Molecular Diversity and Genetic Relatedness of Candida albicans Isolates from Birds in Hungary.

The molecular epidemiology of Candida albicans infections in animals has been rarely studied. In this study, multilocus sequence typing was used to characterise the genetic diversity and population structure of 24 avian origin C. albicans isolates collected from different birds with candidiasis and compared to human isolates. Fourteen diploid sequence types (DSTs) including six new DSTs were determined. Cluster analysis revealed that isolates grouped into 8 clades. Bird isolates mainly belonged to minor clades and Clade 15 with DST 172 was the most common (11 isolates; 45.8%). The remaining isolates were clustered into Clade 7 (5 isolates; 20.8%), Clade 10 (4 isolates; 16.6%), Clade 8 (2 isolates; 8.3%), Clade 4 (1 isolate; 4.2%) and Clade 16 (1 isolate; 4.2%). Unweighted pair group method with arithmetic averages (UPGMA) and eBURST analyses showed that the genetic construction of avian origin C. albicans population is fairly diverse. Although species-specific lineages were not found, some degree of separation in the evolution of bird and human strains could be observed.

Country-wide distribution of bluetongue virus with expanding host spectrum and evidence of vector competence in Hungary.

Following the introduction of bluetongue virus type 4 (BTV-4) in 2014, country-wide monitoring of bluetongue (BT) disease was performed to see whether the virus has become enzootic in Hungary. To analyse the epizootiology of BT, over 110,000 samples collected from domestic and wild ruminants were screened for the presence of BTV RNA and virus-specific antibodies using real-time RT-PCR assay and commercial ELISA kit, respectively. During laboratory analysis, specimens collected from 333 (0.8%) cattle, 79 (2.2%) sheep, 4 (0.9%) goats, and 1 (2.3%) mouflon were found to be positive by viral RNA-detection assay. In addition, antibody to BTV was detected in 5.5% (3158/57,250) of cattle, 10.1% (517/5120) of sheep, 40% (116/290) of goat, and 5.6% (16/284) of buffalo origin samples. The majority of positive samples originated from south-western counties; however, 18 out of 19 counties reported cases or antibody prevalence in the examined animals. Genome sequencing of a representative BTV-4 strain from 2015 was also performed. When comparing this strain with the isolate BTV4-HUN2014 detected only a year earlier in Hungary, mutations at 14 sites were identified within the amplified and sequenced genome. Our findings reinforce the need for continued surveillance of BT disease in Hungary. Keywords: reoviridae; orbivirus; cattle; sheep; goat; biting midge.

The Anopheles gambiae adult midgut peritrophic matrix proteome.

Malaria is a devastating disease. For transmission to occur, Plasmodium, the causative agent of malaria, must complete a complex developmental cycle in its mosquito vector. Thus, the mosquito is a potential target for disease control. Plasmodium ookinetes, which develop within the mosquito midgut, must first cross the midgut's peritrophic matrix (PM), a thick extracellular sheath that completely surrounds the blood meal. The PM poses a partial, natural barrier against parasite invasion of the midgut and it is speculated that modifications to the PM may lead to a complete barrier to infection. However, such strategies require thorough characterization of the structure of the PM. Here, we describe for the first time, the complete PM proteome of the main malaria vector, Anopheles gambiae. Altogether, 209 proteins were identified by mass spectrometry. Among them were nine new chitin-binding peritrophic matrix proteins, expanding the list from three to twelve peritrophins. Lastly, we provide a model for the putative interactions among the proteins identified in this study.

Driving midgut-specific expression and secretion of a foreign protein in transgenic mosquitoes with AgAper1 regulatory elements.

The Anopheles gambiae adult peritrophic matrix protein 1 (AgAper1) regulatory elements were used to drive the expression of phospholipase A2 (PLA2), a protein known to disrupt malaria parasite development in mosquitoes. These AgAper1 regulatory elements were sufficient to promote the accumulation of PLA2 in midgut epithelial cells before a blood meal and its release into the lumen upon blood ingestion. Plasmodium berghei oocyst formation was reduced by approximately 80% (74-91% range) in transgenic mosquitoes. Blood-seeking behaviour and survival of AgAper1-PLA2 transgenic mosquitoes were comparable to sibling wild-type mosquitoes, while fertility was substantially lower. Ultrastructural studies suggest that decreased fitness is a consequence of internal damage to midgut epithelial cells.

Storage and secretion of Ag-Aper14, a novel peritrophic matrix protein, and Ag-Muc1 from the mosquito Anopheles gambiae.

The gene Ag-Aper14 encodes a novel peritrophic matrix (or peritrophic membrane; PM) protein in the mosquito Anopheles gambiae. The Ag-Aper14 protein is merely 89 amino acids long and has a single putative chitin-binding domain. Prior to blood feeding, the Ag-Aper14 protein is stored in secretory vesicles next to the epithelial cell lumenal surface. Immunoelectron microscopy has revealed that Ag-Aper14 co-localizes to the same secretory vesicles as another PM protein, Ag-Aper1, indicating a common mode of regulated secretion. Conversely, Ag-Muc1, an epithelial cell-surface protein, does not co-localize to these secretory vesicles and is detected only on the cell surface. After blood feeding, Ag-Aper14 is secreted and incorporated into the PM that surrounds the ingested blood.

