Bilateral simultaneous femoral neck fracture mimicking abdominal pain in a cerebral palsy patient.

Simultaneous bilateral femoral neck fractures are unusual lesions, generally associated with an underlying condition which causes impaired bone mineralization, triggered by an increased bone stress. We present a 24-year-old cerebral palsy patient, who was previously evaluated in another institution due to inability to walk, interpreted as abdominal pain. No alteration in blood analysis or abdominal X-rays was found. As no response to treatment was observed, a new abdominal X-ray was taken, which incidentally depicted bilateral medial femoral neck fracture. He was referred to our practice after a resection arthroplasty was offered in another institution. After admission, bilateral one-stage THA was performed. Several reports emphasize bone disease as a major precipitating factor, and there is an increased incidence of hip fractures in chronic epilepsy, renal osteodystrophy, and chronic steroid use. Femoral head resection has been proven to be effective in immobilized patients, whereas this was not a reasonable option in this patient who presented walking ability. Despite the treatment election, primary care physicians should be aware of and alert to the possibility of fractures in patients with neurological disorders and calcium metabolism alterations. Late diagnosis of orthopedic injuries in this type of patients may lead to permanent disability.

PCR-RFLP genotyping protocol for chicken Mx gene G/A polymorphism associated with the S631N mutation.

The Mx (myxovirus resistance) gene codes for a protein with antiviral activity. Non-synonymous G/A polymorphism at position 2032 of chicken Mx cDNA results in a change at amino acid 631 of the Mx protein. This mutation has been shown to affect the antiviral activity of the Mx molecule, although recent studies have not confirmed this effect in response to some influenza strains. Nevertheless, the G/A polymorphism could be important for the chicken's response to other viruses. A robust PCR-RFLP protocol for genotyping chicken Mx gene polymorphism associated with the S631N mutation was developed. The F primer anneals to the last intron of the Mx gene, and the R primer anneals to the last exon of the gene, with an expected PCR product of 299 bp. PCR products were digested with Hpy8I. This enzyme cuts the sequence 5'-GTN|NAC-3', 2 bp downstream of the Mx polymorphism for the G allele, whereas the fragment containing the A allele is not cleaved. One hundred and twenty-seven chickens (commercial broilers, White Leghorn and New Hampshire) were genotyped using this protocol, and genotyping data were validated by sequencing. Full identity of results between the two genotyping methods was observed for all 127 samples, proving the reliability and robustness of this PCR-RFLP protocol.

Effect of consanguinity on Argentinean Angus beef DNA traceability.

Since the 1990s several authors have envisaged the use of DNA to certify meat origin. Two major parameters must be assessed before a DNA based traceability protocol can be implemented in the food chain: (i) the information content of a DNA marker set in a specific livestock breed or group of breeds; (ii) the minimum number of DNA markers needed to obtain a statistically acceptable match probability. The objective of the present work was to establish the effect of different levels of inbreeding in the matching efficiency, and the minimum number of microsatellite markers needed, in a DNA based meat traceability program, starting from an 11-microsatellite marker panel. Samples were obtained from beef production farms in South America, where animals are typically bred under pasture-based extensive conditions. Three groups of animals with different consanguinity rates were sampled. Exclusion power (Q) was higher than 0.999998 and match probability lower than 3.01E-08, for the whole set of markers within each group. Both values were affected by consanguinity. To reach a two mismatch criteria exclusion power (Q(2)) of 99.99, six markers were needed in unrelated animals whereas seven markers were needed in related animals. To reach Q(2)=99.9999, 8 and 10 microsatellite markers, respectively, were needed. In general, one or two more microsatellite markers were needed to identify consanguineous animals. This study proved the DNA marker set used to be suitable for the identification of the meat from all slaughtered animals in Argentina, per week, month, and year.

Pig KALRN, MYH1, MLC2V, SNX13, AK1, and PPIA loci RH mapping and chromosome position refining.

The suppressive subtractive hybridization technique was previously used by the authors to identify candidate genes for meat quality in pig. A set of ESTs homologous (>95%) to genes involved in muscle metabolism is reported in the present paper. Four ESTs homologous to MYH1, KALRN, MLC2V, and SNX13 genes plus two genes (AK1, PPIA) used as housekeeping for muscle tissue were assigned to porcine chromosomes using the INRA-Minnesota 7000 rads radiation hybrid panel (IMpRH). Our data confirm and refine the cytogenetic position of the KALRN, AK1, PPIA genes, improve the existing physical map of MYH1 and assign two new genes (MLC2V and SNX13) to swine chromosomes.

Avian TAP genes: detection of nucleotide polymorphisms and comparative analysis across species.

