[Translated article] Preoperative assessment and surgical indications: Separation surgery.

Neurological compression occurs in 10%-20% of patients who develop spinal metastases. In the last decade, the evolution of oncological diagnostic and medical techniques, the change from conventional external radiation to radiosurgery and the new surgical instruments have meant that the treatment of these patients must be indicated in a personalized manner and by consensus, multidisciplinary way, in specific commissions. Today, the biological state of the patient, the presence of mechanical instability, the neurological assessment and degree of epidural compression, as well as the best prognostic categorization of the tumor, are established as decision factors prior to the indication of surgical treatment, treatment that has passed from a cytoreductive concept to that of a spinal cord release from tumor in order to ensure safe radiosurgery.

Preoperative assessment and surgical indications: Separation surgery. / Valoración e indicaciones quirúrgicas: cirugía separación.

Neurological compression occurs in 10%-20% of patients who develop spinal metastases. In the last decade, the evolution of oncological diagnostic and medical techniques, the change from conventional external radiation to radiosurgery and the new surgical instruments have meant that the treatment of these patients must be indicated in a personalized manner and by consensus, multidisciplinary way, in specific commissions. Today, the biological state of the patient, the presence of mechanical instability, the neurological assessment and degree of epidural compression, as well as the best prognostic categorization of the tumor, are established as decision factors prior to the indication of surgical treatment, treatment that has passed from a cytoreductive concept to that of a spinal cord release from tumor in order to ensure safe radiosurgery.

Biocerâmica de fosfato de cálcio nanoestruturada micro-macroporosa em grânulos de absorção rápida no preenchimento de defeito crítico em rádio de coelhos (Oryctolagus cuniculus) / Bioceramics of calcium phosphate nano-structured micro-macro porous granules rapidly absorbed in filling critical radial defect in rabbits (Oryctolagus cuniculus)

O objetivo do presente trabalho foi avaliar, por radiografia, histologia e densitometria óssea, o efeito da HA/ßTCP em grânulos de absorção rápida em defeito ósseo crítico em rádio de coelhos. Foram utilizados 35 coelhos machos, da raça Nova Zelândia, e realizou-se um defeito crítico nos rádios direito e esquerdo. Os animais foram distribuídos em GI, enxerto autólogo e GII, HA/ßTCP em grânulos de absorção rápida. Avaliações radiográficas foram feitas antes da cirurgia, após, aos oito, 15, 30, 45 e 60 dias e avaliações histológicas e de densitometria. Verificou-se diferença significativa ao se comparar a densidade mineral óssea obtida ao longo do tempo de estudo. Observou-se formação de rede vascular entre os poros da biocerâmica desde o primeiro tempo de avaliação, (oito dias). Foram observados tecido ósseo primário e trabéculas em tecido ósseo preexistente a partir de 30 dias da implantação. Aos 60 dias, constatou-se presença de matriz óssea em segmentos ósseos preexistentes, caracterizando a formação óssea centrípeta. A biocerâmica HA/ßTCP nanoestruturada micro-macroporosa em grânulos de absorção rápida não causa alterações microscópicas indicativas de rejeição, permite a invasão e a multiplicação celular, bem como propicia a regeneração óssea, constituindo um implante apropriado para preenchimento de falhas ósseas críticas.(AU)

The objective of this study was to evaluate the effect of HA/ ßTCP on rapid absorption granules in rabbit radiography, histology, and bone densitometry. Thirty - five male rabbits of the New Zealand breed were used and a critical defect was performed on the right and left radios. The animals were distributed in GI, autologous graft and GII HA / ßTCP in rapidly absorbed granules. Radiographic, histological, and densitometry evaluations were performed before surgery, then after eight, 15, 30, 45 and 60 days. A significant difference was found when comparing the bone mineral density obtained over the study time. Formation of vascular network between the bioceramic pores was observed by the first evaluation time, (eight days). Primary bone tissue and trabeculae were observed from preexisting bone tissue after 30 days of implantation. At 60 days, the presence of bone matrix was observed from the preexisting bone segments, characterizing the centripetal bone formation. The micro-macroporous nanocomposite HA / ßTCP of rapidly absorbing granules do not cause microscopic changes indicative of rejection, allows invasion, cell multiplication, and promotes bone regeneration, constituting an appropriate implant for filling of critical bone failures.(AU)

