Cryoablation of breast lesions: our experience.

Cryoablation is a therapeutic technique that uses very low temperatures to destroy cells; transcutaneous cryoablation can be performed under imaging guidance. This simple procedure is done on an outpatient basis with local anesthesia. Cryoablation has proven effective in the treatment of benign breast lesions, such as fibroadenomas. Nowadays, its main use in malignant breast lesions is to reduce the size of the mass in cases where surgery is contraindicated. Nevertheless, other uses in malignant breast disease are being investigated. Cryoablation can strengthen the tumor response to immunotherapy. Likewise, in the near future, it may become an alternative to surgery for the local control of early-stage neoplasms. This article aims to describe our experience using cryoablation with liquid nitrogen to treat benign and malignant breast lesions and to review the fundamentals of the technique and its indications. Since 2019 we have used cryoablation to treat 7 benign and 9 malignant lesions without significant complications, achieving excellent esthetic outcomes. Cryoablation has been incorporated into our institution's breast unit's protocol.

Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis

BACKGROUND: Lawsonia intracellularis remains a problem for the swine industry worldwide. Previously, we designed and obtained a vaccine candidate against this pathogen based on the chimeric proteins: OMP1c, OMP2c, and INVASc. These proteins formed inclusion bodies when expressed in E. coli, which induced humoral and cellular immune responses in vaccinated pigs. Also, protection was demonstrated after the challenge. In this study, we established a production process to increase the yields of the three antigens as a vaccine candidate. RESULTS: Batch and fed-batch fermentations were evaluated in different culture conditions using a 2 L bioreactor. A fed-batch culture with a modified Terrific broth medium containing glucose instead of glycerol, and induced with 0.75 mM IPTG at 8 h of culture (11 g/L of biomass) raised the volumetric yield to 627.1 mg/L. Under these culture conditions, plasmid-bearing cells increased by 10% at the induction time. High efficiency in cell disruption was obtained at passage six using a high-pressure homogenizer and a bead mill. The total antigen recovery was 64% (400 mg/L), with a purity degree of 70%. The antigens retained their immunogenicity in pigs, inducing high antibody titers. CONCLUSIONS: Considering that the antigen production process allowed an increment of more than 70-fold, this methodology constitutes a crucial step in the production of this vaccine candidate against L. intracellularis.

Período de adaptação a dietas com baixa ou alta fibra sobre a digestibilidade e as características fecais em cães / Adaptation period to diets with low or high fiber on digetibility and faecal characteristics in dogs

Objetivou-se avaliar três períodos de adaptação, cinco, 10 e 15 dias, a dietas com baixa ou alta fibra sobre a digestibilidade e características fecais. Foram utilizadas duas dietas, uma contendo 0% de casca de soja (0%CS) e outra com 15% de casca de soja (15%CS), fornecidas a 12 cães adultos. Ao final de cada período, houve coleta total de fezes para avaliação dos coeficientes de digestibilidade aparente (CDA), energia metabolizável (EM) e características fecais. Além disso, foi avaliada a palatabilidade das dietas. Como resultado, não houve efeito do período de adaptação sobre a digestibilidade das dietas (P>0,05). A dieta 15%CS apresentou menor CDA da matéria seca (MS) e EM, em relação à dieta 0%CS (P<0,05). A consistência fecal dos cães não diferiu entre as dietas e os períodos (P>0,05). Houve redução no pH e na amônia fecal dos cães alimentados com a dieta 15%CS (P<0,05), após 10 dias de alimentação. Não houve diferença na palatabilidade das dietas (P>0,05). Períodos de adaptação à dieta superiores a cinco dias não alteram a digestibilidade e a maioria das características fecais de cães, exceto a amônia fecal. A inclusão de 15%CS na dieta reduz a digestibilidade da MS e a EM, bem como o pH e a amônia fecal.(AU)

