Pseudo-Fringe sign in frontal fibrosing alopecia-like cutaneous lupus erythematosus.

BACKGROUND: Marginal alopecia presents itself frequently in dermatological practice. For patients who present peripheral scalp hair loss, cicatricial and non-cicatricial alopecia should be considered, such as ophiasis, traction alopecia and frontal fibrosing alopecia. In spite of the several types of hair loss that have been described in cutaneous lupus erythematosus, the 'pseudo-fringe sign' had never been observed in such patents. OBJECTIVE: This study aims at including the 'pseudo-fringe sign' as another possibility of clinical manifestation in patients with cutaneous lupus erythematosus. METHODS: A case report of a 67-year-old lady with the diagnosis of lupus erythematosus and peripheral hair loss is shown in this study; the respective differential diagnosis have also been discussed. RESULTS: The diagnosis of a scarring alopecia caused by lupus erythematosus was established with dermoscopy and histopathology. CONCLUSION: Cutaneous lupus erythematosus is another diagnostic possibility for patients who display the 'pseudo-fringe sign'.

A phylogenetic and functional perspective on the origin and evolutionary shifts of growth ring anatomical markers in seed plants.

We reconstruct the evolutionary changes in different anatomical markers in order to understand the evolution and functional aspects of growth rings during the diversification of seed plants (spermatophytes), one of the largest and most diverse lineages of the tree of life. We carried out a wide revision of the anatomy of secondary xylem in spermatophytes and reconstructed the evolution of the different anatomical markers in a time-calibrated phylogeny. By embodying a functionally and evolutionarily significant concept in growth rings we reveal a new panorama for their frequency and show how common they are in diverse lineages of tropical plants. In this context, the principal anatomical markers of growth rings are identified in the evolutionary history of plants and their association with climate-related ecological characteristics. We discuss the function of these anatomical markers, especially for thick-walled and/or radially flattened latewood fibres, fibre zone and dilated rays. Despite the high evolutionary lability of the anatomical markers evidenced by our analyses, they appear to represent deep homologies.

The growth ring concept: seeking a broader and unambiguous approach covering tropical species.

The concept of growth rings is little discussed in the literature and their treatment remains somewhat confusing in terms of the diversity of structures described. This situation has a major impact on the study of growth rings in tropical species, in which variations and complexity are greater and accuracy of identification less good. The rigid conceptual delimitations used by dendrochronologists and wood anatomists of temperate regions cannot be applied to the study of growth rings in most tropical species, which has led to neglect of this subject. With the objective of discussing the concept of growth rings, the present study consists of a survey of anatomical, periodicity, causal and evolutionary aspects of the treatment of these structures as evidenced in previous studies, and the evaluation of their application and limitations to the development of this concept. Anatomical aspects arise through radial integrity, or the presence of early and late wood; the degree of distinction of the rings, which may vary from well to poorly defined; and tangential continuity, meaning that rings may form a complete circle in the transverse section of the trunk, or instead be tangentially discontinuous, lens-shaped or in wedges. In addition there are a diversity of anatomical markers which enable us to recognize growth rings. Regarding periodicity, the rings may be annual, infra-annual or supra-annual. Causal aspects include genetic, endogenous and environmental components. Evolutionary aspects continue to be insufficiently investigated, and although most comparative studies treat growth rings recognized by different markers as though they were a single character, it remains questionable whether they can be regarded as homologous or the result of homoplasy. We conclude that the elaboration of a robust but broad definition which can include all the variation observed in growth rings of tropical species is a complex task, which is only possible by overcoming the restrictions adopted by dendrochronologists and wood anatomists of temperate regions for whom growth rings are essentially annual, strongly demarcated, tangentially continuous and restricted to the most pronounced markers.

The European hare (Lepus europaeus) as a potential wild reservoir for ruminant pestiviruses.

