Candida parapsilosis Resistance to Fluconazole: Molecular Mechanisms and In Vivo Impact in Infected Galleria mellonella Larvae

Candida parapsilosis is the main non-albicans Candida species isolated from patients in Latin America. Mutations in the ERG11 gene and overexpression of membrane transporter proteins have been linked to fluconazole resistance. The aim of this study was to evaluate the molecular mechanisms in fluconazole-resistant strains of C. parapsilosis isolated from critically ill patients. The identities of the nine collected C. parapsilosis isolates at the species level were confirmed through molecular identification with a TaqMan qPCR assay. The clonal origin of the strains was checked by microsatellite typing. The Galleria mellonella infection model was used to confirm in vitro resistance. We assessed the presence of ERG11 mutations, as well as the expression of ERG11 and two additional genes that contribute to antifungal resistance (CDR1 and MDR1), by using real-time quantitative PCR. All of the C. parapsilosis (sensu stricto) isolates tested exhibited fluconazole MICs between 8 and 16 μg/ml. The in vitro data were confirmed by the failure of fluconazole in the treatment of G. mellonella infected with fluconazole-resistant strains of C. parapsilosis. Sequencing of the ERG11 gene revealed a common mutation leading to a Y132F amino acid substitution in all of the isolates, a finding consistent with their clonal origin. After fluconazole exposure, overexpression was noted for ERG11, CDR1, and MDR1 in 9/9, 9/9, and 2/9 strains, respectively. We demonstrated that a combination of molecular mechanisms, including the presence of point mutations in the ERG11 gene, overexpression of ERG11, and genes encoding efflux pumps, are involved in fluconazole resistance in C. parapsilosis.     

Severe Asthma in Adults : An Orphan Disease?

Severe asthma affects fewer than 10% of patients with asthma, is associated with a severe risk of death and disability, has a great impact on health and quality of life, and represents a huge cost to patients and society. Given the poor response to treatment and the side effects associated with medications for severe asthma, more efficient, cost-effective, and phenotype-specific medications are needed. Considering severe asthma as an orphan disease could encourage the pharmaceutical industry to stratify studies based on a more detailed characterization of study subjects at baseline, resulting in the development of novel therapeutic approaches.     

CRP evolution pattern in CPAP-treated obstructive sleep apnea patients. Does gender play a role?

Background??aim

C-reactive protein (CRP) is directly implicated in atherogenesis and associated cardiovascular morbidity in patients with obstructive sleep apnea (OSA). Effective continuous positive airway pressure (CPAP) treatment has been shown to gradually decrease CRP levels and thus consequently improve disease-related cardiovascular morbidity. However, the influence of gender on the CRP evolution pattern has never been assessed before. The aim of our study was to investigate possible gender differences in CRP evolution in OSA patients 3 and 6?months after the start of effective CPAP treatment.

Methods

The study population consisted of 436 patients (252 males/184 females) with newly diagnosed moderate to severe OSA and good CPAP compliance assessed by a thorough follow up. High-sensitivity C-reactive protein (hs-CRP) was assessed before CPAP initiation and at the third and sixth month of the follow-up period.

Results

C-reactive protein values showed a statistically significant decrease at the third and sixth month of CPAP therapy [initial values 0.79?±?0.65?mg/dL versus 0.70?±?0.52?mg/dL (p?p?p?p?>?0.05). After 6?months?? treatment, CRP decreased significantly in both genders (males from 0.74?±?0.53?mg/dL to 0.28?±?0.32?mg/dL, p?p?Conclusion Our results suggest a delay in the normalization of CRP levels in females despite effective CPAP treatment. A time period of at least 6?months appeared to be required in women in order to reduce CRP levels and consequent cardiovascular risk. In contrast, CPAP??s protective role in males is achieved at an earlier time point. Gender-related hormonal and genetic factors may influence the above CRP evolution pattern.     

Comparison of minimally invasive fusion and instrumentation versus open surgery for severe stenotic spondylolisthesis with high-grade facet joint osteoarthritis

Purpose

The object of this study was to compare minimally invasive surgery (MIS) with open surgery in a severely affected subgroup of degenerative spondylolisthetic patients with severe stenosis (SDS) and high-grade facet osteoarthritis (FJO).

