc, nickel, cobalt, iron and cadmium were found in run-off from composting areas, compared to areas containing stored and exposed scrap metal. This suggests that factors other than the total amount of exposed material affect the concentration of metals in run-off, such as binding to organic compounds and hydrological transport efficiency. The pollutants transported by stormwater represent a significant environmental threat, comparable to leachate. Careful design, monitoring and maintenance of stormwater run-off drainage systems and infiltration elements are needed if infiltration is to be used as an on-site treatment strategy.

Environmental Monitoring↗

Use of enhanced green fluorescent protein to monitor retroviral-mediated gene therapy in human keratinocytes.

Keratinocytes have great promise as targets for gene therapy involving both skin as well as for systemic disorders due to their availability and potential long life span. Improvement of gene transfer into keratinocytes will be greatly facilitated by markers that will allow both rapid detection and efficient selection of transduced cells. For these purposes, a recombinant version of the Aequorea victoria green fluorescent protein that is enhanced for high-level expression in mammalian cells (EGFP) was placed into a replication-deficient retroviral vector. High-titer retrovirus was used to transduce both primary cultures of neonatal foreskin-derived human keratinocytes (HK) as well as the immortalized keratinocyte-derived cell line HaCaT. Both cell types stably expressed the EGFP, and this marker allowed rapid purification of transduced cells by fluorescence-activated cell sorting. EGFP expression was seen in HaCaT keratinocytes for at least 40 passages, and the presence of this construct did not effect cell growth, or apoptosis in response to UVB or etoposide. Transduced populations of HK were grafted into SCID mice, resulting in a functional epidermis. EGFP expression was readily seen in vivo by exposing the xenografts to an ultraviolet light source. These studies demonstrate the feasibility of using EGFP as a convenient and rapid marker to monitor keratinocyte gene transfer both in vitro and in vivo.

Animals↗

Detection of prognostic significant translocations in childhood acute lymphoblastic leukaemia by one-step multiplex reverse transcription polymerase chain reaction.

The search for chromosomal translocations in de novo cases of childhood acute lymphoblastic leukaemia (ALL) is crucial for the selection of the appropriate therapeutic protocol. In this work, we describe a new method - one-step multiplex reverse transcription polymerase chain reaction (RT-PCR) - to screen for prognostic significant translocations in childhood ALL. Our approach involves a single PCR reaction for the simultaneous detection of the molecular rearrangements resulting from the t(9;22), t(12;21), t(4;11) and t(1;19), with a turnaround time of less than 24 h. This assay proved to be highly sensitive, specific, reproducible and easy to implement in a routine genetics laboratory.

Child, Preschool↗

Distribution of glycogen phosphorylase and cytochrome oxidase in the central nervous system of the turtle Trachemys dorbigni.

Glycogen phosphorylase (GP) and cytochrome oxidase (CO) activities were mapped histochemically in the brain of the turtle Trachemys dorbigni. In the telencephalon, both activities occurred in the olfactory bulb, in all cortical areas, in the dorsal ventricular ridge, striatum, primordium hippocampi and olfactory tubercle. In the diencephalon, they were identified in some areas of the hypothalamus, and in rotundus and geniculate nuclei. Both reactions were detected in the oculomotor, trochlear, mesencephalic trigeminal nuclei, the nucleus of the posterior commissure, torus semicircularis, substantia nigra and ruber and isthmic nuclei of the mesencephalon. In all layers of the optic tectum GP activity was found, but CO only labelled the stratum griseum centrale. In the medulla oblonga both enzymes appear in the reticular, raphe and vestibular nuclei, locus coeruleus and nuclei of cranial nerves. In the cerebellum, the granular and molecular layers, and the deep cerebellar nuclei were positive for both enzymes. The Purkinje cells were only reactive for CO. In the spinal cord, motor and commissural neurones exhibited a positive reaction for the two enzymes. However, CO also occurred in the marginal nucleus and in the lateral funiculus. These results may be useful as a basis for subsequent studies on turtle brain metabolism.

