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The objective of this study was to investigate the potential application of cell-seeded biomaterials for revision arthroplasty and the reconstruction of major joints using the impaction grafting technique. Using morselized cancellous bone graft as a porous scaffold, MG63 cells were seeded on the scaffold and impacted into an acetabulum cup model using a mechanical device constructed from data obtained during impaction grafting by an orthopedic surgeon. Immediately after impaction, cells were trypsinized from the scaffold and processed for cell survival rates using the double-stranded DNA PicoGreen assay. Significant reductions in viable cells were observed between the fifth impact and both the first and second impacts (p < 0.01 and p < 0.05, respectively). Cell survival rate was 21.5% after five impacts. The biological performance of cell-seeded biomaterials may be enhanced by these surviving cells. Compared to allograft bone that is not osteogenic, a cell-seeded biomaterial might also be a suitable substitute for allograft bone for major joint reconstruction at revision arthroplasty., (Copyright 2006 Orthopaedic Research Society.)

Experiments were conducted to determine if Metarhizium anisopliae isolates which are capable of growth at cattle surface temperatures could produce pathogenicity to Boophilus microplus in laboratory and field studies. The diurnal temperature fluctuation on the surface of cattle was monitored. The temperature tolerance of M. anisopliae isolates (ARSEF3297 and IMI386697) was determined and their pathogenicity to B. microplus compared at a standard bioassay temperature (28 degrees C) and at a temperature similar to the cattle surface (31-35 degrees C). The effect of the two isolates on the B. microplus population on cattle under field conditions was determined. The temperature of the fore udder, rear udder, ribs and neck regions of the mixed Holstein cattle fluctuated between 30 and 35 degrees C, in a similar pattern to the prevailing environmental temperature. However, wider fluctuations were obtained on the ears (28-35 degrees C) and spine (30-41 degrees C). The colony radius of both isolates declined as temperature increased, however, the growth of IMI386697 was five times greater than ARSEF3297 at 34 degrees C. At 28 degrees C, the pathogenicity of both isolates to B. microplus was similar, however, at 31-35 degrees C, IMI386697 was more pathogenic than ARSEF3297. Both isolates reduced the B. microplus population on cattle in comparison to the control formulation. However, IMI386697 (8.5+/-0.64 ticks/animal) produced a greater reduction in tick numbers than ARSEF3297 (19.1+/-0.64 ticks/animal). M. anisopliae was re-isolated from 8.9% of the ticks collected from IMI386697 treated cattle as compared to 2.8% of ticks from ARSEF3297 treated cattle.

Studies were conducted to identify oil-based formulating agents (paraffinic oil, palm oil and emulsifiable adjuvant oils (EAOs)) for Metarhizium anisopliae that were superior to water with simple surfactants using a germination test and a bioassay against Boophilus microplus. Germination of conidia in all formulations, except 10% coconut EAO, produced more than 68% germination at 24 h and nearly 100% at 48 h. Coconut oil (average survival time (AST) = 4.6 +/- 0.28 days) and 10% liquid paraffin EAO (AST = 4.4 +/- 0.15 days) enhanced the pathogenicity of M. anisopliae to B. microplus relative to water (AST = 8.4 +/- 0.42 days). M. anisopliae in 10% liquid paraffin EAO was the most effective formulation having a moderately high germination after 24 h and a low AST as well as a high AST in the control. In the second experiment, germination of conidia in 2% liquid paraffin EAO and 2% Cropspray was higher than in 2% Codacide oil at 24 h, however, all treatments reached 100% germination after 48 h. The ASTs of the EAO based M. anisopliae formulations (Average AST = 6.4 +/- 0.54 days) were similar but lower that the ASTs of the controls (Average AST = 9.6 +/- 0.28 days).