This study aimed to evaluate three periods of adaptation, 5, 10 and 15 days, to diets with low or high fiber on digestibility and fecal characteristics. Two diets were used, one containing 0% (0% SH) and one containing 15% soybean hulls (15% SH). Diets were provided to 12 adult dogs. At the end of each period there was total collection of feces to evaluate the apparent digestibility coefficients (ADC), metabolizable energy (ME) and fecal characteristics. We also evaluated diet palatability. There was no effect of the adaptation period on digestibility of diets (P> 0.05). The 15% SH diet reduced ADC of dry matter (DM) and ME, compared to the 0% SH diet (P< 0.05). Fecal consistency of the dogs did not differ among diets and periods (P> 0.05). There was a decrease in fecal pH and ammonia in dogs fed the diet with 15% SH (P< 0.05) after 10 days of feeding (P< 0.05). There was no difference in diet palatability (P> 0.05). Diet adaptation longer than 5 days did not alter the digestibility and most fecal characteristics of dogs, except for fecal ammonia. The inclusion of 15% SH in diet reduces DM digestibility and fecal pH and ammonia in dogs.(AU)

Epidemiological study of intraosseous lesions of the stomatognathic or maxillomandibular complex diagnosed by a Reference Centre in Brazil from 2006-2017.

This epidemiological study was designed to find out the incidence and factors associated with the occurrence of intraosseous lesions diagnosed at a Reference Centre in Brazil. We included all patients diagnosed with intraosseous lesions (cyst, tumour, bone-associated lesion, and periapical disease) during the period 2006-2017, and analysed the association between some sociodemographic and clinical variables and the types of lesion. There was a total of 290 intraosseous lesions, the most common being odontogenic cysts. There was a significant association between age and odontogenic tumours (p=0.001). In relation to the histopathological diagnosis, root cysts were the most common (n=57), followed by dentigerous cysts (n=26). The lesions studied were seen most often in women between the second and fourth decades of life, odontogenic cysts being the most common type found. We know of few publications of similar epidemiological work, either in Brazil or in the rest of the world, so we suggest that more such studies are made.

Requirements and energy efficiency of Pelibuey and Katahdin non pregnant, non lactating ewes in Yucatan, Mexico / Requerimientos y eficiencia energética de ovejas Pelibuey y Katahdin no gestantes, no lactantes en Yucatán, México

Abstract Objective. To estimate metabolizable energy requirements for maintenance (MEm), energy efficiency of weight gain (EEWG) and fat thickness, in Pelibuey and Katahdin ewes in Yucatan, Mexico. Material and method. Eight non pregnant, non lactating, multiparous ewes, were feed at three intake levels with a diet containing 2.0 Mcal/Kg-1 of ME and 11 % of CP. Feed intake was measured daily, ewes weight and thickness of subcutaneous fat (SF) were determined every 14 days. The EMm was estimated by regressing values of live weight against metabolizable energy intake (MEI), EEWG was estimate as gram of weight gain per Mcal of MEI. Results. There were no differences between breeds in MEm (97±4 and 110±4 kcal/Kg0.75, for Pelibuey and Katahdin) and EEWG (58±8 and 63±8 g/Mcal of MEI, for Pelibuey and Katahdin); differences were found in SF (6.1±0.2 and 4.9±0.2 mm, for Pelibuey and Katahdin). Conclusions. The energy requirements for maintenance were similar in Pelibuey and Katahdin ewes in Yucatan, Mexico.

Resumen Objetivo. Estimar los requerimientos de energía metabolizable para el mantenimiento (MEm), le eficiencia energética del incremento de peso (EEWG) y el grosor de grasa, en ovejas Pelibuey y Katahdin en Yucatán, México. Materiales y método. Ocho ovejas multíparas no gestantes, no lactantes, fueron alimentadas a tres niveles de consumo con una dieta que contenía 2.0 Mcal/kg-1 de EM y 11% de PC. El consumo de alimento fue medido diariamente, el peso de ovejas y el grosor de grasa subcutánea (SF) se determinó cada 14 días. La MEm fue estimada por regresión de los valores de peso vivo contra el consumo de EM (MEI), la EEWG fue estimada como los gramos de ganancia de peso por Mcal de MEI. Resultados. No existieron diferencias entre razas en EMm (97±4 y 110±4 kcal/Kg0.75, para Pelibuey y Katahdin respectivamente) y EEWG (58±8 y 63±8 g/Mcal de MEI, para Pelibuey y Katahdin respectivamente); se encontraron diferencias en SF (6.1±0.2 y 4.9±0.2 mm, para Pelibuey y Katahdin respectivamente). Conclusiones. Los requerimientos de energía para mantenimiento fueron similares en las ovejas Pelibuey y Katahdin en Yucatán, México.

