Practical experience in a surgical unit when changing from scrub to rub.

Concurring with the move to a new facility at the end of 2008, alcohol-based hand rub (AHR) dispensers were allocated in all anterooms of operating theatres in a tertiary care hospital in Madrid, Spain. Routine educational sessions on surgical hand disinfection (SHD) were performed by infection control personnel, emphasizing the preferred method of AHR. Our aim was to evaluate the knowledge about SHD, as well as the performance of SHD and perceptions concerning the use of AHR or antiseptic soap hand scrub (ASHS) by surgeons and operating room nurses (ORNs). An anonymous survey on SHD was circulated to the surgeons, resident surgeons and nurses who perform SHD in our hospital. In all, 70 surveys were completed, corresponding to 27 ORNs (38.6%), 20 resident surgeons (29%), and 23 staff surgeons (33%), 41 (59%) of them being females. Forty percent of them perform mostly disinfectant soap hand scrub (DSHS) and 19% mostly hand rubs; 41% perform both methods. AHR rub is believed to be more efficacious than DSHS in reducing hand contamination (mean 6.5 vs 5.6; P > 0.001). Almost half of the personnel (49%) perform AHR >50% of the time (AHR users). AHR use is associated with ORNs (odds ratio: 3.4; 95% confidence interval [CI] 1.2-9.3; P = 0.018) but not with resident surgeons (odds ratio: 4.1; 95% CI 1.3-13.2; P = 0.017). When comparing both methods of SHD, personnel favour AHR for its colour (P > 0.01) and speed of drying (P > 0.01). Compared with surgeons, ORNs scored DSHS as more skin-irritating (4.1 vs 5.2; P = 0.013), more associated with skin dryness (3.2 vs 4.7; P > 0.001), and inferior overall score (4.7 vs 5.6; P > 0.005). Most of the personnel agree that AHR (85%) improves compliance with SHD. Nevertheless ASHS is the method preferred for surgeons (63%) whereas AHR is the method preferred by nurses (70%; P< 0.001). According to auto-evaluation of hand skin after use, AHR has better outcomes in terms of lack of dryness (5.3 vs 4.6; P > 0.05). In addition, nurses scored ASHS lower than did surgeons in terms of skin redness (4.7 vs 6.0; P > 0.001), lack of skin integrity (4.8 vs 5.9; P = 0.004), skin dryness (3.6 vs 5.4; P > 0.001), and overall score (4.0 vs 5.2; P > 0.002). Change of perceptions and behaviours about SHD can be accomplished, and the AHR method is better adhered to by nursing personnel.

Shank vessel injuries: the forgotten vascular injuries.

INTRODUCTION: Vascular injuries in branch vessels of the popliteal artery, such as the tibioperoneal trunk, and shank vessels, such as anterior, posterior tibial, and peroneal vessels, occur in both blunt and penetrating trauma. Their management has evolved significantly in the past few decades. While their incidence is variable, limb loss and morbidity remain significant. MATERIAL AND METHODS: Physical examination, along with measuring an Ankle-Brachial Index (ABI), is still sometimes all that is required for diagnosis and can expeditiously triage those that require urgent operation. Despite our technological advancements and newer algorithms for lower extremity vascular trauma, operative intervention and exposure still remain difficult and pose a great challenge for surgeons that normally do not operate on this area. CONCLUSIONS: Shank vessel injuries still comprise a significant proportion of combat and civilian vascular injuries, and modern advances have led to a dramatic decrease in amputation rates.

Expression analysis and chromosomal assignment of PRA1 and RILP genes.

PRA1 (prenylated Rab acceptor) is a general regulator of Rab proteins, while RILP (Rab interacting lysosomal protein) is a specific effector for Rab7. It has been shown that PRA1 interacts with Rab proteins and with VAMP2. Therefore PRA1 is probably an important factor for membrane traffic, linking together the function of Rab proteins and SNAREs. RILP has a key role in the control of transport to degradative compartments together with Rab7 and probably links Rab7 function to the cytoskeleton. Here we have studied by Northern blot the expression of the two genes in several different human tissues. The 0.8-kb mRNA for human PRA1 is ubiquitously expressed, while the two mRNAs for RILP are differentially expressed. In addition, we have assigned the human PRA1 gene to chromosome 19q13.13-q13.2 and the human RILP gene to chromosome 17p13.3.

The spectrum of inherited mutations causing HPRT deficiency: 75 new cases and a review of 196 previously reported cases.