Ruminant pestiviruses cause important economic losses in livestock and the epidemiological role of free-ranging sympatric wildlife is of special interest for the implementation of pestivirus eradication plans. Moreover, the emergence of high mortality outbreaks of pestivirus in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) since 2001 in the border between Spain and France has increased the value of knowing the hosts that role pestivirus infection. In the present study, pestivirus infection was assessed in 94 sera from wild hunted European hares (Lepus europaeus) collected in two different areas: Pyrenees (alpine and subalpine ecosystems) versus Non Pyrenees (non alpine and subalpine ecosystems). The presence of antibodies against Border Disease Virus (BDV) and Bovine Viral Diarrhea Virus (BVDV) was evaluated by means of the Virus Neutralization Test and the presence of viral RNA in sera samples was assessed by Reverse transcription-polymerase chain reaction (RT-PCR). A total of 34 out of 94 (36.2%; CI95 0.26-0.46) sera presented neutralizing antibodies against ruminant pestiviruses, and significant differences between BDV4 and BVDV1 titres were found in 7 hares. In the Pyrenean area not statistically significant seroprevalence was observed when comparing with the Non Pyrenean area. RT-PCR analysis of sera samples resulted all negative. The results of the present study indicate that the European hare is susceptible to pestivirus infection and that could be involved in the epidemiology of ruminant pestiviruses. To the authors' knowledge, this is the third wild non-artiodactyl with reported antibodies against ruminant pestivirus after the rabbit and Bennet's wallaby.

Identifying the major bacteria causing intramammary infections in individual milk samples of sheep and goats using traditional bacteria culturing and real-time polymerase chain reaction.

Use of DNA-based methods, such as real-time PCR, has increased the sensitivity and shortened the time for bacterial identification, compared with traditional bacteriology; however, results should be interpreted carefully because a positive PCR result does not necessarily mean that an infection exists. One hundred eight lactating dairy ewes (56 Manchega and 52 Lacaune) and 24 Murciano-Granadina dairy goats were used for identifying the main bacteria causing intramammary infections (IMI) using traditional bacterial culturing and real-time PCR and their effects on milk performance. Udder-half milk samples were taken for bacterial culturing and somatic cell count (SCC) 3 times throughout lactation. Intramammary infections were assessed based on bacteria isolated in ≥2 samplings accompanied by increased SCC. Prevalence of subclinical IMI was 42.9% in Manchega and 50.0% in Lacaune ewes and 41.7% in goats, with the estimated milk yield loss being 13.1, 17.9, and 18.0%, respectively. According to bacteriology results, 87% of the identified single bacteria species (with more than 3 colonies/plate) or culture-negative growth were identical throughout samplings, which agreed 98.9% with the PCR results. Nevertheless, the study emphasized that 1 sampling may not be sufficient to determine IMI and, therefore, other inflammatory responses such as increased SCC should be monitored to identify true infections. Moreover, when PCR methodology is used, aseptic and precise milk sampling procedures are key for avoiding false-positive amplifications. In conclusion, both PCR and bacterial culture methods proved to have similar accuracy for identifying infective bacteria in sheep and goats. The final choice will depend on their response time and cost analysis, according to the requirements and farm management strategy.

New molecular variants of epsilon and beta IncP-1 plasmids are present in estuarine waters.

In this work the presence of broad-host-plasmids in an estuary in Portugal has been investigated. Pseudomonas putida KT2442 was used as model recipient bacteria in biparental matings with tetracycline and mercury to select for resistance phenotypes. As a result, 7 transconjugants were shown to carry broad-host-plasmids from the IncP-1 group, as seen by PCR amplification of the trfA gene. Sequence analysis confirmed the isolation of 4 plasmids from ß-1 subgroup and 3 assigned to the recently described ε subgroup. To our knowledge this is the first report concerning the detection and isolation of IncP-1ß and ε plasmids in estuarine waters. Moreover it is shown that, even though the retrieved plasmids are phylogenetically close to previously characterized plasmids, such as pB10 and pKJK5, respectively, they constitute new molecular variants.

Endovascular treatment of carotid-cavernous fistulas: review of 12 cases.

