Biomechanical comparison of nitinol compression staples versus fully threaded lag screws for talonavicular arthrodesis.

Background: Talonavicular arthrodesis (TNA) is indicated for treatment of disorders that require immobilization of the hindfoot. Lag screw fixation is considered the reference standard technique for TNA. Despite consistently favorable clinical results using lag screw fixation, it is still associated with higher than desired complication and failure rates. Nitinol compression staples have been used for TNA based on potential advantages over lag screw fixation. However, functional biomechanical data comparing lag screw and nitinol compression staples for TNA are lacking. Therefore, the objective of this study was to compare nitinol compression staples to fully threaded lag screws for use in TNA with respect to their biomechanical properties during functional robotic testing. Methods: TNA was performed on cadaveric feet (n = 12; 6 matched pairs) using either two nitinol compression staples (Arthrex, Naples, FL) or two fully threaded lag screws (Arthrex, Naples, FL) in random order, alternating between paired left and right feet. After instrumentation, specimens were mounting in a robotic testing system and loaded at 89 N/sec from 30 N to 445 N for 1 min. Then, continuous compressive load of 445 N was applied while cycling from 30° plantarflexion to 15° dorsiflexion for 10 cycles. Optical tracking markers attached to the talus and navicular bone tracked displacements. Translation data were recorded along the X, Y, Z planes. Rotation data were recorded for roll, pitch, and yaw. Significant (p < 0.05) differences between fixation methods were determined using paired t-Tests for each measured variable. Results: There were no statistically significant differences between staples and screws for translation in X, Y, or Z planes. When comparing rotation (roll, pitch, and yaw), there were no statistically significant differences with the exception of increased roll rotation for staple fixation versus lag screw fixation during static compression testing (p = 0.009). Conclusion: Based on comparison to the reference standard lag screw fixation for clinically relevant biomechanical properties measured during functional robotic testing of the hindfoot, nitinol compression staples are a viable option for talonavicular arthrodesis.

A Rare Phenomenon of Isaacs Syndrome: A Case Report.

We illustrate the case of a 71-year-old male who initially presented with sudden onset muscle weakness and ambulation difficulty. Following medication discontinuation and additional clinical studies, he failed to improve and was admitted to the hospital 11 weeks later. He had an associated 20-pound weight loss, sudorrhea, and muscle stiffness only when weight-bearing. A complete connective tissue cascade and a paraneoplastic panel were obtained. Clinical diagnosis of acquired neuromyotonia, or Isaacs syndrome (IS), was made, and he began experiencing significant improvement after intravenous steroid infusion. IS is a rare disease that has been poorly documented in the literature. There have only been a limited number of cases which are globally documented. One of the difficulties is a lack of definite autoantibody with which to correlate the disease; however, there has been some correlation linking the disease to voltage-gated potassium channels. Ultimately, the diagnosis should be driven by history and clinical presentation. The aim of this case report is to highlight a rare disease process and increase awareness among clinicians. We also describe the associated evaluation and recommended treatment for an optimal patient outcome.

Evaluating Acute Bilateral Foot Drop: A Case Report.

We illustrate the case of an 84-year-old Caucasian female who presented with complaints of bilateral lower extremity weakness and ambulation difficulties complicated by a unilateral deep venous thrombosis. Physical examination on hospital admission revealed an acute onset of bilateral foot drop with pes cavus deformity. Bilateral foot drop has been associated with a more chronic presentation due to metabolic, neurologic, and musculoskeletal etiologies. Acute onset of bilateral foot drop has been poorly defined in the literature and is considered a rare pathologic phenomenon, requiring additional investigation into the underlying cause of the presentation. We hypothesize that a spinal cord compression at the T12-L1 level resulted in L5 nerve root compression, resulting in our patient's presentation. Definitive treatment has not been established for this condition; however, studies have been completed to evaluate surgical versus conservative approaches to help restore patients' ambulatory function. Our aim is to incorporate this case report into the limited current literature on acute bilateral foot drop as well as outline possible treatment methods to restore impaired functionality.

Study of liquid scintillation alpha spectral properties.

Alpha spectra were acquired with Quantulus 1220 liquid Scintillation Counter (LSC) and correlations among parameters, such as alpha particle energy, sample quenching, peak's centroid and resolution were established. The effect of quenching and factors such as types of counting vials, extractive reagents and composition of extractive scintillation cocktail on alpha spectral resolution, was experimentally studied.