Characterization of a carboxypeptidase A gene from the mosquito, Aedes aegypti.

A gut-specific carboxypeptidase A gene (AeCPA) from the mosquito, Aedes aegypti, was cloned and characterized. The gene has an open reading frame that predicts a protein of 427 amino acids, 61% of which are identical to an Anopheles gambiae carboxypeptidase A sequence. AeCPA messenger RNA (mRNA) was not detected during larval and pupal development. In situ hybridization experiments indicated that AeCPA mRNA is expressed by posterior midgut epithelial cells. In sharp contrast to An. gambiae carboxypeptidase A gene expression, AeCPA mRNA accumulates to high levels only late ( approximately 16-24 h) after ingestion of a blood meal. The temporal profile of AeCPA gene induction is similar to that of Ae. aegypti late trypsin, suggesting the existence of common regulatory elements.

Haematology outreach clinics in the Free State and northern Cape.

OBJECTIVE: Evaluation of haematology outreach clinics in the Northern Cape and Free State. DESIGN: Retrospective analysis of records from March 1994 to February 1996. SETTING: Central South Africa is sparsely populated. Consultants from Bloemfontein held outpatient clinics in hospitals (with laboratories) in Bethlehem, Kimberley and Kroonstad. SUBJECTS: 117 patients with suspected haematological disease. MAIN OUTCOME MEASURES: Input measures (population, number of clinics and costs), process measures (patient numbers, patients per clinic, new consultations per clinic, patients' domicile, how they were referred, types of diagnoses and number of patients with non-haematological disorders) and output measures (attrition, changes in attendance and savings). MAIN RESULTS: The 84 clinics that were held, with 636 consultations, did not cost the State anything. Only 6% of the 117 patients had no haematological problem. Sixty-eight per cent had chronic haematological neoplasms. In Kimberley most of the patients came from Kimberley Hospital, while most of the patients at the other clinics were referred via Bloemfontein. There was only a 10% attrition rate and only one-third of patients were referred to Bloemfontein. We saved paying patients an estimated R21,260 in transport costs, while saving the State R172,992 by seeing patients at secondary, instead of tertiary, hospitals. CONCLUSIONS: It is cheaper to send a doctor to an outreach clinic than to refer patients to a central facility, provided there is enough work for a doctor at the clinic. It costs the State much less for patients to be seen at a secondary than a tertiary hospital. Positive spin-offs include academic stimulation of doctors and laboratories in the periphery, with more appropriate referrals to teaching hospitals. Weaknesses include poor availability of expensive drugs at the clinics and lack of standardised records. By commuting to outreach clinics, specialists can greatly reduce health expenditure and spread it from tertiary to lower levels. At the same time more patients have access to their services.

Radiation dose estimates of 186Re-hydroxyethylidene diphosphonate for palliation of metastatic osseous lesions: an animal model study.

A common complication in patients with breast or prostate cancer is bone metastases causing pain. New radionuclide therapy methods have recently been proposed for palliation, including 186Re-hydroxyethylidene diphosphonate (186Re-HEDP). This paper reports on the local development of 186Re-HEDP and the biodistribution studied in animals for eventual use in patients. Adult dose was computed assuming a 70 kg standard man. The 186Re was labelled to HEDP using standard techniques. The biodistribution in five Chacma baboons (Papio ursinus) was studied. Doses ranging from 39.4 to 44.9 MBq kg(-1) (mean 43.6 +/- 2.8 MBq kg[-1]) were administered, corresponding to an adult human dose of 2960 MBq (80 mCi). Whole-body images of the animals were obtained with a dual-headed scintillation camera on an hourly basis for 6 h post-injection and then daily for 3 days. The bone, soft tissue, kidneys and urinary bladder were considered source organs and data from these organs were used in a compartmental model to obtain the mean residence times of the radionuclide in the different source organs. Radiation dose estimates for 186Re-HEDP were subsequently obtained with the MIRDOSE 3 program. The estimated absorbed radiation doses to some of the organs (expressed in mGy MBq[-l]) were as follows: bone surface 1.69; kidneys 0.09; liver 0.04; ovaries 0.04; red marrow 0.75; total body 0.12; urinary bladder wall 0.43. 186Re-HEDP yielded an effective dose of 0.17 mSv MBq(-1). The radiation dose delivered to the bone marrow in this study did not cause any detrimental effect to the baboons, indicating that locally produced 186Re-HEDP is suitable for clinical use.

Rapid induction by a blood meal of a carboxypeptidase gene in the gut of the mosquito Anopheles gambiae.