In humans, mutations in the gene encoding the purine salvage enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) are associated with a spectrum of disease that ranges from hyperuricemia alone to hyperuricemia with profound neurological and behavioral dysfunction. Previous attempts to correlate different types or locations of mutations with different elements of the disease phenotype have been limited by the relatively small numbers of available cases. The current article describes the molecular genetic basis for 75 new cases of HPRT deficiency, reviews 196 previously reported cases, and summarizes four main conclusions that may be derived from the entire database of 271 mutations. First, the mutations associated with human disease appear dispersed throughout the hprt gene, with some sites appearing to represent relative mutational hot spots. Second, genotype-phenotype correlations provide no indication that specific disease features associate with specific mutation locations. Third, cases with less severe clinical manifestations typically have mutations that are predicted to permit some degree of residual enzyme function. Fourth, the nature of the mutation provides only a rough guide for predicting phenotypic severity. Though mutation analysis does not provide precise information for predicting disease severity, it continues to provide a valuable tool for genetic counseling in terms of confirmation of diagnoses, for identifying potential carriers, and for prenatal diagnosis.

An unexpected affected female patient in a classical Lesch-Nyhan family.

Lesch-Nyhan disease is a genetic disorder of purine metabolism caused by defective activity of the enzyme hypoxanthine-guanine phosphoribosyl transferase (HPRT), resulting from mutation in the corresponding gene on the long arm of the X chromosome (Xq26). The classical phenotype, which includes spasticity, involuntary movements, developmental disability, and self-injurious behavior, occurs exclusively in males, while heterozygous, carrier females are clinically normal. We analyzed an Argentine family in which there were male and female siblings with clinically identical classic features of Lesch-Nyhan disease. The mother and an older daughter were carriers and had normal phenotypes. We identified the HPRT mutation in the family. It is a C --> T transition at position 508 of the cDNA (c.508 C --> T) that changes the CGA codon for Arg(169) to the TGA stop codon (R169X). The female patient was karyotypically normal and heterozygous for the mutation. She inherited the HPRT mutation from her mother, but she also had unexpected nonrandom inactivation of the paternal X chromosome carrying the normal HPRT gene. This additional genetic alteration is the cause of the clinical expression of disease in this female patient.

Predisposition to lung tumorigenesis.

Mouse inbred strains with inherited predisposition and resistance to lung cancer provide an essential tool for the dissection of the genetics of this complex disease. We have previously mapped a major locus (Pulmonary adenoma susceptibility 1, Pas1) affecting inherited predisposition to lung cancer in mice on chromosome 6, near Kras2. Appropriate crosses that include susceptible mice (Pas1(s)) provide a model system for identifying loci that can modify the lung cancer predisposition phenotype caused by Pas1. Using this approach we have mapped the Pulmonary adenoma resistance 1 (Par1) locus that behaves like a modifier locus of Pas1. More recently, we mapped additional lung tumor resistance loci (Par2, and Par4), and a locus specifically involved with lung tumor progression (Papg1). The mapping of Pas1 in mice stimulated us to test the possible association of genetic markers located in the homologous human region (12p12) with risk and prognosis of lung adenocarcinomas in man. In the Italian population, we carried out an association study by genotyping lung adenocarcinoma patients and healthy controls for genetic markers located in the putative region of interest. Homozygosity of the A2 allele at a Kras2/RsaI polymorphism, and allele 2 at a VNTR polymorphism in the PTHLH gene showed borderline statistically significant associations with lung cancer risk. Furthermore, the same alleles were significantly associated with tumor prognosis. Studies on association were then performed in the Japanese and in European populations. In the Japanese population, the KRAS2/RsaI marker was significantly associated with prognosis of lung adenocarcinoma, whereas the European study did not confirm this association. Our results may provide evidence for the existence of the human PAS1 locus, suggesting that the mouse model of inherited predisposition to lung tumorigenesis is predictive of a human genetic mechanism of susceptibility to lung cancer.

Linkage disequilibrium and physical mapping of Pas1 in mice.

By using linkage disequilibrium (LD) analysis in 21 strains of known susceptibility to lung cancer and by assembling a YAC contig, we mapped to a approximately 1.5-Mb region on distal mouse chromosome 6 the Pas1 locus, the major determinant of lung cancer predisposition in mice. Our results, on the basis of haplotype and phenetic analysis, suggest that the Pas1(s) susceptibility allele is shared by several mouse-inbred strains of independent origin, which show either high or intermediate predisposition to lung tumorigenesis. Therefore, the Pas1(s) allele is probably derived from an ancestral mouse rather than from independent mutations of the same gene. We showed the feasibility of LD in common inbred strains for the fine mapping of disease loci, and provided the biological basis and the reagents for the cloning of the Pas1 gene.

Isolation and characterization of the murine zinc finger coding gene, ZT2: expression in normal and transformed myogenic cells.

