Breast Cancer Patient's Outcomes after Neoadjuvant Chemotherapy and Surgery at 5 and 10 Years for Stage II-III Disease.

Introduction: Neoadjuvant chemotherapy in breast cancer offers the possibility to facilitate breast and axillary surgery; it is a test of chemosensibility in vivo with significant prognostic value and may be used to tailor adjuvant treatment according to the response. Material and Methods: A retrospective single-institution cohort of 482 stage II and III breast cancer patients treated with neoadjuvant chemotherapy based on anthracycline and taxans, plus antiHEr2 in Her2-positive cases, was studied. Survival was calculated at 5 and 10 years. Kaplan-Meier curves with a log-rank test were calculated for differences according to age, BRCA status, menopausal status, TNM, pathological and molecular surrogate subtype, 20% TIL cut-off, surgical procedure, response to chemotherapy and the presence of vascular invasion. Results: The pCR rate was 25.3% and was greater in HER2 (51.3%) and TNBC (31.7%) and in BRCA carriers (41.9%). The factors independently related to patient survival were pathology and molecular surrogate subtype, type of surgery, response to NACT and vascular invasion. BRCA status was a protective prognostic factor without reaching statistical significance, with an HR 0.5 (95%CI 0.1-1.4). Mastectomy presented a double risk of distant recurrence compared to breast-conservative surgery (BCS), supporting BCS as a safe option after NACT. After a mean follow-up of 126 (SD 43) months, luminal tumors presented a substantial difference in survival rates calculated at 5 or 10 years (81.2% compared to 74.7%), whereas that for TNBC was 75.3 and 73.5, respectively. The greatest difference was seen according to the response in patients with pCR, who exhibited a 10 years DDFS of 95.5% vs. 72.4% for those patients without pCR, p < 0001. This difference was especially meaningful in TNBC: the 10 years DDFS according to an RCB of 0 to 3 was 100%, 80.6%, 69% and 49.2%, respectively, p < 0001. Patients with a particularly poor prognosis were those with lobular carcinomas, with a 10 years DDFS of 42.9% vs. 79.7% for ductal carcinomas, p = 0.001, and patients with vascular invasion at the surgical specimen, with a 10 years DDFS of 59.2% vs. 83.6% for those patients without vascular invasion, p < 0.001. Remarkably, BRCA carriers presented a longer survival, with an estimated 10 years DDFS of 89.6% vs. 77.2% for non-carriers, p = 0.054. Conclusions: Long-term outcomes after neoadjuvant chemotherapy can help patients and clinicians make well-informed decisions.

Infecciones invasivas debidas a Saprochaete capitata en pacientes con neoplasias hematológicas: informe de cinco casos y revisión del tratamiento antimicótico / Invasive infections caused by Saprochaete capitata in patients with haematological malignancies: report of five cases and review of the antifungal therapy

Background. Saprochaete capitata (formerly known as Geotrichum capitatum and Blastoschizomyces capitatus) is a ubiquitous fungus found in soil, water, air, plants and dairy products. It colonizes the skin, and bronchial and intestinal tract of healthy people producing serious opportunistic infections in patients with haematological malignancies, especially in those with acute leukaemia. Since 1960s its presence is being increasingly recognized in this group of patients. The clinical spectrum of S. capitata disseminated infections is very similar to that produced by Candida, being easily misinterpreted. The associated high mortality and low susceptibility to fluconazole and echinocandins of S. capitata require the acknowledgement of this emergent infection so that it can be properly treated. Case report. We report 5 new cases of S. capitata disseminated infection in patients with advanced haematological malignancies observed in the haematology unit between the years 2004 and 2010, and review the state-of-the-art for diagnosis and treatment of this infection. Conclusions. Based on our experience, the prophylactic use of or the empirical antifungal treatment with fluconazole and/or echinocandins would not be adequate for oncohaematological patients in those hospitals where S. capitata infection may be highly prevalent (AU)

Invasive infections caused by Saprochaete capitata in patients with haematological malignancies: report of five cases and review of the antifungal therapy.

BACKGROUND: Saprochaete capitata (formerly known as Geotrichum capitatum and Blastoschizomyces capitatus) is a ubiquitous fungus found in soil, water, air, plants and dairy products. It colonizes the skin, and bronchial and intestinal tract of healthy people producing serious opportunistic infections in patients with haematological malignancies, especially in those with acute leukaemia. Since 1960s its presence is being increasingly recognized in this group of patients. The clinical spectrum of S. capitata disseminated infections is very similar to that produced by Candida, being easily misinterpreted. The associated high mortality and low susceptibility to fluconazole and echinocandins of S. capitata require the acknowledgement of this emergent infection so that it can be properly treated. CASE REPORT: We report 5 new cases of S. capitata disseminated infection in patients with advanced haematological malignancies observed in the haematology unit between the years 2004 and 2010, and review the state-of-the-art for diagnosis and treatment of this infection. CONCLUSIONS: Based on our experience, the prophylactic use of or the empirical antifungal treatment with fluconazole and/or echinocandins would not be adequate for oncohaematological patients in those hospitals where S. capitata infection may be highly prevalent.

