First molecular detection of Leishmania infantum in Sergentomyia minuta (Diptera, Psychodidae) in Alentejo, southern Portugal.

Protozoan parasites, such as Leishmania spp., are the causative agents of many insect-borne infectious diseases with medical and veterinary importance. Leishmaniasis, caused by Leishmania spp., is transmitted by female phlebotomine sand flies. In the Alentejo region of Portugal, located at the north of Algarve, cases of human and canine leishmaniasis caused by Leishmania infantum have been notified. However, no recent studies regarding the sand fly fauna in the region are available. We therefore aimed to explore the phlebotomine sand fly species found in both, Évora and Beja Districts, to gain an insight about the leishmaniasis epidemiology in these areas. After the identification of the insect species, PCR molecular tests were used to assess L. infantum infection rate in the sand fly captured females, together with the analysis of blood meal sources of the insect vectors. One Sergentomyia minuta female was positive for L. infantum infection and another for human blood as a meal source. The occurrence of this phlebotomine species infected with L. infantum may suggest that, in the Mediterranean basin, leishmaniasis epidemiology is changing. Also, if the importance of S. minuta for the zoonotic and anthroponotic cycle of leishmaniasis is later proven, the strategies to control its vector will inevitably to be rethought.

Human cysticercosis in Portugal: long gone or still contemporary?

Cysticercosis, a leading cause of acquired epilepsy in developing countries, has been controlled or eradicated in industrialized countries. This paradigm has recently been challenged, with human neurocysticercosis (NCC) being increasingly diagnosed in these countries. In order to assess the NCC burden in Portugal, a retrospective study on NCC hospitalizations (2006-2013) was conducted based on the national database on hospital morbidity: 357 hospitalized cases were detected. NCC was most frequent in the following age groups: 20-64 years (n = 197, 55·2%) >64 years (n = 111, 31·1%), and <20 years (n = 49, 13·7%). In the Norte and Centro regions cases tended to be older than in the Lisboa and Vale do Tejo Region. The results raise concerns for imported and autochthonous disease, suggesting the Lisboa and Vale do Tejo Region, due to its higher frequency of cases at younger ages, as a priority for research and intervention, and further suggest that NCC should be under surveillance (notifiable). The National Observatory of Cysticercosis and Taeniasis has been established and will define NCC cases as well as monitoring and surveillance.

The role of Rhipicephalus sanguineus sensu lato saliva in the dissemination of Rickettsia conorii in C3H/HeJ mice.

Animal models have been developed for the study of rickettsial pathogenesis. However, to understand what occurs during the natural route of rickettsial transmission via the tick bite, the role of tick saliva should be considered in these models. To address this, we analysed the role of tick saliva in the transmission of Rickettsia conorii (Rickettsiales: Rickettsiaceae) in a murine host by intradermally (i.d.) inoculating two groups of susceptible C3H/HeJ mice with this Rickettsia, and infesting one group with nymphal Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae) ticks. Quantification of bacterial loads and mRNA levels of interleukin-1ß (IL-1ß), IL-10 and NF-κB was performed in C3H/HeJ lung samples by real-time quantitative polymerase chain reaction (PCR) and real-time reverse transcriptase PCR, respectively. Lung histology was examined to evaluate the pathological manifestations of infection. No statistically significant difference in bacterial load in the lungs of mice was observed between these two groups; however, a statistically significant difference was observed in levels of IL-1ß and NF-κB, both of which were higher in the group inoculated with rickettsiae but not infected with ticks. Lung histology in both groups of animals revealed infiltration of inflammatory cells. Overall, this study showed that i.d. inoculation of R. conorii caused infection in the lungs of C3H/HeJ mice and tick saliva inhibited proinflammatory effects.

Human and porcine Taenia solium infections in Mozambique: identifying research priorities.

The objective of this paper is to critically review and summarize available scientific and lay literature, and ongoing studies on human and porcine cysticercosis in Mozambique to identify knowledge gaps and direct immediate and long-term research efforts. Data on the spatial distribution and prevalence of the disease in human and swine populations are scarce and fragmented. Human serological studies have shown that 15-21% of apparently healthy adults were positive for cysticercosis antibodies or antigen, while in neuropsychiatric patients seroprevalence was as high as 51%. Slaughterhouse records indicate a countrywide occurrence of porcine cysticercosis, while studies have shown that 10-35% of pigs tested were seropositive for cysticercosis antibodies or antigen. Current research in Mozambique includes studies on the epidemiology, molecular biology, diagnosis and control of the disease. Future research efforts should be directed at better understanding the epidemiology of the disease in Mozambique, particularly risk factors for its occurrence and spread in human and swine populations, documenting the socio-economic impact of the disease, identifying critical control points and evaluating the feasibility and epidemiological impact of control measures and development of local level diagnostic tools for use in humans and swine.

