Synthesis of a novel ß-cyclodextrin-functionalized Fe3O4/BaMoO4:Dy3+ magnetic luminescent hybrid nanomaterial and its application as a drug carrier.

Magnetic luminescent hybrid nanoparticles Fe3O4/BaMoO4:Dy3+ (MLHNPs) were successfully synthesized by the co-precipitation method using ethylene glycol. The MLHNPs are functionalized with ß-cyclodextrin (CD) using 3-aminopropyl triethoxysilane (APTES). The surface functionalization of nanoparticles and their conjugation have been confirmed via powder XRD, FTIR, HRTEM, and PL analyses. The feasibility of functionalized MLHNPs as nanocarriers for hydrophobic drugs (triazole derivatives) was verified by studying the uptake and release profile using a triazole derivative as a hydrophobic drug. In addition, the photoluminescence spectroscopy results confirmed the optical imaging capability of MLHNPs. The outcomes of loading and release disclose that this system is likely to be a suitable tool as a hydrophobic drug carrier.

Inhalational Ciclesonide found beneficial in prevention of fat embolism syndrome and improvement of hypoxia in isolated skeletal trauma victims.

BACKGROUND: Many studies have established intravenous corticosteroid as an effective prophylactic therapy in fat embolism syndrome (FES). However, its use is limited among surgeons because of systemic side effects. Inhalational steroids have least systemic effects and are widely used for several chest conditions (i.e., asthma), but their effectiveness in FES has not been established. QUESTION/PURPOSE: This study was sought to evaluate the (1) efficacy and (2) safety of inhalational Ciclesonide (CIC) in prevention of FES and treatment of hypoxemia in isolated skeletal trauma victims. METHODS: A nonrandomized prospective control trial was designed in which all patients between 18 and 40 years with isolated skeletal injury who presented within 8 h of injury were allocated to either Trial group or control group. Trial group patients received 640 mcg of inhalational CIC with a metered-dose inhaler at the time of admission, and at 24 h. Control group patients did not receive any prophylactic therapy. Both groups were evaluated for development of FES (Gurd's criteria) and hypoxemia (PaO2 <70 mmHg) for 72 h. The complications related to CIC administration were evaluated in trial group patients during their hospital stay. RESULTS: Of 35 patients in each group, two patients in Trial group and nine patients in control group developed FES (P = 0.022). Eight patients in Trial group had hypoxemia at the time of admission, six of them improved and one additional patient developed hypoxemia after inhalational CIC administration. In control group, ten patients had hypoxia at the time of admission, only one of them improved and remaining nine patients had persistent hypoxemia even after 72 h. Additionally, three patients developed hypoxemia. A significant improvement in hypoxemia and a significant decrease in the incidence of FES were observed in Trial group (P < 0.05) compared to control group. None of the patients presented with any complications or adverse effects of steroid in Trial group. CONCLUSION: Inhalational CIC is a safe and effective therapy for prevention of FES and also an effective drug for treatment of hypoxemia in orthopedic trauma victims. LEVEL OF EVIDENCE: Level III, therapeutic study.

Nonunions and malunions of the pelvis.

INTRODUCTION: Neglected pelvic fractures manifesting as pelvic nonunion or malunion are usually due to inadequate initial fixation or negligence of the injury because of increased attention towards other associated life-threatening conditions. The management of such injuries is complex. A systematic review was conducted to spot the clinical manifestations, evaluation, management and outcome of pelvic nonunion and malunion. MATERIALS AND METHODS: Two databases ("Pubmed" and "Google scholar") were searched to look for relevant literature on pelvic non-union and malunion.  The search was limited to 'English language' and 'Human being'. RESULTS: A total of 500 articles found, of which 10 articles were only reviewed which met the inclusion criteria. These articles discussed the clinical management and treatment of pelvic malunion and non-union following trauma without associated acetabular injury. CONCLUSION: The usual presentations of pelvic non-union and malunion are pain, deformity, gait abnormality or instability. A detailed preoperative evaluation is essential as a majority of them have associated hip and spine injury which may be the cause of symptoms. Radiographs and 3D CT scans have helped surgeons in deciding the best way of management. The surgeries are usually complex and may need multiple-staged procedures. Soft tissue release, multiple osteotomies to achieve anatomical or near-anatomical reduction, augmentation of healing process using bone graft and stabilizing the nonunion/ osteotomy site using plates/screws/rods is the basic principle of surgery. Per-operative use of somato-sensory evoked potential evaluation helps the surgeon in preventing iatrogenic nerve injury. Despite these precautions and surgeries, most of the patients do not regain their preinjury functional activity.

