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J Wells

Publications and source records attributed to J Wells.

At least 55 records · Page 3Linked to original sources
PubMed1998

Blood brain barrier endothelial cells express candidate amyloid precursor protein-cleaving secretases.

Proteolytic cleavage of the amyloid precursor protein (A beta PP) results in the generation of the amyloidogenic fragment known as amyloid beta peptide (A beta). Deposition of A beta in the brain parenchyma and cerebrovasculature is a feature of Alzheimer's disease (AD). To date, the process whereby A beta is generated and deposited remains unclear. We have previously established that activated platelets from AD patients retain more A beta PP on their surface than control platelets. We report here that an endothelial cell-derived enzyme can cleave this surface platelet A beta PP. Human blood brain barrier endothelial cells from brains of AD patients were assayed for potential A beta PP-cleaving enzymes using synthetic peptide substrates encompassing the A beta N-terminus cleavage site. A protease activity capable of cleaving A beta PP on the surface of AD platelets was noted. The A beta PP cleavage is partially inhibited by EDTA, by ZincOV, as well as by a specific inhibitor of the Zn metalloprotease E.C.3.4.24.15. Furthermore, the protease is recognized by an antibody directed against it, using immunohistochemistry, Western blot analysis and flow cytometry. The protease is not secreted, but rather resides intracellularly as well as on the surface of the endothelial cells. The data suggest that E.C.3.4.24.15 synthesized by brain endothelial cells may process the platelet-derived A beta PP, yielding fragments which could contribute to cerebrovascular A beta deposits.

Alzheimer Disease↗
PubMed1997

Microtubule-mediated transport of organelles and localization of beta-catenin to the future dorsal side of Xenopus eggs.

The dorsal-ventral axis in frog embryos is specified during the first cell cycle, when the cortex rotates relative to the cytoplasmic core along parallel microtubules associated with the core. Cytoplasmic transfer experiments suggest that dorsal determinants are transported 90 degrees from the vegetal pole to the dorsal equator, even though the cortex rotates only 30 degrees. Here we show that, during rotation, small endogenous organelles are rapidly propelled along the subcortical microtubules toward the future dorsal side and that fluorescent carboxylated beads injected into the vegetal pole are transported at least 60 degrees toward the equator. We also show that deuterium oxide, which broadens the zone of dorsalization even though it reduces the extent of rotation and is known to randomize the microtubules, also randomizes the direction of organelle transport. Moreover, beta-catenin, a component of the Wnt signaling pathway that possesses dorsalizing activity in Xenopus, colocalizes with subcortical microtubules at the dorsal side of the egg at the end of rotation. We propose that cortical rotation functions to align subcortical microtubules, which then mediate the transport of dorsal determinants toward their plus ends on one side of the egg.

Animals↗
PubMed1997

Comparison of calcium mobilization in response to noradrenaline and acetylcholine in submandibular cells of newborn and adult rats.

The response of mature and immature rat submandibular cells to alpha-receptor stimulation was compared in terms of the generation of inositol triphosphate (IP3) and Ca2+ mobilization, and of how the calcium mobilization response affects acetycholine (ACh)-induced Ca2+ mobilization. In mature cells, noradrenaline (NA) caused much smaller IP3 and Ca2+ responses than ACh. However, the Ca2+ release induced by NA was enough to partially discharge an agonist-sensitive store and to reduce Ca2+ release by a subsequent ACh stimulus. Exposure to NA also caused an influx of Ca2+ in the mature cells, which was largely associated with Ca2+ entry induced by store depletion (i.e. capacitative entry). In the immature submandibular cells of newborn rats, NA caused essentially no IP3 response and a small Ca2+ release, which only partially affected the Ca2+ released by a subsequent exposure to ACh. In contrast to adult cells, immature cells did not show an increased Ca2+ influx after exposure to NA. However, prestimulation with this agonist potentiated the Ca2+ influx activated by ACh in the cells of newborn rats, but not in cells of adult rats. As both mature and immature submandibular cells have a well-developed phosphoinositide turnover response to ACh, the findings in mature cells suggest a less efficient coupling between alpha-receptors and phospholipase C, while those in immature cells suggest that this coupling is even less functional in the early stages of postnatal development. In permeabilized and 45Ca(2+)-loaded mature cells, cyclic ADP-ribose (cADPR) released 13.4% of loaded 45Ca2+ and this release was significantly reduced by pre-exposure to IP3. Similarly, pre-exposure to cADPR also reduced the IP3-induced 45Ca2+ release. It is concluded that: (1) stimulation with NA induces a smaller Ca2+ release in mature and immature submandibular cells than ACh; (2) the mediator for this small Ca2+ mobilization may be cADPR; and (3) NA stimulates capacitative Ca2+ entry in mature cells, but not in immature cells, and it also activates a Ca2+ entry pathway distinct from the one induced by store depletion, particularly in immature cells.

