Pre-treatment quality of life as a predictor of distant metastasis-free survival and overall survival in patients with head and neck cancer who underwent free flap reconstruction.

PURPOSE: This study examined the prognostic associations of pre-treatment quality of life (QoL) with overall survival (OS) and distant metastasis-free survival (DFMS) among patients with head and neck cancer (HNC) who underwent free flap reconstruction. METHODS: A cohort of 127 HNC patients who received free flap reconstruction between November 2010 and June 2014 at a hospital were recruited. Pre-treatment QoL was measured by the University of Washington Quality of Life Questionnaire, which contains six physical domains, including speech, swallowing, appearance, saliva, taste and chewing, as well as the six social-emotional domains of pain, activity, recreation, shoulder, mood, and anxiety. Cox regression analyses were performed. RESULTS: Results showed that pre-treatment QoL was predictive of OS and DMFS. Of the domains, swallowing, chewing, speech, taste, saliva, pain and shoulder were demonstrated to be significant predictors of OS. Additionally, swallowing, chewing, speech, pain and activity were demonstrated making significant contributions to DMFS. CONCLUSION: Our data supported that physical domains of pre-treatment QoL were predictors for OS and DFMS in HNC patients with free-flap reconstruction. Longitudinal studies are warranted to clarify the prognostic abilities of social-emotional domains. Information on pre-treatment QoL should be taken into account to individualize care plan for these patients, and hence prolong their survival.

Mononeuropathy multiplex and chylothorax as earlier manifestations of pulmonary tuberculosis.

Mononeuropathy multiplex (MNM) and chylothorax are rare clinical disorders. The concurrence of these two disorders with Mycobacterium tuberculosis infection has not been reported. We herein report a patient who was initially diagnosed with fever of unknown origin and MNM, and then developed chylothorax. Pulmonary tuberculosis was proved 1 month after chylothorax appeared. With low-dose prednisolone 15 mg day(-1) and anti-tuberculosis drugs, all these disorders completely resolved 1 year later.

Protein binding and antimicrobial activity of ceftriaxone: comparative assessments by gradient plate technique and time-kill study.

In conjunction with initial bactericidal rate measurements, this study evaluated the impact of protein binding on the antimicrobial activity of ceftriaxone employing the gradient plate technique. Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 were employed as test organisms. At various albumin concentrations (0 to < or = 16% w/v), the effects of albumin binding were estimated by the shift in concentration-responses (via initial bactericidal rate) and the distance of inhibition zone (DIZ) on gradient agar plates. Supplementation with 4% albumin reduced the antimicrobial activity of ceftriaxone against E. coli as suggested by the similar 1.5-fold shift in both initial bactericidal rate and DIZ when compared to those with no albumin. As with S. aureus, the inclusion of albumin increased the initial bactericidal rate but the DIZ assessments showed a reduction in antimicrobial activity. The reason for the difference observed on the initial bactericidal effect between the two species is unknown. However, the similar patterns of change in the DIZ data at increasing albumin concentrations from 0 to 4% for both organisms suggest consistency and reproducibility of the gradient plate technique. This technique appears to be an attractive alternative for the assessment of the effect of protein binding on antibiotics.

Clinical characteristics of Group G streptococcal bacteremia in Taiwan.

The results of this retrospective study showed that Group G streptococcal bacteremia was an acute febrile disease with low mortality. Most patients were > 60 y old but there was a strong association between age < 60 y and malignancy. Cases clustered in summer and the most common port of entry was the skin.

Mice with a targeted disruption of the Fanconi anemia homolog Fanca.