Probable enfermedad de Kawasaki diagnosticada en fase de secuelas en un hombre adulto de 56 años de edad / Possible Kawasaki disease in a 56 years-old male patient diagnosed in sequelae phase

Se presenta el caso de un paciente de 56 años edad que sufre un evento isquémico agudo con arterias coronarias con una importante dilatación en los segmentos proximales izquierdos y una dilatación de hasta 10 mm de diámetro en la coronaria derecha proximal y todo el segmento vertical, compatibles con enfermedad de Kawasaki en fase de secuelas crónicas.

We present a 56 years-old male patient with an acute ischemic episode. The patient showed significant dilation of the left main coronary artery and proximal segments of the left descending artery, as well as a right coronary artery with a 10 mm diameter in the proximal and medial segments. These findings are compatible with a Kawasaki's disease in a chronic phase.

Use of Triticum aestivum in open wound healing: a clinical, pathological, and tensiometric assessment in the rabbit model / Uso de Triticum aestivum na cicatrização de feridas: um ensaio clínico, histopatológico e tensiométrico em coelhos

While Triticum sp. has been shown to act in wound healing, stimulating collagen synthesis by fibroblasts, the use of this plant extract has yet to be assessed in vivo, in commercially viable presentations. This study used rabbits and assessed, on days seven, 14, and 21, the presence or absence of granulation tissue and epithelialization, histopathological structures, and scar quality through the breaking and tension strength. Treatments, performed for 21 days, were aqueous extract of T. aestivum at a concentration of 2mg/mL (group I) and 10mg/mL (group II) and a nonionic cream (control group). We demonstrate that the formation of granulation tissue was not significantly different between treatments. In the analysis of epithelial tissue, wounds in group II differed from other treatments by day 7. On days 14 and 21 there was no significant clinical difference between groups. In the histopathological evaluation, scar quality and rupture strength did not differ between the groups in the studied period. In the tension strength evaluation, group I differed from the others, presenting a higher tension strength overall. The studied treatments did not differ regarding healing evolution of the skin wounds, but T. aestivum extract, at 2mg/mL, presents better results in the tension strength evaluation...

O extrato de trigo (Triticum sp.) vem sendo usado na cicatrização de feridas por estimular a síntese de fibroblastos, entretanto a sua aplicabilidade in vivo em apresentações comercialmente viáveis ainda tem de ser demonstrada. Neste estudo, avaliaram-se feridas cutâneas de coelhos tratadas com extrato aquoso de T. aestivum quanto à presença de tecido de granulação e epitelização, estruturas histológicas, qualidade cicatricial, além de ensaio tensiométrico. As feridas foram tratadas diariamente, por 21 dias, com diferentes concentrações do extrato (grupo I = 2mg/mL; grupo II = 10mg/mL) ou apenas o veículo (grupo controle = creme não iônico), e avaliadas nos dias sete, 14 e 21. A formação de tecido de granulação não diferiu entre os tratamentos. A epitelização aconteceu em menor tempo em feridas do grupo II, mas aos 14 dias já não havia diferença neste parâmetro. Na avaliação histopatológica, a qualidade cicatricial e a força de ruptura não diferiram no período estudado, entretanto a resistência tensiométrica das feridas do grupo I foi maior que a dos demais tratamentos. Dessa forma, conclui-se que, mesmo não havendo diferença na evolução cicatricial de feridas tratadas ou não com extrato aquoso de T. aestivum, o uso desse composto, a 2mg/mL, resultou em tecidos cicatriciais mais resistentes à tração...

[Sentinel node biopsy after neoadjuvant chemotherapy in breast cancer. Its relation with molecular subtypes]. / La biopsia del ganglio centinela después de quimioterapia neoadyuvante en el cáncer de mama. Relación con los subtipos moleculares.