Carotid-cavernous fistulas (CCFs) are an uncommon pathology in current neurosurgical practice. Furthermore, they generally present a challenge to the neurosurgeon because of both their deep location and the major neurovascular structures that surround them. CCFs consist of a vascular anomaly in which blood flows from meningeal branches of the internal and external carotid arteries, or directly from the internal carotid artery, into the venous circulation around and in the cavernous sinus. Twelve patients with direct and indirect/dural CCFs who underwent examination and treatment between July 2003 and February 2006 are reported here. All patients of this retrospective evaluation were symptomatic at the time of diagnosis, which was confirmed by cerebral angiography. The patients were treated by endovascular approaches and the CCFs were occluded. From our evaluation, we concluded that the endovascular procedure is safe, effective and minimally invasive in patients with CCF.

Application of on-line OUR measurements to detect actions points to improve baculovirus-insect cell cultures in bioreactors.

The continuous monitoring of a process based on the culture of Sf9 insect cells and infection by a baculovirus as a vector to obtain recombinant VP2 protein is studied. On-line OUR determination is based on the direct oxygen measurement in the cell culture vessel and the application of the dynamic method. This approximation allows a proper description of cell growth, with precise identification of the balanced cell growth end and the most important action times in the process, as virus infection time and final cell harvesting. A detailed study of the OUR profiles allows on-line monitoring of the effects of infection and expression protein process, a tool enabling the automatisation of the protein production process in a baculovirus-insect cell system. These parameters have been defined as time of action (TOAs), and include the most relevant actions to take in these type of processes: time of infection (TOI), time of feeding (TOF) and time of harvesting (TOH).

Endonasal endoscopic repair of cerebrospinal fluid rhinorrhea.

Cerebrospinal fluid (CSF) rhinorrhea implies a communication between the subarachnoid space and the upper respiratory tract. Trauma and postoperative defects are the more common causes of CSF rhinorrhea. The authors review their results with endoscopic repair of skull base defects associated with CSF rhinorrhea involving the paranasal sinuses. A total of 10 patients, 7 males and 3 females, were treated under endoscopic vision from 1997 to 2001. The causes of CSF rhinorrhea were traumatic (7), postoperative (2) and spontaneous (1). Four patients had the diagnosis and the site confirmed after intrathecal fluorescein-saline injection. The obliteration of the CSF leak was achieved with fat free, mucoperichondrial, or mucoperiostal free grafts taken from the middle or inferior turbinate and kept in place by fibrin glue. Primary closure was obtained in 10 patients and one patient developed a recurrence 14 months later. The repair of the CSF rhinorrhea by endonasal endoscopic surgery is safe, very effective and is a valid alternative to the cranial approach.

[Identification of the allergenic taxa of pollen in patients with pollinosis to determine the risk season]. / Identificación de los taxones de pólenes alergénicos en pacientes polínicos para conocer la temporada de riesgo.

BACKGROUND: Determining the risk season for the presence of pollen in the atmosphere aids primary care physicians in the diagnosis and treatment of allergic diseases. Our objective was to identify the taxa of pollen that cause allergic rhinoconjunctivitis in a sample of patients from a health center who presented seasonal symptoms. METHODS: We designed an observational, cross-sectional, non-randomized study to be carried out in the Cazoña Health Center in Santander, Spain. We selected 30 volunteers of both sexes, aged between 13 and 69 year old, who suffered seasonal rhinoconjunctivitis symptoms and who had always lived in Santander. Patients underwent skin-prick tests with the 25 pollen extracts routinely used in Spain, house dust mite (HDM), cat dander and Alternaria extracts by means of the Prick-Film system. The test result was expressed as the percentage of the papule area caused by histamine. Measurement was performed by scanning the copied papule area with the Prick-Scan program for PC. RESULTS: All patients were positive to grass pollen, 26 % were sensitized to grass pollen only and 23 % were also sensitized to other non-pollen allergens. Twenty-seven percent tested positive to Plantago, 20 % to Quercus and 13 % to Morus; the remaining pollen extracts were positive in less than 10 %. Fifty-six percent of the patients were positive to HDM and 6 % to cat dander. CONCLUSIONS: A warning period for pollinosis patients in the city of Santander can be defined. In our case, the important period is the grass pollen season, since the remaining pollen taxa sensitized few patients. Most of our patients were also sensitized to HDM.