Entomopathogenic fungal isolates of Arachnid origin were assessed for their ability to produce mortality and inhibit egg hatching in Boophilus microplus with the aim of selecting an isolate for development into a myco-acaricide for control of cattle ticks. The ability of the most promising isolate to target developmental stages of more than one tick species and the optimum concentration of fungal inoculum to be used for future studies were determined. Metarhizium anisopliae was the most pathogenic of the three fungal species tested on B. microplus, producing shorter average survival times (ASTs) for engorged adults (AST = 5.2 +/- 0.1 days) and larvae (AST = 9.3 +/- 0.4 days), and a longer average hatching times (AHT; AHT = 19.8 +/- 0.5 days) in comparison to Simplicillium lamellicola and Paecilomyces farinosus. In comparative studies on two tick species with similar life cycles, M. anisopliae produced a shorter AST in engorged adult B. microplus (AST = 8.8 +/- 0.3 days) than Rhipicephalus sanguineus (AST = 10.3 +/- 0.3 days). M. anisopliae was pathogenic to larvae of B. microplus (AST = 7.7 +/- 0.4 days), however, had no effect on larvae of R. sanguineus (AST = 14.6 +/- 0.3 days) as the AST of this treatment was similar to its untreated control (AST = 14.1 +/- 0.4 days). M. anisopliae lengthened the AHTs in both B. microplus (AHT = 16.4 +/- 0.3 days) and R. sanguineus (AHT = 16.7 +/- 0.3 days) in comparison to the controls. The ASTs of engorged adult B. microplus treated with M. anisopliae shortened as the concentration was increased from 1 x 10(7) to 5 x 10(8) conidia/ mL. A further increase in concentration, 1 x 10(9) conidia/mL (AST = 10.2 +/- 0.4 days) did not shorten or lengthen the AST in comparison to 5 x 10(8) conidia/mL (AST = 9.4 +/- 0.3 days).

Although the mechanisms of osteoinduction by bone morphogenic proteins (BMPs) are increasingly understood, the most appropriate time to administer BMPs exogenously is yet to be clarified. The purpose of this study was to investigate when BMP may be administered to a fracture arena to maximise the enhancement of healing. Forty mice with externally fixed left femoral fractures were randomised into four groups: Group I, the control group was given a placebo of 30 microl saline at day 0; Groups II, III and IV were given 30 microl saline plus 2.5 microg rhBMP-2, at post-operative days 0, 4 or 8, respectively. Sequential radiographs were taken at days 0, 8, 16. On day 22 the mice were sacrificed and both femora were harvested for biomechanical assessment in 3-point bending and histological evaluation. Radiographic analysis indicated that healing of fractures in Groups II and III was significantly greater (p < 0.05) than those in Groups I and IV, at both 16 and 22 days post-fracture. The highest median bone mineral content at the fracture site was evidenced in Group III and II. Furthermore, Group III also had the highest relative ultimate load values, followed by Groups II, IV and I. Greater percentage peak loads were observed between Group I and both Groups II and III (p < 0.05). Histological examination confirmed that at 22 days post-fracture, only fractures in Groups II and III had united with woven bone, and Groups I and IV still had considerable amounts of fibrous tissue and cartilage at the fracture gap. Data presented herein indicates that there is a time after fracture when rhBMP administration is most effective, and this may be at the time of surgery as well as in the early fracture healing phases.

Administration of phthalates is known to cause toxicity and liver cancer in rodents through the activation of peroxisome proliferator-activated receptors (PPARs), and the monoesters appear to be the active metabolites that function as ligands of PPARs. There is evidence that PPARs exhibit significant species differences in response to ligand activation. In this study, the activation of mouse and human PPARalpha, PPARbeta, and PPARgamma by a broad class of phthalate monoesters was investigated using a trans-activation assay, functional analysis of PPARalpha target gene expression, and a PPARgamma-mediated differentiation assay. These studies demonstrated a range in the ability of various phthalate monoesters to activate PPARalpha, with the mouse PPARalpha generally being activated at lower concentrations and exhibiting a greater response than human PPARalpha. Similarly, a range in the trans-activation of mouse PPARbeta by phthalate monoesters was also observed, but this effect was not found with human PPARbeta. A number of phthalate monoesters activated both mouse and human PPARgamma, with similar sensitivity being exhibited by both receptors. These studies show that the potency and efficacy of phthalate monoesters for the activation of PPARalpha and PPARgamma increase with increasing side-chain length. These studies also show that mouse PPARalpha and PPARbeta are generally activated at lower concentrations of phthalate monoesters than human PPARalpha and PPARbeta, and that both mouse and human PPARgamma exhibit similar sensitivity to phthalate monoesters. Lastly, there is a good relationship between the relative ability of phthalate monoesters to trans-activate PPARalpha and PPARgamma, and the relative induction of PPARalpha target gene mRNA and PPARgamma-mediated adipocyte differentiation, respectively.