Acetylcholine↗
PubMed1997

Post-exercise cutaneous hyperaemia resulting from local exercise of an extremity.

Large changes in skin blood flow occur after exercise. Most studies have concentrated on the systemic effects of vigorous exercise on skin blood flow. We were interested in the post-exercise response in the neighbourhood of focal exercise. We used a painless neuromuscular electronic stimulator to exercise the muscles of the forearm, producing flexion of the fingers. There was no change in blood pressure and only a small increase in heart rate during this exercise. We measured blood flow during a 5-min pre-exercise period and a 5-min post-exercise period at the forearm, at the dorsum of the index finger and on the pad of the index finger. We also measured values on the contralateral non-exercised extremity during exercise as well as during matched time periods in control experiments with no exercise. Exercise did elicit an increased blood flow in the post-exercise period at all three sites compared with the control experiments with no exercise and on the contralateral extremity. For example, the increase in blood flow at the finger dorsum was 2.1 +/- 0.1 ml (min 100 g)-1 after exercise compared with -0.08 +/- 0.09 ml min-1 100 g-1 during the control experiment and 0.1 +/- 0.1 ml (min 100 g)-1 on the contralateral arm (all P < 0.01). The local application of heat at the site of blood flow monitoring produced a substantial increase in the post-exercise response at the two finger locations [27.4 +/- 0.4 ml (min 100 g)-1 at the finger dorsum], but not at the arm. This is the first demonstration that highly focal exercise, unaccompanied by a systemic haemodynamic response, can elicit a post-exercise cutaneous hyperaemia. Local heating produced a large synergistic increase in the post-exercise hyperaemia at sites with arteriovenous microvascular perfusion but not at sites with primarily nutritive perfusion. These findings show that local vasoregulatory changes occur in response to exercise, even in the absence of whole-body haemodynamic and thermal change.

Arm↗
PubMed1997

Circadian relationships between interleukin (IL)-6 and hypothalamic-pituitary-adrenal axis hormones: failure of IL-6 to cause sustained hypercortisolism in patients with early untreated rheumatoid arthritis.

Systemic symptoms in rheumatoid arthritis (RA) are mediated, at least in part, by elevated levels of circulating interleukin (IL)-6, and this cytokine is also a potent stimulus of the hypothalamic-pituitary-adrenal axis. To evaluate the 24-h circadian secretory dynamics of ACTH, cortisol, and IL-6 and their interactions in patients with early untreated RA, we recruited and studied five newly diagnosed, untreated RA patients early in the course of their disease and five age-, gender-, and race-matched control subjects. We collected serial blood samples over 24 h and measured plasma ACTH and cortisol every 30 min and IL-6 every hour. The 24-h collection was followed by administration of ovine CRH (oCRH) and post-oCRH serial blood samples over 2 h. We analyzed the 24-h overall levels of these hormones and their circadian variations and performed time-lagged cross-correlation analyses among them. The untreated RA patients had 24 h time-integrated plasma ACTH, plasma cortisol levels, and urinary free cortisol excretion that were not significantly different from control subjects, in spite of their disease activity. However, an earlier morning surge of plasma ACTH and cortisol in the patients was suggested. Plasma ACTH and cortisol responses to oCRH were similar in RA patients and controls. IL-6 levels were significantly increased in the RA patients compared with control subjects during the early morning hours (P < 0.05). There was pronounced circadian variation of plasma Il-6 levels. In the RA patients, we detected a positive temporal correlation between plasma levels of IL-6 and ACTH/cortisol, with elevated levels of IL-6 before the elevations of ACTH and cortisol by 1 and 2 h, respectively. In the same patients, we detected a negative effect of cortisol upon IL-6 exerted with a delay of 5 h. The data presented here suggest that although endogenous IL-6 may stimulate secretion of ACTH and cortisol, overall activity of the hypothalamic-pituitary-adrenal axis remains normal and apparently is insufficient to inhibit ongoing inflammation in early untreated RA patients.

Adrenocorticotropic Hormone↗
PubMed1997

Differences in intracellular CA2+ stores of submandibular cells of adult and newborn rats.