Animals↗

Pharmacokinetics of radioiodinated growth hormones in the turtle Chrysemys dorbigni.

Growth hormone binding proteins (GHBP) have been identified in the blood of many species. The aim of the present work is to study the physiological role of the GHBP in the turtle serum which we recently described. Binding studies were carried out using in vivo pharmacokinetic and chromatographic techniques as well as in vitro methods. When (125)I-GH was injected in physiological concentration into Chrysemys dorbigni turtles, the first step of pharmacokinetics was the binding of a significant fraction of the labeled GH by the GHBPs present in serum. The decay curve followed a three compartments model and gave the equation: Ae(-alphat) + Be(-betat) + Ce(-gammat). The fast compartment with t(1/2) of 14.4 min or 25.2 min, for hGH and bGH represents 30.3% and 18.9% of total radioactivity, respectively, at hypothetical time zero (not experi mental). Chromatographic studies reveal that this rapid compartment represents free GH. The second and third compartments represent complex forms between GH and GHBPs present in the turtle serum, and represent 70% and 80% of total radioactivity for hGH and bGH, respectively. In vitro chromatographic studies showed direct evidence of the presence of GHBPs in the turtle serum. The presence of these GHBPs changed the pharmacokinetics of labeled GH in plasma and the subsequent liver uptake of GH. The labeled hGH or bGH binds to turtle serum in similar proportion, but maximal liver uptake of these hormones are completely different (L/B ratio of 9.2 +/- 0.6 (n = 5) for ( 125)I-hGH and 4.8 +/- 0.3 (n = 7) for (125)I-bGH). The reasons for these differences could be that human GH binds to lactogenic and somatotropic receptors and bovine GH binds only to somatotropic receptors.

Animals↗

Axotomy increases NADPH-diaphorase activity in the dorsal root ganglia and lumbar spinal cord of the turtle Trachemys dorbigni.

Seven days after transection of the sciatic nerve NADPH-diaphorase activity increased in the small and medium neurons of the dorsal root ganglia of the turtle. However, this increase was observed only in medium neurons for up to 90 days. At this time a bilateral increase of NADPH-diaphorase staining was observed in all areas and neuronal types of the dorsal horn, and in positive motoneurons in the lumbar spinal cord, ipsilateral to the lesion. A similar increase was also demonstrable in spinal glial and endothelial cells. These findings are discussed in relation to the role of nitric oxide in hyperalgesia and neuronal regeneration or degeneration.

Animals↗

Functional glomerular reserve in recipients of en bloc pediatric transplant kidneys.

BACKGROUND: The transplantation of an adequate renal mass is increasingly recognized to be of importance. The improved graft survival is probably due to a lesser risk of developing hyperfiltration-associated lesions. METHODS: We have reviewed the glomerular reserve in our recipients of en bloc pediatric transplant kidneys after an intravenous amino acid overload and compared them to single adult kidney transplant recipients. RESULTS: En bloc transplants evidenced increased glomerular filtration rate as compared with baseline as from the second hour of amino acid infusion (from 71+/-14 to 84.9+/-17 ml/min, 1.73 m2, P<0.05) and increased renal plasma flow as from the third hour (from 335+/-116 to 402+/-155 ml/min, 1.73 m2, P<0.05). In the single adult kidney recipient group, no change was seen either in the glomerular filtration rate (from 62.5+/-13 to 58.1+/-13 ml/min, 1.73 m2, P=NS) nor in renal plasma flow (from 354+/-125 to 304+/-98 ml/min, 1.73 m2, P=NS). CONCLUSIONS: These results show that patients receiving en bloc pediatric kidney transplantations have a greater renal functional reserve and show a lesser risk of hyperfiltration.

Adult↗

[Uveitis].