Avaliação histológica e por microscopia eletrônica de varredura da biocerâmica de fosfato de cálcio nano-estruturada micromacro porosa em grânulos em defeito crítico de rádio de coelhos / Histological and nano-structured calcium phosphate bioceramic micro-macro porous granules in critical defect rabbits radius scanning electron microscopy evaluation

Objetivou-se avaliar, histologicamente e por microscopia eletrônica de varredura (MEV), a evolução de defeitos críticos experimentais em rádio de coelhos preenchidos ou não com biocerâmica fosfocálcica nanoestruturada micromacro porosa em grânulos. Utilizaram-se 70 coelhos, Nova Zelândia, adultos jovens, machos, e realizou-se um defeito crítico nos rádios. Os membros constituíram os grupos: GI, biocerâmica lenta, GII, biocerâmica moderada e GIII, controle negativo. Após cada período experimental, os animais foram sacrificados, e os rádios coletados. As avaliações histológicas foram realizadas aos oito, 15, 30, 45, 60, 90 e 120 dias, e as análises de MEV aos 60, 90 e 120 dias. Histologicamente, observou-se processo de reparação óssea mais adiantado nos grupos GI e GII comparando-se ao GIII. Na MEV, constatou-se maior proporção de osso maduro e presença de ósteons secundários nos GI e GII, sendo mais evidente no GII, confirmando os achados histológicos. As cerâmicas promoveram preenchimento completo do defeito, enquanto no grupo controle houve preenchimento no centro do defeito, permanecendo espaços vazios nas laterais. Conclui-se que o emprego das biocerâmicas de absorção lenta e moderada favorece a regeneração óssea completa em defeitos críticos sendo indicadas como substituto ósseo. A maturação óssea ocorre mais precocemente quando se emprega a cerâmica de absorção moderada.(AU)

The objective was to evaluate, through histologic exam and by scanning electron microscopy (SEM), the evolution of experimental critical defects in radio or not filled with nano-structured calcium phosphate micro-macro porous bioceramic granules rabbits. We used 70 rabbits, New Zealand, young adults, males, there was a critical defect in radio. The members were the groups: GI, slow bioceramic, GII, GIII and bioceramic moderate, negative control. After each experimental period, the animals were sacrificed and the radios harvested. Histological evaluations were performed at eight, 15, 30, 45, 60, 90 and 120 days and SEM analyzes at 60, 90 and 120 days. Histologically there was bone healing process earlier in GI and GII compared to GIII. In SEM we observed a higher proportion of mature bone and presence of secondary osteons in GI and GII, being more evident in the GII, confirming the histological findings. Ceramic promoted complete filling of the defect, while the control group was filling in the center of the defect, with empty spaces remaining on the sides. In conclusion, the use of bioceramics, slow and moderate absorption favor complete bone regeneration in critical defects being indicated as a bone substitute. The maturation occurs earlier when employing the ceramic moderate absorption.(AU)

Risk factors for proximal neck complications after endovascular aneurysm repair using the endurant stentgraft.

