Evaluation of the usage and influence of groin ultrasound in primary and secondary healthcare settings.

AIM: Groin ultrasound scan (USS) is used to aid the diagnosis of inguinal hernias. Our radiology department offers access to image symptomatic patients for general practitioners (GPs) as well as surgeons. We examined the utilisation of groin USS in primary and secondary healthcare settings, and investigated its influence on proceeding to surgery. METHODS: A retrospective data analysis was performed for patients seen in the surgical outpatient clinics (January 2010-January 2011). Clinical, radiological, and surgical findings were compared. RESULTS: 267 USS were performed by musculoskeletal radiology specialists; patients were referred for USS by GPs in 98 cases (36.7 %), compared to 169 cases (63.3 %) where the referral for USS was organised by surgeons. Clinical examination by surgeons detected inguinal hernias in 105 groins (39.3 %), and USS detected inguinal hernias in 154 groins (57.7 %). Of 162/267 (60.7 %) cases where clinical examination was negative, 98/162 (60.4 %) also had a negative USS; only five of these patients (5.1 %) underwent surgery. In the 64/162 (39.6 %) cases where only the USS findings were positive, 19/64 underwent surgery (29.7 %). When hernia was detected on both USS and clinical examination (n = 90), 68/90 underwent surgery (75.6 %). For patients who underwent surgery, sensitivity for hernia detection was 80 % for clinical examination versus 96.3 % for USS. CONCLUSION: Groin USS is highly sensitive, and patients could be referred for USS by GPs when clinical examination findings are equivocal or negative. If USS is also negative, patients may be managed conservatively in primary care setting if they remain asymptomatic. Positive clinical examination findings appear to have a greater influence in the decision to treat surgically.

A new mechanism of injury in ankylosing spondylitis: non-traumatic hyperextension causing atlantoaxial subluxation.

Ankylosing spondylitis (AS) is a progressive multisystem chronic inflammatory disorder. The hallmark of this pathological process is a progressive fusion of the zygapophyseal joints and disc spaces of the axial skeleton, leading to a rigid kyphotic deformity and positive sagittal balance. The ankylosed spine is unable to accommodate normal mechanical forces, rendering it brittle and susceptible to injury. Traumatic hyperextension injury of the cervical spine leading to atlantoaxial subluxation (AAS) in AS patients can often be fatal. We report a non-traumatic mechanism of injury in AS progressing to AAS attributable to persistent hyperextension, which resulted in fatal migration of C2 through the foramen magnum.

Optimization of Fermentation Medium for the Production of Glucose Isomerase Using Streptomyces sp. SB-P1.

The combination of medium ingredients has a profound influence on the metabolic pathways running in the microorganism which regulates the production of numerous metabolites. Glucose isomerase (GI), an enzyme with huge potential in the market, can isomerise glucose into fructose. GI is used widely for the production of High-Fructose Corn Syrup (HFCS). HFCS is used as a sweetener in food and pharmaceutical industries. Streptomyces are well-known producers of numerous enzymes including glucose isomerase. An array of 75 isolates was screened for the production of glucose isomerase. The isolate Streptomyces sp. SB-P1 was found to produce maximum amount of extracellular GI. Sucrose and raffinose among pure carbon sources and corn cob and wheat husk among crude agro residues were found to yield high enzyme titers. Potassium nitrate among pure nitrogen sources and soy residues among crude sources gave maximum production. Quantitative effect of carbon, nitrogen, and inducer on GI was also determined. Plackett-Burman design was used to study the effect of different medium ingredients. Sucrose and xylose as carbon sources and peptone and soy residues as nitrogen sources proved to be beneficial for GI production.

Janus kinase 2 regulates Bcr-Abl signaling in chronic myeloid leukemia.