Uveitis is a general term that refers to the inflammation of uveal tract, which is an important cause of blindness in young people. It is well known that uveitis can be the initial manifestation of a systemic disease (S.D.), and may appear years before the diagnosis of the primary disease. Uveitis should be integrated in a systemic study with proper testing. Therefore, the diagnosis is a matter for the ophthalmologist and the Specialist in internal medicine. We have made a retrospective study of 71 patients with chronic uveitis or panuveitis. We found 54.9% of primary uveitis and 45.1% of S.D. associated uveitis, most of them with Behçet's disease (16/71) and Ankylosing Spondilytis (7/71). HLA typing of the patients showed a decreased frequency of HLA A1 and HLA A3 antigens and an increased frequency of the HLA B27 antigen, when compared to a Portuguese control population. We confirmed the important role of HLA B27 as an independent susceptibility factor for anterior uveitis. The lowest HLA A3 frequency was observed in the group of S.D. associated uveitis, which could suggest that this antigen may play a role as a factor of resistance to uveitis.

Adolescent↗

Insulin binding sites in gills of the estuarine crab Chasmagnathus granulata.

Bovine 125I-insulin was injected into the estuarine crab Chasmagnathus granulata in order to study its distribution and specific uptake by tissues. The highest radioactivity uptake occurred in both anterior and posterior gills, which reached maximum values at 30-60 min following labeled insulin administration. Heart and hepatopancreas concentrated a very low amount of radioactivity (only 9 and 3%, respectively, of that shown by gills). A significant reduction of the uptake was observed in the gills when an excess of unlabeled insulin was injected together with the labeled hormone. In vitro studies also showed specific uptake of 125I-insulin by the gills incubated at 25 degrees C, which reached a plateau after 120-min incubation, suggesting a saturable process. The inhibition of 125I-insulin uptake was dose dependent on unlabeled insulin. Glucagon did not compete with radioactivity uptake by gills in vivo and in vitro. Further characterization of insulin-binding sites was performed in gill membrane. The amount of unlabeled insulin that prevented 50% of the 125I-insulin uptake was 7.78 micrograms/ml, and the Scatchard plot analysis established the presence of binding site with Kd of 3.11 microM and Bmax of 0.14 microM (r = 0.99). Ovine prolactin was not able to prevent. 125I-insulin binding to gill membrane. These findings seem to indicate the presence of specific binding sites for insulin or insulin-like substance in crab gills, which deserves further studies.

Animals↗

Identification and initial characterization of serum growth hormone binding protein in the turtle Chrysemys dorbigni.

Proteins that bind growth hormone (GHBP) have been identified in the blood of many mammalian and avian species, but not in reptilian species. We carried out binding studies with the serum of turtles using chromatographic techniques as well as the dextran-charcoal separation method. As in other species, we found at least two different GHBPs: one with high MW and low affinity and the other with lower MW and higher affinity. The high affinity GHBP was partially purified using gel filtration and affinity chromatography, reaching a degree of purification of 11,000 times (0.17 nmol/g of serum protein in the serum vs 1900 nmol/g protein in the purified material). When the high affinity GHBP was characterized, it was found to have a dissociation constant (Kd: 2.6 +/- 0.7 nM) similar to those described for mouse or rat, but lower than those for chicken, rabbit or man. The binding capacity (Bmax) was 120 +/- 43 fmoles/mg of protein, which can be also expressed as 1.08 +/- 0.38 pmol/ml of serum. A preliminary MW estimation of 50-60 kDa was obtained for turtle higher affinity GHBP. The specificity of this high affinity GHBP is somatogenic, since bovine GH competes as well as human GH for 125I-hGH bound to binding protein, while ovine PRL competes only partially and with low affinity. Unrelated hormones, as insulin and glucagon, can not displace the 125I-hGH bound to turtle GHBP. A very important seasonal variation in turtle GHBP activity was observed: maximum binding was found in November (springtime), followed by a continuous decline over March and May.