The identification of tumor-suppressor genes in solid tumors by classical cancer genetics methods is difficult and slow. We combined nonsense-mediated RNA decay microarrays and array-based comparative genomic hybridization for the genome-wide identification of genes with biallelic inactivation involving nonsense mutations and loss of the wild-type allele. This approach enabled us to identify previously unknown mutations in the receptor tyrosine kinase gene EPHB2. The DU 145 prostate cancer cell line, originating from a brain metastasis, carries a truncating mutation of EPHB2 and a deletion of the remaining allele. Additional frameshift, splice site, missense and nonsense mutations are present in clinical prostate cancer samples. Transfection of DU 145 cells, which lack functional EphB2, with wild-type EPHB2 suppresses clonogenic growth. Taken together with studies indicating that EphB2 may have an essential role in cell migration and maintenance of normal tissue architecture, our findings suggest that mutational inactivation of EPHB2 may be important in the progression and metastasis of prostate cancer.

Here we report the identification of a novel transcript containing SNF2, PHD-finger, RING-finger, helicase, and linker histone domains mapping to the q24 band region of human chromosome 6. These domains are characteristic of several DNA repair proteins, transcription factors, and helicases. We have cloned both human and mouse homologs of this novel gene using interexon PCR and RACE technologies. The human cDNA, termed SHPRH, is 6018 bp and codes for a putative protein of 1683 amino acids. The mouse cDNA, termed Shprh, is 7225 bp and codes for a putative protein of 1616 amino acids. The deduced amino acid sequences of the two proteins share 86% identity. Both genes are expressed ubiquitously, with a transcript size of approximately 7.5 kb. Mapping of this gene to 6q24, a region reported to contain a tumor suppressor locus, prompted us to evaluate SHPRH by mutation analysis in tumor cell lines. We have identified one truncating and three missense mutations, thus suggesting SHPRH as a possible candidate for the tumor suppressor gene.

We report here the identification of a gene associated with the hyperparathyroidism-jaw tumor (HPT-JT) syndrome. A single locus associated with HPT-JT (HRPT2) was previously mapped to chromosomal region 1q25-q32. We refined this region to a critical interval of 12 cM by genotyping in 26 affected kindreds. Using a positional candidate approach, we identified thirteen different heterozygous, germline, inactivating mutations in a single gene in fourteen families with HPT-JT. The proposed role of HRPT2 as a tumor suppressor was supported by mutation screening in 48 parathyroid adenomas with cystic features, which identified three somatic inactivating mutations, all located in exon 1. None of these mutations were detected in normal controls, and all were predicted to cause deficient or impaired protein function. HRPT2 is a ubiquitously expressed, evolutionarily conserved gene encoding a predicted protein of 531 amino acids, for which we propose the name parafibromin. Our findings suggest that HRPT2 is a tumor-suppressor gene, the inactivation of which is directly involved in predisposition to HPT-JT and in development of some sporadic parathyroid tumors.