The intracellular Ca2+ stores of submandibular acinar cells of adult and newborn rats were compared by measuring changes in cytosolic Ca2+ ([Ca2+]i) with the Ca2+-sensitive fluorescent probe, fura-2, in intact cells and 45Ca2+ release in permeabilized cells. In cells of adult rats, acetylcholine (ACh) and thapsigargin (TG) elicited, respectively, a 105% and 125% net increase in [Ca2+]i in Ca2+-free medium. In cells of newborn animals, the ACh-induced [Ca2+]i increase (218%) was larger; but the TG-induced increase was significantly smaller (77%), suggesting that the TG-sensitive Ca2+ pump of the store is less developed, abundant, or responsive in these cells. After ACh or TG, ionomycin elicited a further Ca2+ release. Ionomycin by itself completely discharged the ACh- and TG-sensitive store in cells of mature glands but not in cells of immature glands, suggesting a further difference in the distribution of the agonist and ionomycin sensitivities in the two types of cells. After exposure to ACh + ionomycin or TG + ionomycin, collapsing the intracellular pH gradient with monensin caused a third and large Ca2+ release (262% and 289%, respectively) in cells of adult rats, but a significantly smaller release (96%, P < 0.001, and 57%, P < 0.001, respectively) in cells of newborn rats, suggesting that a third, IP3-insensitive acidic store is significantly smaller in cells of immature glands. In permeabilized and 45Ca2+-loaded cells, inositol-1,4,5-trisphosphate (IP3, 5 microM) induced an identical 45Ca2+ release (34% and 33%, respectively) in both types of cells, but TG induced a significant smaller 45Ca2+ release in cells of immature glands (15%) than in cells of mature glands (33%). Monensin discharged <10% of loaded 45Ca2+ in both types of cells, indicating that the radiotracer was not loaded into the monensin-sensitive store. These results suggest that in cells of immature glands: (1) the TG-sensitive Ca2+ pump of the IP3-sensitive store is not well developed; (ii) the IP3-sensitive store is not completely discharged by ionomycin; and (iii) an acidic store likely to be associated with secretory granules is also underdeveloped and of a smaller size than in cells of mature glands.

Acetylcholine↗
PubMed1996

Evidence for a Ca2+ pool associated with secretory granules in rat submandibular acinar cells.

Intracellular Ca2+ stores in rat submandibular acinar cells were characterized using the Ca(2+)-sensitive fluorescent indicator fura 2 and the radiotracer 45Ca2+. Acetylcholine induced a rapid Ca2+ release from a store sensitive to inositol 1,4,5-trisphosphate (IP3) and to thapsigargin (TG). After this store was presumably depleted, ionomycin caused a further increase in cytosolic free Ca2+ concentration ([Ca2+]i), suggesting the presence of an IP3-insensitive Ca2+ release from a store that is more extensive and heterogeneous than the IP3-sensitive one and includes a small mitochondrial component. After both of these stores had been discharged, exposure to monensin caused an additional release of Ca2+ from a third store. This store appears to be associated with secretory granules, since Ca2+ release was significantly reduced when degranulation was induced by isoprenaline. This third store appears to be insensitive to IP3, discharges Ca2+ when the pH gradient across the limiting membrane is collapsed with monensin and only in the presence of both ionomycin and monensin. Ca2+ release from this store is not by Na+/Ca2+ exchange, since simply altering [Na+]i did not cause significant Ca2+ release. In permeabilized cells, IP3 and TG released approx. 35% of 45Ca2+, and ionomycin released an additional 57%, whereas monensin only caused a small additional release, suggesting that only IP3- and ionomycin-sensitive stores are loaded with 45Ca2+ under these conditions. The absence of significant isotope uptake into the ionomycin+monensin-sensitive store may result from a low rate of tracer accumulation or from the lack of Ca2+ pumps in the store. The pattern of response was similar in the presence and absence of mitochondrial inhibitors, indicating that the store is not located in mitochondria. In summary, these results suggest that a substantial IP3-insensitive Ca2+ store is present in secretory granules in rat submandibular acinar cells.

Acetylcholine↗
PubMed1996

Evidence for reduced capacity for Ca2+ removal from the cytosol in submandibular cells of newborn rats.