Concentration of 3H in ground water and estimation of committed effective dose due to ground water ingestion in some places in the Maharashtra state, India.

The measurement of tritium in environmental samples requires highest possible sensitivity. In the present study, the authors have optimised the counting window for the analysis of (3)H in environmental samples using the recently installed Ultra Low Level Quantulus 1220 Liquid Scintillation Counting at BARC to improve the detection limit of the system. The optimised counting window corresponding to the highest figure of merit of 883.8 was found to be 20-162 channels. Different brands of packaged drinking waters were analysed to select a blank that would define the system background. The minimum detectable activity (MDA) achieved was 1.5 Bq l(-1) for a total counting time of 500 min. The concentration of tritium in well and bore well water samples collected from the villages of Pune, villages located at 1.8 km from Tarapur Atomic Power Station, Kolhapur and Ratnagiri, was analysed. The activity concentration ranged from 0.55 to 3.66 Bq l(-1). The associated age-dependant dose from water ingestion in the study area was estimated. The effective committed dose recorded for different age classes is negligible compared with World Health Organization and US Environmental Protection Agency dose guidelines.

Standardization of ¹³¹I: implementation of CIEMAT/NIST method at BARC, India.

The CIEMAT/NIST efficiency tracing method using ³H standard was implemented at Radiation Safety Systems Division, Bhabha Atomic Research Centre (BARC) for the standardization of ¹³¹I radioactive solution. Measurements were also carried out using the 4π ß-γ coincidence counting system maintained as a primary standard at the laboratory. The implementation of the CIEMAT/NIST method was verified by comparing the activity concentration obtained in the laboratory with that of the average value of the APMP intercomparison (Yunoki et al., in progress, (APMP.RI(II)-K2.I-131)). The results obtained by the laboratory is linked to the CIPM Key Comparison Reference Value (KCRV) through the equivalent activity value of National Metrology Institute of Japan (NMIJ) (Yunoki et al., in progress, (APMP.RI(II)-K2.I-131)), which was the pilot laboratory for the intercomparison. The procedure employed to standardize ¹³¹I by the CIEMAT/NIST efficiency tracing technique is presented. The activity concentrations obtained have been normalized with the activity concentration measured by NMIJ to maintain confidentiality of results until the Draft-A report is accepted by all participants. The normalized activity concentrations obtained with the CIEMAT/NIST method was 0.9985 ± 0.0035 kBq/g and using 4π ß-γ coincidence counting method was 0.9909 ± 0.0046 kBq/g as on 20 March 2009, 0 h UTC. The normalized activity concentration measured by the NMIJ was 1 ± 0.0024 kBq/g. The normalized average of the activity concentrations of all the participating laboratories was 1.004 ± 0.028 kBq/g. The results obtained in the laboratory are comparable with the other international standards within the uncertainty limits.

Validation of efficiency tracing and zero detection threshold techniques using liquid scintillation analyser TriCarb.

Efficiency tracing with unquenched 14C and zero detection threshold with unquenched 3H as tracers are practical and simple techniques which have been implemented to quantify the activity of various beta emitters using liquid scintillation analyser. These techniques are used to study the influence of quench level on activity quantification and the activity levels up to which these techniques are applicable. The results indicate that, for an activity level of 166.67 Bq, both the techniques are in good agreement with the reference activity with a relative discrepancy of ≤4.6 %. The relative discrepancy of ~10 % is observed for extreme quench values of ~111. For all the radionuclides with the activity level of 1.67 Bq, the uncertainty in activity quantification raises to ~8 % and for the activity level from 8.33 to 100 Bq, the uncertainty reduces to 1 %.

Comparative study of different methods for the activity quantification of (3)H and (14)C radionuclides in dual labeled samples using liquid scintillation analyzer.

Different methods such as full spectrum DPM (FSDPM), dual isotope estimation and inclusion methods were studied for the activity quantification of (3)H and (14)C in dual beta labeled samples using liquid scintillation analyzer. A standardized Packard tritiated water was used as tritium source and (14)C activity was standardized by CIEMAT/NIST method and compared with the results obtained by the above three methods. Minimum detectable activity was 2100dpm/l for (3)H and 1200dpm/l for (14)C with a counting time of 300min. The accuracy of the results obtained was found to be within +/-10% for (3)H: (14)C activity ratios 1:1, 1:2, 2:1, 6:1, 9:1, 13:1 and 18:1.