Management of neglected acetabular fractures.

Management of neglected acetabular fractures is a difficult task. Osteosynthesis in such cases may not be an ideal solution because of the femoral head damage due to pressure by the fractured acetabular edge, avascular necrosis, difficulty in mobilizing the fragments due to callus formation, difficulty in indirect reduction of the fracture fragments and macerated acetabular fragments all contributing to inadequate fracture reduction. Majority of such fractures are now treated with total hip replacement. While treating such fractures with THR, problems associated with neglected acetabular fractures such as fracture non-union, hip dislocation, protrusio, cavitary bone defect or peripheral bone defect must be considered. 3D computed tomography scan provides a clear view about the acetabular and periacetabular bony anatomy. Impaction grafting and antiprotrusio cage or ring with a cemented acetabular cup can address most of the hip protrusio and cavitary bone defects. Segmental bone defect needs cortical strut-bone graft fixation and subsequent implantation of a cemented or uncemented acetabular cup implantation. Fracture non-union needs approximate reduction and fixation with plates followed by bone grafting and implantation of an acetabular cup. Despite these efforts, the outcome of THR in neglected acetabular fracture is considerable worse than after conventional hip replacement.

Human bioequivalence evaluation of two losartan potassium tablets under fasting conditions.

The bioequivalence of two different tablet formulations containing losartan potassium 100 mg was determined in healthy volunteers after a single oral dose in a randomized crossover study. Test and reference products were administered to 60 volunteers with 240 ml water after overnight fasting. Plasma concentrations of losartan and its active carboxylic acid metabolite were monitored over a period of 36 h after drug administration by validated LC/MS/MS analytical method. The pharmacokinetic parameters Cmax, AUC0-t, AUC0-∞, AUC0-t/AUC0-∞, tmax, Kel and t½ were determined from plasma concentration time profile of both formulations for losartan and its active metabolite losartan carboxylic acid and were found to be in good agreement. The carboxylic acid metabolite was considered for profiling purpose only. The analysis of variance did not show any significant difference between the two formulations and 90% confidence intervals for the ratio of Cmax (84.89-104.09%), AUC0-t (95.84-102.84%) and AUC0-∞ (96.43-103.25%) values for losartan between the test and reference products were within the 80-125% interval, satisfying the bioequivalence criteria of the US FDA guidelines. These results indicate that the test and the reference products of losartan potassium are bioequivalent and, thus, may be prescribed interchangeably.

Predictors of postoperative outcome for acetabular fractures.

BACKGROUND: The outcomes of surgically treated acetabular fractures are dependent on many factors. The purpose of this retrospective study is to evaluate these factors in a group of patients operated on by a single surgeon in one institute. METHODS: One hundred and eighteen patients, treated surgically for their displaced acetabular fracture and who had completed two years follow-up, were evaluated clinically with Modified Postel Merle d'Aubigné score and radiologically with Matta's radiological outcome grading. The effect of age (≤ 55 or >55 years), gender, fracture displacement (≤ 20mm or >20mm), hip dislocation, delay in surgery (≤ 2 weeks or >2 weeks), associated injury and length of follow-up (≤ 5 years or >5 years) on the functional outcome was evaluated. RESULTS: There were 99 (83.9%) males and 19 (16.1%) females with mean age of 38.75 years (16 to 65 years). The mean duration of follow-up was 3.95 years (range 2 to 14 years). The mean Modified Postel Merle d'Aubigné score was 15.7 ± 2.2 (range, 8 to 18). The clinical outcome was excellent in 27 (22.9%), good in 52 (44.2%), fair in 20 (16.9%), and poor in 19 (16.1%, 10 patients who underwent THR for secondary arthritis were considered as poor outcome) patients. The Modified Postel Merle d'Aubigné score was significantly affected by quality of reduction (P=0.0001), presence of associated injuries (P=0.0001), initial fracture displacement of >20mm (P=0.018), joint dislocation (P=0.015) and delay in surgery (P=0.001). However, age, gender, fracture type and length of follow-up did not have any effects on the clinical outcome. CONCLUSION: Poor reduction, associated injuries, fracture displacement of >20mm, joint dislocation and late surgery definitely carry poor prognosis in predicting the outcome of surgically treated acetabular fractures.

Effectiveness of a community-based observation of anti-tuberculosis treatment in Bangalore City, India, 2010-2011.