OBJECTIVE: To assess the incidence and risk factors for proximal aneurysm neck related complications with a late generation device for endovascular abdominal aneurysm repair (EVAR). METHODS: Data were retrieved from a prospective registry (Endurant Stent Graft Natural Selection Global Postmarket Registry) involving 79 institutions worldwide. The risk factors tested were age, gender, surgical risk profile, proximal neck length (<10 mm), diameter (>30 mm), supra- and infrarenal angulation (>60° and 75°), mural thrombus/calcification (>50%) and taper (>10%), and AAA diameter (>65 mm). Two neck related composite endpoints were used, for intra-operative (type-1a endoleak, conversion, deployment/retrieval complication or unintentional renal coverage) and post-operative (type-1a endoleak or migration) adverse events. Independent risk factors were identified using multivariable backwards modeling. RESULTS: The study included 1263 patients (mean age 73, 10.3% female) from March 2009 to May 2011. Twenty three (1.8%) intra-operative adverse events occurred. Neck length <10 mm (OR 4.9, 95% CI 1.1-22.6) and neck thrombus/calcification >50% (OR 4.8, 95% CI 1.7-13.5) were risk factors for intra-operative events. The planned 1 year follow up visit was reached for the entire cohort, and the 2 year visit for 431 patients. During this time, 99 (7.8%) events occurred. Female gender (HR 1.9, 95% CI 1.1-3.2), aneurysm diameter >65 mm (HR 2.8, 95% CI 1.9-4.2), and neck length <10 mm (HR 2.8, 95% CI 1.1-6.9) were significant post-operative risk factors. Neck angulation, neck taper, large diameter neck, and presence of thrombus/calcification were not predictors of adverse outcome in this study. CONCLUSION: These results support the adequacy of this device in the face of adverse neck anatomy, and confirm neck length as the most relevant anatomical limitation for EVAR. Additionally, the study confirms the decline in early to mid-term intervention rates with a newer generation device in a large patient sample. Lastly, it suggests that neck related risk factors affect outcome and impact on prognosis in varying degrees.

Real-time PCR in clinical practice: a powerful tool for evaluating Leishmania chagasi loads in naturally infected dogs.

The performance of the less expensive SYBR-Green-based PCR assay, for quantifying Leishmania chagasi in smears of bone-marrow aspirates from naturally infected, mongrel dogs, was recently compared with that of a similar PCR based on TaqMan chemistry. Aspirates were obtained from 36 infected dogs and examined for parasites by direct examination, culture, and quantitative PCR (qPCR) using specific primers (based on the parasite's kinetoplast DNA), DNA extracted from a smear, and either the SYBR-Green or TaqMan chemistries. Every aspirate smear was found PCR-positive for L. chagasi (whether the assay employed SYBR Green or TaqMan) but only 74% of the aspirates were found positive by culture and only 33% by direct, microscopical examination. There was no evidence of PCR inhibition when the DNA was collected from smears, and the parasite loads estimated using the SYBR-Green PCR were almost identical to those estimated using the TaqMan PCR (r=0.99). As a method for quantifying parasite loads in dogs infected with L. chagasi (and, probably, other mammals infected with other leishmanial parasites), PCR based on SYBR Green may therefore be an appropriate and inexpensive alternative to PCR based on TaqMan, and a reliable clinical tool.

[Hybrid infrainguinal revascularization for critical limb ischemia]. / Revascularização infrainguinal híbrida em isquémia crítica.

The introduction of endovascular procedures in the routine of vascular surgery allowed for the expansion of therapeutic options in the diverse areas of vascular disease. Endoluminal revascularization can be regarded as a usefull complement of conventional surgical techniques. An above-knee femoro-popliteal bypass surgery ePTFE graft has been performed, followed by a sheath introduction on the graft body. This allows a simplified access to crural vessels, while providing efficient revascularization of the femoro-popliteal sector. Distal revascularization was performed using angioplasty and stenting when appropriate, as to insure continuous flux in at least one of the tibial or the peroneal arteries. The aim of this kind of procedure is to insure efficient femoro-popliteal revascularization and to achieve a run-off to the foot in at least one vessel. This type of intervention may be particularly attractive in Leriche-Fontaine's grade IV patients with no available autologous venous graft.

Rat ventral prostate xanthine oxidase-mediated metabolism of acetaldehyde to acetyl radical.

Alcohol drinking is known to lead to deleterious effects on prostate epithelial cells from humans and experimental animals. The understanding of the mechanisms underlying these effects is relevant to intraprostatic ethanol treatment of benign prostatic hyperplasia and to shed some light into the conflictive results linking alcohol consumption to prostate cancer. In previous studies, we provided evidence about the presence in the rat ventral prostate of cytosolic and microsomal metabolic pathways of ethanol to acetaldehyde and 1-hydroxyethyl radical and about the low levels of alcohol dehydrogenase and aldehyde dehydrogenase. Acetaldehyde accumulation in prostate tissue and oxidative stress promotion were also observed. In this study, we report that in the ventral prostate cytosolic fraction, xanthine oxidoreductase is able to metabolize acetaldehyde to acetyl radical. The identification of the acetyl was performed by GC-MS of the silylated acetyl-PBN adduct. Reference adduct was generated chemically. Formation of acetyl was also observed using pure xanthine oxidase. The generation of acetyl by the prostate cytosol was inhibited by allopurinol, oxypurinol, diphenyleneiodonium chloride, folate, and ellagic acid. Results suggest that metabolism of ethanol to acetaldehyde and to 1-hydroxyethyl and acetyl radicals could be involved in the deleterious effects of alcohol drinking on prostate epithelial cells.