OBJECTIVE: To evaluate the influence of the molecular subtype (MS) in the Sentinel Node Biopsy (SNB) technique after neoadjuvant chemotherapy (NAC) in women with locally advanced breast cancer (BC) and a complete axillary response (CR). MATERIAL AND METHODS: A prospective study involving 70 patients with BC treated with NAC was carried out. An axillary lymph node dissection was performed in the first 48 patients (validation group: VG), and in case of micro- or macrometastases in the therapeutic application phase (therapy group:TG). Classified according to MS: 14 luminal A; 16 luminal B HER2-, 13 luminal B HER2+, 10HER2+ non-luminal, 17 triple-negative. RESULTS: SNB was carried out in 98.6% of the cases, with only one false negative result in the VG (FN=2%). Molecular subtype did not affect SN detection. Despite the existence of axillary CR, statistically significant differences were found in the proportion of macrometastasis (16.7% vs. 35.7%, p=0.043) on comparing the pre-NAC cN0 and cN+. Breast tumor response to NAC varied among the different MS, this being lowest in luminal A (21.5%) and highest in non-luminal HER2+ group (80%). HER2+ and triple-negative were the groups with the best axillary histological response both when there was prior clinical involvement and when there was not. CONCLUSIONS: Molecular subtype is a predictive factor of the degree of tumor response to NAC in breast cancer. However, it does not affect SNB detection and efficiency. SNB can also be used safely in women with prior node involvement as long as a complete clinical and radiological assessment is made of the node response to NAC.

Intraoperative ultrasound in conservative surgery for non-palpable breast cancer after neoadjuvant chemotherapy.

AIMS: A complete clinical response after neoadjuvant chemotherapy (NACT) in breast cancer patients hinders the localization of the residual lesion and the removal of a minimum amount of breast tissue. The aim of the present work is to report our single-centre experience with intraoperative ultrasound-guided (IOUS) excision performed by surgeons in these patients. PATIENTS AND METHODS: From January 2008 to December 2012, IOUS excisions were performed on 58 patients with a previous intralesional ultrasound-detectable metallic marker and non-palpable breast cancer after NACT. The specimen margins were estimated by ultrasonography and macroscopic pathologic examination. Successful lesion removal, specimen weight, and analysis of the results as regards margins were evaluated, and the need for breast-conserving re-excision and mastectomy was considered. RESULTS: After NACT the average ultrasound/mammography and MRI diameters were 11.7 mm (0-30) and 9.1 mm (0-40) respectively. In all cases, the residual lesion or tissue around the marker was removed. The average weight of the specimens was 26.4 g (6-84), being lower in cases of complete response according to ultrasound (p < 0.05). In 4 patients (6.8%), breast-conserving re-excision was carried out, and in 3 patients (5.2%) a secondary mastectomy was performed, two of which had invasive lobular carcinoma. CONCLUSIONS: The emplacement of a readily echodetectable metal marker before NACT makes IOUS excision feasible in an increasing number of complete clinical responses, with the excision of small amounts of breast tissue and a high percentage of conservative breast surgery. This technique requires surgeons to be trained, but has the advantage of a reduced use of other hospital services, better planning of operating theatres, and less discomfort for patients, which means that it is attractive and indeed recommendable.

Protection of red-legged partridges (Alectoris rufa) against West Nile virus (WNV) infection after immunization with WNV recombinant envelope protein E (rE).

West Nile virus (WNV) is maintained in nature in an enzootic transmission cycle between birds and mosquitoes, although it occasionally infects other vertebrates, including humans, in which it may result fatal. To date, no licensed vaccines against WNV infection are available for birds, but its availability would certainly benefit certain populations, as birds grown for restocking, hunting activities, or alimentary purposes, and those confined to wildlife reservations and recreation installations. We have tested the protective capability of WNV envelope recombinant (rE) protein in red-legged partridges (Alectoris rufa). Birds (n=28) were intramuscularly immunized three times at 2-weeks interval with rE and a control group (n=29) was sham-immunized. Except for 5 sham-immunized birds that were not infected and housed as contact controls, partridges were subcutaneously challenged with WNV. Oropharyngeal and cloacal swabs and feather pulps were collected at several days after infection and blood samples were taken during vaccination and after infection. All rE-vaccinated partridges elicited anti-WNV antibodies before challenge and survived to the infection, while 33.3% of the sham-immunized birds succumbed, as did 25% of the contact animals. Most (84%) unvaccinated birds showed viremia 3 d.p.i., but virus was only detected in 14% of the rE vaccinated birds. WNV-RNA was detected in feathers and swabs from sham-immunized partridges from 3 to 7 d.p.i., mainly in birds that succumbed to the infection, but not in rE vaccinated birds. Thus, rE vaccination fully protected partridges against WND and reduced the risk of virus spread.

Effects of MSC coadministration and route of delivery on cord blood hematopoietic stem cell engraftment.