Fosfoglicerato quinasa y fructosa bifosfato aldolasa de Candida albicans como nuevos antígenos reconocidos por la IgA salival humana / Phosphoglycerate kinase and fructose bisphosphate aldolase of Candida albicans as new antigens recognized by human salivary IgA

Fundamento. Candida albicans es un hongo dimórfico oportunista que, con frecuencia, está presente en la cavidad oral del ser humano donde da lugar a infecciones en pacientes inmunocomprometidos. Influida por las condiciones de crecimiento, la variabilidad antigénica es un factor de patogenicidad. Objetivos. Determinar el efecto del estrés nutricional y térmica en la expresión antigénica de C. albicans, e identificar los principales antígenos reconocidos por la inmunoglobulina secretora A (sIgA) salival humana. Métodos. En diferentes condiciones nutricionales, se indujo un choque térmico en células de C. albicans en fase de crecimiento estacionario y exponencial. La expresión de los determinantes proteicos de C. albicans se analizó mediante inmunotransferencia frente a la saliva humana. Los antígenos se purificaron y caracterizaron mediante electroforesis bidimensional y se identificaron mediante microsecuenciación de proteínas. Resultados. Se caracterizaron cinco antígenos reconocidos por la IgA salival como manoproteínas debido a su reactividad con la concanavalina A. Ninguno manifestó reactividad con los anticuerpos monoclonales anti-proteína de choque térmico. Se encontró que el choque térmico y el estrés nutricional regulaban dos de ellos (de 42 y 36 kDa) identificados como fosfoglicerato quinasa y fructosa bifosfato aldolasa, respectivamente. Conclusión. Estas enzimas glucolíticas son antígenos mayores de C. albicans, y su expresión diferencial y el reconocimiento por el sistema de la respuesta inmunitaria de la mucosa podrían participar en la protección frente a las infecciones orales(AU)

Background. Candida albicans is an opportunistic dimorphic fungus commonly present in the human oral cavity that causes infections in immunocompromised patients. The antigen variability, influenced by growth conditions, is a pathogenicity factor. Aims. To determine the effect of nutritional and heat stress on the antigen expression of C. albicans, and to identify major antigens recognized by human salivary secretory immunoglobulin A (sIgA). Methods. Under various different nutritional conditions, heat shock was induced in C. albicans cells in stationary and exponential growth phases. The expression of protein determinants of C. albicans was assessed by Western blot analysis against human saliva. The antigens were purified and characterized by two-dimensional electrophoresis and identified by protein microsequencing. Results. Five antigens recognized by salivary IgA were characterized as mannoproteins due to their reactivity with concanavalin A. They did not show reactivity with anti-heat shock protein monoclonal antibodies. Two of them (42 and 36kDa) were found to be regulated by heat shock and by nutritional stress and they were identified as phosphoglycerate kinase and fructose bisphosphate aldolase, respectively. Conclusions. These glycolytic enzymes are major antigens of C. albicans, and their differential expression and recognition by the mucosal immune response system could be involved in protection against oral infection(AU)

Clinical, tomographic, and histological assessment of periosteal guided bone regeneration with cortical perforations in advanced human critical size defects.

BACKGROUND: Large osseous defects that fail to heal spontaneously require ridge augmentation prior to implant placement. The periosteum can act as an effective barrier membrane. Little is known about the influence of bone decortication in enhancing guided bone regeneration outcomes. PURPOSE: The aim of the present study was a clinical, tomographic, and histological evaluation of bone healing in large defect sites treated with cortical perforations without the use of other membranes but the periosteum. MATERIAL AND METHODS: Ten consecutive patients undergoing ridge augmentation on the pre-maxilla due to severe bone loss were followed for an average of 35 months. Recipient sites were cortico-perforated and augmented using a combination of autogenous particulate and block grafts. The periosteal membrane was preserved and it fully covered the autografts. Histological analysis was performed in four sites from a trephine core taken at the time of implant osteotomy preparation. Tomographic assessment (computed tomography [CT] scan) at baseline and post-augmentation evaluated graft volume maintenance. RESULTS: Recipient sites were re-entered for implant placement showing good incorporation of the grafts with minimal volume loss. Biopsy specimens showed viable bone rich in osteoblast-like cells with little or no inflammatory cells. Clinical exam revealed absence of implant transparency, mucosal recession, mobility, bleeding on probing, or suppuration at follow-up. CT scan evaluation showed an average increased bucco-lingual width at the recipient site of 8.1 mm ± 0.9 (2.5 fold) versus a 3.2 ± 0.9 at baseline (p < .0001; CI 95%: 4.04-5.71 mm), maintaining on average 98% of the augmented width at 2.9 years. CONCLUSIONS: Periosteal preservation seems to be sufficient as a barrier membrane to protect particulate or block osseous grafts provided that good primary closure is achieved. Bone decortication may enhance clinical and histological outcomes. Graft viability (biopsy specimens) and volume maintenance (CT evaluation) remained stable 35 months post-augmentation.