Human and porcine taenia solium infections in mozambique: identifying research priorities

The objective of this paper is to critically review and summarize available scientific and lay literature, and ongoing studies on human and porcine cysticercosis in Mozambique to identify knowledge gaps and direct immediate and long-term research efforts. Data on the spatial distribution and prevalence of the disease in human and swine populations are scarce and fragmented. Human serological studies have shown that 15­21% of apparently healthy adults were positive for cysticercosis antibodies or antigen, while in neuropsychiatric patient's seroprevalence was as high as 51%. Slaughterhouse records indicate a countrywide occurrence of porcine cysticercosis, while studies have shown that 10­35% of pigs tested were seropositive for cysticercosis antibodies or antigen. Current research in Mozambique includes studies on the epidemiology, molecular biology, diagnosis and control of the disease. Future research efforts should be directed at better understanding the epidemiology of the disease in Mozambique, particularly risk factors for its occurrence and spread in human and swine populations, documenting the socio-economic impact of the disease, identifying critical control points and evaluating the feasibility and epidemiological impact of control measures and development of local level diagnostic tools for use in humans and swine.

Intestinal parasites in dogs and cats from the district of Évora, Portugal.

Intestinal parasites, both helminths and protozoa, are commonly found in domestic animals, and the possible transmission of enteric parasites from dogs and cats to humans may constitute a global potential health risk worldwide. In the present study, we analysed 148 stool samples from dogs (n=126) and cats (n=22) collected from animal shelters and veterinary clinics, in the district of Évora, Portugal. Microscopic examination confirmed that Giardia was the most frequent parasite in the studied population (34/148; 23%). Other parasites such as Ancylostoma sp., Isospora spp., Toxocara, Trichuris spp., Toxascaris and Toxoplasma were also found. Furthermore, molecular characterization of Giardia duodenalis analysis targeting the small subunit ribosomal RNA (ssu-rRNA) was performed revealing the presence of host-specific (C and D) and zoonotic assemblages (A and B). This work points out to the importance of protozoan parasites in companion animals, and reanalyses the need for parasite prophylaxis.

Long-term follow-up in Stuve-Wiedemann syndrome: a clinical report.

Stuve-Wiedemann syndrome (SWS) is an autosomal recessively inherited disorder that is usually associated with high mortality in the neonatal period. Eleven cases have been published with prolonged survival, the oldest being 16 years. This phenotype is characterized by progressive skeletal anomalies including short stature, severe spinal deformities, bowing of the long bones, contractures and spontaneous fractures, and by neurological features that resemble dysautonomia. Here we report on the natural history of a Portuguese girl from birth till 12 years. The diagnosis was molecularly confirmed by the detection of a homozygous 4 bp deletion (167_170 del TAAC) in exon 3 of LIFR. We compare the findings in this patient to other patients with prolonged survival from the literature.

A novel apical midpiece defect in the spermatozoa of a bull without an apparent decrease in motility and fertility. A case study.

Despite some limitations as predictors of fertility, evaluation of sperm morphology and progressive motility is the commonest method to assess viability of frozen/thawed semen. In this article we describe by light and transmission electron microscopy a novel midpiece structural defect observed in 24-36% of frozen/thawed sperm cells from a Charolais bull, used in artificial insemination programs without any apparent ill effect to the fertility. After thawing, the sperm progressive motility ranged from 65 to 80% and the pregnancy rate for all artificial inseminations performed (43%) did not differ (p>0.05) from results obtained with insemination with semen of other bulls (40%). The defect consisted in mitochondrial aplasia at the neck region, mitochondrial segmental elongation and gaps and thickening of the outer dense fibers at the apical region of the midpiece, and loss of the cementing substance and development of plasma membrane extensions in the entire midpiece. No structural abnormalities were found in the capitulum, proximal centriole, striated columns, axoneme, annulus and fibrous sheath. The thickness of the outer fibers returned to normal at the distal region of the midpiece. Based on the examination it is suggested that the alterations might be originally caused by loss of the cementing substance that links mitochondria to the plasma membrane in association with mitochondrial aplasia at the neck region of the midpiece. The abnormality appeared not related to other described sperm defect syndromes, although it shared particular characteristics with the dag defect, segmental aplasia of the mitochondrial sheath, corkscrew defect and pseudodroplet defect.