Despite the success of imatinib mesylate (IM) in the early chronic phase of chronic myeloid leukemia (CML), patients are resistant to IM and other kinase inhibitors in the later stages of CML. Our findings indicate that inhibition of Janus kinase 2 (Jak2) in Bcr-Abl+ cells overcomes IM resistance although the precise mechanism of Jak2 action is unknown. Knocking down Jak2 in Bcr-Abl+ cells reduced levels of the Bcr-Abl protein and also the phosphorylation of Tyr177 of Bcr-Abl, and Jak2 overexpression rescued these knockdown effects. Treatment of Bcr-Abl+ cells with Jak2 inhibitors for 4-6 h but not with IM also reduced Bcr-Abl protein and pTyr177 levels. In vitro kinase experiments performed with recombinant Jak2 showed that Jak2 readily phosphorylated Tyr177 of Bcr-Abl (a Jak2 consensus site, YvnV) whereas c-Abl did not. Importantly, Jak2 inhibition decreased pTyr177 Bcr-Abl in immune complexes but did not reduce levels of Bcr-Abl, suggesting that the reduction of Bcr-Abl by Jak2 inhibition is a separate event from phosphorylation of Tyr177. Jak2 inhibition by chemical inhibitors (TG101209/WP1193) and Jak2 knockdown diminished the activation of Ras, PI-3 kinase pathways and reduced levels of pTyrSTAT5. These findings suggest that Bcr-Abl stability and oncogenic signaling in CML cells are under the control of Jak2.

Inhibition of Grb2 expression demonstrates an important role in BCR-ABL-mediated MAPK activation and transformation of primary human hematopoietic cells.

Chronic myeloid leukemia (CML) results from the expression of the BCR/ABL oncogene in a primitive hematopoietic cell. However, BCR/ABL-activated signaling mechanisms are dependent on the cellular context in which it is expressed, and mechanisms underlying primitive human hematopoietic cell transformation by BCR-ABL are not well understood. Our previous studies have shown that BCR/ABL-Y177 has an essential role in Ras activation and human hematopoietic progenitor transformation in CML. The adapter protein growth factor receptor-binding protein-2 (Grb2) can bind phosphorylated BCR/ABL-Y177, induce Grb2-SoS complex formation and activate Ras signaling. We investigated the role of Grb2 in CML progenitor transformation by cotransducing human CD34+ cells with lentivirus vectors expressing short hairpin RNA to Grb2 and retrovirus vectors expressing BCR/ABL. We show that Grb2 knockdown significantly inhibits proliferation and survival of BCR-ABL-expressing CD34+ cells, but not control CD34+ cells. Grb2 knockdown reduced mitogen-activated protein kinase (MAPK) activity in BCR-ABL-expressing hematopoietic cells. We conclude that inhibition of Grb2 expression demonstrates an important role in BCR-ABL-mediated MAPK activation and transformation of primary human hematopoietic cells.These results support further investigation of downstream effectors of Grb2-mediated signals and targeting of Grb2 interactions in the treatment of CML.

Transcorporeal tunnel approach for unilateral cervical radiculopathy: a 2-year follow-up review and results.

BACKGROUND: The long-term complications of traditional discectomy and fusion surgery have led to the need for minimally invasive procedures that do not require a complete discectomy and fusion. Jho developed an anterior unco-foraminotomy that we have modified, with the approach being more medial than that of Jho, into an anterior transcorporeal tunnel approach which we use for cervical spondylotic unilateral radiculopathy. METHODS: A retrospective analysis was carried out in 30 patients who underwent a transcorporeal "tunnel" anterior micro-foraminotomy for unilateral radicular symptoms with a follow-up more than 2 years. All were operated by a single surgeon using the same technique from the vertebral body proximal to the lesion and proceeding downwards to the herniation. At final follow-up we reviewed the clinical and radiological results. RESULTS: All patients in the immediate post-operative period showed relief of their symptoms, and there were major complications. 3 patients complained about the numbness in the immediate postoperative period which resolved within 3 months. There was a significant improvement in NDI from pre-operative 55.16% to postoperative 5.82% ( P <0.001). Average pre-operative VAS scores for arm and neck were 8.15 and 4.05, respectively; which improved to 1.05 and 1.23 ( P <0.001) postoperatively. There was an average 9% decrease (from 7.8 mm to 7.3 mm) in the post-operative disc height compared to the preoperative disc height; however, it was clinically and radiologically insignificant. The long-term results were favourable and there were no major complications. CONCLUSION: The transcorporeal tunnel approach can be used as an alternative treatment for cervical spondylotic radiculopathy.

Percutaneous endoscopic lumbar herniectomy for high-grade down-migrated L4-L5 disc through an L5-S1 interlaminar approach: a technical note.