Hematopoietic stem cell transplantation (HSCT) using umbilical cord blood (UCB) progenitors is increasingly being used. One of the problems that may arise after UCB transplantation is an impaired engraftment. Either intrabone (IB) injection of hematopoietic progenitors or mesenchymal stem cell (MSC) coadministration has been proposed among the strategies to improve engraftment. In the current study, we have assessed the effects of both approaches. Thus, NOD/SCID recipients were transplanted with human UCB CD34+ cells administered either intravenously (IV) or IB, receiving or not bone marrow (BM)-derived MSCs also IV or IB (in the right femur). Human HSC engraftment was measured 3 and 6 weeks after transplantation. Injected MSCs were tracked weekly by bioluminescence. Also, lodgment within the BM niche was assessed at the latter time point by immuno-fluorescence. Our study shows regarding HSC engraftment that the number of BM human CD45+ cells detected 3 weeks after transplantation was significantly higher in mice cotransplanted with human MSCs. Moreover, these mice had a higher myeloid (CD13+) engraftment and a faster B-cell (CD19+) chimerism. At the late time point evaluated (6 weeks), human engraftment was higher in the group in which both strategies were employed (IB injection of HSC and MSC coadministration). When assessing human MSC administration route, we were able to track MSCs only in the injected femurs, whereas they lost their signal in the contralateral bones. These human MSCs were mainly located around blood vessels in the subendosteal region. In summary, our study shows that MSC coadministration can enhance HSC engraftment in our xenogenic transplantation model, as well as IB administration of the CD34+ cells does. The combination of both strategies seems to be synergistic. Interestingly, MSCs were detected only where they were IB injected contributing to the vascular niche.

[Post-traumatic supraorbital neuralgia: a benign condition]. / Neuralgia supraorbitaria postraumática: una entidad benigna.

INTRODUCTION: Supraorbital neuralgia has only recently been described. Most of the cases reported involve patients suffering from chronic idiopathic neuralgias that are difficult to treat and sometimes require surgery to release the nerve. We present our experience in patients with a variant of this neuralgia which has a known causation, is commonly seen and has a benign prognosis. CASE REPORTS: We studied five patients, four females and one male, with a mean age of 55 years (range: 29-69 years). They had all suffered direct banal traumatic injury to the frontal region due to different causes. Four of them developed continuous, piercing or burning-type pain; three of them had paroxysmal pain and one had itching. There were no autonomic manifestations. All of them were found to be abnormally sensitive in the affected area, with tactile hypaesthesia, hyperalgesia or allodynia and a positive Tinel's sign. Neuroimaging tests were normal. Two patients were treated with gabapentin and amitriptyline. One was treated with an anaesthetic blockade, which afforded temporary relief. Three of them received no treatment at all. After one year of follow-up, all of them had improved and three were no longer in pain, although sensory alterations persisted in all cases. CONCLUSIONS: Post-traumatic supraorbital neuralgia is a frequent condition, although it is probably underdiagnosed. It has its own characteristic clinical and developmental features that distinguish it from idiopathic supraorbital neuralgia. Progress is usually good and it responds favourably to symptomatic treatment, if needed.

Diagnostic efficacy and safety of 99mtc-labeled monoclonal antibody ior c5 in patients with colorectal and anal carcinomas: final report clinical trial phase I/II.

UNLABELLED: Monoclonal antibody (mAb) ior c5 is an IgG1, which recognizes a glycoprotein tumor specific antigen IOR C2 over expressed in the surface of colon and ovarian cancer cells. The aim of the present work was to evaluate the diagnostic efficacy of the 99mTc-labeled mAb ior c5 for the detection of colorectal tumors, its metastases and recurrences. METHODS: Eighty six patients with colorectal or anal cancer, mean age 57 +/- 13 yrs, were involved in a phase 1/11 multicentric, open clinical trial to assess the ability of Radioimmunoscintigraphy (RIS) with 99mTc- mAb ior c5 to detect those tumors. Seventy-four patients received 1 mg of c5 labeled with 1480-1850 MBq to determinate diagnosis efficacy and safety of murine mAb by intravenously (i.v.) bolus injection (group 1). In order to evaluate pharmacokinetic, bio distribution and dosimetry of this radiolabel molecule, 12 patients received 3 mg of labeled ior c5. Planar anterior and posterior images of the lesion sites and suspected metastases were acquired at 2, 4 and 18-24 hours after injection using a matrix size 128 x128 and 500 Kcounts per view. SPECT were scanned at 5 hr post-injection, using a 360' circular orbit with 64 images. HAMA response was measured in serum at 2, 4, 8 and 12 week post-administration. RESULTS: Labeling efficiency was (97.8 - 0.6) %. No adverse reactions or side effects were observed. Overall sensitivity, specificity, accuracy, positive and negative predictive values of the immunoscintigraphy were 95.80%, 100%, 96.51%, 100.0% and 82.35%, respectively. Unknown metastases were detected in 37 of 86 cases (43.02%). No HAMA response was found. CONCLUSIONS: Immunoscintigraphy with 99mTc-labeled mAb ior c5 could be a useful procedure for the diagnosis and follow-up of the patients with colorectal tumors, its metastasis and recurrences.