Nitroreductive metabolic activation of some carcinogenic nitro heterocyclic food contaminants in rat mammary tissue cellular fractions.

Several nitrofurans and nitroimidazoles have been widely used in veterinary medicine. Some of these compounds are breast carcinogens in rodents and their mechanism of action is hypothesized to be related to reactive metabolites generated by nitroreduction and/or via oxygen-dependent redox cycling. The present work describes the nitroreductive metabolism of nitrofurazone, nitrofurantoin, furazolidone, and metronidazole by the cytosolic and microsomal fractions of mammary tissue from female Sprague-Dawley rats. The data obtained were compared with those obtained with nifurtimox and benznidazole, two well-known rodent carcinogen/mutagens nitroheterocycles. The nitroreductase activity of pure milk xanthine-oxidoreductase (XOR) was evaluated for screening purposes. All the nitrofurans were nitroreduced either by the pure XOR or the cytosolic fraction in the presence of hypoxanthine, and these activities were inhibited by allopurinol. Furthermore, they were nitroreduced by the microsomal fraction in the presence of NADPH, except for the nitrofurazone, suggesting the participation of cytochrome P450 reductase. Nitrofurans metabolism was significantly more intense than that of NFX. No equivalent nitroreductase activity was observed in either subcellular fraction using nitroimidazolic compounds as substrates. These results suggest that the nitroreductive metabolism of nitrofurans and the subsequent redox cycling might be involved in the associated mammary tissue carcinogenic effects.

Deleterious effects induced by oxidative stress in liver nuclei from rats receiving an alcohol-containing liquid diet.

Highly purified rat-liver nuclei were previously shown to have nuclear ethanol (EtOH) metabolizing system able to bioactivate alcohol to acetaldehyde and 1-hydroxyethyl radicals. These reactive metabolites were able to covalently bind to nuclear proteins and lipids potentially being able to provoke oxidative stress of nuclear components. In this study, the above-mentioned possibility was explored. Sprague Dawley male rats (125-150 g) were fed a standard Lieber and De Carli liquid diet for 28 days. Controls were pair-fed with a diet, in which EtOH was isocalorically replaced with carbohydrate. The presence of a chlorzoxazone hydroxylase activity inducible by the repetitive EtOH drinking further suggested the presence of CYP2E1 in the highly purified nuclei. Nuclei from EtOH-drinking rats evidenced significantly increased susceptibility to a t-butyl hydroperoxide challenge as detected by chemiluminescence emission, increased formation of protein carbonyls, and decreased content of protein sulfhydryls. In contrast, no significant changes in the nuclear lipid hydroperoxides formation or even decreases in the 8-oxo-7,8-dihydro-2-deoxyguanosine were observed. No significant differences were observed in different parameters of the alkaline Comet assay. In immunohistochemical studies performed, no expression of p53 was observed in the livers of the animals under the experimental conditions tested. Since nuclear proteins and lipids are known to play a role in cell growth, differentiation, repair and signaling, their alterations by either oxidative stress, or by covalent binding might be of relevance to liver tumor promotion.

Impact of marked weight loss induced by bariatric surgery on bone mineral density and remodeling.