TAP1 and TAP2 genes code for the two subunits of the transporter associated with antigen processing (TAP), and in chicken they are located between the two MHC class I genes. Using primers based on chicken sequences, the genomic regions corresponding to chicken TAP1 exons 6 to 7 and TAP2 exons 4 to 6 (which encode portions of the chicken TAP1 and TAP2 molecules corresponding to the human peptide-binding regions) were amplified and sequenced from chicken (70 birds), turkey (24), pheasant (6), and guinea fowl (7). A total of 80 within-species single nucleotide polymorphisms (SNPs) were identified. None of the chicken SNPs detected here was present in public databases. The SNP frequencies in chicken were 9.57 SNP/kb in TAP1 and 19.16 SNP/kb in TAP2, while turkey showed similar SNP frequencies in the two genes. Putative amino acid sequences were inferred to identify non-synonymous substitutions. The alignment of the consensus polypeptide sequences showed that most of the amino acid variations were conserved or semi-conserved substitutions. In conclusion, a high variability in the level of nucleotide polymorphism was observed within the two genes, with chicken showing the highest polymorphism rate in both genes. Most of the SNPs identified were within introns, and a general conservation of both amino acid numbers and characteristics of residues among and within the species was found. These data underline the functional importance of these molecules, but also suggest their capacity to bind different antigenic peptides.

Reconstruction of large defects postmandibulectomy for oral cancer using plates and myocutaneous flaps: a long-term follow-up.

A series of 28 consecutive cases of mandibular reconstruction by means of reconstructive plates and myocutaneous flaps were reviewed. In all cases mandibular resection was indicated for treatment of squamous cell carcinoma of the oral cavity: 25 pelviglossomandibulectomies (resulting in large defects from the angle of the mandible), 2 Commando operations (resulting in lateral defects) and 1 anterior sectional mandibulectomy (resulting in an anterior defect). Tumour stages were T1-T2 (4 cases) and T3-T4 (24 cases). Success was defined as plate maintenance 6 months' postoperatively/postradiotherapy. The overall success rate was 32.2%. Lateral-centre-lateral (L-C-L) defects had 32% success, L (lateral) defects had 50% success and in the single case of a C (centre) defect, the plate was not maintained. Stainless steel reconstruction plates showed a similar success rate as titanium plates (30% versus 34%). In cases not submitted to radiotherapy there were more maintained plates than in cases that received radiotherapy (45.5% versus 23.6%). Reconstruction plates are not effective in bridging large defects of the resected mandible. Only in selected cases that are not eligible for microvascular free flaps should plates and myocutaneous flaps be considered as an option for mandibular reconstruction.

Temporomandibular joint ankylosis: report of 14 cases.

The authors present a review of 14 patients with temporomandibular joint ankylosis treated between March 1992 and February 1997. Etiology of the ankylosis was trauma in four patients, ear infection in two, systemic infection in one case, congenital in another, and unknown in six. Patients were divided into two groups, according to their age: 16 years and under and over 16 years of age. The basic principle of surgical treatment in both groups is ample access for osseous resection and coronoidectomy. Costochondral grafts were used in group one (nine patients), while interposition of a silicone block, was performed in the second group (five patients). Follow-up evaluations were from twelve to 53 months (average 28.2 months). One case of recurrence occurred in the first group and no recurrences in the second group. The average long-term mouth opening in both groups was 32.8 mm.

A biolistic process for in vitro gene transfer into chicken embryos

Chicken embryos kept in culture medium were bombarded using a high helium gas pressure biolistic device. To optimize the factors that affect transformation efficiency, the lacZ gene under control of the human cytomegalovirus immediate early enhancer/promoter was used as a reporter gene. There was an inverse relationship between survival rate and transformation efficiency. The best conditions obtained for high embryo survival and high transformation efficiency were achieved with 800 psi helium gas pressure, 500 mmHg vacuum, gold particles, an 8 cm DNA-coated microparticle flying distance to the embryo and embryo placement 0.5 cm from the center of the particle dispersion cone. Under these conditions, transformation efficiency was 100 percent, survival rate 25 percent and the number of expression units in the embryo body cells ranged from 100 to 1,000. Expression of green fluorescent protein was also detected in embryos bombarded under optimal conditions. Based on the results obtained, the biolistic process can be considered an efficient method for the transformation of chicken embryos and therefore can be used as a model system to study transient gene expression and tissue-specific promoters

A biolistic process for in vitro gene transfer into chicken embryos.

Chicken embryos kept in culture medium were bombarded using a high helium gas pressure biolistic device. To optimize the factors that affect transformation efficiency, the lacZ gene under control of the human cytomegalovirus immediate early enhancer/promoter was used as a reporter gene. There was an inverse relationship between survival rate and transformation efficiency. The best conditions obtained for high embryo survival and high transformation efficiency were achieved with 800 psi helium gas pressure, 500 mmHg vacuum, gold particles, an 8 cm DNA-coated microparticle flying distance to the embryo and embryo placement 0.5 cm from the center of the particle dispersion cone. Under these conditions, transformation efficiency was 100%, survival rate 25% and the number of expression units in the embryo body cells ranged from 100 to 1,000. Expression of green fluorescent protein was also detected in embryos bombarded under optimal conditions. Based on the results obtained, the biolistic process can be considered an efficient method for the transformation of chicken embryos and therefore can be used as a model system to study transient gene expression and tissue-specific promoters.