BACKGROUND: L4-L5 disc herniations can be treated with percutaneous endoscopic lumbar discectomy (PELD) using a transforaminal posterolateral approach. Although PELD has some distinct advantages over conventional open discectomy, inadequate decompression is a major cause of failure of the procedure, especially with high-grade migrations. The objective of this technical note is to present a new surgical approach for treating high-grade, down-migrated, L4-L5 disc herniations through an L5-S1 interlaminar endoscopic approach. METHOD: This technical report presents 4 consecutive patients with high-grade, down-migrated, L4-L5 disc herniations, who were treated with PELD through an L5-S1 interlaminar approach under local anesthesia and conscious sedation. All patients were evaluated clinically using both the visual analogue scale (VAS) for back and leg pain and the Oswestry disability index (ODI) and radiologically using MR imaging postoperatively. RESULTS: All 4 patients experienced improvement in their preoperative symptoms and signs immediately postoperatively. The mean VAS scores for back and leg pain improved from 3.75 to 1.75 and from 8.5 to 0.75, respectively. The mean ODI score improved from 65% to 3%. Postoperative MR imaging also depicted L5 root decompression. There were no complications during the procedure. CONCLUSION: This technical note presents a new technique for treating high-grade, down-migrated, L4-L5 disc herniations with PELD using an L5-S1 interlaminar approach.

The prevalence and radiological findings in 1347 elderly patients with scoliosis.

In order to determine the epidemiology of adult scoliosis in the elderly and to analyse the radiological parameters and symptoms related to adult scoliosis, we carried out a prospective cross-sectional radiological study on 1347 adult volunteers. There were 615 men and 732 women with a mean age of 73.3 years (60 to 94), and a mean Cobb angle of 7.55 degrees (sd 5.95). In our study, 478 subjects met the definition of scoliosis (Cobb angle > or = 10 degrees ) showing a prevalence of 35.5%. There was a significant difference in the epidemiological distribution and prevalence between the age and gender groups. The older adults showed a larger prevalence and more severe scoliosis, more prominent in women (p = 0.004). Women were more affected by adult scoliosis and showed more linear correlation with age (p < 0.001). Symptoms were more severe in those with scoliosis than in the normal group, but were similar between the mild, moderate and severe scoliosis groups (p = 0.224) and between men and women (p = 0.231). Adult scoliosis showed a significant relationship with lateral listhesis, vertebral rotation, lumbar hypolordosis, sagittal imbalance and a high level of the L4-5 disc (p < 0.0001, p < 0.0001, p = 0.002, p = 0.002, p < 0.0001 respectively). Lateral listhesis, lumbar hypolordosis and sagittal imbalance were related to symptoms (p < 0.0001, p = 0.001, p < 0.0001 respectively).

Decolorization of water soluble azo dyes by bacterial cultures, isolated from dye house effluent.

The aim of this work is to isolate and characterize bacterial isolates form dye house effluent, and to check their ability of decolorizing sulfonated azo dyes, and also to study influence of various environmental parameters on same process. Among seven Gram positive bacterial isolates obtained form dye house effluent, M1 (Bacillus cereus) and M6 were proved to be more potent for decolorizing sulfonated azo dyes under aerobic conditions. Maltose as carbon source and peptone as nitrogen source enhanced decolorization efficiency of M1 (B. cereus). HPTLC studies proved that more than 97% of the dye (Reactive Red 195) was degraded by bacteria after 72 h of incubation. These results along with spectrophotometric data prove the efficiency of bacteria suggesting their possible use in treating dye containing effluents.

Femoral lengthening in achondroplasia: magnitude of lengthening in relation to patterns of callus, stiffness of adjacent joints and fracture.

Extensive limb lengthening may be indicated in achondroplastic patients who wish to achieve a height within the normal range for their population. However, increasing the magnitude of lengthening is associated with further complications particularly adjacent joint stiffness and fractures. We studied the relationship between the magnitude of femoral lengthening and callus pattern, adjacent joint stiffness and fracture of the regenerate bone in 40 femoral lengthenings in 20 achondroplastic patients. They were divided into two groups; group A had lengthening of less than 50% and group B of more than 50% of their initial femoral length. The patterns of radiological callus formation were classified according to shape, type and features. The incidence of callus features, knee stiffness and regenerate bone fracture were analysed in the two groups. Group B was associated with an increased incidence of concave, lateral and central callus shapes, adjacent joint and stiffness and fracture. Statistically, the incidence of stiffness in adjacent joints and regenerate bone fracture was significantly associated with the magnitude of lengthening. We suggest that careful radiological assessment of the patterns of callus formation is a useful method for the evaluation and monitoring of regenerate bone.