The increase in cytosolic free Ca2+ concentration ([Ca2+]i) seen in submandibular cells of early postnatal rats following exposure to acetylcholine (ACh) is larger than in cells of adult rats. To elucidate possible reasons for this difference, we compared Ca2+ movements through Ca2+ pumps in both types of cells using Ca(2+)-sensitive fluorescent probe fura-2 and the radiotracer 45Ca2+. Ca2+ release induced by endoplasmic reticulum (ER) Ca(2+)-pump inhibitor thapsigargin (TG) was significantly smaller in neonatal cells than in adult cells, whereas the inositol 1,4,5-trisphosphate (IP3)-elicited Ca2+ release was comparable in both cell types. This suggests that although the size of the IP3-sensitive Ca2+ pool is adequate in immature cells, the activity of TG-sensitive Ca2+ pump in this pool is lower. The activity of the plasma membrane (PM) Ca(2+)-pump, measured by extrusion of 45Ca2+, was also significantly lower in immature cells. These results indicate that both ER and PM Ca2+ pumps may be functionally underdeveloped in immature cells, and that the enhanced increase of [Ca2+]i seen in response to ACh in immature cells may be partially, if not completely, due to a reduced capacity for removal of Ca2+ from the cytosol by active mechanisms.

Acetylcholine↗
PubMed1996

Genetic susceptibility to experimental autoimmune uveoretinitis in the rat is associated with an elevated Th1 response.

This study examines whether genetic susceptibility vs genetic resistance to experimental autoimmune uveoretinitis (EAU) are connected to a predisposition to mount a Th1-dominated (IFN-gamma high, IL-4 low) vs a Th2-dominated (IL-4 high, lFN-gamma low) response. Lewis rats developed disease with high incidence after immunization with the uveitogenic peptide R16, whereas F344 rats were resistant. Primed lymph node cells from both strains proliferated in culture in response to R16. However, while the Lewis cultures transferred EAU to syngeneic recipients, those of F344 did not. The Lewis cultures produced substantially more IFN-gamma mRNA and protein in response to R16, than did those of F344. Both strains made low levels of IL-10 mRNA and IL-4 mRNA. Unlike the primary cultures, long-term (R16-specific) T cell lines derived from each of the strains transferred EAU equally well to their respective recipients, and produced similar, high levels of IFN-gamma mRNA and protein. Treatment of F344 with Bordetella pertussis toxin concurrently with immunization abrogated its resistance, enhanced Ag-specific IFN-gamma production in culture, and yielded a primed cell population capable of transferring EAU. Conversely, immunization of Lewis rats with R16 in IFA induced little or no disease; the primed cells produced minimal amounts of IFN-gamma and did not transfer EAU. However, addition of IL-12 into the culture resulted in a highly pathogenic, IFN-gamma-producing cell population. We conclude that genetic susceptibility to ocular autoimmunity in this model is connected to an elevated Th1 response. Genetic resistance, however, does not seem to involve an elevated Th2 response, but rather an inhibited development of Th1-like effector cells.

Amino Acid Sequence↗
PubMed1996

A simplified, competitive RT-PCR method for measuring rat IFN-gamma mRNA expression.

We describe an adaptation of competitive RT-PCR to quantitate rat IFN-gamma mRNA expression. An IFN-gamma DNA mimic that shared the same primers and had an identical sequence to the target mRNA except for deletion of 66 nucleotides was created by a simple PCR amplification from target cDNA. To reduce variations of initial RNA concentrations, beta-actin cDNAs from each target RNA sample were normalized using the densitometric data. A known amount of pretitrated DNA competitor was then used to analyze the relative levels of target cDNA in different samples by PCR co-amplification. The amplification efficiency for both target and competitor remained constant throughout the PCR reaction, and the ratio of target to competitor PCR product remained proportional to the initial ratio of target to competitor. Relative mRNA levels among samples determined by this method were comparable to levels determined by northern blot analysis. They were also comparable to levels of IFN-gamma protein estimated by ELISA. We conclude that this method can be used to estimate the relative abundance of the target mRNA. This method is adaptable to quantitation of other cytokines and is particularly valuable if there are numerous samples or if the amount of initial mRNA is limited.

Animals↗
PubMed1996

Management of thyroid carcinoma with radioactive 131I.