Animals↗

Acromesomelic dwarfism: a new variation.

The rate skeletal disorder, acromesomelic dwarfism, is characterized by short stature and short limbs. Bone dysplasia is evident. We report two cases of a variation of this disorder in a Portuguese woman and her son. The clinical features of these two cases differ from those of cases previously reported in the literature.

Adult↗

Decreased basal and acute insulin-stimulated effect on the uptake of glucose and amino acid in vitro by thyroid glands from streptozotocin-diabetic rats.

Since experimental diabetes in rats and mice is associated with impairment of several aspects of thyroid function, we determined glucose and amino acid uptake in vitro by isolated thyroid glands from normal and streptozotocin-diabetic rats. Adult male Wistar rats weighing 150-200 g were used. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ, 65 mg/kg body weight) and after five days only rats with blood glucose levels higher than 250 mg/dl were used. The thyroid glands were preincubated in Krebs-Ringer bicarbonate buffer in the presence or absence of insulin (0.7 nM to 7 muM) for 90 min and then incubated with the same concentration of the hormone or its vehicle plus 0.2 microCi of [1-14C]-2-deoxy-D-glucose ([14C]DG) or [1-14C] methylaminoisobutyric acid ([14C]MeAIB) for 15 to 180 min. The uptake of [14C]DG or [14C]MeAIB by the thyroid glands of normal rats increased as a function of incubation time, and the presence of insulin (7 microM) induced a significant increase of labelled DG from 3.30 +/- 0.11 to 4.16 +/- 0.12 and of labelled meAIB from 1.79 +/- 0.06 to 3.10 +/- 0.17 tissue/medium ratio (T/M) and after 45 min of incubation. The lowest concentration of insulin that increased both [14C]DG and [14C]MeAIB transport was 7 nM. Thyroid glands from STZ rats exhibited lower basal values of [14C]DG (4.03 +/- 0.11 T/M) or [14C]MeAIB uptake (1.05 +/- 0.05 T/M) than glands from normal rats (4.62 +/- 0.13 and 1.70 +/- 0.08 T/M, respectively). Insulin produced a stimulatory effect on the transport of both substrates in STZ rats. However, the maximal stimulating concentration of the hormone did not restore [14C]DG and [14C]MeAIB uptake to control values (4.89 +/- 0.17 in STZ rats versus 5.44 +/- 0.17 T/M in controls for [14C]DG, and 1.51 +/- 0.11 in STZ rats versus 2.19 +/- 0.10 T/M in controls for [14C]MeAIB). These results indicate that insulin exerts a direct action on the thyroid gland, and its absence or reduction affects thyroid metabolism, contributing, at least in part, to the abnormality in thyroid function associated with diabetes mellitus.

Amino Acids↗

[Primary lymphomas of the brain not related to immunodeficiency].

Primary brain lymphoma (PBL) is a rare disease, the study of which is based on small series and case reports. The AA review new cases of PBL not associated to immunodeficiency at the Portuguese Institute of Oncology of Porto since 1978. Five male patients were found with lymphoma primarily located in the brain, with aggressive histology, IE staging, detected by CAT, NMR or brain scintigraphy and with histologic material collected by stereotaxic biopsy (3 cases) or by craniotomy (2 cases). Three patients were in complete remission (25,5 and 1 year) and 2 were dead with a survival of 12 and 17 months. The cases and literature are discussed.

Adult↗

Fat-tissue injection versus graft: experimental study in rabbits.

An experimental study of fat-tissue implants in the ears and frontal regions of rabbits was conducted, starting with blocks of tissue for surgical implantation or with multifragmented pieces for injection. The implants were observed from day 7 to month 12. Low integration levels were observed in all animals, except for 1 case that had 20% and 42% of integration, respectively, for the block grafts and for the injected multifragmented tissue.

Adipocytes↗