In the context of a project aimed at the identification of zinc finger proteins involved in skeletal muscle histogenesis and differentiation, we isolated a murine gene, named ZT2. The 2.44kb partial cDNA clone corresponds to the 3' region of the gene, and contains a 0.54kb open reading frame encoding four C2H2-like zinc finger domains, organized in tandem. This cDNA hybridizes with multiple transcripts (2, 4.5 and 7kb), whose expression levels vary in different tissues and at different developmental stages in the same tissue. At least in skeletal muscle we observed differences in the polyadenylation state of the transcripts at different stages of development. Moreover, ZT2 expression is correlated with cell proliferation and transformation. Sequence analysis and genetic mapping indicate that ZT2 is the homologue of ZNF125, one of the linked zinc finger encoding genes localized on human Chr 11q23. In humans, a high frequency of tumor-associated translocations is found in this chromosome region. As expected, ZT2 maps to the corresponding region on chromosome 9 in the mouse.

Prognostic value of loss of heterozygosity and KRAS2 mutations in lung adenocarcinoma.

The prognostic values of loss of heterozygosity (LOH) at loci frequently involved in non-small cell lung cancer and of KRAS2 gene mutations were studied in lung adenocarcinoma patients. LOHs were relatively common, ranging from 24% at chromosome 10p to 55% at chromosome 17p13. KRAS2 mutations at codon 12 were present in 26% of cases, were more common in male than in female patients and were associated with a smoking habit. No association between LOH at any site and clinical stage or survival rate was found. On the other hand, we observed a trend between the presence of any type of KRAS2 mutations and poor survival. Analysis of specific KRAS2 mutations showed a strong effect of Asp (GAT) and Val (GTT) mutations, resulting in a very poor survival compared with wild type group, whereas the most common mutation (Cys, TGT) was not associated with prognosis. Our results indicate the importance of specific activating mutations of the KRAS2 gene as genetic markers of clinical outcome for patients with lung adenocarcinoma.

Genetic mapping of the mouse Rab7 gene and pseudogene and of the human RAB7 homolog.

Rab proteins are small GTP-ases localized to distinct membrane compartments in eukaryotic cells and regulating specific steps of intracellular vesicular membrane traffic. The Rab7 protein is localized to the late endosomal compartment and controls late steps of endocytosis. We have isolated, by library screening, the 5' region, including the promoter, of the mouse Rab7 gene and a Rab7 pseudogene. We have mapped, by genetic linkage analysis, the mouse Rab7 gene on Chromosome (Chr) 6 and the Rab7-ps1 pseudogene on Chr 9, where the Rab7 gene has been previously reported to map. By radiation hybrid mapping, we have located the human RAB7 gene on Chr 3, in a region homologous to the mouse Chr 6, where the Rab7 gene maps.

Analysis of the retinoic acid receptor alpha gene as a candidate for the pulmonary adenoma resistance 1 gene.

The retinoic acid receptor alpha (Rara) gene, which maps in the same region as the pulmonary adenoma resistance (Par1) locus on mouse chromosome 11 (Manenti G et al., Nature Genet 12:455-457, 1996), was tested as a candidate gene for Par1. We report here the analysis of loss of heterozygosity, nucleotide sequence comparison, gene expression, and biochemical activity of the Rara gene from the Mus spretus(Par1/+) and A/J (Par1/-) mouse strains. The two Rara alleles were distinguished by two amino-acid variations but had similar biochemical activity and expression levels, leading to the exclusion of Rara as a candidate Par1 gene.

The gene encoding the transcriptional repressor BERF-1 maps to a region of conserved synteny on mouse chromosome 16 and human chromosome 3 and a related pseudogene maps to mouse chromosome 8.

We have recently identified and characterized a Kruppel-like zinc finger protein (BERF-1), that functions as a repressor of beta enolase gene transcription. By interspecific backcross analysis the gene encoding BERF-1 was localized 4.7 cM proximal to the Mtv6 locus on mouse chromosome 16, and an isolated pseudogene was localized to mouse chromosome 8, about 5.3 cM distal to the D8Mit4 marker. Nucleotide sequence identity and chomosome location indicate that the gene encoding BERF-1 is the mouse homologue (Zfp148) of ZNF148 localized to human chromosome 3q21, a common translocation site in acute myeloid leukemia patients.

Genetic mapping of cancer susceptibility/resistance loci in the mouse.

Genetic linkage experiments using crosses between mouse inbred strains with an inherited predisposition and resistance to lung cancer make it possible to investigate the genetics of the complex inheritance of susceptibility and resistance to lung cancer. We have previously mapped a major locus (pulmonary adenoma susceptibility 1, Pas1) affecting inherited predisposition to lung cancer in mice onto chromosome 6, near Kras2. Appropriate crosses that include Pas1/+ mice provide a model system for identifying loci that can modify the lung cancer predisposition phenotype caused by Pas1. Using this approach, we mapped the pulmonary adenoma resistance 1 (Par1) locus on to mouse chromosome 11; this locus selectively inhibits lung tumor development in Pas1/+ animals and therefore behaves like a modulator gene of Pas1. More recently, we have mapped lung tumor modifier loci specifically affecting the initiation and progression of lung cancer. Thus experimental models provide an essential tool for the mapping of lung cancer susceptibility/resistance genes and for the subsequent cloning of candidate genes.