A search for genes induced rapidly (< 3 h) after a blood meal in the gut of the human malaria vector Anopheles gambiae led to the identification of a carboxypeptidase gene (AgCP). We report the sequence of the 1302 nt AgCP transcribed sequence, 710 nt of upstream and 585 nt of downstream DNA. The AgCP open reading frame is 60.4% identical at the nucleotide level to a blackfly, Simulium vittatum, carboxypeptidase gene. The transcriptional start site of AgCP was determined by primer extension. Expression of AgCP mRNA is detectable in the guts of pupae and sugar-fed adult female mosquitoes and is induced (approximately 10-fold) within 3 h of a blood meal. By 24 h after a blood meal, mRNA abundance returns to a level close to that present before a blood meal. Whole-mount in situ hybridization shows that AgCP mRNA expression is restricted to most or all cells of the posterior midgut. Expression of the AgCP and trypsin genes were compared and shown to differ in two fundamental ways: (1) the peak of AgCP expression after a blood meal occurs approximately 20 h before that of trypsin; and (2) induction of the AgCP gene is independent of the composition of the ingested meal whereas trypsin induction requires the presence of protein. The potential use of the AgCP promoter for driving the expression of genes that hinder the development of parasites in the mosquito gut is discussed.

Peyronie's disease--a perspective on the disease and the long-term results of radiotherapy.

From 1966 to 1988, 98 of 108 patients with symptomatic Peyronie's disease received radiotherapy at our institution. In 11 of 61 patients (18%) who attended the clinic regularly for follow-up for longer than a year, new lesions distinct from the original lesions developed. This confirms that there is progression of the disease in a substantial number of cases after treatment. Long-term follow-up over an average of 111.5 months was achieved by means of a questionnaire in 47 of the 98 cases (48%). Forty-one of these patients (87.2%) had sexual intercourse after radiation. Twenty-eight of the 41 (68.3%) still have intercourse. Their average age at present is 59.6 years while the average age of the 13 patients (31.7%) not having intercourse is 70.9 years. The decline in sexual activity is thus age-related. Twenty-one of 25 patients (84%) experienced relief from pain, and angulation of the penis improved in 17 of 44 patients (38.6%) after radiotherapy. Radiotherapy may therefore be of benefit to patients with active Peyronie's disease and should be investigated in a randomised controlled study.

Racial differences in patients with adenocarcinoma of the endometrium.

OBJECTIVE: To determine the differences between white and black women with regard to the presentation and behavior of adenocarcinoma of the endometrium. METHOD: Records of 273 (68%) white patients and 117 (32%) black patients with endometrial adenocarcinoma were reviewed in Bloemfontein, South Africa. Survival data was calculated according to the direct method where losses in follow-up were regarded as tumor deaths. RESULTS: Most patients (82%) were treated by pre-operative radium followed by total abdominal hysterectomy and bilateral salpingo-oophorectomy, with post-operative external irradiation where indicated. Pre-operatively, fewer black women had reached FIGO stage I, while a larger number had advanced to stages II-IV (P = 0.0024). In addition, the tumor differentiation was more often poor in the black group (P < 0.0001). Ten-year follow-up was achieved in 84% of the white patients and 51% of the black patients and the 10-year survival figures were 67% for white patients and 28% for blacks (P < 0.0001). CONCLUSION: Endometrial adenocarcinoma is a more aggressive disease in black women than it is in whites.

Identification of species-specific DNA sequences in North American blackflies.

Blackflies are vectors for Onchocerca volvulus, a filarial parasite that infects tens of millions of people in Africa and Latin America. Different species of the insect vectors, varying greatly in their ability to transmit the infection, may populate overlapping geographical regions and be morphologically similar. A rapid and reliable method for distinguishing among blackfly species is presently not available. We have isolated cloned DNAs coding for repetitive sequences that can distinguish between two species of North American blackflies, Simulium pictipes and Simulium vittatum. These DNAs do not cross hybridize to three African blackfly species. The assay is very sensitive, less than 1/1000th of the amount of DNA in a single fly being sufficient for detection with a radioactive probe. Diagnosis with nonradioactive probes by a procedure that is amenable for use in the field was also demonstrated.

[Palliative radiation schedules with and without chemotherapy in advanced carcinoma of the esophagus]. / Palliërende bestralingskedules met en sonder chemoterapie in gevorderde karsinoom van die esofagus.

In a prospective randomized study, 272 patients with advanced oesophageal cancer were treated with radiation and bleomycin or vinblastine or a combination of both cytostatics. Two radiation schedules were tested, the one involving larger fractions in a shorter period of time. Evaluation of the treatment results showed that 67% of patients experienced significant relief of dysphagia. The median survival was 3 months and the 1-year survival 7%. No statistical difference in survival was noted among the different chemotherapy or radiation groups (level of significance 5%). It was concluded that monochemotherapy as given in this study made no difference in the treatment result. The larger radiation fraction and shorter treatment time schedule was as effective as the more conventional schedule and is the preferred treatment because of the shorter hospitalization period. A pharyngostomy tube proved effective in feeding the patients. Oesophageal cancer is a systemic disease and polychemotherapy will have to play a prominent part in a multimodality treatment approach if results are to improve.