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Biomedical subjects

M Castillo

Publications and source records attributed to M Castillo.

At least 19 recordsLinked to original sources

Binding site on pea chloroplast fructose-1,6-bisphosphatase involved in the interaction with thioredoxin.

When we compare the primary structures of the six chloroplast fructose-1,6-biophosphatases (FBPase) so far sequenced, the existence of a poorly conserved fragment in the region just preceeding the redox regulatory cysteines cluster can be observed. This region is a good candidate for binding of FBPase to its physiological modulator thioredoxin (Td), as this association shows clear differences between species. Using a cDNA clone for pea chloroplast FBPase as template, we have amplified by PCR a DNA insert coding for a 19 amino acid fragment (149Pro-167Gly), which was expressed in pGEMEX-1 as a fusion protein. This protein strongly interacts with pea Td m, as shown by ELISA and Superose 12 gel filtration, depending on pH of the medium. Preliminary assays have shown inhibition of FBPase activity in the presence of specific IgG against the 19 amino acid insert. Surprisingly the fusion protein enhances the FBPase activation in competitive inhibition experiments carried out with FBPase and Td. These results show the fundamental role played by this domain in FBPase-Td binding, not only as docking point for Td, but also by inducing some structural modification in the Td molecule. Taking as model the structural data recently published for spinach photosynthetic FBPase, this sequence from a tertiary and quaternary structural point of view appears available for rearrangement.

Amino Acid Sequence

Adenocarcinoma of the endolymphatic sac: imaging features and preoperative embolization.

Adenocarcinoma arising from the endolymphatic sac is very rare. Its imaging features are nonspecific and the differential diagnosis involves mainly paraganglioma or aggressive meningioma. In our case, the blood supply to the tumor was identical to that found in glomus jugulotympanicum tumors and the patient benefitted from preoperative embolization.

Adenocarcinoma

Predictive value of flow cytometric analysis in DNA contents in patients with locally advanced head and neck carcinoma.

A retrospective study was performed on 61 eligible patients with stage III and IV (AJC/UICC Staging System) squamous carcinomas of the head and neck region who were treated with definitive radiotherapy with, or without, surgery. DNA contents were measured by flow cytometric analysis of archival paraffin blocks and were correlated with clinicopathological findings, tumour response and patient survival. Comparison of variables including treatment modality was performed for identification of significant prognostic factors. There were 28 diploid, 27 aneuploid tumours and the remaining six were questionable. All patients were followed-up for at least two years or until death. Aneuploid tumours had a significantly higher S-phase fraction (percentage S-phase) (p < 0.001). Neither ploidy nor percentage S-phase were found to have predictive value in tumour response or patient survival within the power of a sample size of 61. Twenty of the 27 (74 per cent) aneuploid tumours had a complete response (CR) whereas 19 out of 28 (68 per cent) diploid tumours achieved CR. Five-year survival by the Kaplan-Meier method was 33 per cent for both aneuploid and diploid tumours. However, nodal stage (N stage) was found to have significant predictive value in both tumour response and patient survival. The complete response for stage N0 patients was 96 per cent, N1 patients 61 per cent, N2 patients 60 per cent and 43 per cent for N3 patients (p < 0.002). Similarly, the five year survival for the N0 and N3 groups of patients was 53 per cent and 29 per cent respectively (p < 0.05).

Adult

Coconut oil affects lipoprotein composition and structure of neonatal chicks.

Supplementation of 10 or 20% coconut oil in the diet for 1-2 weeks produced a significant hypercholesterolemia in neonatal chicks. Plasma triacylglycerol concentration significantly increased after the addition of 20% coconut oil for 2 weeks. These results show that newborn chicks are more sensitive to saturated fatty acids from coconut oil than adult animals. The effects of this saturated fat on lipoprotein composition were studied for the first 1-2 weeks of neonatal chick life. Coconut oil supplementation in the diet (20%) for 2 weeks increased cholesterol concentration in all the lipoprotein fractions, while 10% coconut oil only increased cholesterol in low-density and very-low-density lipoproteins, an increase that was significant after 1 week of treatment. Similar results were obtained for triacylglycerol concentration after 2 weeks of treatment. Changes in phospholipid and total protein levels were less profound. Coconut oil decreased low-density and very-low-density lipoprotein fluidity, measured as total cholesterol/phospholipid ratio. Changes in esterified cholesterol/phospholipid and triacylglycerol/phospholipid ratios suggest that coconut oil affects the distribution of lipid components in the core of very-low-density particles. Likewise, the esterified cholesterol/triacylglycerol ratio was clearly increased in the low-density, and especially in the very-low-density, fraction after the first week of coconut oil feeding. Our results show that neonatal chick provides a suitable model in which to study the role of very-low-density lipoproteins in atherogenesis and the rapid response to saturated fatty acids with 12-14 carbons.

Age Factors

Squamous cell carcinomas that arise in the oral cavity and tongue base: can CT help predict perineural or vascular invasion?