[Brachial presentation of spinal pseudochoreoathetosis. The result of proprioceptive information being processed in parallel]. / Seudocoreoatetosis espinal de presentación braquial. Resultado del procesamiento paralelo de la información propioceptiva.

INTRODUCTION: The term pseudochoreoathetosis is used to refer to the choreoathetoid movements that are produced by alterations in the proprioceptive sensitivity due to damage it has suffered at some point along its course. Proprioceptive sensitivity is considered to go up as far as the cortex along the posterior cords of the spinal cord, which means that if they are injured in the cervical region there should be a sensory deficit in both the upper (UL) and lower limbs (LL). CASE REPORTS: We describe five cases of transverse myelitis with localised cervical injury that selectively and mainly affected the posterior cords of the spinal cord. In the five patients there was selective involvement of the proprioceptive sensitivity in the UL respecting the LL and pseudochoreoathetoid movements of the limb that has been deafferented for proprioceptive sensitivity. The dissociation between the UL and the LL occurs because the spinocerebellar and spinocervical tracts take the proprioceptive information from the LL in parallel to the posterior cords, which receive the proprioceptive sensitivity from the UL. CONCLUSIONS: At present, the most widely accepted physiopathological mechanism explaining pseudochoreoathetosis consists in a failure in the integration of the sensory-motor afferences in the striatum, which causes faulty sensory-motor integration at this level and gives rise to pseudochoreoathetosis.

Assessing microleakage of different class V restorations after Er:YAG laser and bur preparation.

This study assessed in vitro marginal leakage of class V cavities prepared by turbine and Er:YAG laser and restored with different materials. Sixty cavities with enamel and dentine margins were prepared and assigned to six groups: I, II, III by turbine and IV, V, VI by Er:YAG laser. The following restorative systems were used: groups I and IV: Bond 1 + Alert; II and V: Fuji II LC; III and VI: SBMP + Dispersalloy. After finishing, specimens were thermocycled for 8 h and 45 min (500 cycles), isolated, immersed in a 0.2% Rhodamine B solution, sectioned oro-facially and analysed for leakage. The dye penetration means (%) were: occlusal I: 10.09 (+/- 21.28), II: 3.25 (+/- 10.27), III: 0, IV: 41.77 (+/- 42.48), V: 23.37 (+/- 33.79), VI: 12.66 (+/- 24.06); cervical I: 16.49 (+/- 26.67), II: 4.34 (+/- 13.71), III: 0, IV: 37.71 (+/- 30.47), V: 39.56 (+/- 43.35) and VI: 72.53 (+/- 37.79). The use of Er:YAG laser for cavity preparation yielded higher degree of marginal leakage, as compared with the use of conventional air-turbine. The enamel interface provided better marginal sealing, comparing with dentine/cementum margin. As to the cavity preparation device (i.e. laser or bur), the analysis of the results showed that bonded amalgam and Fuji II LC provided less infiltration, than Alert. On the other hand, for lased cavities, Alert provided the best results, similar to those of Fuji II LC and superior to those reached by bonded amalgam.

Localization and identification of phenolic compounds in Theobroma cacao L. somatic embryogenesis.

Cocoa breeders and growers continue to face the problem of high heterogeneity between individuals derived from one progeny. Vegetative propagation by somatic embryogenesis could be a way to increase genetic gains in the field. Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. This study was conducted to investigate the phenolic composition of cocoa flowers (the explants used to achieve somatic embryogenesis) and how it changes during the process, by means of histochemistry and conventional chemical techniques. In flowers, all parts contained polyphenolics but their locations were specific to the organ considered. After placing floral explants in vitro, the polyphenolic content was qualitatively modified and maintained in the calli throughout the culture process. Among the new polyphenolics, the three most abundant were isolated and characterized by 1H- and 13C-NMR. They were hydroxycinnamic acid amides: N-trans-caffeoyl-l-DOPA or clovamide, N-trans-p-coumaroyl-l-tyrosine or deoxiclovamide, and N-trans-caffeoyl-l-tyrosine. The same compounds were found also in fresh, unfermented cocoa beans. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. Given the antioxidant nature of these compounds, they could reflect the stress status of the tissues.