Fanconi anemia (FA) is a hereditary chromosomal instability syndrome with cancer predisposition. Bone marrow failure resulting in pancytopenia is the main cause of death of FA patients. Diagnosis of FA is based on their cellular hypersensitivity to DNA crosslinking agents and chromosome breakages. Somatic complementation experiments suggest the involvement of at least eight genes in FA. The gene for complementation group A (FANCA) is defective in the majority of FA patients. We show here that mice deficient of FANCA: are viable and have no detectable developmental abnormalities. The hematological parameters showed a slightly decreased platelet count and a slightly increased erythrocyte mean cell volume in mice at young age, but this did not progress to anemia. Consistent with the clinical phenotype of FA patients, both male and female mice showed hypogonadism and impaired fertility. Furthermore, embryonic fibroblasts of the knock-out mice exhibited spontaneous chromosomal instability and were hyper-responsive to the clastogenic effect of the crosslinker mitomycin C.

Cloning and characterization of murine fanconi anemia group A gene: Fanca protein is expressed in lymphoid tissues, testis, and ovary.

Fanconi anemia (FA) is an autosomal recessive disorder in humans characterized by bone marrow failure, cancer predisposition, and cellular hypersensitivity to cross-linking agents such as mitomycin C and diepoxybutane. FA genes display a caretaker function essential for maintenance of genomic integrity. We have cloned the murine homolog of FANCA, the gene mutated in the major FA complementation group (FA-A). The full-length mouse Fanca cDNA consists of 4503 bp and encodes a protein with a predicted molecular weight of 161 kDa. The deduced Fanca mouse protein shares 81% amino acid sequence similarity and 66% identity with the human protein. The nuclear localization signal and partial leucine zipper consensus motifs found in the human FANCA protein were also present in the murine homolog. In spite of the species difference, the murine Fanca cDNA was capable of correcting the cross-linker sensitive phenotype of human FA-A cells, suggesting functional conservation. Based on Northern as well as Western blots, Fanca was mainly expressed in lymphoid tissues, testis, and ovary. This expression pattern correlates with some of the clinical symptoms observed in FA patients. The availability of the murine Fanca cDNA now allows the gene to be studied in experimental mouse models.

Unusual pulmonary manifestations of disseminated Penicillium marneffei infection in three AIDS patients.

Penicillium marneffei is a rare fungal pathogen which can cause human infections in people predominantly living in South-east Asia and the southern portion of China. We report three cases of systemic P. marneffei infection in patients infected with HIV who lived in or had travelled to endemic areas. The clinical manifestation includes high fever, chills, weight loss, general malaise, chronic cough, haemoptysis, multiple skin lesions, abnormal liver function, etc. Chest X-ray showed single or multiple cavitary lesions with smooth or irregular thin wall. P. marneffei is cultured from blood, sputum, skin biopsy, sono-guide aspiration and bronchoscopic biopsy. After antifungal therapy with intravenous amphotericin B or oral fluconazole, skin lesions resolved completely within 2 weeks and cavitary lesions in the lungs changed to chronic fibrotic and interstitial processes after several months to a few years later. Our two cases had been treated as either pulmonary tuberculosis or suspected malignancy. A definite diagnosis and early treatment are important because this fungal infection is a marker of AIDS in South-east Asia.

Expression cloning of a cDNA for the major Fanconi anaemia gene, FAA.

Fanconi anaemia (FA) is an autosomal recessive disorder characterized by a diversity of clinical symptoms including skeletal abnormalities, progressive bone marrow failure and a marked predisposition to cancer. FA cells exhibit chromosomal instability and hyper-responsiveness to the clastogenic and cytotoxic effects of bifunctional alkylating (cross-linking) agents, such as diepoxybutane (DEB) and mitomycin C (MMC). Five complementation groups (A-E) have been distinguished on the basis of somatic cell hybridization experiments, with group FA-A accounting for over 65% of the cases analysed. A cDNA for the group C gene (FAC) was reported and localized to chromosome 9q22.3 (ref.8). Genetic map positions were recently reported for two more FA genes, FAA (16q24.3) and FAD (3p22-26). Here we report the isolation of a cDNA representing the FAA gene, following an expression cloning method similar to the one used to clone the FAC gene. The 5.5-kb cDNA has an open reading frame of 4,368 nucleotides. In contrast to the 63-kD cytosolic protein encoded by the FAC gene, the predicted FAA protein (M(r) 162, 752) contains two overlapping bipartite nuclear localization signals and a partial leucine zipper consensus, which are suggestive of a nuclear localization.