Seroprevalence of human cysticercosis in Maputo, Mozambique.

We carried out a serosurvey for cysticercosis among people visiting the Central Hospital of Maputo, the capital of Mozambique, between January and June 1993. A standardized questionnaire was designed to obtain information on demographic, socioeconomic, environmental, and behavioral characteristics related to the transmission of the infection. Four hundred eighty-nine individuals were tested for anti-cysticercosis antibodies: 222 blood donors and patients from the Department of Orthopedics, 148 patients from the Department of Neurology, and 119 patients from the Department of Psychiatry. The overall positivity rate was 12.1% (59 of 489). Anti-cysticercus antibodies was detected in 14.9% of the blood donors and patients from the Department of Orthopedics, 11.5% of the patients from the Department of Neurology, and 7.6% of the patients from the Department of Psychiatry. Living in poor sanitary conditions seems to be an important factor related to human cysticercosis in Maputo, Mozambique.

The EORTC randomized trial on three fractions per day and misonidazole in advanced head and neck cancer: prognostic factors.

In trial no. 22811 on a randomized comparison of multiple fractions per day (MFD), with or without misonidazole, to conventional fractionation in advanced head and neck cancer, a large number (523) of patients was entered in a short period of time. No differences in treatment results were obtained, but the study created an important database, allowing for detailed evaluation of the most important factors influencing prognosis. In univariate analysis, factors significantly influencing survival and locoregional control were: performance status, histological differentiation, tumor site, tumor and nodal staging, and tumoral and nodal volume. In multivariate analysis, significant factors for survival were nodal involvement, tumor stage, performance status, and tumor site. Significant factors for locoregional control were nodal involvement and total tumor burden. This analysis suggests that total tumor burden (volume) should be included in the interpretation of treatment results in head and neck cancer.

The EORTC randomized trial on three fractions per day and misonidazole (trial no. 22811) in advanced head and neck cancer: long-term results and side effects.

From 1981 to 1984, a randomized study was done by the EORTC Radiotherapy Group comparing a fractionation schedule with three fractions per day (multiple fractions per day, MFD), with or without misonidazole, to conventional fractionation. The aim of the study was to obtain improved local and regional control and survival by shortening of the treatment time in the first 2 weeks of irradiation. Three fractions of 1.6 Gy/day (4-h interval) were given during 10 irradiation days to a total of 48 Gy. After 3-4-weeks interval, a boost was given to 67.2 or 72 Gy also in three fractions per day. This schedule was compared to an identical arm with misonidazole 1 g/m2/day and a third arm with conventional fractionation (70 Gy in 35 fractions, 7 weeks or 75 Gy in 44 fractions, 9 weeks). A total number of 523 patients was included in the study. Acute mucositis was much heavier in patients treated with three fractions per day (Van den Bogaert et al. Int. J. Radiat. Oncol. Biol. Phys. 8: 1649-1655, 1982). Early results, communicated in 1986 (Van den Bogaert et al. Int. J. Radiat. Oncol. Biol. Phys. 12: 587-591, 1986) showed no differences in treatment outcome between the three treatment arms. Long-term results and data on late effects are now available. Survival at 5 years was 18% (SE 1.9%) and locoregional control was 27% (SE 2.9%). No statistically significant differences could be observed between the three treatment arms.(ABSTRACT TRUNCATED AT 250 WORDS)

Uptake of lead and its influence in the alga Selenastrum capricornutum printz.

The influence of nutrient level, hours of fight and the flux of air by volume of solution on uptake of lead has been examined from batch growth experiments carried out with the alga Selenastrum capricornutum Printz. The organic matter released by the alga has been characterized in terms of absorbance at 285 nm, dissolved organic carbon and maximum intensity of fluorescence at the excitation peak. The lability of lead complexes formed with the alga exudates has been investigated in terms of differential pulse anodic stripping voltammetry (DPASV). It has been noticed that lead inhibits the division of the cells which become bigger in size but with less pigment per cell.

