Syringe loading introduces macromolecules into living mammalian cell cytosol.
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We describe a simple, efficient, gentle and inexpensive technique for the introduction of normally impermeant macromolecules into the cytosol of living mammalian cells growing in suspension or attached to the culturing substratum. Loading is achieved by the production of transient, survivable plasma membrane disruptions as cells are passed back and forth through a standard syringe needle or similar narrow orifice. The loading volume required, which contains cells and the macromolecule to be loaded, can be as little as 5 microliters, thus minimizing the use of valuable reagents. In addition, we report that the surfactant molecule, Pluronic F-68, is capable of altering the physical properties of the plasma membrane in such a way as to increase loading efficiency and the long-term survivability of cells loaded by this and other mechanically based cell-loading techniques. (+info)
A simple method for bone-graft handling during spine surgery.
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Morsellized bone-graft handling during spine surgery to get vertebral fusion can be a slow, laborious and time-consuming procedure. It is not absolutely exempt from complication risk. An easy, quick and inexpensive alternative technique is described. (+info)
Optimum method for administration of biosynthetic human growth hormone: a randomised crossover trial of an Auto Injector and a pen injection system.
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The use of optimum conventional growth hormone administration, using a growth hormone vial combined with an Auto Injector, was compared with a pen injection system using a cartridge of growth hormone. In both methods of administration the concentration of growth hormone was 16 IU/ml. Thirty patients (22 boys, eight girls) who had all previously been treated with growth hormone (4 IU/ml) administered using needles and syringes (without an Auto Injector) were randomised into receiving one of either treatment for three months and then crossed over for a further three months. Fourteen patients (10 boys, four girls) initially received KabiVial 16 IU/ml combined with an Auto Injector while 16 patients (12 boys, four girls) were treated with KabiPen 16 IU/ml. Mean age in both groups was 9.6 years. The majority of patients in both groups were treated with a regimen of either 15 or 20 units/m2/week as a daily subcutaneous injection. Of the 30 patients who started in this trial, two who commenced using an Auto Injector refused to change to a pen system and were excluded from further analysis. When scored on a scale of -5 to +5 general convenience when changing from an Auto Injector to the KabiPen decreased from +4.7 to +1.0. When assessed for pain, the Auto Injector group scored +4.7, which decreased to -0.2 (more painful) for the pen. At the end of the trial 23 patients (82%) chose to continue with the KabiVial/Auto Injector combination as they found this less painful and the child did not see the needle or need to insert the needle manually. Five patients (18%) continued with the KabiPen as they considered the device smaller and easier to use. The accuracy of dosing using KabiVial was 100% compared with the range of 88% to 111% using KabiPen as the latter was available only in 0.5 unit increments. No growth hormone was wasted using KabiVial, although a mean of 0.6 units was wasted with every 16 IU cartridge in the KabiPen system. It is concluded that patients should be able to contribute to the choice of growth hormone delivery systems and that newer methods need careful assessment. (+info)
Incidence and risk factors of HCV and HIV infections in a cohort of intravenous drug users in the North and East of France.
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In order to evaluate the incidence and risk factors of infection by hepatitis C virus (HCV) among injecting drug users (IDUs), we conducted a prospective cohort study of HCV- and human immunodeficiency virus (HIV)-negative IDUs in the North and East of France. A total of 231 HCV and HIV IDUs who had injected drugs at least once in their lifetime were followed up every 3 months over a 12-month period. Serum anti-HCV and anti-HIV were tested at inclusion in the study and at the end of the follow-up. Data on injecting practices were collected at inclusion and at each visit. Of the 231 participants included, 165 (71.4%) underwent a final HCV and HIV serum test. The incidence was nil for HIV infection and 9/100 person-years (95% CI 4.6-13.4) for HCV infection. In a multivariable analysis, we found that syringe and cotton sharing were the only independent predictive factors of HCV seroconversion. (+info)
The glycan-rich outer layer of the cell wall of Mycobacterium tuberculosis acts as an antiphagocytic capsule limiting the association of the bacterium with macrophages.