Although prostate cancer is the most common non-cutaneous malignancy diagnosed in men in the United States, little is known about inherited factors that influence its genetic predisposition. Here we report that germline mutations in the gene encoding 2'-5'-oligoadenylate(2-5A)-dependent RNase L (RNASEL) segregate in prostate cancer families that show linkage to the HPC1 (hereditary prostate cancer 1) region at 1q24-25 (ref. 9). We identified RNASEL by a positional cloning/candidate gene method, and show that a nonsense mutation and a mutation in an initiation codon of RNASEL segregate independently in two HPC1-linked families. Inactive RNASEL alleles are present at a low frequency in the general population. RNASEL regulates cell proliferation and apoptosis through the interferon-regulated 2-5A pathway and has been suggested to be a candidate tumor suppressor gene. We found that microdissected tumors with a germline mutation showed loss of heterozygosity and loss of RNase L protein, and that RNASEL activity was reduced in lymphoblasts from heterozyogous individuals compared with family members who were homozygous with respect to the wildtype allele. Thus, germline mutations in RNASEL may be of diagnostic value, and the 2-5A pathway might provide opportunities for developing therapies for those with prostate cancer.

Surface ordering in a homologous series of alkyl cyanobiphenyl (nCB) liquid crystals having a direct isotropic-smectic-A (I-A) transition was investigated using evanescent-wave ellipsometry. The liquid crystal was bounded by a solid substrate treated with a silane surfactant which induced homeotropic (perpendicular) ordering of the liquid crystal molecules in the smectic-A phase. In the isotropic phase, one of the liquid crystals (10CB) partially wet the interface with an orientationally ordered, homeotropically aligned layer. The ordered interfacial layer grew without layering transitions but remained finite in thickness as the bulk I-A transition was approached. The interfacial layer has significantly lower orientational order than is observed in the smectic phase, indicating the possibility that the surface region of 10CB may be in a surface-induced, nonspontaneous nematic phase. The other liquid crystals (11 and 12CB) showed no surface ordering behavior whatsoever. Models describing the ordered surface layer of 10CB are presented. The results can be interpreted as a sharp transition in the surface ordering behavior as the chain length of the liquid crystal is varied, at the I-A transition of a liquid crystal.

To investigate the relationship between HPC2/ELAC2 and prostate cancer risk, we performed the following analyses: (1) a linkage study of six markers in and around the HPC2/ELAC2 gene at 17p11 in 159 pedigrees with hereditary prostate cancer (HPC); (2) a mutation-screening analysis of all coding exons of the gene in 93 probands with HPC; (3) family-based and population-based association study of common HPC2/ELAC2 missense variants in 159 probands with HPC, 249 patients with sporadic prostate cancer, and 222 unaffected male control subjects. No evidence for linkage was found in the total sample, nor in any subset of pedigrees based on characteristics that included age at onset, number of affected members, male-to-male disease transmission, or race. Furthermore, only the two previously reported missense changes (Ser217Leu and Ala541Thr) were identified by mutational analysis of all HPC2/ELAC exons in 93 probands with HPC. In association analyses, family-based tests did not reveal excess transmission of the Leu217 and/or Thr541 alleles to affected offspring, and population-based tests failed to reveal any statistically significant difference in the allele frequencies of the two polymorphisms between patients with prostate cancer and control subjects. The results of this study lead us to reject the three alternative hypotheses of (1) a highly penetrant, major prostate cancer-susceptibility gene at 17p11, (2) the allelic variants Leu217 or Thr541 of HPC2/ELAC2 as high-penetrance mutations, and (3) the variants Leu217 or Thr541 as low-penetrance, risk-modifying alleles. However, we did observe a trend of higher Leu217 homozygous carrier rates in patients than in control subjects. Considering the impact of genetic heterogeneity, phenocopies, and incomplete penetrance on the linkage and association studies of prostate cancer and on the power to detect linkage and association in our study sample, our results cannot rule out the possibility of a highly penetrant prostate cancer gene at this locus that only segregates in a small number of pedigrees. Nor can we rule out a prostate cancer-modifier gene that confers a lower-than-reported risk. Additional larger studies are needed to more fully evaluate the role of this gene in prostate cancer risk.

This qualitative study examines residential child care workers' styles of engaging adolescent clients. Analysis focuses on staff practices and behavior that facilitate a treatment context conducive to healthier client attachments. Findings demonstrate the complexity inherent in residential treatment and illuminate barriers to optimal staff-client relationships. Suggestions are offered for improvement of agency policies and for future research.