Data about the impact of bariatric surgery (BS) and subsequent weight loss on bone are limited. The objective of the present study was to determine bone mineral density (BMD), bone remodeling metabolites and hormones that influence bone trophism in premenopausal women submitted to BS 9.8 months, on average, before the study (OGg, N = 16). The data were compared to those obtained for women of normal weight (CG, N = 11) and for obese women (OG, N = 12). Eight patients in each group were monitored for one year, with the determination of BMD, of serum calcium, phosphorus, magnesium, parathyroid hormone, 25-hydroxyvitamin D, insulin-like growth factor-I (IGF-I) and osteocalcin, and of urinary calcium and deoxypyridinoline. The biochemical determinations were repeated every three months in the longitudinal study and BMD was measured at the end of the study. Parathyroid hormone levels were similar in the three groups. IGF-I levels (CG = 332 +/- 62 vs OG = 230 +/- 37 vs OGg = 128 +/- 19 ng/mL) were significantly lower in the operated patients compared to the non-operated obese women. Only OGg patients presented a significant fall in BMD of 6.2% at L1-L4, of 10.2% in the femoral neck, and of 5.1% in the forearm. These results suggest that the weight loss induced by BS is associated with a significant loss of bone mass even at sites that are not influenced by weight overload, with hormonal factors such as IGF-I being associated with this process.

Impact of marked weight loss induced by bariatric surgery on bone mineral density and remodeling

Data about the impact of bariatric surgery (BS) and subsequent weight loss on bone are limited. The objective of the present study was to determine bone mineral density (BMD), bone remodeling metabolites and hormones that influence bone trophism in premenopausal women submitted to BS 9.8 months, on average, before the study (OGg, N = 16). The data were compared to those obtained for women of normal weight (CG, N = 11) and for obese women (OG, N = 12). Eight patients in each group were monitored for one year, with the determination of BMD, of serum calcium, phosphorus, magnesium, parathyroid hormone, 25-hydroxyvitamin D, insulin-like growth factor-I (IGF-I) and osteocalcin, and of urinary calcium and deoxypyridinoline. The biochemical determinations were repeated every three months in the longitudinal study and BMD was measured at the end of the study. Parathyroid hormone levels were similar in the three groups. IGF-I levels (CG = 332 ± 62 vs OG = 230 ± 37 vs OGg = 128 ± 19 ng/mL) were significantly lower in the operated patients compared to the non-operated obese women. Only OGg patients presented a significant fall in BMD of 6.2 percent at L1-L4, of 10.2 percent in the femoral neck, and of 5.1 percent in the forearm. These results suggest that the weight loss induced by BS is associated with a significant loss of bone mass even at sites that are not influenced by weight overload, with hormonal factors such as IGF-I being associated with this process.

Biochemical and ultrastructural alterations in the rat ventral prostate due to repetitive alcohol drinking.

Previous studies showed that cytosolic and microsomal fractions from rat ventral prostate are able to biotransform ethanol to acetaldehyde and 1-hydroxyethyl radicals via xanthine oxidase and a non P450 dependent pathway respectively. Sprague Dawley male rats were fed with a Lieber and De Carli diet containing ethanol for 28 days and compared against adequately pair-fed controls. Prostate microsomal fractions were found to exhibit CYP2E1-mediated hydroxylase activity significantly lower than in the liver and it was induced by repetitive ethanol drinking. Ethanol drinking led to an increased susceptibility of prostatic lipids to oxidation, as detected by t-butylhydroperoxide-promoted chemiluminiscence emission and increased levels of lipid hydroperoxides (xylenol orange method). Ultrastructural alterations in the epithelial cells were observed. They consisted of marked condensation of chromatin around the perinuclear membrane, moderate dilatation of the endoplasmic reticulum and an increased number of epithelial cells undergoing apoptosis. The prostatic alcohol dehydrogenase activity of the stock rats was 4.84 times lower than that in the liver and aldehyde dehydrogenase activity in their microsomal, cytosolic and mitochondrial fractions was either not detectable or significantly less intense than in the liver. A single dose of ethanol led to significant acetaldehyde accumulation in the prostate. The results suggest that acetaldehyde accumulation in prostate tissue might result from both acetaldehyde produced in situ but also because of its low aldehyde dehydrogenase activity and its poor ability to metabolize acetaldehyde arriving via the blood. Acetaldehyde, 1-hydroxyethyl radical and the oxidative stress produced may lead to epithelial cell injury.