Periodontopathogen and Epstein-Barr virus contamination affects transplanted bone volume in sinus augmentation.

BACKGROUND: Bone microbial contamination can impair osteogenesis. Human herpesviruses-associated vasculitis can cause vascular damage within the osseous graft and host. This study is conducted to substantiate specific contamination and assess the impact 6 months after sinus augmentation. METHODS: Culture- and polymerase chain reaction (PCR)-based identification were done on harvested bone particles and unstimulated whole saliva in a group of 30 patients undergoing maxillary sinus augmentation. Patients were divided into two groups: those with and those without a history of periodontitis. Radiographic evaluation was done to assess and compare bone healing and volume gain at baseline and 6 months post-transplantation. RESULTS: Seventeen patients had a history of periodontitis, and 13 did not. Ten showed culture- and PCR-negative results and belonged to the periodontally healthy group. The 17 patients with periodontitis showed culture- or PCR-positive results for the targeted periodontal pathogens. Patients with periodontitis were 2.3 times more likely to have positive salivary Epstein-Barr virus type 1 (EBV-1) than those with no history of periodontitis. The likelihood of having moderate to pronounced bone volume loss 6 months postaugmentation was 7.5 times greater in those patients presenting contamination with ≥3 specific pathogens (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, or Prevotella intermedia) versus those with only one (P <0.05). The odds ratio (OR) of pronounced volume loss was 16.3 times higher in those contaminated with a combination of salivary EBV-1 and ≥3 of the previously mentioned species versus only EBV-1 (P <0.05). Individuals showing positive salivary EBV-1 had bone bacterial contamination associated 57% of the time. The OR of having bone microbial contamination in patients with a history of periodontitis was 37.5 times higher than in individuals without periodontitis. CONCLUSIONS: This study confirms contamination of bone, harvested intraorally, with key periodontopathogens in individuals undergoing sinus augmentation. Specific microbial contamination can impair osteogenesis. Saliva may act as a vehicle to transport EBV and other pathogens into the sinus. Increased bone volume loss seems to be associated with the occurrence of specific periodontal anaerobic species, salivary EBV-1, or the combination of both.

Phosphoglycerate kinase and fructose bisphosphate aldolase of Candida albicans as new antigens recognized by human salivary IgA.

BACKGROUND: Candida albicans is an opportunistic dimorphic fungus commonly present in the human oral cavity that causes infections in immunocompromised patients. The antigen variability, influenced by growth conditions, is a pathogenicity factor. AIMS: To determine the effect of nutritional and heat stress on the antigen expression of C. albicans, and to identify major antigens recognized by human salivary secretory immunoglobulin A (sIgA). METHODS: Under various different nutritional conditions, heat shock was induced in C. albicans cells in stationary and exponential growth phases. The expression of protein determinants of C. albicans was assessed by Western blot analysis against human saliva. The antigens were purified and characterized by two-dimensional electrophoresis and identified by protein microsequencing. RESULTS: Five antigens recognized by salivary IgA were characterized as mannoproteins due to their reactivity with concanavalin A. They did not show reactivity with anti-heat shock protein monoclonal antibodies. Two of them (42 and 36 kDa) were found to be regulated by heat shock and by nutritional stress and they were identified as phosphoglycerate kinase and fructose bisphosphate aldolase, respectively. CONCLUSIONS: These glycolytic enzymes are major antigens of C. albicans, and their differential expression and recognition by the mucosal immune response system could be involved in protection against oral infection.

Prevalence of Candida species in necrotic pulp with chronic periapical processes.

The aim of this study was to identify species of the genus Candida in mucosa of oral cavity and in single-rooted teeth with pulp necrosis with chronic endodontic periapical processes, with radiographic images 2+/-4 mm and without clinical symptomatology, in immunocompetent patients. The study included 82 immunocompetent patients of both sexes aged 18-70 years with a clinical dental diagnosis of septic pulp necrosis. Samples were taken from root canals with sterile # 25 paper points and from oral mucosa with a sterile swab. Seven different Candida species were identified (C. albicans, C. dubliniensis, C. guilliermondii, C. krusei, C. parapsilopsis, C. tropicalis and C. glabrata). All of them were present in oral mucosa, while two of them (C. parapsilopsis and C. glabrata) were not identified in the periapical zone of necrotic canals. Considering all the samples isolated from oral mucosa, there was a significantly greater frequency of C. albicans than there was in the periapical zone of necrotic canals.

Long-term onlay graft volume maintenance in aggressive periodontitis patients.

Treatment planning in the esthetic zone has classically presented some of the greatest challenges to the practitioner. The purpose of this article is to describe a staged, multidisciplinary approach and follow-up to a case of aggressive periodontitis. Microbial sampling for suspected periodontopathogens was taken before and after treatment. Qualitative polymerase chain reaction analysis was done to detect the presence of cytomegalovirus and Epstein-Barr virus type 1 6 years after active periodontal therapy. A control computed tomography scan taken 5.5 years postaugmentation showed stable bone levels and excellent volume maintenance of the transplanted block graft.