Grade 4 spondylolisthesis of the L5 vertebra associated with dural ectasia in neurofibromatosis.

Spondylolisthesis associated with neurofibromatosis is rare, and only 12 cases have been reported so far. However, only one report of grade 4 spondylolisthesis with neurofibromatosis has been reported in the literature. A 15-year-old boy with neurofibromatosis was admitted for back pain and neurological claudication. Radiograph showed grade 4 spondylolisthesis of the L5 vertebra with scalloping of the L4-L5 vertebrae. L4-L5 laminectomy, reduction, L3-S1 posterior instrumentation and fusion were performed. The reduction of the spondylisthesis was done entirely from the posterior approach using pedicle screws. Radiography at four months showed a broken S1 screw with a loss of reduction. The patient was re-operated on, to provide additional stability with pelvic fixation. He was pain-free with a good fusion at the two-year follow-up. Adequate posterior stabilisation with fusion gives good results in grade 4 spondylolisthesis associated with neurofibromatosis and dural ectasia.

Enhanced BCR-ABL kinase inhibition does not result in increased inhibition of downstream signaling pathways or increased growth suppression in CML progenitors.

The therapeutic success of imatinib in chronic myeloid leukemia (CML) is hampered by persistence of malignant stem cells. We investigated whether nilotinib, a more potent BCR-ABL kinase inhibitor could target CML primitive progenitors more effectively than imatinib. CML and normal progenitor cells were cultured with nilotinib or imatinib in growth factor supplemented medium. Nilotinib inhibited BCR-ABL kinase activity at lower concentrations than imatinib. Nilotinib inhibited mitogen-activated protein kinase (MAPK), AKT and STAT5 phosphorylation in CML CD34(+) cells in the absence of growth factors (GFs), but did not suppress AKT and STAT5 activity, and resulted in increased MAPK activity, in the presence of GFs. Nilotinib and imatinib resulted in similar suppression of CML primitive and committed progenitors in long-term culture-initiating cell and colony-forming cell assays. Inhibition of progenitor growth was related to marked reduction in proliferation, but only a modest increase in apoptosis. Nilotinib did not show increased efficacy in reducing nondividing CML progenitors compared with imatinib. These results indicate that more potent tyrosine kinase inhibitors by themselves will not be more effective in eliminating CML progenitors than imatinib and that additional mechanism required for maintenance of malignant stem cells need to be identified to improve targeting of leukemia stem cells.

Degradation of xenobiotic compounds by lignin-degrading white-rot fungi: enzymology and mechanisms involved.

White-rot fungi (WRF) are ubiquitous in nature with their natural ability to compete and survive. WRF are the only organisms known to have the ability to degrade and mineralize recalcitrant plant polymer lignin. Their potential to degrade second most abundant carbon reserve material lignin on the earth make them important link in global carbon cycle. WRF degrade lignin by its unique ligninolytic enzymatic machinery including lignin peroxidase, manganese peroxidase, laccase, cellobiose dehydrogenase, H2O2-generating enzymes, etc. The ligninolytic enzymes system is non-specific, extracellular and free radical based that allows them to degrade structurally diverse range of xenobiotic compounds. Lignin peroxidase and manganese peroxidase carry out direct and indirect oxidation as well as reduction of xenobiotic compounds. Indirect reactions involved redox mediators such as veratryl alcohol and Mn2+. Reduction reactions are carried out by carboxyl, superoxide and semiquinone radicals, etc. Methylation is used as detoxification mechanism by WRF. Highly oxidized chemicals are reduced by transmembrane redox potential. Degradation of a number of environmental pollutants by ligninolytic system of white rot fungi is described in the present review.

DNA analysis with multiplex microarray-enhanced PCR.