Health in early adulthood: the contribution of the 1978/79 Ribeirão Preto birth cohort.

The increase in non-communicable chronic diseases of adults is due to demographic changes and changes in the risk factors related to physical activity, smoking habits and nutrition. We describe the methodology for the evaluation of persons at 23/25 years of age of a cohort of individuals born in Ribeirão Preto in 1978/79. We present their socioeconomic characteristics and the profile of some risk factors for chronic diseases. A total of 2063 participants were evaluated by means of blood collection, standardized questionnaires, anthropometric and blood pressure measurements, and methacholine bronchoprovocation tests. The sexes were compared by the chi-square test, with alpha = 0.05. Obesity was similar among men and women (12.8 and 11.1%); overweight was almost double in men (30.3 vs 17.7%). Weight deficit was higher among women than among men (8.6 and 2.6%). Women were more sedentary and consumed less alcohol and tobacco. Dietary fat consumption was similar between sexes, with 63% consuming large amounts (30 to 39.9 g/day). Metabolic syndrome was twice more frequent among men than women (10.7 vs 4.8%), hypertension was six times more frequent (40.9 vs 6.4%); altered triglyceride (16.1 vs 9.8%) and LDL proportions (5.4 vs 2.7%) were also higher in men, while women had a higher percentage of low HDL (44.7 vs 39.5%). Asthma and bronchial hyper-responsiveness were 1.7 and 1.5 times more frequent, respectively, among women. The high prevalence of some risk factors for chronic diseases among young adults supports the need for investments in their prevention.

Health in early adulthood: the contribution of the 1978/79 Ribeirão Preto birth cohort

The increase in non-communicable chronic diseases of adults is due to demographic changes and changes in the risk factors related to physical activity, smoking habits and nutrition. We describe the methodology for the evaluation of persons at 23/25 years of age of a cohort of individuals born in Ribeirão Preto in 1978/79. We present their socioeconomic characteristics and the profile of some risk factors for chronic diseases. A total of 2063 participants were evaluated by means of blood collection, standardized questionnaires, anthropometric and blood pressure measurements, and methacholine bronchoprovocation tests. The sexes were compared by the chi-square test, with alpha = 0.05. Obesity was similar among men and women (12.8 and 11.1 percent); overweight was almost double in men (30.3 vs 17.7 percent). Weight deficit was higher among women than among men (8.6 and 2.6 percent). Women were more sedentary and consumed less alcohol and tobacco. Dietary fat consumption was similar between sexes, with 63 percent consuming large amounts (30 to 39.9 g/day). Metabolic syndrome was twice more frequent among men than women (10.7 vs 4.8 percent), hypertension was six times more frequent (40.9 vs 6.4 percent); altered triglyceride (16.1 vs 9.8 percent) and LDL proportions (5.4 vs 2.7 percent) were also higher in men, while women had a higher percentage of low HDL (44.7 vs 39.5 percent). Asthma and bronchial hyper-responsiveness were 1.7 and 1.5 times more frequent, respectively, among women. The high prevalence of some risk factors for chronic diseases among young adults supports the need for investments in their prevention.

Reactions of nifurtimox with critical sulfhydryl-containing biomolecules: their potential toxicological relevance.

Nifurtimox (Nfx) is a drug used in the treatment of Chagas' disease, an endemic parasitic disease from Latin American countries. It produces undesirable side-effects in patients, frequently forcing the treatment to be stopped. Its toxic mechanism is not fully understood. In this work we describe purely chemical reactions of Nfx with relevant cellular sulfhydryl (SH) compounds. The compounds tested were glutathione (GSH), cysteine (RSH), lipoic acid (LA) and coenzyme A (CoA). All reacted with Nfx to give nitrite (NO(-) (2)). The relative reaction rates were CoA>LA>GSH>RSH. In studies with GSH and RSH the formation of nitrite was accompanied by decreases in Nfx concentration and increases in the formation of a reaction product revealed by HPLC. We failed to show the presence of liver cytosolic GST (GSH transferase activity)-mediated formation of NO2- from Nfx. These NO(-) (2)-releasing processes occurred under in vivo conditions in Nfx-treated Sprague-Dawley male rats (240-260 g body weight) at a dose of 100 mg Nfx kg(-1) p.o. In urine samples NO(-) (2) excretion was accompanied by unchanged drug and two unidentified more polar metabolites detectable by HPLC. The Nfx reactions with critical SH from molecules such as GSH, RSH, LA and CoA, and potentially others containing SH residues (e.g. enzymes or structural proteins), might have toxicological relevance not only for the Nfx side-effects but also for the chemotherapeutic effects on Trypanosoma cruzi. In addition, Nfx reactions with GSH might be crucial in Nfx detoxification.