PURPOSE: To evaluate the role of radioactive 131I in the management of patients with well differentiated carcinoma of the thyroid. METHODS AND MATERIALS: Between 1965 and 1995, a total of 117 patients with well-differentiated carcinoma of the thyroid underwent either lobectomy or thyroidectomy followed by 100-150 mCi of 131I. RESULTS: With a median follow-up of 8 years, only four patients (3%) developed a recurrence of their disease. The 5-year actuarial survival was 97% with a 10-year survival of 91%. There were no severe side effects noted after 131I therapy. CONCLUSIONS: Radioactive 131I is a safe and effective procedure for the majority of patients with well-differentiated thyroid carcinoma. We currently recommend that all patients undergo a subtotal or total thyroidectomy followed by 131I thyroid scanning approximately 4 weeks after surgery. If the thyroid scan shows no residual uptake and all disease is confined to the thyroid, we recommend following patients with annual thyroid scans and serum thyroglobulin levels. If there is any residual uptake detected in the neck or if the tumor extends beyond the thyroid, we recommend routine thyroid ablation of 100-150 mCi of radioactive 131I.

Adolescent↗
PubMed1996

Transneuronal changes of the inhibitory circuitry in the macaque somatosensory thalamus following lesions of the dorsal column nuclei.

The inhibitory circuitry of the ventroposterolateral nucleus (VPL) of the macaque somatosensory thalamus was analyzed in normal animals and in those surviving for a few days or several weeks following a unilateral lesion of the cuneate nucleus, the source of medial lemniscal (ML) axons carrying information from the contralateral upper extremity. Inhibitory synaptic terminals in the VPL were defined as those that contain flattened or pleomorphic synaptic vesicles and that can be shown to be immunoreactive for gamma-aminobutyric acid (GABA). There are two types of these profiles: F axon terminals that arise from neurons of the thalamic reticular nucleus, and perhaps from VPL local circuit neurons (LCNs); and the dendritic appendages of LCNs that form presynaptic dendrites (PSDs). ML terminals normally have extensive synaptic interactions with PSDs but not with F axon terminals. Electron microscopic analyses revealed that cuneatus lesions resulted in a rapid loss of ML terminals and a statistically significant reduction in both F and PSD synaptic profiles. Confocal scanning microscopy also demonstrated a profound loss of GABA immunoreactivity in the deafferented VPL. These changes persisted for more than 20 weeks, without any evidence of reactive synaptogenesis of surviving sensory afferents or of inhibitory synapses. The changes in GABA circuitry are transneuronal, and the possible mechanisms that may underlie them are discussed. It is suggested that the altered GABAergic circuitry of the VPL in the monkey may serve as a model for understanding changes in somatic sensation in the human following peripheral or central deafferentation.

Afferent Pathways↗
PubMed1996

One hundred and fifty hepatic resections: evolution of technique towards bloodless surgery.

A technique of hepatic resection is described and the results of 150 resections are reviewed. Hepatic transection was performed, under intermittent portal inflow occlusion, using ultrasonic aspiration to skeletonize portal branches and venous tributaries. Control of venous haemorrhage during resection was optimized by argon beam coagulation and lowering central venous pressure to between 0 and 4 cmH2O by extradural blockade and systemic nitroglycerine infusion. One patient with jaundice died in hospital, giving a mortality rate of 0.7 per cent. There were no deaths in patients without jaundice and cirrhosis. Fifteen patients (10.0 per cent) had significant complications, nine medical and six surgical, including three bile leaks (2.0 per cent). Mean blood loss was 814 ml for the whole study but only 434 ml in the last 4 years. During this latter period mean blood transfusion in hospital was 0.5 units and mean postoperative haemoglobin value fell by 0.7 g per 100 ml. Hepatic resection can be performed with the same degree of confidence and similar low morbidity as any other major surgical procedure.

Adolescent↗
PubMed1996

Analysis and comparison of the mouse and human immunoglobulin heavy chain JH-Cmu-Cdelta locus.

We report here 23,686 bases of contiguous DNA sequences from the mouse germline immunoglobulin heavy chain (H) constant (C) mu delta region. The sequence spans the joining (JH) regions, the mu constant region (C mu), the delta constant region (C delta) coding regions, a domain relic, the mu switch region (S mu), seven blocks of simple sequence repeats, a large unique sequence inverted repeat, a large unique sequence forward repeat, and all of the intervening material. A comparison of this 23.7-kb region with the corresponding human C mu/C delta region reveals clear homology in the coding and introns of C mu but not in the 5' flanking J gene segments nor in the intergenic and C delta regions. This mixed pattern of similarity between the human and the mouse sequences contrasts with high levels of similarity found in the T-cell receptor C alpha/C delta region and alpha and beta myosin genes and the very low levels found in the gamma-crystallin, XRCC1, and beta-globin gene clusters. The human and mouse comparison further suggests the incorporation of novel sequences into expressed genes of IgD.

Animals↗