Fungicidal monoclonal antibody C7 interferes with iron acquisition in Candida albicans.

We have developed a monoclonal antibody (MAb), C7, that reacts with the Als3p and enolase present in the Candida albicans cell wall and exerts three anti-Candida activities: candidacidal activity and inhibition of both adhesion and filamentation. To investigate the mode of action of MAb C7 on fungal viability, we examined changes in the genome-wide gene expression profile of C. albicans grown in the presence of a subinhibitory concentration of MAb C7 (12.5 µg/ml) by using microarrays. A total of 49 genes were found to be differentially expressed upon treatment with MAb C7. Of these, 28 were found to be upregulated and 21 were found to be downregulated. The categories of upregulated genes with the largest number of variations were those involved in iron uptake or related to iron homeostasis (42.86%), while the energy-related group accounted for 38.10% of the downregulated genes (8/21). Results were validated by real-time PCR. Since these effects resembled those found under iron-limited conditions, the activity of MAb C7 on C. albicans mutants with deletions in key genes implicated in the three iron acquisition systems described in this yeast was also assessed. Only mutants lacking the TPK1 gene and, to a lesser extent, the TPK2 gene were less sensitive to the candidacidal effect of MAb C7. FeCl(3) or hemin at concentrations of ≥ 7.8 µM reversed the candidacidal effect of MAb C7 on C. albicans in a concentration-dependent manner. The results presented in this study provide evidence that the candidacidal effect of MAb C7 is related to the blockage of the reductive iron uptake pathway of C. albicans.

Long-term block graft stability in thin periodontal biotype patients: a clinical and tomographic study.

PURPOSE: To assess the esthetic and functional outcome, as well as the volume maintenance, of autogenous block grafts placed in anterior sextants of thin periodontal biotype subjects over a long-term period. MATERIALS AND METHODS: Fifteen consecutive patients were followed up yearly for an average of 40 months after autogenous block grafting. Preoperative and postoperative cone beam computed tomographic scans were analyzed to evaluate bone volume maintenance around the implants placed in the grafted sites. Clinical parameters (mucosal recession and implant transparency through the soft tissue) were assessed at prosthesis delivery and follow-up to evaluate the esthetic outcome. Digital photographs were used to confirm clinical outcomes. RESULTS: The average augmentation per site was 2.2 times the initial buccolingual (BL) width, and 97% of the augmented width was maintained after 3.3 years. The difference between preaugmentation and postaugmentation BL width, 3.3 versus 7.4 mm, was statistically significant (P < .0001; CI 95%: 3.4 to 4.9 mm). There was a lack of implant transparency or mucosal recession around the implants in all 15 patients after an average of 40 months. CONCLUSIONS: Autogenous block grafting seems to be a predictable treatment modality to reconstruct alveolar ridge defects in the long term. A thin periodontal biotype did not seem to affect the volume of transplanted bone for the population studied.

Clinical factors associated with a Candida albicans Germ Tube Antibody positive test in Intensive Care Unit patients.

BACKGROUND: Poor outcomes of invasive candidiasis (IC) are associated with the difficulty in establishing the microbiological diagnosis at an early stage. New scores and laboratory tests have been developed in order to make an early therapeutic intervention in an attempt to reduce the high mortality associated with invasive fungal infections. Candida albicans IFA IgG has been recently commercialized for germ tube antibody detection (CAGTA). This test provides a rapid and simple diagnosis of IC (84.4% sensitivity and 94.7% specificity). The aim of this study is to identify the patients who could be benefited by the use of CAGTA test in critical care setting. METHODS: A prospective, cohort, observational multicentre study was carried out in six medical/surgical Intensive care units (ICU) of tertiary-care Spanish hospitals. Candida albicans Germ Tube Antibody test was performed twice a week if predetermined risk factors were present, and serologically demonstrated candidiasis was considered if the testing serum dilution was ≥ 1:160 in at least one sample and no other microbiological evidence of invasive candidiasis was found. RESULTS: Fifty-three critically ill non-neutropenic patients (37.7% post surgery) were included. Twenty-two patients (41.5%) had CAGTA-positive results, none of them with positive blood culture for Candida. Neither corrected colonization index nor antifungal treatment had influence on CAGTA results. This finding could corroborate that the CAGTA may be an important biomarker to distinguish between colonization and infection in these patients. The presence of acute renal failure at the beginning of the study was more frequent in CAGTA-negative patients. Previous surgery was statistically more frequent in CAGTA-positive patients. CONCLUSIONS: This study identified previous surgery as the principal clinical factor associated with CAGTA-positive results and emphasises the utility of this promising technique, which was not influenced by high Candida colonization or antifungal treatment. Our results suggest that detection of CAGTA may be important for the diagnosis of invasive candidiasis in surgical patients admitted in ICU.