Lack of class I HLA expression in neuroblastoma is associated with high N-myc expression and hypomethylation due to loss of the MEMO-1 locus.

Class I HLA expression is low in neuroblastoma tumours and cell lines. We have recently mapped a modifier of methylation for HLA-C (MEMO-1) to chromosomal bands 1p35-36.1, a region deleted in many neuroblastomas. Hypomethylation of HLA-C is strongly correlated with allelic loss of the MEMO-1 locus. Here, we show that loss of MEMO-1 is associated with hypomethylation of both the 5' and 3' regions of class I HLA loci. We next investigated the relationship between methylation and expression of class I HLA in 28 cell lines of neuroectodermal tumours. Cell lines with hypermethylated HLA-C and HLA-A loci have relatively high expression, while most cell lines with hypomethylated loci have no or a reduced expression. It was reported earlier that high expression of c- or N-myc can suppress class I HLA expression. Remarkably, also N-myc amplification in neuroblastomas is associated with allelic loss of 1p35-36. Therefore, we have analysed the relationships between allelic loss of the MEMO-1 locus, class I HLA methylation and expression, and N-myc amplification and expression. This study shows a tight inter-relationship between these phenomena. Our data suggest a model in which hypomethylation of class I HLA due to loss of the MEMO-1 locus and high N-myc expression could collaborate in the down-regulation of class I HLA expression.

No mutations found by RET mutation scanning in sporadic and hereditary neuroblastoma.

Neuroblastoma occasionally occurs in diseases associated with abnormal neurocrest differentiation, e.g. Hirschsprung disease. Expression studies in developing mice suggest that the proto-oncogene RET plays a role in neurocrest differentiation. In humans expression of RET is limited to certain tumor types, including neuroblastoma, that derive from migrating neural crest cells. Mutations of RET are found associated with Hirschsprung disease. These data prompted us to investigate expression of RET and to search for gene mutations in neuroblastoma. Out of 16 neuroblastoma cell lines analyzed, 9 show clear expression of RET in a Northern blot analysis. In a single strandt conformation polymorphism (SSCP) analysis of all exons, no mutations were detected other than neutral polymorphisms. In a patient with neuroblastoma, from a family in which different neurocrestopathies, including neuroblastoma and Hirschsprung disease, had occurred, we also failed to detect RET mutations. Possibly, expression of RET in neuroblastoma merely reflects the differentiation status of the tumor cells. The absence of mutations suggests that RET does not play a crucial role in the tumorigenesis of neuroblastoma.

A human modifier of methylation for class I HLA genes (MEMO-1) maps to chromosomal bands 1p35-36.1.

Class I HLA genes are expressed in almost all tissues, but expression is low or undetectable in many neuroblastomas. We analysed class I HLA methylation in normal tissues and in 28 neuroectodermal tumour cell lines. HLA-C is hypermethylated in normal adult tissues and 13 cell lines, while 15 cell lines show the hypomethylated phenotype. Hypomethylation of HLA-C strongly correlates with hemizygous deletion of a 9 cM interval on 1p35-36.1, suggesting that this region encodes a modifier of methylation for HLA-C. To test whether hypomethylation of class I HLA genes results from loss of a modifier gene, we fused a hypomethylating neuroblastoma cell line with a hypermethylating cell line. Methylation of class I HLA genes was induced in the hybrids. Furthermore, methylation of HLA-C, -E and -A genes, which are encoded in a 1.4 Mb region on 6p21, is correlated in most cell lines. Our results suggest that 1p35-36.1 encodes a modifier of methylation for class I HLA genes, that is deleted in many neuroblastomas.

