Evaluation of epoetin supplemented with oral iron in patients with solid malignancies and chronic anemia not receiving anticancer treatment

OBJECTIVE: To evaluate the effectiveness and improvement in quality of life (QOL) of epoetin alfa administration supplemented with oral iron as a therapeutic regimen for patients with solid malignancies and anemia of chronic disease (ACD), not receiving chemotherapy and/or radiotherapy. PATIENTS AND METHODS: A total of 100 patients with cancer-related anemia, not subjected to chemotherapy and/or radiotherapy, were randomized to receive for a maximum of 24 weeks either oral iron, equivalent to 200 mg elemental iron once daily, or epoetin alfa 40,000 IU subcutaneously once weekly plus oral iron once daily. RESULTS: Patients in the epoetin alfa group had, from baseline to study end, a mean increase in hemoglobin (Hb) levels of 2.4 g/dL, whereas in the control group the mean Hb level decreased by 0.1 g/dL, (p<0.001). Improvement in QOL as assessed by the LASA and the FACT-An questionnaire were greater in patients in the epoetin alfa group than in the control group (mean change, LASA-energy level: 30.4 mm vs. 0.4 mm, -daily activities: 31.7 mm vs. 0.4 mm, -overall well-being. 32.4 mm vs. 4.9, FACT-An: 43.3 vs. 13.4, respectively). As for ECOG score, patients in the epoetin alfa group had a mean improvement of 0.16 from baseline to study end (control group: 0.06). Improvement in QOL parameters and in ECOG scores correlated positively with increased hemoglobin levels. CONCLUSION: Our results suggest that weekly epoetin alfa therapy supplemented with daily oral iron increases Hb levels and improves QOL in patients with solid malignancies and ACD who are not receiving chemotherapy and/or radiotherapy. This regimen offers optimal therapy in this population taking into consideration physician's convenience and patient's compliance.     

A Golgi localization signal identified in the Menkes recombinant protein

Menkes disease arises from a genetic impairment in copper transport. The gene responsible for the phenotype has been identified as a copper transporting ATPase ( ATP7A ). Recently, the protein encoded by the ATP7A gene has been localized to the Golgi complex. In order to investigate the role of the Menkes disease protein in copper transport, recombinant constructs containing both the full-length open reading frame and an alternatively spliced form have been successfully expressed and localized in mammalian cells. Other studies of a patient with occipital horn syndrome, an allelic variant of Menkes disease, have demonstrated that only this alternatively spliced isoform and not the full-length form is expressed in this patient. The milder form of this patient's phenotype suggests that the alternatively spliced isoform has some functional role in copper transport. In the present study the full-length recombinant Menkes protein was shown by immunofluorescence to localize to the Golgi apparatus and the alternatively spliced form, lacking sequences for transmembrane domains 3 and 4 encoded by exon 10, was shown to localize to the endoplasmic reticulum. Using sequences from exon 10 fused to a non-Golgi reporter molecule, a 38 amino acid sequence containing transmembrane domain 3 of the Menkes protein was found to be sufficient for localization to the Golgi complex. Therefore, the protein sequence encoded by exon 10 may be responsible for this differential localization and both isoforms may be required for comprehensive transport of copper within the cell.      

HIP-I: a huntingtin interacting protein isolated by the yeast two- hybrid system

We report the discovery of the huntingtin interacting protein I (HIP-I) which binds specifically to the N-terminus of human huntingtin, both in the two-hybrid screen and in in vitro binding experiments. For the interaction in vivo, a protein region downstream of the polyglutamine stretch in huntingtin is essential. The HIP1 cDNA isolated by the two- hybrid screen encodes a 55 kDa fragment of a novel protein. Using an affinity-purified polyclonal antibody raised against recombinant HIP-I, a protein of 116 kDa was detected in brain extracts by Western blot analysis. The predicted amino acid sequence of the HIP-I fragment exhibits significant similarity to cytoskeleton proteins, suggesting that HIP-I and huntingtin play a functional role in the cell filament networks. The HIP1 gene is ubiquitously expressed in different brain regions at low level. HIP-I is enriched in human brain but can also be detected in other human tissues as well as in mouse brain. HIP-I and huntingtin behave almost identically during subcellular fractionation and both proteins are enriched in the membrane containing fractions.      