PURPOSE: To determine the ability to use computed tomography (CT) to predict invasion of adjacent nerves or vessels by oral cavity tumors. MATERIALS AND METHODS: Contrast material-enhanced CT scans and histopathologic reports were retrospectively reviewed in 48 patients (36 men, 12 women) aged 38-75 years who underwent gross total resection of squamous cell carcinomas arising in the tongue, the base of the tongue, and the floor of the mouth. CT criteria for diagnosis of perineural or vascular invasion were aggressive tumor margins, invasion of the sublingual space, and direct adjacency of the tumor to the enhanced lingual vasculature in the sublingual space. CT and histopathologic findings of perineural and/or vascular invasion by tumor were correlated in all patients. RESULTS: With the above criteria, CT findings predictive of perineural or vascular invasion had a sensitivity of 88%; specificity, 83%; positive predictive value, 85%; and negative predictive value, 84%. CONCLUSION: CT findings can be used to predict perineural or vascular invasion by oral cavity tumors.

Adult

Orbital involvement by Wegener's granulomatosis: imaging findings.

OBJECTIVE: The purpose of this study was to define patterns of Wegener's granulomatosis (WG) orbital disease involvement. SUBJECTS AND METHODS: We evaluated the orbital examinations of 14 WG patients (CT for nine, MR imaging for three, and both CT and MR imaging for two) for the number of involved orbits, intraconal or extraconal location, involvement of the orbital apex, sinus involvement, bony erosion, and optic nerve compression. MR images were analyzed for signal characteristics and enhancement patterns. RESULTS: We found orbital disease to be unilateral in 12 patients (86%) and bilateral in two patients (14%) (total of 16 orbits). In 11 orbits (69%), coexistent orbital and sinus diseases with bony erosion were present. Orbital disease without sinus disease was seen in five orbits (31%). Intraorbital disease distribution was as follows: solely extraconal--seven orbits (44%) (four with involvement of muscle); intraconal with muscular involvement--one orbit (6%); combined intra- and extraconal--seven orbits (44%); and primarily orbital apex--one orbit (6%). Involvement of the apex was seen in six orbits. The WG mass was hypointense relative to orbital fat on three standard T2-weighted MR images. We saw homogeneous MR contrast enhancement in all four orbits studied on enhanced T1-weighted images. CONCLUSION: Intraorbital WG involvement is usually accompanied by paranasal sinus disease. A hypointense signal on T2-weighted MR images is helpful in suggesting the diagnosis.

Aged

Hb Costa Rica or alpha 2 beta 2 77(EF1)His --> Arg: the first example of a somatic cell mutation in a globin gene.

We have identified a minor hemoglobin component (approximately 5%) in the blood of a healthy Costa Rican female, but not in her mother and two brothers (father not studied), that has an His --> Arg replacement at position beta 77 (Hb Costa Rica). No other amino acid replacements were observed and no beta- or gamma-chain-like peptides were present. Hb Costa Rica has abnormal stability. Sequence analyses of numerous polymerase chain reaction (PCR)-amplified segments of DNA that contain exon 2 of the beta gene failed to identify a CAC --> CGC (His --> Arg) mutation. The same was the case when cDNA was sequenced, indicating that a beta-Costa Rica-mRNA could not be detected with this procedure. Gene mapping of genomic DNA with Bg/II, BamHI, and HindIII gave normal fragments only and with the same intensity as observed for the fragments of a normal control. The quantities of the beta chain variants Hb J-Iran and Hb Fukuyama with related mutations at beta 77 vary between 30% and 45% in heterozygotes, whereas that of Hb F-Kennestone with the same His --> Arg mutation but in the G gamma-globin gene, is a high 40%-45% (as percentage of total G gamma) in a heterozygous newborn. These different observations exclude a heterozygosity of the A --> G mutation at codon beta 77, as well as a deletion comparable to that of Hbs Lepore or Kenya, or a beta-globin gene duplication, and point to a nontraditional inheritance of Hb Costa Rica. Allele-specific amplification of cDNA with appropriate primers identified the presence of a low level of mutated mRNA in the reticulocytes of the patient, which was confirmed by dotblot analysis of the same material with 32P-labeled probes. Comparable amplification products were not observed in genomic DNA. The A --> G mutation apparently occurred in a somatic cell at a relatively early stage in the development of the hematopoietic cell system, and Hb Costa Rica accumulated through rapid cell divisions in patchy areas in the bone marrow (somatic mosaicism). An unequal distribution of Hb Costa Rica over the red cells supports this possibility.

Adult

Magnetic resonance imaging of the olfactory apparatus.

We review the normal anatomy of the olfactory system with special emphasis on its extraaxial portions. Pathological processes affecting this cranial nerve are divided into those that do not require imaging and those that do. Processes in which patients benefit from imaging, mostly with magnetic resonance, include suspected tumors and congenital abnormalities.

Brain Neoplasms

Magnetic resonance imaging of cranial nerves IX, X, XI, and XII.