Recomendaciones sobre el diagnóstico de la enfermedad fúngica invasora de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC). Actualización 2010 / Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) guidelines for the diagnosis of invasive fungal infections. 2010 update

Este texto incluye una actualización de las recomendaciones sobre el diagnóstico de la infección fúngica invasora de la Sociedad Española de Microbiología Clínica y Enfermedades Infecciosas (SEIMC) elaboradas en 2004 (Enferm Infecc Microbiol Clin. 2004, 22:32-9). En la actualización de 2010 se incluye una revisión exhaustiva de las novedades tecnológicas de los últimos años, así como los niveles de evidencia para recomendar cada una de las técnicas de diagnóstico. En primer lugar, se analizan los métodos convencionales, examen microscópico y cultivo, con sus limitaciones, lo que ha llevado a desarrollar métodos alternativos como la cuantificación de antígenos y de ADN fúngico. Las indicaciones de los métodos alternativos se analizan para las diferentes infecciones fúngicas, candidiasis, aspergilosis y micosis invasoras por otras especies. Por último, se incluye una breve descripción de los métodos de identificación molecular y se revisan las pruebas para realizar estudios de sensibilidad a los antifúngicos, los procedimientos de referencia, las técnicas comerciales y sus indicaciones (AU)

These guidelines are an update of recommendations for the diagnosis of invasive fungal infections by the Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) published in 2004 (Enferm Infecc Microbiol Clin. 2004, 22:32-9). In this updated version of the guidelines, a comprehensive review of the most recent diagnostic innovations and levels of evidence to recommend those diagnostic procedures are included. We first analyse conventional diagnostic methods, microscopic examination and culture, underlining their limitations which have led to the development of alternative methods, such as fungal antigen and DNA quantification. Those alternative methods of diagnosis are analysed by fungal infection. We also briefly review the methods for molecular identification of fungal species and recommendations for carrying out susceptibility tests for antifungal drugs, including reference procedures, commercial techniques and their indications (AU)

Bone microbial decontamination agents in osseous grafting: an in vitro study with fresh human explants.

BACKGROUND: Establishing a safe prophylactic antimicrobial protocol in bone grafting may enhance osseous volume outcomes. The purpose of this in vitro study is to assess human osteoblast response and safety after explant antimicrobial exposure. METHODS: Fresh human bone explants were exposed to three antimicrobials: povidone-iodine (PovI; 0.05%, 1%, and 5%), chlorhexidine (CHX; 0.2% and 1%), and sodium hypochlorite (NaOCl; 2.5%, 4.5%, and 5.25%) at different times (15, 30, 45, and 60 seconds) and concentrations to assess cellular toxicity. Explants were washed three times with saline after exposure. Controls, explants cultured in the absence of antimicrobials, were performed for all experimental situations tested. Trials were conducted in triplicate. Particle size influence on osteoblast growth was determined between bone fragments with a diameter <2 and ≥2 to 5 mm. Test and control groups were monitored by light microscopy to evaluate cellular growth. Osteoblast differentiation and morphology was assessed by alkaline phosphatase activity and scanning electron microscopy (SEM). RESULTS: Osteoblast growth was similar for particles <2 and ≥2 to 5 mm. Alkaline phosphatase control reference values were not significantly different from test groups (0.35 mU/mL ± 0.004 versus 0.34 mU/mL ± 0.009; P >0.05). Light microscopy showed on average 97% osteoblastic growth for bone particles exposed to PovI 5% and CHX 0.2% for all times and CHX 1% up to 30 seconds. The odds ratio of positive osteoblastic growth after a 30-second 2.5% NaOCl exposure was 2.4 times higher than after 5.25%. On average, one of two replicas yielded positive growth with 2.5% NaOCl and one of three with 5.25%. After 60-second explant exposure, positive osteoblastic growth was 7.7 times more likely to occur with 5% PovI or 0.2% CHX than with 5.25% NaOCl (P <0.05). SEM analysis confirmed light microscopy similar cellular adhesion and osteoblast phenotypic features between test and control groups. CONCLUSIONS: Best osteoblastic growth occurred after bone PovI exposure and CHX 0.2%. Cellular toxicity seems to be influenced by the type of antimicrobial, concentration, and exposure time. SEM analysis confirmed absence of osteoblast phenotypic alterations after exposure. Decontamination agents can safely be used in bone transplantation using up to 5% PovI and 0.2% CHX for 1 minute and CHX 1% for 30 seconds.

[Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) guidelines for the diagnosis of invasive fungal infections. 2010 update]. / Recomendaciones sobre el diagnóstico de la enfermedad fúngica invasora de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC). Actualización 2010.

These guidelines are an update of recommendations for the diagnosis of invasive fungal infections by the Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) published in 2004 (Enferm Infecc Microbiol Clin. 2004, 22:32-9). In this updated version of the guidelines, a comprehensive review of the most recent diagnostic innovations and levels of evidence to recommend those diagnostic procedures are included. We first analyse conventional diagnostic methods, microscopic examination and culture, underlining their limitations which have led to the development of alternative methods, such as fungal antigen and DNA quantification. Those alternative methods of diagnosis are analysed by fungal infection. We also briefly review the methods for molecular identification of fungal species and recommendations for carrying out susceptibility tests for antifungal drugs, including reference procedures, commercial techniques and their indications.