Inhibition by salmeterol and cilomilast of fluticasone-enhanced IP-10 release in airway epithelial cells.

The CXC chemokines, IP-10/CXCL10 and IL-8/CXCL8, play a role in obstructive lung disease by attracting Th1/Tc1 lymphocytes and neutrophils, respectively. Inhaled corticosteroids (ICS) and long acting beta 2-agonists (LABA) are widely used. However, their effect(s) on the release of IP-10 and IL-8 by airway epithelial cells are poorly understood. This study examined the effects of fluticasone, salmeterol, and agents which raise intracellular cAMP (cilomilast and db-cAMP) on the expression of IP-10 and IL-8 protein and mRNA. Studies were performed in cultured human airway epithelial cells during cytokine-stimulated IP-10 and IL-8 release. Cytokine treatment (TNF-alpha, IL-1beta and IFN-gamma) increased IP-10 and IL-8 protein and mRNA levels. Fluticasone (0.1 nM to 1 microM) increased IP-10 but reduced IL-8 protein release without changing IP-10 mRNA levels assessed by real time RT-PCR. The combination of salmeterol (1 micro M) and cilomilast (1-10 mu M) reduced IP-10 but had no effect on IL-8 protein. Salmeterol alone (1 micro M) and db-cAMP alone (1 mM) antagonised the effects of fluticasone on IP-10 but not IL-8 protein. In human airway epithelial cells, inhibition by salmeterol of fluticasone-enhanced IP-10 release may be an important therapeutic effect of the LABA/ICS combination not present when the two drugs are used separately.

Determination of 243Am by pulse shape discrimination liquid scintillation spectrometry.

Alpha specific activity of 243Am was determined using pulse shape discrimination in liquid scintillation spectrometry. 238Pu, 36Cl and 239Np (purified from 243Am) were used for obtaining the spillover of alpha/beta particles into the beta/alpha channels, respectively. Synthetic mixtures of 241Am/243Am were prepared. Using the alpha-specific activity, weights of the stock solutions used and the half-life of 241Am and 243Am isotopes, the expected 241Am/243Am atom ratios in the mixtures were determined and compared with those obtained by thermal ionization mass spectrometry (TIMS). An agreement of about 1% was obtained between the 241Am/243Am atom ratios determined by the two methods. This shows that liquid scintillation counting with pulse shape discrimination can be used for 243Am determination with an accuracy better than 1%.

CXCR3 surface expression in human airway epithelial cells: cell cycle dependence and effect on cell proliferation.

We recently demonstrated that human bronchial epithelial cells (HBEC) constitutively express the CXC chemokine receptor CXCR3, which when activated, induces directed cell migration. The present study in HBEC examined the relative expression of the CXCR3 splice variants CXCR3-A and -B, cell cycle dependence of CXCR3 expression, and the effects of the CXCR3 ligand, the interferon-gamma-inducible CXC chemokine I-TAC/CXCL11, on DNA synthesis and cell proliferation. Both CXCR3-A and -B mRNA, assessed by real-time RT-PCR, were expressed in normal HBEC (NHBEC) and the HBEC line 16-HBE. However, CXCR3-B mRNA was 39- and 6-fold greater than CXCR3-A mRNA in NHBEC and 16-HBE, respectively. Although most HBEC (>80%) assessed by flow cytometry and immunofluorescence microscopy contained intracellular CXCR3, only a minority (<40%) expressed it on the cell surface. In this latter subset of cells, most (>75%) were in the S + G(2)/M phases of the cell cycle. Stimulation of CXCR3 with I-TAC enhanced thymidine incorporation and cell proliferation and increased p38 and ERK1/2 phosphorylation. These data indicate that 1) human airway epithelial cells primarily express CXCR3-B mRNA, 2) surface expression of CXCR3 is largely confined to the S + G(2)/M phases of the cell cycle, and 3) activation of CXCR3 induces DNA synthesis, cell proliferation, and activation of MAPK pathways. We speculate that activation of CXCR3 exerts a mitogenic effect in HBEC, which may be important during airway mucosal injury in obstructive airway diseases such as asthma and chronic obstructive pulmonary disease.

Regulation of IL-1-induced gingival collagenase gene expression by activator protein-1 (c-Fos/c-Jun).