Antigenic properties and diagnostic potential of baculovirus-expressed infectious bursal disease virus proteins VPX and VP3.

The routine technique for detecting antibodies specific to infectious bursal disease virus (IBDV) is a serological evaluation by enzyme-linked immunosorbent assay (ELISA) with preparations of whole virions as the antigens. To avoid using complete virus in the standard technique, we have developed two new antigens through the expression of the VPX and VP3 genes in insect cells. VPX and especially VP3 were expressed at high levels in insect cells and simple to purify. The immunogenicity of both proteins was similar to that of the native virus. VPX was able to elicit neutralizing antibodies but VP3 was not. Purified VPX and VP3 were tested in an indirect ELISA with more than 300 chicken sera. There was an excellent correlation between the results of the ELISA using VPX and those of the two commercial kits. VP3 did not perform as well as VPX, and the linear correlation was significantly lower. A comparison with the standard reference technique, seroneutralization, showed that the indirect ELISA was more sensitive. Therefore, VPX-based ELISA is a good alternative to conventional ELISAs that use whole virions.

Viability and DNA maintenance in nonculturable spiral Campylobacter jejuni cells after long-term exposure to low temperatures.

Survival of Campylobacter jejuni at 4 and 20 degrees C was investigated by using cellular integrity, respiratory activity, two-dimensional (2D) protein profile, and intact DNA content as indicators of potential viability of nonculturable cells. Intact DNA content after 116 days, along with cellular integrity and respiring cells, was detected for up to 7 months at 4 degrees C by pulsed-field gel electrophoresis. Most changes in 2D protein profiles involved up- or down-regulation.

Double staining (CTC-DAPI) for detection and enumeration of viable but non-culturable Campylobacter jejuni cells.

Direct microscopic enumeration of viable Campylobacter jejuni cells (ie, respiring bacteria) were performed in both culturable and non-culturable states. Five different C jejuni strains were used, including a reference strain, ATCC 33291. Cells from all five strains were incubated alone with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), a redox dye. It was reduced by an electron transport chain to an insoluble red fluorescent CTC formazan salt, which accumulated intracellularly. The presence of these red CTC crystals in the bacteria cells was indicative of cellular respiratory activity. Counterstaining with 4'-6 diamino-2 phenylindole (DAPI), which fluoresces in blue, made a suitable contrast and allowed simultaneous enumeration of total and viable bacteria on a single filter. Four hours of incubation with 5 mM CTC under a microaerobic atmosphere was found to be the optimal condition yielding the maximum number of respiring cells (both culturable and non-culturable). When used in combination with standard culture techniques, double staining makes it possible to monitor the viable but non-culturable cells of jejuni obtained by starvation more easily than with the direct viable count procedure.

Effect of the pre-treatments for milk samples filtration on direct viable cell counts.

Escherichia coli O25:H-42 was selected to study the effect of pre-treatments on the enumeration of direct viable cells from milk samples. Before and after inducing cell elongation by cellular division inhibitors, three pre-treatments for milk-filtration were used. One involved a pre-treatment with trypsin (1.5 min at 50 degrees C), addition of hot Triton X-100 after heating and filter rinses with phosphate saline buffer. The other two involved pre-treatment with trypsin and Triton X-100 (10 min at 50 degrees C), filter rinses with hot Triton X-100 and organic solvents. Pre-treatments applied after inducing cell elongation had an effect on cell recovery from milk samples depending on the pre-treatment used. The most suitable, on the basis of the number and percentage of enlarged cells obtained was the first described. The others selectively affected recovery of elongated cells. Pre-treatments applied before inducing the cell elongation, negatively affected viability with enumeration in milk samples being significantly (P < 0.001) lower than those found in controls. However, the negative effects of first pre-treatment on viability was lower than that produced by the pre-treatments involving organic solvents.