Association of chromosome 12p genetic polymorphisms with lung adenocarcinoma risk and prognosis.

The mapping near Kras2 of pulmonary adenoma susceptibility 1 (Pas1), a major locus affecting inherited predisposition to lung cancer in mice prompted us to test the homologous human region (12p12) for association with lung adenocarcinoma, by a population-based study. We genotyped 213 lung adenocarcinoma patients and 219 healthy blood donor subjects for five polymorphic markers mapping in the putative region of interest. Three marker polymorphisms, located in a region spanning approximately 700 kb, were significantly associated with lung adenocarcinoma risk. Furthermore, polymorphisms in KRAS2 and PTHLH loci were also associated with tumor prognosis. These results suggest the existence of a human Pas1 homologous locus on chromosome 12p12.

Mapping of the mouse Tdgf1 gene and Tdgf pseudogenes.

Teratocarcinoma-derived growth factor-1 (Tdgf1), a member of the "EGF family" of growth factors, is expressed during mouse gastrulation in the forming mesoderm and later in the truncus arteriosus of the developing heart. In humans, TDGF1 is highly expressed in germ cell tumors and in colon and mammary carcinomas. In mouse, one gene (Tdgf1) and two pseudogenes (Tdgf1-ps1 and Tdgf1-ps2) have been isolated and characterized. Tdgf1 corresponds to the gene expressed in F9 teratocarcinoma cells. Tdgf1-ps1 and Tdgf1-ps2 are two intronless sequences with all the characteristics of retroposons. In the present paper, we assign the chromosomal location for Tdgf1, Tdgf1-ps1, and Tdgf1-ps2 sequences to Chromosomes (Chrs) 9, 16, and 17, respectively. Two previously described mouse mutants, scant hair (sch) and fur deficient (fd), map near the Tdgf1 gene. Analysis of their DNA coding region provided no evidence that Tdgf1 could be the responsible gene for these phenotypes. Finally, analysis of the DNA from several Mus musculus strains and from Mus spretus mice revealed a highly variable restriction pattern and the absence of the Tdgf1-ps1 genomic sequence from the Mus spretus genome.

Association between cigarette smoking and FHIT gene alterations in lung cancer.

Epidemiologic data have strongly indicated that cigarette smoking is linked to the development of lung cancer. However, little is known of the molecular targets of carcinogens contained in tobacco smoke. To identify genetic lesions characteristic of tobacco damage, we undertook a molecular analysis of microsatellite alterations within the FHIT gene and FRA3B, as well as at an independent locus on chromosome 10, D10S197, in lung tumors from heavy smokers and in tumors from never smokers. Loss of heterozygosity affecting at least one locus of the FHIT gene was observed in 41 of 51 tumors in the smokers group (80%) but in only 9 of 40 tumors in nonsmokers (22%). The comparison between the frequency of losses in FHIT in smokers and nonsmokers was statistically significant (P = 0.0001), whereas no difference in loss of heterozygosity rate was observed at D10S197 locus. These findings suggest that FHIT is a candidate molecular target of carcinogens contained in tobacco smoke.

Identification of a novel human kinesin-related gene (HK2) by the cDNA differential display technique.

We have used the cDNA differential display technique to isolate genes regulated by the synthetic retinoid N-(4-hydroxyphenyl)-all-trans-retinamide (HPR), a cancer chemopreventive agent in vivo and a powerful inducer of apoptotic cell death in vitro. Here we report the identification of a novel gene, the expression of which is markedly up-regulated in tumor cells after treatment for 30-60 min with HPR. The full-length cDNA of this gene, determined by screening of a human placenta cDNA, is 3.5 kb long and contains an open reading frame of 2037 nt. The gene is > 90% homologous to the mouse KIF2, a gene belonging to the family of kinesin-related motor proteins, and we therefore named it HK2 (human kinesin 2). A shorter form of the HK2 mRNA (HK2s), containing a 57-nt deletion in the open reading frame, has also been detected. Northern analysis revealed that HK2 is widely expressed among hemopoietic and nonhemopoietic cell lines and tissues. By the use of radiation hybrids, HK2 has been localized to chromosome 5q12-q13. Kinesins constitute a superfamily of motor proteins that use energy liberated from ATP hydrolysis to move cargo along microtubules and are implicated in mechanisms of mitosis or meiosis. The role of HK2 in the growth-inhibitory and apoptotic responses elicited by HPR remains to be established.