Matrix metalloproteinase-1 is probably involved in the progression of periodontal disease. The aim of this study was to investigate whether IL-1beta stimulates the expression of the activator protein 1 (AP-1) transcription factor and, consequently, if the AP-1 transcription factor participates in the regulation of collagenase gene expression in human gingival fibroblast cells. In this study, we demonstrate that the concentration of the protein components of AP-1 transcription factor, c-Fos and c-Jun, is enhanced by IL-1beta both at mRNA and protein levels, utilizing Northern blot analysis, electrophoretic mobility gel shift assay and Western blot analysis. The IL-1beta stimulated the collagenase-CAT and AP-1-CAT activities in a dose dependent manner with respect to the amount of DNA used in transfections. Further, overexpression of c-Fos and c-Jun proteins revealed a dose-dependent transcriptional activation of the collagenase promoter. These findings, coupled with the existence of AP-1 consensus DNA binding sites on the collagenase gene promoter, show that regulation of collagenase gene expression by IL-1beta involves the transcription factor AP-1 in gingival fibroblasts.

Construction of attenuated HIV-1 accessory gene immunization cassettes.

Delivery of genetic expression cassettes into animals can effectively induce both humoral and cellular immunity to the expressed gene product. Previously, we used this strategy to immunize against HIV-1 structural and enzymatic proteins in mice, non-human primates and in humans. In contrast, the use of the accessory genes including vif, vpr, vpu and nef as immunotherapeutic vaccine targets has not been well characterized. Our goal is to design an effective genetic HIV vaccine, which includes the accessory genes as part of a multi-component immunogen. In order to develop accessory genes as genetic vaccines, we have molecularly cloned and analysed the sequence variation and immunogenic potential present in these genes derived from viral isolates obtained from HIV-1 infected patients and laboratory isolates. Prototype genetic variants were selected and their ability to induce humoral and cellular immune responses was studied in animal models. We observed that attenuated accessory genes can effectively induce both humoral and cellular responses in mice and the resulting immune response is directly correlated with DNA concentrations delivered and the number of boosts. This strategy can be used generally to develop an effective, safe DNA vaccine for any pathogen.

Inhibition of gingival collagenase gene expression by dexamethasone.

Glucocorticoids are potent immunosuppressants shown to be effective in the treatment of inflammatory diseases. Reportedly, they work, in part, by inhibiting cytokines and other transcription factors including AP-1. In this study we investigated the mechanisms of efficient repression of collagenase gene expression by dexamethasone in the human gingival fibroblast. Northern analyses showed that IL-1-dependent collagenase mRNA production was significantly decreased in the presence of dexamethasone. The influence of dexamethasone on the transcription factor NF-kappaB, STAT3, and AP-1 was investigated by using the gel mobility shift assay with nuclear extracts prepared from the cells grown in the presence of dexamethasone. We observed that in addition to AP-1, binding of NF-kappaB and STAT3 to DNA was also decreased significantly. Additionally, dexamethasone induced the transcription of the I kappaB-alpha gene suggesting that in the presence of dexamethasone, NF-kappaB quickly reassociates with newly synthesized I kappaB-alpha and markedly reduces the amount of NF-kappaB. CAT transfection studies utilizing collagenase promoter demonstrated a dose-dependent transcriptional inhibition of IL-1-induced gingival collagenase gene expression by dexamethasone. These data reveal that collagenase gene expression can be regulated by the impairment of IL-1-stimulated NF-kappaB, STAT3, and AP-1 activities, and can highlight a possible molecular mechanism for the anti-inflammatory effects of glucocorticoids.

IL-1 beta alters beta-adrenergic receptor adenylyl cyclase system function in human airway epithelial cells.