Bipolar disorder is a severe mental illness characterized by mood swings of elation and depression. Family, twin, and adoption studies suggest a complex genetic etiology that may involve multiple susceptibility genes and an environmental component. To identify chromosomal loci contributing to vulnerability, we have conducted a genome-wide scan on approximately 396 individuals from 22 multiplex pedigrees by using 607 microsatellite markers. Multipoint nonparametric analysis detected the strongest evidence for linkage at 13q32 with a maximal logarithm of odds (lod) score of 3.5 (P = 0. 000028) under a phenotype model that included bipolar I, bipolar II with major depression, schizoaffective disorder, and recurrent unipolar disorder. Suggestive linkage was found on 1q31-q32 (lod = 2. 67; P = 0.00022) and 18p11.2 (lod = 2.32; P = 0.00054). Recent reports have linked schizophrenia to 13q32 and 18p11.2. Our genome scan identified other interesting regions, 7q31 (lod = 2.08; P = 0. 00099) and 22q11-q13 (lod = 2.1; P = 0.00094), and also confirmed reported linkages on 4p16, 12q23-q24, and 21q22. By comprehensive screening of the entire genome, we detected unreported loci for bipolar disorder, found support for proposed linkages, and gained evidence for the overlap of susceptibility regions for bipolar disorder and schizophrenia.

/ This paper describes the fundamental design features, and construction methods and sequence, of a rehabilitation project on a small suburban creek in Moscow, Idaho, USA. A meandering channel pattern was reestablished for approximately 280 m of straightened, dredged channel, a new floodplain was excavated, and the new riparian zone was replanted. The new stream channel was sized to accommodate an estimated natural bankfull discharge ( approximately 5.6 cms), and floodplain design attempted to match the conveyance of the old enlarged channel (14-20 cms). The project was coordinated by a local nonprofit environmental organization, and the design and construction were tailored to donated materials and a largely volunteer labor force. A high-magnitude flood event (ca. 50-year recurrence interval) six months after construction had no significant impact on the newly constructed channel and revetments, but underscored the need for important detailing of the structures. The use of volunteer labor, while entailing certain benefits, complicates project planning and construction. The most general lesson learned from this project is that sponsoring agencies and clients need to be informed of the many steps and sequencing of properly constructed, complex stream rehabilitation projects as well as the high time and cost requirements for these tasks. KEY WORDS: Stream corridor restoration; Channel design; Streambank revetments

Thirty-two children with femoral shaft fractures were treated conservatively with initial skin traction followed by an additional period in a spica cast. After 12 to 20 months of follow up, none had any pain and all of them were attending school without problems. Shortening of more than 2 cm occurred in 6 (19%) of the 32 patients. The most important factor associated with shortening was an overlap of more than 2 cm of shortening of the fracture ends at the time of cast fitting. The average compensatory overgrowth at final assessment was 7 mm. Angular deformity did not pose a problem. This is a safe, simple and practical method to treat childhood femoral shaft fractures.

An initial genome scan was performed on 540 individuals from 97 families segregating bipolar disorder, collected through the National Institutes of Mental Health Genetics Initiative. We report here affected-sib-pair (ASP) data on 126 marker loci (approximately 68,000 genotypes) mapping to chromosomes 4, 7, 9, 18, 19, 20, and 21q, under three affection status models. Modest increases in identical-by-descent (IBD) allele sharing were found at the following loci: D4S2397 and D4S391 (P < 0.05) on 4p, D4S1647 (P < 0.05) on 4q, D7S1802 and D7S1869 (low P = 0.01) on 7p, D9S302 (P = 0.004) on 9q, and D20S604 on 20p and D20S173 on 20q (P < 0.05). In addition, five markers on 7q displayed increased IBD sharing (P = 0.046-0.002). Additional ASP analyses on chromosomes 18 and 21q marker data were performed using disease phenotype models defined previously. On chromosome 18, only D18S40 on 18p and D18S70 on 18q yielded a slight elevation in allele sharing (P = 0.02), implying that the reported linkages in these regions were not confirmed. On chromosome 21q, a cluster of markers within an approximately 9 cM interval: D21S1254, D21S65, D21S1440, and D21S1255 exhibited excess allele sharing (P = 0.041-0.008). Multilocus data on overlapping marker quartets, from D21S1265 to D21S1255, which were consistent with increased IBD sharing (P < 0.01, with a low of 0.0009), overlapped a broad interval of excess allele sharing reported previously, increasing support for a susceptibility locus for bipolar disorder on 21q.