Ewing's sarcoma in children: twenty-five years of experience at the Instituto Portugês de Oncologia de Francisco Gentil (I.P.O.F.G.).

Fifty children with Ewing's sarcoma were consecutively treated from 1962 to 1987 and retrospectively analyzed at the I.P.O.F.G. of Lisbon. At first diagnosis, 10 cases had distant metastases. The remaining 40 patients had clinically localized disease, and different protocols were followed over the years. The best results were obtained with chemotherapy and radiotherapy with or without surgery; and for these children the two-year survival rate was 42.8% vs. 8.3% for the group of patients submitted to local treatment alone. Besides the treatment modality, other factors influenced the prognosis, such as inflammatory signs, sex, tumor volume, and tumor site as well as evidence of distant metastases.

Morphologic and morphometric studies on tumor necrosis produced by radiotherapy, and hyperthermia singly and in combination.

The purpose of the present study was to evaluate the therapeutic effects of hyperthermia associated with radiotherapy on neoplasia. Two transplanted experimental tumors (undifferentiated carcinoma of mouse breast and sarcoma 37) were used. A protocol was followed that included, for both models, four groups of animals: 1) control group; 2) radiotherapy group; 3) hyperthermia group; 4) radiotherapy associated with hyperthermia group. The animals were sacrificed after therapy according to a different schedule for each type of tumor. The morphology of the neoplasia in every group of treated tumors was compared with that of the control group. For quantitative evaluation of the necrosis, we studied the ratio of the tumor necrotic areas (N) to a tumor standard area (T). From the results obtained, the following conclusions were made: 1) Necrosis produced by combined treatment was significantly greater than that obtained by using one only; 2) necrosis appeared early after treatment and remained relatively unchanged; 3) The damaging effect of the hyperthermia occurred earlier than that produced by radiotherapy when these methods were used separately, thus suggesting distinct mechanisms of tumor necrosis.

Metal complexes of 1-oxa-4,7,10-triazacyclododecane-N,N',N''-triacetic acid.

The macrocyclic complexone 1-oxa-4,7,10-triazacyclododecane-N,N',N''-triacetic acid (cODTA) has been synthesized and its protona constants, stability constants of metal complexes and enthalpy changes for the formation of alkaline-earth complexes have been determined. Although it is not so powerful a complexing agent as the N-acetate derivative of the corresponding tetra-aza macrocycle, cDOTA, this is still one of the strongest complexones known, particularly towards the alkaline-earth metals. The complexes of the transition metals are also very stable and there is an inversion of the Irving-Williams order of stability for the complexes of cobalt and nickel.

Haemostatic and antialgic effects of the 25 MV photon beam concentrated dose in the treatment of carcinoma of the cervix.

The authors present the results of a preliminary study on the use of a concentrated dose of 13 Gy in 48 hours in the treatment of cancer of the cervix, especially for haemostatic and antialgic purposes and mostly as a first part of a split course of radiotherapy. In 13 patients with early stages of the disease, the method was used as an emergency treatment with good response in 84.6% of the cases without serious complications. In 43 patients with advanced disease, the concentrated dose was generally repeated for palliation and relieved symptoms in about 72% of the cases but with complications in 16.3%. The authors recommend this method only when absolutely necessary to stop severe haemorrhage and suggest that patients with good response to the first flash course should continue treatment with conventional fractionation, bearing in mind the tolerance of the normal tissues.

Palliative telecobalt-60 with concentrated dose in TNM classification T4 of the breast.

A method is presented of rapid, palliative radiotherapy for carcinoma of the breast, composed of two sessions of 6.5 or 8.5 Gy delivered at a 48-hour interval. The radiobiological equivalence of this unconventional technique is presented. Our preliminary results in a series of 80 patients were published in 1978. The present report is based on the analysis of 112 patients with T4 of the breast submitted to the "flash" dose, from March 1978 to December 1981. The method was used not only for relief of symptoms such as pain and hemorrhage, but also as part of intensive radiotherapy in all patients with good response and with no manifestation of distant dissemination so that they were able to resume treatment 2.5 or 3 weeks later, with conventional fractionation. Some of these patients became operable. About half of the cases had castration and/or hormone therapy and/or chemotherapy. In the group of patients with T4M0, 59.8% were alive after more than 1 year, and of them, 43.7% had no evidence of disease. These encouraging results suggest that this method be advocated in T4 of the breast because of its rapidity and good tolerance with no significant complications.