SETTING: The Revised National Tuberculosis Control Programme in an urban setting of Bangalore City, India. OBJECTIVES: To compare treatment outcomes and smear conversion rates among new smear-positive tuberculosis (TB) patients undergoing treatment administered by community directly observed treatment (DOT) providers with those undergoing treatment administered by institutional DOT providers in Bangalore City in 2010-2011. METHOD: Cohort study of routine data recorded from treatment cards of TB patients undergoing treatment under the public health services from 1 October 2010 to 30 September 2011. RESULT: Treatment records of 1864 new smear-positive TB patients registered during this period were evaluated. Among those evaluated, 604 (32%) had been administered treatment by community DOT providers and the remainder by institutional DOT providers. The treatment success rate in those undergoing community DOT was 93% (n = 564) and that of those undergoing institutional DOT was 75% (n = 951; RR 1.23, 95%CI 1.19-1.28). The sputum smear conversion rate of patients who underwent community DOT was 92% and that of those who underwent institutional DOT was 71% at the end of 2 months. CONCLUSION: We conclude that community DOT for treatment supervision of TB patients is more effective than institutional DOT and that it should be reinforced.

Comparison of accuracy and safety of computed tomography guided and unguided transthoracic fine needle aspiration biopsy in diagnosis of lung lesions.

OBJECTIVE: To study the accuracy and safety of CT guided and unguided transthoracic fine needle aspiration biopsy in diagnosis of lung lesions. METHOD: The study was carried out in 79 hospitalised patients during the period 1997-1999. In 52 patients having peripheral and large sized lung lesion (> 5 cm in diameter) in chest X-ray unguided FNAB was performed and in the rest 27 patients having relatively central and small sized lesion (< 5 cm). CT guided FNAB was performed. Also in 15 patients having two times failed unguided aspiration, CT guided FNAB was performed. RESULTS: The diagnostic yield of unguided aspiration was 71.1% (37 out of 52). Out of 37 patients 29 (78.3%) had malignant lesion and eight (21.6%) non-malignant lesion. Sensitivity and specificity for detecting malignancy was 90.6% and 100% respectively. Complications were seen in 10 patients (19.3%). Diagnostic yield of CT guided FNAB was 95.2% (40 out of 42), 33 (82.5 %) had malignant lesion, seven (17.5%) had benign lesion. Sensitivity and specificity for detecting malignancy was 97.1% and 100% respectively. Minor complications were seen in three patients (7.1%). CONCLUSION: It was concluded that CT guided

Raised serum thiobarbituric acid reactive substance levels in malaria.

To study the extent of serum lipid peroxidation in malaria, 62 patients of falciparum malaria (18 uncomplicated and 44 complicated), 15 patients of vivax malaria and 25 healthy controls were enrolled in this study. The extent of serum lipid peroxidation was evaluated by estimating serum thiobarbituric acid reactive substances (TBARS) colorimetrically. The mean serum TBARS levels were 1.5 +/- 0.29, 1.21 +/- 0.2 and 3.58 +/- 1.35 nmol/ml in controls, vivax malaria and falciparum malaria patients respectively. The TBARS level was significantly more in complicated falciparum malaria patients (4.2 +/- 1.03 nmol/ml) than uncomplicated falciparum malaria patients (2.01 +/- 0.61 nmol/ml). The TBARS level was also more in patients who died (4.82 +/- 0.64 nmol/ml) when compared to the survivors (2.92 +/- 1.05 nmol/ml).

Long-term erythropoietin expression in rodents and non-human primates following intramuscular injection of a replication-defective adenoviral vector.

Erythropoietin (Epo)-responsive anemia is a debilitating complication of chronic renal failure and human immunodeficiency virus (HIV) infection that effects more than 150,000 Americans. Patients with Epo-responsive anemias are currently treated with repeated injections of recombinant human Epo. In the studies described in this report, we have examined the safety and efficacy of using a single intramuscular (i.m.) injection of replication-defective adenoviral vectors (RDAd) encoding Epo for the treatment of Epo-responsive anemias in both mice and non-human primates. Our results demonstrate that there is a threshold dose of virus (2.5-8 x 10(7) pfu/gram of body weight) which is required to obtain long-term Epo expression and polycythemia in both species. A single i.m. injection of mice with 10(9) pfu of an RDAd encoding murine Epo (AdmEpo) resulted in elevations in hematocrits from control values of 49 +/- 0.9% to treated values of 81 +/- 3%, which were stable for more than 1 year. Similarly, a single i.m. injection of a monkey with 4 x 10(11) pfu of an RDAd-encoding simian Epo (AdsEpo) resulted in elevations of hematocrits from control levels of 40% to treated levels of > or =70%, which were stable for 84 days. Intramuscular injection of monkeys with AdsEpo appeared to be safe in that we did not detect abnormalities in chest X-rays, serum chemistries, hematologic, or clotting profiles (apart from elevated hematocrits) or organ histologies during the 84-day time course of the experiment. Taken together, these results suggest the feasibility of using i.m. injection of RDAd for the treatment of Epo-responsive anemias in humans.