Prevalence of Candida species in necrotic pulp with chronic periapical processes

El propósito de este estudio fue identificar especies del géneroCandida en mucosa de cavidad bucal y dientes unirradiculares con necrosis pulpar con procesos periapicales crónicos de origen endodóntico con imágenes radiográficas entre 2±4 mm sin sintomatología clínica en pacientes inmunocompetentes. El estudio incluyó 82 pacientes inmunocompetentes de ambos sexos con edades entre 18-70 años con diagnóstico clínico dentario de necrosis pulpar séptica. Las muestras fueron obtenidas del conducto radicular con conos de papel estéril # 25 y de lamucosa bucal mediante hisopo estéril. Se identificaron 7 especies diferentes de Candida (C. albicans, C. dubliniensis,C. guilliermondii, C. krusei, C. arapsilopsis, C. tropicalis y C. glabrata). Todas estuvieron presentes en mucosa bucal, mientras que dos de ellas (C. parapsilopsis y C. glabrata) nose identificaron en zona periapical de conductos con necrosis Del total de las muestras aisladas de mucosa bucal hubo una frecuencia significativamente mayor de C. albicans que la proporción de estas levaduras en la zona periapical en conductos con necrosis.

What makes Aspergillus fumigatus a successful pathogen? Genes and molecules involved in invasive aspergillosis

Dos secciones incluyen los genes y moléculas relacionadas con la absorción de nutrientes, la señalización y las regulaciones metabólicas implicadas en la virulencia, incluyendo enzimas, como las serin-proteasas (alp/asp f 13, alp2 y asp f 18), metaloproteasas (mep/asp f 5, mepB y mep20), aspártico-proteasas (pep/asp f 10, pep2 y ctsD), dipeptidilpeptidasas (dppIV y dppV) y fosfolipasas (plb1-3 y fosfolipasa C); sideróforos y la adquisición de hierro (sidA-G sreA, ftrA, fetC, mirB-C y amcA); adquisición de zinc (zrfA-H, zafA, y pacC); biosíntesis de aminoácidos, absorción de nitrógeno, y regulación por Cross-pathway Control (areA, rhbA, mcsA, lysF, cpcA/gcn4p y cpcC/gcn2p); vías de biosíntesis generales (pyrG, hcsA, y pabaA) y biosíntesis de trehalosa (tpsA y tpsB); otras vías de regulación, como MAP quinasas (sakA/hogA, mpkA-C, ste7, pbs2, mkk2, steC/ste11, bck1, ssk2 y sho1), proteínas G (gpaA, sfaD y cpgA), AMPc-PKA (acyA, gpaB, pkaC1 y pkaR), histidin-quinasas (fos1 y tcsB), señalización de Ca2+(calA/cnaA, crzA, gprC y gprD), familia Ras (rasA, rasB y rhbA), y otros (ace2, medA, y srbA). Por último, también se comentan los efectos de los alérgenos de A. fumigatus (Asp f 1 a Asp f 34) en la AI. Los datos obtenidos generan un complejo rompecabezas, cuyas piezas serían factores de virulencia o diferentes actividades del hongo, que se deben reunir para obtener una visión conjunta de la virulencia de A. fumigatus. Los estudios de expresión mediante microarrays de ADN podrían ser útiles para entender esta compleja virulencia, y para detectar dianas para desarrollar métodos rápidos de diagnóstico y nuevos agentes antifúngicos. Aspergillus fumigatus es un patógeno oportunista que causa el 90% de las aspergilosis invasoras (AI) con un 50–95% de mortalidad. Se ha postulado la existencia de factores de virulencia característicos, pero en A. fumigatus existe una gran variabilidad de factores de virulencia «no clásicos». Todos los estudios han demostrado que la virulencia de este hongo es multifactorial, asociada a su estructura, su capacidad de crecimiento y adaptación a condiciones de estrés, sus mecanismos de evasión del sistema inmune y su capacidad de causar daños en un huésped. En esta revisión se pretende dar una visión general de los genes y moléculas que intervienen en el desarrollo de la AI. La sección de termotolerancia incluye cinco genes relacionados con la capacidad de que el hongo crezca a más de 30°C (thtA, cgrA, afpmt1, kre2/afmnt1 y hsp1/asp f 12). En las siguientes secciones se discuten las moléculas y los genes relacionados con la interacción con el huésped y con la respuesta inmune. Estas secciones incluyen el β-glucano, el α-glucano, la quitina, el galactomanano, galactomanoproteinas (afmp1/asp f 17 y afmp2), hidrofobinas (rodA/hyp1 y rodB), la DHN-melanina, sus respectivas enzimas sintasas (fks1, rho1-4, ags1-3, chsA-G, och1-4, mnn9, van1, anp1, glfA, pksP/alb1, arp1, arp2, abr1, abr2 y ayg1) y enzimas modificantes (gel1-7, bgt1, eng1, ecm33, afpigA, afpmt1-2, afpmt4, kre2/afmnt1, afmnt2-3, afcwh41 y pmi), varias enzimas relacionadas con la protección del estrés oxidativo como catalasas (catA, cat1/catB, cat2/katG, catC y catE), superóxido dismutasas (sod1-2, sod3/asp f 6 y sod4), oxigenasas de ácidos grasos (ppoA-C), glutatión transferasas (gstA-E) y otros (afyap1, skn7 y pes1), y los transportadores de moléculas (mdr1-4, atrF, abcA-E y msfA-E)...(AU)