Cranial nerves IX (glossopharyngeal), X (vagus), and XI (spinal accessory) are intimately related. Therefore, for imaging purposes they are best considered as a unit rather than as individual structures. The XIth nerves are not considered true cranial nerves because they are formed by cephalic (bulbar) and caudal (spinal) divisions. The XIIth (hypoglossal) nerves arise inferiorly to the above mentioned cranial nerves and exit the skull via a different set of foramina. Because the course of these four pairs of cranial nerves extends from the base of the skull to the upper thorax, a combination of both magnetic resonance imaging (MRI) and computed tomography is needed for optimal evaluation. This article will emphasize the use of MRI when appropriate.

Accessory Nerve

Baseline and follow-up MRI of the internal auditory canal after suboccipital resection of acoustic schwannoma: appearances and clinical correlations.

Prospective baseline MRI was obtained on 31 patients who had "total" removal of acoustic schwannoma 6 months to 9 years previously. Follow-up MRI was performed after 1-2 years on patients with questionable abnormalities. We found four patterns (1) internal auditory canals (IAC) with nonenhancing soft-tissue strands, possibly scars or distorted residual nerves (8); (2) IAC with marginal enhancement-reactive dura mater (16); (3) IAC with contrast-enhancing globular tissues suggesting residual or recurrent tumour (5); (4) high-signal intensity in the IAC before contrast medium administration, probably related to graft with fat/fascia/muscle (2). Prospective 1-to 2-year follow-up studies were available in 8, 5, and 1 patients in groups 2, 3, and 4 respectively. In group 2, dural enhancement remained unchanged in 5 patients and decreased in 3. In group 3 follow-up showed 1 tumour recurrence (surgically confirmed) and 4 stable abnormalities. In group 4, follow-up in 1 of the 2 patients was stable. In groups 1 and 2, the MRI features correlate well with complete tumor removal. Whether follow-up in these groups is indicated needs to be determined. In group 3 residual or recurrent tumor cannot be excluded. In group 4, grafts may prevent adequate visualization of the IAC.

Adult

Disposition and covalent binding of ibuprofen and its acyl glucuronide in the elderly.

Ibuprofen is an over-the-counter nonsteroidal anti-inflammatory drug with a low incidence of severe adverse reactions. It is metabolized by oxidation to carboxyibuprofen and hydroxyibuprofen and by conjugation to an acyl glucuronide. In vitro studies have indicated that ibuprofen glucuronide is labile and reactive, forming covalent adducts with proteins. To verify the formation of ibuprofen-protein adducts in vivo, the pharmacokinetics of ibuprofen glucuronide and its covalent binding to plasma proteins were studied in five elderly patients who received long-term administration of oral doses of ibuprofen. Plasma levels of ibuprofen glucuronide were low relative to those of ibuprofen; the ratio of area under the plasma concentration versus time curve for the glucuronide relative to the parent drug was only 4%. Covalent binding of ibuprofen to plasma protein was observed in all patients, correlating well with the area under the plasma concentration versus time curve of ibuprofen glucuronide (r = 0.966). Compared with reports for other nonsteroidal anti-inflammatory drugs that form acyl glucuronides, plasma levels of ibuprofen-protein adduct are low during long-term administration. The observed lower reactivity in vivo is probably attributable to the greater stability of ibuprofen glucuronide relative to other acyl glucuronides.

Administration, Oral

Gap junction turnover, intracellular trafficking, and phosphorylation of connexin43 in brefeldin A-treated rat mammary tumor cells.

Intercellular gap junction channels are thought to form when oligomers of connexins from one cell (connexons) register and pair with connexons from a neighboring cell en route to forming tightly packed arrays (plaques). In the current study we used the rat mammary BICR-M1Rk tumor cell line to examine the trafficking, maturation, and kinetics of connexin43 (Cx43). Cx43 was conclusively shown to reside in the Golgi apparatus in addition to sites of cell-cell apposition in these cells and in normal rat kidney cells. Brefeldin A (BFA) blocked Cx43 trafficking to the surface of the mammary cells and also prevented phosphorylation of the 42-kD form of Cx43 to 44- and 46-kD species. However, phosphorylation of Cx43 occurred in the presence of BFA while it was still a resident of the ER or Golgi apparatus yielding a 43-kD form of Cx43. Moreover, the 42- and 43-kD forms of Cx43 trapped in the ER/Golgi compartment were available for gap junction assembly upon the removal of BFA. Mammary cells treated with BFA for 6 h lost preexisting gap junction "plaques," as well as the 44- and 46-kD forms of Cx43 and functional coupling. These events were reversible 1 h after the removal of BFA and not dependent on protein synthesis. In summary, we provide strong evidence that in BICR-M1Rk tumor cells: (a) Cx43 is transiently phosphorylated in the ER/Golgi apparatus, (b) Cx43 trapped in the ER/Golgi compartment is not subject to rapid degradation and is available for the assembly of new gap junction channels upon the removal of BFA, (c) the rapid turnover of gap junction plaques is correlated with the loss of the 44- and 46-kD forms of Cx43.

Animals