Comparative Study of Teicoplanin vs Vancomycin for the Treatment of Methicillin-Resistant Staphylococcus aureus Bacteraemia.

Forty patients with methicillin-resistant Staphylococcus aureus (MRSA) bacteraemia were randomised to receive either teicoplanin or vancomycin therapy to compare the clinical efficacy and safety of these glycopeptides. Treatment was successful in 17 (85%) of 20 patients who were randomised to the teicoplanin group, with 6 cures and 11 improvements, and in 15 (75%) of 20 patients randomised to the vancomycin group, with 8 cures and 7 improvements (p = 0.69). Microbiologically, all MRSA pathogens isolated were susceptible to both glycopeptides by the disc diffusion test. The mean zone of inhibition for teicoplanin was 18 ± 2mm (range 16 to 20mm) and 20 ± 2mm (range 16 to 24mm) for vancomycin. The minimum inhibitory concentration required to inhibit the growth of 90% of organisms in culture (MIC90) for all MRSA isolates was 2.0 mg/L (range 0.5 to 4 mg/L) for teicoplanin and 2.0 mg/L (range 0.5 to 2 mg/L) for vancomycin. The microbiological eradication rate was 85% (17 of 20 isolates) for teicoplanin and 75% (15 of 20 isolates) for vancomycin. None of the failures were due to the emergence of resistant pathogens. Adverse reactions occurred in 19% of patients treated with teicoplanin and 60% of patients treated with vancomycin. There was no significant difference in the occurrence of skin rash (p = 0.60) or in elevation of aminotransferase (p = 0.18). However, nephrotoxicity was significantly greater in the vancomycin group than in the teicoplanin group (50 vs 9.5%, p < 0.05).In conclusion, the results of this study demonstrate that teicoplanin appears to be a valuable alternative to vancomycin because it is as efficacious as vancomycin, has fewer adverse reactions, and is conveniently administered.

Successful treatment of subcutaneous mycoses with fluconazole: a report of two cases.

Lymphocutaneous sporotrichosis and chromoblastomycosis are subcutaneous mycoses caused by traumatic implantation of the fungus into the skin. Medical treatments for chromoblastomycosis has been disappointing, while lymphocutaneous sporotrichosis usually responds well to iodides. Here we present a case of chromomycosis and a case of lymphocutaneous sporotrichosis. Both patients were treated successfully with fluconazole.

Identification of two distinct chromosome 12-derived amplification units in neuroblastoma cell line NGP.

The neuroblastoma cell line NGP contains two homogeneously staining regions (hsr). One of these hsrs contains MYCN sequences. Reverse painting experiments demonstrated that the second HSR consisted of two chromosome 12-derived amplification units, located at 12q14-15 and 12q24. Southern blot and fluorescence in situ hybridization (FISH) analysis showed amplification of genes located at 12q14-15: SAS, MDM2, and CDK4, GLI, CHOP, CDK2, and A2MR were found not to be amplified. FISH further demonstrated amplification of RSN, a gene located at 12q24. The finding of two distinct chromosome 12 amplification units in a neuroblastoma cell line NGP is reminiscent of recent findings in well-differentiated liposarcoma (WDLPS) and other sarcomas. The second amplification unit on chromosome 12 in NGP is located more distal (12q24) than the one observed in WDLPS (12q21). The mechanism and biologic significance of this amplification process in neuroblastoma and WDLPS remain to be elucidated.

Molecular cytogenetic analysis of 1;17 translocations in neuroblastoma.