Two sections cover genes and molecules related with nutrient uptake, signaling and metabolic regulations involved in virulence, including enzymes, such as serine proteases (alp/asp f 13, alp2, and asp f 18), metalloproteases (mep/asp f 5, mepB, and mep20), aspartic proteases (pep/asp f 10, pep2, and ctsD), dipeptidylpeptidases (dppIV and dppV), and phospholipases (plb1–3 and phospholipase C); siderophores and iron acquisition (sidA–G, sreA, ftrA, fetC, mirB–C, and amcA); zinc acquisition (zrfA–H, zafA, and pacC); amino acid biosynthesis, nitrogen uptake, and cross-pathways control (areA, rhbA, mcsA, lysF, cpcA/gcn4p, and cpcC/gcn2p); general biosynthetic pathway (pyrG, hcsA, and pabaA), trehalose biosynthesis (tpsA and tpsB), and other regulation pathways such as those of the MAP kinases (sakA/hogA, mpkA–C, ste7, pbs2, mkk2, steC/ste11, bck1, ssk2, and sho1), G-proteins (gpaA, sfaD, and cpgA), cAMP-PKA signaling (acyA, gpaB, pkaC1, and pkaR), His kinases (fos1 and tcsB), Ca2+ signaling (calA/cnaA, crzA, gprC and gprD), and Ras family (rasA, rasB, and rhbA), and others (ace2, medA, and srbA). Finally, we also comment on the effect of A. fumigatus allergens (Asp f 1–Asp f 34) on IA. The data gathered generate a complex puzzle, the pieces representing virulence factors or the different activities of the fungus, and these need to be arranged to obtain a comprehensive vision of the virulence of A. fumigatus. The most recent gene expression studies using DNA-microarrays may be help us to understand this complex virulence, and to detect targets to develop rapid diagnostic methods and new antifungal agents. Aspergillus fumigatus is an opportunistic pathogen that causes 90% of invasive aspergillosis (IA) due to Aspergillus genus, with a 50–95% mortality rate. It has been postulated that certain virulence factors are characteristic of A. fumigatus, but the “non-classical” virulence factors seem to be highly variable. Overall, published studies have demonstrated that the virulence of this fungus is multifactorial, associated with its structure, its capacity for growth and adaptation to stress conditions, its mechanisms for evading the immune system and its ability to cause damage to the host. In this review we intend to give a general overview of the genes and molecules involved in the development of IA. The thermotolerance section focuses on five genes related with the capacity of the fungus to grow at temperatures above 30°C (thtA, cgrA, afpmt1, kre2/afmnt1, and hsp1/asp f 12)... (AU)

What makes Aspergillus fumigatus a successful pathogen? Genes and molecules involved in invasive aspergillosis.