In 22 multiplex pedigrees screened for linkage to bipolar disorder, by use of 18 markers on chromosome 21q, single-locus affected-sib-pair (ASP) analysis detected a high proportion (57%-62%) of alleles shared identical by descent (IBD), with P values of .049-.0008 on nine marker loci. Multilocus ASP analyses revealed locus trios in the distal region between D21S270 and D21S171, with excess allele sharing (nominal P values <.01) under two affection-status models, ASM I (bipolars and schizoaffectives) and ASM II (ASM I plus recurrent unipolars). In addition, under ASM I, the proximal interval spanned by D21S1436 and D21S65 showed locus trios with excess allele sharing (nominal P values of .03-.0003). These findings support prior evidence that a susceptibility locus for bipolar disorder is on 21q.

A severe form of an extremely unusual injury, ipsilateral fracture dislocation of the radial shaft head associated with elbow dislocation, is described. Radial shaft fracture was irreducible even after the radial head and the elbow were reduced. Open reduction and internal fixation are viable options.

The compartment syndrome is a rare complication of surgery in the lithotomy position, but one that may result in neurovascular damage and permanent disability. Herein, we present a case of bilateral three-compartment syndrome following urethroplasty in the dorsolithotomy position, with review of the literature and discussion of the causes and possible means to prevent this complication.

Nineteen children with circulating anticoagulants or anticardiolipin antibodies and negative or only weakly positive antinuclear factors were reviewed for their clinical manifestations. The relationship between these antibodies and the morbidity in the pediatric age group will be discussed.

Child psychiatry is a relatively new medical profession in Israel. For many years adult psychiatrists treated children and adolescents without official recognition of the profession. The development of child psychiatry in the medical world, the determination of colleagues and support of outstanding child psychiatrists from abroad (Professors Solnit and Noshpitz) led to the acknowledgment and official recognition of child psychiatry by the Scientific Council of the Israel Medical Association in 1981. Almost all graduates in child and adolescent psychiatry in Israel fill responsible positions and prove the necessity for more child psychiatrists and child psychiatry in the medical and psychiatric system both in hospitals and in the community.

Seventy adolescent admissions to a psychiatric unit were evaluated with a Hebrew translation of the Schedule for Affective-Disorders and Schizophrenia for School-Aged Children (K-SADS-P). Interrater and test-retest reliability and mother-child agreement were evaluated for diagnoses, symptom clusters, and 20 affective symptoms. K-SADS diagnoses were also compared with 3-month unit evaluation diagnoses as a measure of consensual validity. The reliability of assessing psychosocial functioning was additionally examined. Reliability and validity of diagnoses were high and reliability of symptoms and syndromes was good to excellent. Mother-child agreement faired less well on all measures. The use of semistructured interviews and DSM-III criteria in an Israeli adolescent psychiatric setting are discussed.

We have analyzed 196 cases of pyloroduodenal obstruction due to peptic ulcer disease. Obstruction occurred in 19% of peptic ulcer patients. Males predominated five to one over females. Diagnosis could usually be made on clinical grounds alone. Air contrast x-ray studies were better than conventional barium meal x-rays and are recommended. Symptoms were relieved with conservative treatment alone in 17% of patients. We suggest that such therapy should take the form of preoperative preparation and that the period between diagnosis of obstruction and surgery should be kept to a minimum. Gastrojejunostomy for drainage was the procedure of choice. Postvagotomy gastric atony was very infrequent.