Epoxy matrix for solid-state dye laser applications.

The preparation and performance of an epoxy-based matrix impregnated with Pyrromethene 580 for solid-state dye laser applications are discussed. The matrix proved to be stable and efficient as a laser medium when pumped by a frequency-doubled, Q-switched Nd:YAG laser with a 10-ns pulse width. Stability measurements were performed on a 1-mm-thick epoxy sample, doped with Pyrromethene 580 at a concentration of 4 x 10(-3) M. When the sample was pumped at millijoule energy levels, the stability was measured to be ~55,000 pulses from a single spot on the sample before the power dropped by a factor of half.

Two-photon-induced fluorescence from the phycoerythrin protein.

Two-photon-induced fluorescence is observed from the photodynamic phycobiliprotein phycoerythrin. Temporal, spectral, and intensity-dependent properties of the two-photon-induced fluorescence emission from phycoerythrin excited by a 1.06-mum laser beam are reported. The measured two-photon absorption cross section of phycoerythrin is an order of magnitude larger than that of Rhodamine 6G. The potential applications of phycobiliproteins for two-photon-induced fluorescence for microscopy of three-dimensional biological samples and three-dimensional optical memory are discussed.

Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of a plasmid DNA vector.

Erythropoietin (Epo)-responsive anemia is a common and debilitating complication of chronic renal failure and human immunodeficiency virus infection. Current therapy for this condition involves repeated intravenous or subcutaneous injections of recombinant Epo. In this report, we describe the development of a novel muscle-based gene transfer approach that produces long-term expression of physiologically significant levels of Epo in the systemic circulation of mice. We have constructed a plasmid expression vector, pVRmEpo, that contains the murine Epo cDNA under the transcriptional control of the cytomegalovirus immediate early (CMV-IE) promoter, the CMV-IE 5' untranslated region, and intron A. A single intramuscular (i.m.) injection of as little as 10 micrograms of this plasmid into immunocompetent adult mice produced physiologically significant elevations in serum Epo levels and increased hematocrits from preinjection levels of 48 +/- 0.4% to levels of 64 +/- 3.3% 45 days after injection. Hematocrits in these animals remained elevated at greater than 60% for at least 90 days after a single i.m. injection of 10 micrograms of pVRmEpo. We observed a dose-response relationship between the amount of plasmid DNA injected and subsequent elevations in hematocrits. Mice injected once with 300 micrograms of pVRmEpo displayed 5-fold increased serum Epo levels and elevated hematocrits of 79 +/- 3.3% at 45 days after injection. The i.m. injected plasmid DNA remained localized to the site of injection as assayed by the PCR. We conclude that i.m. injection of plasmid DNA represents a viable nonviral gene transfer method for the treatment of acquired and inherited serum protein deficiencies.

Immune responses to transgene-encoded proteins limit the stability of gene expression after injection of replication-defective adenovirus vectors.

The use of replication-defective adenoviruses (RDAd) for human gene therapy has been limited by host immune responses that result in transient recombinant gene expression in vivo. It remained unclear whether these immune responses were directed predominantly against viral proteins or, alternatively, against foreign transgene-encoded proteins. In this report, we have compared the stability of recombinant gene expression in adult immunocompetent mice following intramuscular (i.m.) injection with identical RDAd encoding self (murine) or foreign (human) erythropoietin. Our results demonstrate that immune responses direct against foreign transgene-encoded proteins are the major determinants of the stability of gene expression following i.m. injection of RDAd. Moreover, we demonstrate long-term recombinant gene expression in immunocompetent animals following a single i.m. injection of RDAd encoding a self protein. These findings are important for the design of future preclinical and clinical gene therapy trials.

Muscle-based gene therapy: realistic possibilities for the future.

The past five years have witnessed tremendous growth in the field of gene therapy, with pre-clinical and clinical gene therapy trials for diseases as diverse as cancer, AIDS and atherosclerosis. These studies have utilized many different vectors and target organs in order to achieve therapeutic effects. In this review, we examine the rationale for using skeletal muscle as a target tissue for gene therapy, discuss the wide array of vectors that have been used for muscle-based gene therapy, summarize the disease-targets that have been approached using these techniques, and discuss some of the obstacles that remain to be overcome en route to successful muscle-based human gene therapy.