Aspergillus fumigatus is an opportunistic pathogen that causes 90% of invasive aspergillosis (IA) due to Aspergillus genus, with a 50-95% mortality rate. It has been postulated that certain virulence factors are characteristic of A. fumigatus, but the "non-classical" virulence factors seem to be highly variable. Overall, published studies have demonstrated that the virulence of this fungus is multifactorial, associated with its structure, its capacity for growth and adaptation to stress conditions, its mechanisms for evading the immune system and its ability to cause damage to the host. In this review we intend to give a general overview of the genes and molecules involved in the development of IA. The thermotolerance section focuses on five genes related with the capacity of the fungus to grow at temperatures above 30°C (thtA, cgrA, afpmt1, kre2/afmnt1, and hsp1/asp f 12). The following sections discuss molecules and genes related to interaction with the host and with the immune responses. These sections include ß-glucan, α-glucan, chitin, galactomannan, galactomannoproteins (afmp1/asp f 17 and afmp2), hydrophobins (rodA/hyp1 and rodB), DHN-melanin, their respective synthases (fks1, rho1-4, ags1-3, chsA-G, och1-4, mnn9, van1, anp1, glfA, pksP/alb1, arp1, arp2, abr1, abr2, and ayg1), and modifying enzymes (gel1-7, bgt1, eng1, ecm33, afpigA, afpmt1-2, afpmt4, kre2/afmnt1, afmnt2-3, afcwh41 and pmi); several enzymes related to oxidative stress protection such as catalases (catA, cat1/catB, cat2/katG, catC, and catE), superoxide dismutases (sod1, sod2, sod3/asp f 6, and sod4), fatty acid oxygenases (ppoA-C), glutathione tranferases (gstA-E), and others (afyap1, skn7, and pes1); and efflux transporters (mdr1-4, atrF, abcA-E, and msfA-E). In addition, this review considers toxins and related genes, such as a diffusible toxic substance from conidia, gliotoxin (gliP and gliZ), mitogillin (res/mitF/asp f 1), hemolysin (aspHS), festuclavine and fumigaclavine A-C, fumitremorgin A-C, verruculogen, fumagillin, helvolic acid, aflatoxin B1 and G1, and laeA. Two sections cover genes and molecules related with nutrient uptake, signaling and metabolic regulations involved in virulence, including enzymes, such as serine proteases (alp/asp f 13, alp2, and asp f 18), metalloproteases (mep/asp f 5, mepB, and mep20), aspartic proteases (pep/asp f 10, pep2, and ctsD), dipeptidylpeptidases (dppIV and dppV), and phospholipases (plb1-3 and phospholipase C); siderophores and iron acquisition (sidA-G, sreA, ftrA, fetC, mirB-C, and amcA); zinc acquisition (zrfA-H, zafA, and pacC); amino acid biosynthesis, nitrogen uptake, and cross-pathways control (areA, rhbA, mcsA, lysF, cpcA/gcn4p, and cpcC/gcn2p); general biosynthetic pathway (pyrG, hcsA, and pabaA), trehalose biosynthesis (tpsA and tpsB), and other regulation pathways such as those of the MAP kinases (sakA/hogA, mpkA-C, ste7, pbs2, mkk2, steC/ste11, bck1, ssk2, and sho1), G-proteins (gpaA, sfaD, and cpgA), cAMP-PKA signaling (acyA, gpaB, pkaC1, and pkaR), His kinases (fos1 and tcsB), Ca(2+) signaling (calA/cnaA, crzA, gprC and gprD), and Ras family (rasA, rasB, and rhbA), and others (ace2, medA, and srbA). Finally, we also comment on the effect of A. fumigatus allergens (Asp f 1-Asp f 34) on IA. The data gathered generate a complex puzzle, the pieces representing virulence factors or the different activities of the fungus, and these need to be arranged to obtain a comprehensive vision of the virulence of A. fumigatus. The most recent gene expression studies using DNA-microarrays may be help us to understand this complex virulence, and to detect targets to develop rapid diagnostic methods and new antifungal agents.

Potential of anti-Candida antibodies in immunoprophylaxis.

The need for new options for the treatment of invasive candidiasis has fuelled the use of antibodies in combination with conventional antifungal therapy. After a long period of time in which antibodies were considered irrelevant in the resistance against invasive candidiasis, it was demonstrated that a number of antibodies or their engineered derivatives directed against Candida albicans cell-wall polysaccharides and glycopeptides, as well as against some protein epitopes, confer protection against invasive candidiasis. This has confirmed this approach as a new strategy for the prophylaxis of invasive candidiasis. Of particular interest is Mycograb, a human recombinant monoclonal antibody that inhibits heat shock protein 90, and has been administrated in combination with lipid-associated amphotericin B to patients with invasive candidiasis, and the fungicidal anti-beta-glucan antibodies induced by the glycoconjugate vaccine composed of a beta-glucan polysaccharide conjugated with the diphtheria toxoid CRM 197. However, despite the promising data obtained in vitro and in animal models, at present there is very little clinical experience on the use of antibodies in Candida immunoprophylaxis.

Human bone repair after mandibular symphysis block harvesting: a clinical and tomographic study.

BACKGROUND: There are limited data on the healing potential of osseous defects in the human mandible. Animal model studies have shown that defect fill is size dependent. METHODS: Twenty patients who had autogenous block transplants harvested from the mandibular symphysis were included in the study. Computerized tomography (CT) scans were carried out at an average of 26.7 months after augmentation to assess bone healing. Subgroups were compared on the basis of bone volume harvested, healing time, incision design, symphysis midline preservation, age, and gender. Percentage bone fill was calculated by comparing the preoperative and postoperative CT scans using a computer software program. RESULTS: CT scan analysis showed a significant percentage (74.5% +/- 10.36%) of bone fill at an average of 26.7 +/- 22.3 months (range, 4 to 72 months). Healing time and bone volume harvested were significant variables influencing the osteogenic potential of mandibular donor-site defects. Defects <0.5 cc with a healing period of 34.2 months showed 81% +/- 7.4% bone fill, whereas those >0.5 cc and 7.2 months of healing had a repair of 63.8% +/- 12.2% (P <0.05; 95% CI, 2.46 to 31.93). A positive trend in bone fill was observed for subgroups receiving sulcular incisions (80%) and midline preservation (77.5%). CONCLUSIONS: The osteogenic potential of human osseous repair in the mandibular symphysis is size and time dependent. The process of osteogenesis of repair in humans seems to be multifactorial. Such factors as preservation of the periosteum and symphysis cortical midline may positively influence defect fill allowing for reharvesting.