We have developed a highly sensitive method for DNA analysis on 3D gel element microarrays, a technique we call multiplex microarray-enhanced PCR (MME-PCR). Two amplification strategies are carried out simultaneously in the reaction chamber: on or within gel elements, and in bulk solution over the gel element array. MME-PCR is initiated by multiple complex primers containing gene-specific, forward and reverse, sequences appended to the 3' end of a universal amplification primer. The complex primer pair is covalently tethered through its 5' end to the polyacryl- amide backbone. In the bulk solution above the gel element array, a single pair of unattached universal primers simultaneously directs pseudo-monoplex PCR of all targets according to normal solution-phase PCR. The presence of a single universal PCR primer pair in solution accelerates amplification within gel elements and eliminates the problem of primer interference that is common to conventional multiplex PCR. We show 10(6)-fold amplification of targeted DNA after 50 cycles with average amplification efficiency 1.34 per cycle, and demonstrate specific on-chip amplification of six genes in Bacillus subtilis. All six genes were detected at 4.5 pg of bacterial genomic DNA (equivalent to 10(3) genomes) in 60 independent amplification reactions performed simultaneously in single reaction chamber.

Copper in split and daily ejaculates.

The authors studied the copper level in three splits of 21 ejaculates to locate the origin of copper in semen. The copper level was rich in the first split and poor in the second. The results indicated that copper is released from all portions of the genital tract and that the major share is from the prostate gland. Another group of 26 volunteers submitted their semen samples once a day for 10 days for a study of the role of copper in fertility. A gradual increase in sperm motility was observed from the first day. There was no consistency in the level of copper in this study, and it did not correlate with any of the semen parameters studied.

Reduced alveolar macrophage production of tumor necrosis factor during sepsis in mice and men.

BACKGROUND AND METHODS: Tumor necrosis factor (TNF) has been implicated as a major humoral mediator of sepsis and endotoxin shock. TNF is secreted by cells of the reticuloendothelial system, including alveolar macrophages. Alveolar macrophage TNF production has been postulated to play a pathogenetic role in the development of adult respiratory distress syndrome (ARDS) in sepsis. To evaluate alveolar macrophage production of TNF during sepsis and endotoxin shock, we studied the effects of sepsis and/or in vivo lipopolysaccharide on the in vitro production of TNF by pulmonary alveolar macrophages. Human pulmonary alveolar macrophages were obtained by bronchoalveolar lavage from six septic and five nonseptic patients, cultured in the presence or absence of lipopolysaccharide (1 ng/mL), and assayed for TNF activity in a bioassay using fibroblast lysis. A murine model of sepsis was also utilized to study pulmonary alveolar macrophage TNF production under more controlled conditions. Normal mice were given ip injections of either lipopolysaccharide or saline. After 2 hrs, pulmonary alveolar macrophages were obtained and cultured in saline or various concentrations of lipopolysaccharide (0.001 to 10 micrograms/mL). RESULTS: There was no difference in baseline TNF activity, expressed as per cent lysis at 1:10 dilution, between pulmonary alveolar macrophages from control and septic patients (35.7 +/- 5.5% vs. 24.4 +/- 9.3%, respectively) (p greater than .05). However, when stimulated with lipopolysaccharide in vitro, the pulmonary alveolar macrophages from nonseptic patients produced significantly (p less than .01) more TNF (82.8 +/- 3.6%) than did pulmonary alveolar macrophages from patients with the septic syndrome (35.2 +/- 3.8%). Similar findings were obtained using the murine sepsis model. The baseline TNF activity in pulmonary alveolar macrophages from control mice was 22.9 +/- 7.0% (mean +/- SEM) and from lipopolysaccharide-injected mice was 26.8 +/- 3.3% (p greater than .05). Stimulation with 1 ng/mL lipopolysaccharide in vitro produced an increase in TNF activity in both groups, but the increase was greater in the control mice (68.1 +/- 5.7%) than in the lipopolysaccharide-injected mice (47.5 +/- 5.3%) (p less than .01). When the murine pulmonary alveolar macrophages were stimulated with higher concentrations of lipopolysaccharide (0.1 to 10 micrograms/mL), pulmonary alveolar macrophages from lipopolysaccharide-injected mice produced less than 25.5% of the TNF produced by pulmonary alveolar macrophages from control mice. CONCLUSIONS: These studies indicate that sepsis and endotoxin injection result in a rapid decrease in the ability of pulmonary alveolar macrophages from both humans and mice to produce and secrete TNF in response to lipopolysaccharide. We speculate that a downregulation of TNF production or of macrophage responsiveness to lipopolysaccharide has occurred. These results suggest that sustained TNF production by macrophages is not required for lung injury in sepsis.