A biotinylated undecylthiophene copolymer bioconjugate for surface immobilization: creating an alkaline phosphatase chemiluminescence-based biosensor.

Methodology is described for the creation of a molecular assembly consisting of the enzyme alkaline phosphatase immobilized onto a glass surface using a biotinylated conjugated copolymer, poly(3-undecylthiophene-co-3-thiophenecarboxaldehyde) 6-biotinamidohexanohydrazone. The biotinylated polymer is attached to the inside walls of a silanized glass capillary via hydrophobic interactions, and a streptavidin-conjugated alkaline phosphatase is interfaced with the polymer through the classical biotin-streptavidin interaction. Utilizing a simple optical setup, we can detect the activity of as little as approximately 0.1 fmol of alkaline phosphatase with this molecular assembly. The assembly is mechanically robust and retains the majority of bound enzyme activity for up to 30 days. We have utilized this molecular assembly for the detection of organophosphorus-based pesticides. Both paraoxon and methyl parathion inhibit the enzyme-mediated generation of chemiluminescence signal. We are able to detect paraoxon and methyl parathion concentrations down to 500-700 ppb.

Chemiluminescence-based inhibition kinetics of alkaline phosphatase in the development of a pesticide biosensor.

The use and application of the enzyme alkaline phosphatase in a chemiluminescence assay are discussed. The enzyme catalyzes the hydrolysis of a macrocyclic phosphate compound generating a chemiluminescence signal. On the basis of inhibition of this signal, a methodology for the detection and quantitation of organophosphorus-based pesticides has been developed. The methodology is studied with alkaline phosphatase in the bulk aqueous phase, and detection of the signal is accomplished by a simple optical setup. Parts per billion level detection of paraoxon and methyl parathion in bulk solutions is achieved. The technique is rapid and sensitive and is applicable to the detection of most organophosphorus-based pesticides. The results from kinetic studies indicate a mixed type of inhibition of the enzyme by paraoxon and methyl parathion. The detection methodology forms an integral part of a biosensor under development and is adaptable to incorporating optical fibers for remote detection of pesticides.

Molecular assembly of proteins and conjugated polymers: Toward development of biosensors.

A molecular assembly in which a conjugated polymer is interfaced with a photodynamic protein is described. The conjugated polymer, functionalized with biotion, is designed such that it can be physisorbed on or chemically grown off a glass surface. The streptavidin-derivatized protein is immobilized on the biotinylated polymer matrix through the strong biotin-streptavidin interactions. The assembly, built on the surface of an optical fiber or on the inside walls of a glass capillary, forms an integral part of a biosensor for the detection of environmental pollutants such as organophosphorus-based insecticides. The Protein in the system can be replaced by any biological macromolecule of interest. We study one specific case, the enzyme alkaline phosphatase. The enzyme catalyzes a reaction producing an intermediate compound that chemiluminesces, and the chemiluminescence singnal is monitored to detect and quantify insecticides such as paraoxon and methyl parathion. Preliminary results indicate ppb level detection with response time less than 1 minute. (c) 1995 John Wiley & Sons, Inc.

Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replication-defective adenovirus.

A number of inherited and acquired serum protein deficiencies including hemophilias A and B, diabetes mellitus, and the erythropoietin-responsive anemias are currently treated with repeated subcutaneous or intravenous infusions of purified or recombinant proteins. The development of an in vivo gene-transfer approach to deliver physiologic levels of recombinant proteins to the systemic circulation would represent a significant advance in the treatment of these disorders. Here we describe the construction of a replication-defective adenovirus (AdEF1hEpo) containing the human erythropoietin (hEpo) cDNA under the transcriptional control of the cellular elongation factor 1 alpha (EF1 alpha) promoter and the 4F2 heavy chain (4F2HC) enhancer. Neonatal CD-1 and adult SCID mice injected once intramuscularly (i.m.) with 10(7) to 10(9) plaque-forming units (pfu) of this virus displayed significant dose-dependent elevations of serum hEpo levels and increased hematocrits, which were stable over the 4-month time course of these experiments. Adenovirus injected i.m. remained localized at the site of injection and there was no evidence of either systemic infection or a localized inflammatory response. These results suggest that i.m. injection of recombinant replication-defective adenovirus vectors may serve as a paradigm for the treatment of human serum protein deficiencies.