Methods

From January 2009 to February 2010, 49 patients with severe SDS and high-grade FJO were treated using either MIS or open TLIF. Intraoperative and diagnostic data, including perioperative complications and length of hospital stay (LOS), were collected, using retrospective chart review. Surgical short- and long-term outcomes were assessed according to the Oswestry disability index (ODI) and visual analog scale (VAS) for back and leg pain.

Results

Comparing MIS and open surgery, the MIS group had lesser blood loss, significantly lesser need for transfusion (p = 0.02), more rapid improvement of postoperative back pain in the first 6 weeks of follow-up and a shorter LOS. On the other hand, we experienced in the MIS group a longer operative time. The distribution on the postoperative ODI (p = 0.841), VAS leg (p = 0.943) and back pain (p = 0.735) scores after a mean follow-up of 2 years were similar. The overall proportion of complications showed no significant difference between the groups (29 % in the MIS group vs. 28 % in the open group, p = 0.999).

Conclusion

Minimally invasive surgery for severe SDS leads to adequate and safe decompression of lumbar stenosis and results in a faster recovery of symptoms and disability in the early postoperative period.     

Screening performance of different methods defining fetal nasal bone hypoplasia as a single and combined marker for the detection of trisomy 21 in the second trimester

Objective: To evaluate different methods of defining fetal nasal bone hypoplasia in the second trimester for the detection of trisomy 21.

Methods: Prospective study in Greek women undergoing anomaly scan between 18?+?0 and 23?+?6 weeks. The following methods of defining nasal bone hypoplasia were evaluated, either as a single marker or in combination with others: (1) BPD to nasal bone length (NBL) ratio; (2) multiples of the median (MoM) of NBL, according to normal curves from a Greek population; (3–4) NBL?<?2.5 percentile according to normal curves (3) commonly used internationally curves and (4) curves from a Greek population.

Results: In total, 1301 singleton fetuses were evaluated???10 with trisomy 21. The best detection rate of trisomy 21 was achieved when the applied method was nasal bone percentiles adjusted to maternal ethnicity, in combination with other markers (<2.5 percentile according to normal curves from a Greek population; p?<?0.001; sensitivity 50%; specificity 94.8%; false-positive rate 5.2%; positive likelihood ratio 9.6).

Conclusion: Screening performance of fetal nasal bone hypoplasia in detecting trisomy 21 varies according to the method applied. The best screening performance is achieved by using percentiles adjusted to maternal ethnicity in combination with other markers of aneuploidy.     

Conventional radiographs: are they still the standard in localization of projectiles?

The penetration of air gun pellets in facial soft tissue can cause major problems during the removal of foreign bodies, although conventional radiography, computed tomography, image-guided surgical removal, and ultrasound have been applied to facilitate the procedure. It was the aim of the present case report to introduce a modified intraoperative method for the localization of air gun pellets, based on the use of radiopaque markers in conventional radiographs. A 66-year-old patient attempted to commit suicide by using an air gun. The pellet hit the right temporal region. A computed tomographic (CT) scan was acquired to localize the foreign body. The first attempt to remove the pellet through the penetrating wound failed. Because of a dislodgement of the pellet, the CT scan could no longer be used for the localization of the air gun pellet. As the air gun pellet was positioned under the zygomatic arch, ultrasound was unable to identify its position. Successful intraoperative localization of the projectile was performed after fixation of radiopaque markers to the skin in the region of the estimated localization, with conventional radiographs in 2 planes, acquired with a mobile dental x-ray device. Although the markers remained attached to the patient as reference makers, the air gun pellet was removed easily. The use of radiopaque markers in conventional radiographs in 2 planes allows fast, intraoperative localization of radiopaque foreign bodies within soft tissue. The procedure can be carried out with a conventional x-ray device that should be available in every oral and maxillofacial practice. The use of reference markers should be considered a standard procedure for the localization of radiopaque foreign bodies in the head and neck.