HLA-B27: a registry of constitutive peptide ligands.

The very strong association of human leukocyte antigen (HLA)-B27 with spondyloarthritis might be related to its peptide-presenting properties. The natural polymorphism of this molecule influences both peptide specificity and disease susceptibility. In this study, we present a comprehensive compilation of known natural ligands of HLA-B27 arising from endogenous proteins of human cells, together with a statistical assessment of residue usage among constitutive peptide repertoires of multiple HLA-B27 subtypes. This analysis provides evidence that every peptide position, including "non-anchor" ones, may be subjected to selection on the basis of its contribution to HLA-B27 binding and also allows a quantization of residue preferences at known anchor positions. The present registry is intended as a basis on which to build up reliable criteria to assess the effect of HLA-B27 polymorphism on peptide presentation, for T-cell epitope predictions, and for molecular mimicry studies.

Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize.

An amaranth ( Amaranthus hypochondriacus) 11S globulin cDNA, encoding one of the most important storage proteins (amarantin) of the seed, with a high content of essential amino acids, was used in the transformation of CIMMYT tropical maize genotype. Constructs contained the amarantin cDNA under the control of a tissue-specific promoter from rice glutelin-1 ( osGT1) or a constitutive ( CaMV 35S) promoter with and without the first maize alcohol dehydrogenase intron ( AdH). Southern-blot analysis confirmed the integration of the amarantin cDNA, and copy number ranged from one to more than ten copies per maize genome. Western-blot and ultracentrifugation analyses of transgenic maize indicate that the expressed recombinant amarantin precursors were processed into the mature form, and accumulated stably in maize endosperm. Total protein and some essential amino acids of the best expressing maize augmented 32% and 8-44%, respectively, compared to non-transformed samples. The soluble expressed proteins were susceptible to digestion by simulated gastric and intestinal fluids, and it is suggested that they show no allergenic activity. These findings demonstrate the feasibility of using genetic engineering to improve the amino acid composition of grain crops.

Endometrial response to a cyclic regimen of percutaneous 17beta-estradiol and low-dose vaginal micronized progesterone in women with mild-to-moderate hypertension.

Endometrial response to natural estradiol and low-dose vaginal progesterone replacement therapy was evaluated in 20 postmenopausal women with chronic, mild-to-moderate hypertension. A cyclic hormone replacement therapy (HRT) regimen was used (21/28 days) with percutaneous estradiol (1.5 mg/day) and vaginal micronized progesterone (100 mg/day). Menopausal symptoms decreased and estradiol concentrations increased substantially and remained in the physiological range throughout treatment. Serum gonadotropin concentrations decreased significantly (p < 0.001, Friedman's ANOVA). Bone mineral density increased by 2.1% (p = 0.029) only at the lumbar spine. Endometrial thickness remained unchanged. Breakthrough bleeding or spotting occurred in 18% of cycles in the first 3 months of HRT, 30% in months 4-9 and 22% in months 10-12. Withdrawal bleeding occurred in 40% of cycles in the first 3 months and decreased to 25% in months 10-12. At month 12, there were 11 women with amenorrhea due to endometrial atrophy. Nine women had active endometria (proliferative or secretory) and thus reported vaginal bleeding. No severe bleeding, hyperplasia, or carcinoma was found. Vaginal bleeding was tolerated, and no subject withdrew from the study. Results suggest that this regimen confers endometrial protection and is well tolerated, and can therefore safely be used for at least 1 year by postmenopausal women with hypertension and menopausal symptoms.