Combining fetal sonography with genetic and allele pathogenicity studies to secure a neonatal diagnosis of Bardet–Biedl syndrome

Bardet–Biedl syndrome (BBS) is a rare pediatric ciliopathy characterized by marked clinical variability and extensive genetic heterogeneity. Typical diagnosis of BBS is secured at a median of 9 years of age, and sometimes well into adolescence. Here, we report a patient in whom prenatal detection of increased nuchal fold, enlarged echogenic kidneys, and polydactyly prompted us to screen the most commonly mutated genes in BBS and the phenotypically and genetically overlapping ciliopathy, Meckel–Gruber syndrome (MKS). We identified the common Met390Arg mutation in BBS1 in compound heterozygosity with a novel intronic variant of unknown significance (VUS). Testing of mRNA harvested from primary foreskin fibroblasts obtained shortly after birth revealed the VUS to induce a cryptic splice site, which in turn led to a premature termination and mRNA degradation. To our knowledge, this is the earliest diagnosis of BBS in the absence of other affected individuals in the family, and exemplifies how combining clinical assessment with genetic and timely assays of variant pathogenicity can inform clinical diagnosis and assist with patient management in the prenatal and neonatal setting.     

Fenticonazole Activity Measured by the Methods of the European Committee on Antimicrobial Susceptibility Testing and CLSI against 260 Candida Vulvovaginitis Isolates from Two European Regions and Annotations on the Prevalent Genotypes

The activity of fenticonazole was studied against 260 West and Southeast European vulvovaginal candidiasis isolates, and low MICs were displayed. Fenticonazole was assessed by European Committee on Antimicrobial Susceptibility Testing and CLSI microdilution methods for the first time, and the results showed excellent agreement (97%) and significant interclass correlation coefficient (P < 0.0001). Also, the levels of agreement for the results for itraconazole, fluconazole, and ketoconazole were 84%, 90%, and 98% (P < 0.0001), respectively. Multilocus typing by PCR fingerprinting and subsequent cluster analysis delineated geographically associated alignments for Candida albicans and fluconazole resistance-related clusters for Candida glabrata.Uncomplicated vulvovaginal candidiasis (VVC) affects approximately 75% of women at reproductive age (13, 17, 22); Candida albicans is a major cause and Candida glabrata accounts for approximately 5% of cases worldwide (30). The recommended first-line therapy for uncomplicated VVC is topical azoles (4, 7, 25, 27, 28), unless resistance of the isolate is substantiated or azole hypersensitivity is diagnosed (4, 8). Identifying antifungal resistance in vitro is clinically important, but variable host responses to treatment and unpredictable fungal load in the vulvovaginal mucosa (in loco) invariably weaken in vitro with in vivo correlations. However, standardized susceptibility testing of isolates to local antifungals could provide data on the in vitro activity of newer topical antifungals.Recording agreement of the results of the CLSI (24) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) (10) reference methods in determining the susceptibility of VVC isolates from Belgian and Greek patients to fenticonazole, a topical imidazole (8, 12), forms the basis of this report. Subsequently, PCR fingerprinting was used to investigate whether distinct geographical and azole-resistant clinical isolate subpopulations can be recognized.A total of 260 baseline C. albicans and C. glabrata isolates from pregnant, nonpregnant, and diabetic women were tested (Table ​(Table1).1). Isolates were identified in Chromagar medium (Chromagar, Paris, France) and identified with the API ID 32 C system (bioMerieux, Marcy l''Etoile, France). All C. albicans isolates were screened for Candida dubliniensis (3, 18, 31) and C. glabrata strains were screened for Candida nivariensis and Candida bracarensis (1, 19) to ensure that susceptibility testing and PCR fingerprints corresponded only to C. albicans and C. glabrata isolates.

TABLE 1.

Origin of isolates from 260 patients with uncomplicated vulvovaginitis