Loss of chromosome 1 short arm material, resulting from terminal deletions or unbalanced translocations, is a frequent finding in advanced neuroblastoma. In translocations, often relatively small portions of a second chromosome are translocated to the chromosome 1 short arm. The chromosomal origin of this translocated material could often not be identified using banding analysis only. Recent studies, applying fluorescent in situ hybridisation, showed that in the majority of these translocations, chromosome 17 is involved. In this study, the nonrandom occurrence of unbalanced 1;17 translocations is further supported by their presence in 3/7 neuroblastoma cell lines. Analysis of the 1p breakpoints extends our earlier observation of breakpoint heterogeneity. A similar scattering of 17q breakpoints was observed. The 1p and 17q breakpoints of the constitutional 1;17 translocation did not coincide with any of the 1;17 translocation breakpoints found in neuroblastoma cell lines. Cell lines, not containing 1;17 translocations, contained other chromosome 17 rearrangements. As a result, extra copies of 17q are found in all cell lines, suggesting a role for genes on 17q in neuroblastoma development. The possible significance of 1;17 translocations in neuroblastoma is discussed.

Deletion mapping in neuroblastoma cell lines suggests two distinct tumor suppressor genes in the 1p35-36 region, only one of which is associated with N-myc amplification.

Neuroblastoma is characterized by deletions of the short arm of chromosome 1 (1p) and amplification of the N-myc oncogene. We have made somatic cell hybrids of two human neuroblastoma cell lines, one with and one without N-myc expression and amplification. The expression of the amplified N-myc gene is completely switched off in the hybrids. This suggests that N-myc expression results from loss of a repressor function. As N-myc amplification is associated with loss of heterozygosity (LOH) of 1p36, we analysed 1p deletions in 16 neuroblastoma cell lines. The seven cell lines without N-myc amplification have no deletions or relatively small deletions, with an SRO on 1p36.23-33. This suggests that a tumor suppressor gene maps in this region. All nine cell lines with N-myc amplification have larger deletions, with an SRO from 1p35-36.1 to the telomere. This suggests that a second tumor suppressor gene which is associated with N-myc amplification maps more proximally. Fine mapping of 1p36 deletions in the two cell lines of the fusion experiment suggests that the distal locus is not a repressor of N-myc expression, but the more proximal locus could be a candidate for this function.

1;17 translocations and other chromosome 17 rearrangements in human primary neuroblastoma tumors and cell lines.

We report on the finding of a t(1;17) in two primary neuroblastomas. Subsequent fluorescence in situ hybridization (FISH) analysis revealed the presence of 1;17 translocations in four out of nine neuroblastoma cell lines. The chromosome 1 short arm breakpoints were determined using region-specific probes. FISH screening also demonstrated or confirmed the presence of 11;17 translocations in three cell lines and other chromosome 17 rearrangements in those cell lines that did not carry a t(1;17) or t(11;17). Our data extend previous cytogenetic findings and suggest that, in addition to the known involvement of chromosome 1, one or more genes on chromosome 17 also play a role in neuroblastoma development.

Activation of the mas oncogene involves coupling to human alphoid sequences.

We have shown previously that mouse NIH3T3 cells transfected with DNA from a human ovarian carcinoma were rendered tumourigenic by an activated mas oncogene in four independent transfection experiments. In all cases the 5'-noncoding region was rearranged in comparison to the original ovarian tumour DNA. We now report that in all four transfectants the newly acquired sequences consist of human centromeric alpha satellite repeat DNA. In at least three transfectants the alphoid DNA originates from the centromere of chromosome three. Analysis of the sequences of the recombination site in one transfectant revealed that a homologous sequence of five base pairs (CAGCA) is present in both parental strands, and might thus have contributed to the recombinational event. To establish a conclusive role for alphoid DNA in the activation of mas, we performed a co-transfection experiment in NIH3T3 cells with cloned alphoid DNA and the mas coding sequence. We show that the transfectants expressing a transformed phenotype contain amplified mas linked to alphoid DNA. NIH3T3 cells transfected with plasmids that contained alphoid sequences cloned directly upstream of the mas coding sequence, and injected into nude mice, gave rise to tumours with amplified mas sequences (7/7). In six of these tumours the alphoid sequences were amplified as well. Our data suggest a novel mechanism of oncogene activation: recombination with normal alphoid repeat DNA resulting in amplification of the oncogene.