Frequency and determinants of microalbuminuria in mild hypertension: a primary-care-based study.

OBJECTIVE: To evaluate the frequency of microalbuminuria and its relationship with several risk factors and left ventricular mass in a population of mildly hypertensive subjects attended in a primary-care setting. DESIGN: Cross-sectional study. SETTING: Eight primary-care centres. PATIENTS: Two hundred and twenty-three non-diabetic patients recently diagnosed with mild hypertension were included in the study. None of them had clinical evidence of target-organ damage or had received prior antihypertensive treatment. INTERVENTIONS: Subjects included in the study underwent clinical interview, measurement of blood pressure (BP) on three visits, blood analysis, measurement of albumin by immunonephelometry in three overnight urine collections, 24 h BP monitoring and M-mode and Doppler echocardiography. MAIN OUTCOME MEASURES: Tobacco habit, clinic BP, body mass index, serum lipids and uric acid, glycaemia, urinary albumin excretion (UAE), ambulatory BP and left ventricular mass index. RESULTS: The frequency of microalbuminuria was 7.2%. Microalbuminuric patients were more likely to be men and to be characterized by higher ambulatory BP, body mass index and uric acid levels. Regression analysis demonstrated that male sex and 24 h systolic BP were determinants of UAE. Patients with white-coat hypertension showed lower UAE than did subjects with sustained hypertension. Although a certain relationship between UAE and left ventricular mass index was found, these variables were not significantly correlated. CONCLUSIONS: A low proportion of mildly hypertensive patients attended in a primary care setting are microalbuminuric. In this population, UAE is an expression of BP values over 24 h and correlates with several risk factors.

Repeated administration of hepatitis C virus core-encoding plasmid to mice does not necessarily increase the immune response generated against this antigen.

DNA immunization is a promising approach in generating immune responses to infectious pathogens in many different animal models. In an effort to augment the anti-[hepatitis C virus (HCV) core] immune response, generated after DNA immunization, the importance of vaccination regimen regarding dose and boosting was investigated in the present study. Balb/c mice were intramuscularly injected with an expression plasmid encoding a truncated variant comprising amino acids 1-176 of the HCV core protein. The highest anti-core antibody titres (1:3700) were detected in mice inoculated with 50-100 microg of core-encoding plasmid. Additionally, we demonstrated that antibody levels induced by a single injection of DNA could be further increased by boosting with a second injection of DNA three weeks after primary immunization. However, administration of additional doses or lengthening of the resting period between inoculations resulted in similar or even weaker anti-core antibody responses. A similar anti-(HCV core) lymphoproliferative response was also detected in animals that had the highest level of anti-core antibodies. These results indicate that, in clinical trials, vaccination regimen might be a critical factor in generating optimal anti-(HCV core) immune responses after genetic immunization.

Surgery for atrial fibrillation using radiofrequency catheter ablation: assessment of results at one year.

OBJECTIVE: The results obtained in 43 patients using direct intraoperative radiofrequency catheter ablation, as an alternative to surgical incisions, to perform atrial fibrillation surgery, are presented. METHODS: Forty-three patients with ages ranging from 43 to 74 years (x = 59), with chronic atrial fibrillation with an average duration 6+/-5 years were operated. Eleven patients suffered from clinically relevant tachyarrythmia and eight had previous thromboembolic events. All but one patient had concomitant mitral valve surgery. Direct intraoperative radiofrequency catheter ablation was used to perform endocardial bilateral isolation of the pulmonary veins from the left atrium. RESULTS: There were no local or general complications, namely bleeding or thromboembolic events. Of the 33 patients with more than 3 months of follow-up, 36% remained in atrial fibrillation (Santa Cruz score 0); 30% had Score 4; 18% had Score 3; 6% had Score 2; 9% had Score 1. CONCLUSIONS: We conclude that the use of intraoperative radiofrequency catheter ablation is fast and safe. Presently, this is our method of choice for surgical treatment of atrial fibrillation in mitral patients.