Inflammatory cells release a variety of cytokines, including interleukin (IL)-1 beta, into the airway in asthma. This study examined the effects of human IL-1 beta on the function of the beta-adrenergic receptor (beta AR)-adenylyl cyclase (AC) system in BEAS-2B cells, a human airway epithelial cell line. IL-1 beta markedly increased beta AR density (Bmax; P < 0.001) primarily by increasing the percentage of the beta 2AR subtype (from 67 to 91%; P < 0.001). Bmax increased monotonically over time in response to 200 pM IL-1 beta and was approximately 2.5-fold greater than control cells between 36 and 42 h. In contrast, the concentration response of Bmax to IL-1 beta given for 18 h was biphasic. Bmax increased with IL-1 beta concentrations from 2 to 200 pM, but, at > 200 pM, it decreased progressively toward control values. IL-1 beta-induced increases in Bmax with IL-1 beta were associated with approximately threefold increases in beta 2 AR mRNA and were blocked by the protein synthesis inhibitor cycloheximide. Despite the marked increase in Bmax, however, IL-1 beta depressed adenosine 3',5'-cyclic monophosphate (cAMP) responses to isoproterenol and forskolin, a direct activator of AC (P < 0.001 by analysis of variance for both). The inhibitory effect of IL-1 beta on cAMP production appeared to be explained by increases in the activity of an inhibitory GTP binding protein because IL-1 beta treatment increased the activity of a pertussis toxin ADP-ribosylated Gi alpha protein by approximately 2.5-fold; and pretreatment of intact cells with pertussis toxin inhibited the effect of IL-1 beta on cAMP production. These data indicate that IL-1 beta-mediated changes in the beta AR-AC system function in airway epithelial cells are complex and involve expression of receptor protein, GTP binding protein, and possibly AC itself. Increases in IL-1 beta may contribute to abnormalities in airway function in subjects with asthma.

Chronic effects of catecholamines on the beta 2-adrenoreceptor system in cultured human airway epithelial cells.

Chronic catecholamine treatment induces beta-adrenergic receptor (beta AR) downregulation, i.e., a loss of total cell receptors. In the human respiratory tract, the mechanism(s) underlying beta AR downregulation remains poorly understood. The present study, therefore, examined the effects of 24 h of exposure to isoproterenol (Iso; 10 nM or 1 microM) on beta AR density and the rate of beta AR degradation, steady-state beta 2-adrenergic receptor (beta 2 AR) mRNA levels, and the content of Gs alpha and Gi alpha proteins in cultured human bronchial epithelial cells (i.e., the BEAS-2B cell line). beta AR density assessed by binding with [125I]iodopindolol decreased in a dose-dependent fashion with 24 h of Iso exposure. With Iso (1 microM), beta AR density decreased by approximately 82%. In contrast, forskolin (100 microM) and dibutyryl adenosine 3',5'-cyclic monophosphate (1 mM), agents that also increase adenosine 3',5'-cyclic monophosphate (cAMP) levels, had no significant effect on beta AR density. Iso exposure also elicited a concomitant decrease in Iso-stimulated cAMP but had no significant effect on the content of the G proteins G alpha i2 and Gs alpha assessed by immunoblotting and toxin-catalyzed ADP ribosylation. Of note, Iso exposure (1 microM) had no effect on steady-state levels of beta 2 AR mRNA measured both by Northern analysis and by reverse transcriptase-polymerase chain reaction. However, beta AR half-life assessed in the presence of the protein synthesis inhibitor cycloheximide was reduced by approximately 60% in Iso-treated cells (i.e., from 37 h in control to 16 h in 1 microM Iso). These results suggest that, in human airway epithelial cells, beta 2 AR downregulation 1) is not primarily driven by intracellular cAMP levels, 2) is not associated with significant decreases in steady-state levels of beta 2 AR mRNA, and 3) is largely posttranslationally regulated by increases in the rate of receptor protein degradation.

Colorimetric determination of reserpine in polyherbal formulations.

A simple spectrophotometric method is reported her for the estimation of reserpine in polyherbal formulations. Estimation is based on the reaction with3-methylbenzolinone-2hydrazone (MBTH) reagent in presence of cerric ammonium sulphate to yield a violet coloured chromogen, which exhibits and absorption maxima at 580 nm. The chromogen is stable for 10 minutes.

Rule-based system for the fast identification of species of Indian Anopheline mosquitoes.

In a developing country like India, classification and identification of the species of Anopheline mosquitoes in control operations of mosquito-borne diseases is of paramount importance. The WHO monograph, which describes the taxonomic data in the form of a pictorial key is generally difficult to understand by a non-taxonomist. Utilizing the principles of ID3 algorithm, a novel rule-based system, for the fast identification of unknown species of Indian Anopheline mosquitoes, is developed. The rule-based system is user-friendly, menu-driven and even a novice can make use of it in the identification of the unknown species with little practice. The above software is available on floppy disk and can be obtained with a minimum cost. This program can be ported on 5 1/4" or 3 1/2" floppy disk.