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Mycobacterium tuberculosis, the causative agent of tuberculosis, is a facultative intracellular pathogen that infects macrophages and other host cells. We show that sonication of M. tuberculosis results in the removal of material from the surface capsule-like layer of the bacteria, resulting in an enhanced propensity of the bacteria to bind to macrophages. This effect is observed with disparate murine and human macrophage populations though, interestingly, not with freshly explanted alveolar macrophages. Enhanced binding to macrophages following sonication is significantly greater within members of the M. tuberculosis family (pathogens) than within the Mycobacterium avium complex (opportunistic pathogens) or for Mycobacterium smegmatis (saprophyte). Sonication does not affect the viability or the surface hydrophobicity of M. tuberculosis but does result in changes in surface charge and in the binding of mannose-specific lectins to the bacterial surface. The increased binding of sonicated M. tuberculosis was not mediated through complement receptor 3. These results provide evidence that the surface capsule on members of the M. tuberculosis family may be an important virulence factor involved in the survival of M. tuberculosis in the mammalian host. They also question the view that M. tuberculosis is readily ingested by any macrophage it encounters and support the contention that M. tuberculosis, like many other microbial pathogens, has an antiphagocytic capsule that limits and controls the interaction of the bacterium with macrophages. (+info)
A multistate trial of pharmacy syringe purchase.
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Pharmacies are a potential site for access to sterile syringes as a means for preventing human immunodeficiency virus (HIV), but the type and extent of their utility is uncertain. To examine pharmacy syringe purchase, we conducted a standardized, multistate study in urban and rural areas of four states in which attempts to purchase syringes were documented. Of 1,600 overall purchase attempts, 35% were refused. Colorado (25%) and Connecticut (28%) had significantly lower rates of refusal than Kentucky (41%) and Missouri (47%). Furthermore, urban settings had higher rates of refusal (40%) than rural settings (31%, P < .01). Race and gender did not have a consistent impact on rates of refusal. Despite potential advantages of pharmacies as sites for access to sterile syringes, pharmacy purchase of syringes faces significant obstacles in terms of the practices in different jurisdictions. (+info)
Legal needle buying in St. Louis.
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This study sought to determine if and why barriers to the over-the-counter purchase of syringes in the St. Louis metropolitan area might exist, given that no ordinance prohibits such a sale there. Two male research assistants (one African American, one White) approached 33 of the area's pharmacies to buy syringes. In 14 of those pharmacies, either the purchase was refused or the minimum number of syringes that could be bought was so large (at least 100) that the sale was not practical. Racial bias in rates of refusal and implications for prohibiting or restricting legal availability of syringes are discussed. (+info)
Anesthetic efficacy of the anterior middle superior alveolar (AMSA) injection.
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The purpose of this prospective, randomized, blinded study was to determine the anesthetic efficacy of the anterior middle superior alveolar (AMSA) injection using the computer-assisted Wand Plus injection system versus a conventional syringe. The authors, using a crossover design, randomly administered in a blind manner 2 AMSA injections utilizing the computer-assisted injection system and a conventional syringe to 40 subjects during 2 separate appointments. A pulp tester was used to test for anesthesia, in 4-minute cycles for 60 minutes, of the central and lateral incisors, canine, and first and second premolars. Anesthesia was considered successful when 2 consecutive no responses (80 readings) with the pulp tester were obtained. For all teeth, except the central incisor, the use of the computer-assisted injection system was significantly (P < .05) more likely to result in pulpal anesthesia than the use of the conventional syringe technique. For the computer-assisted injection system, successful pulpal anesthesia ranged from 35 to 58%, and for the conventional syringe, successful pulpal anesthesia ranged from 20 to 42%. For both techniques, the onset of pulpal anesthesia was slow, and duration of pulpal anesthesia declined steadily over 60 minutes. We conclude that although the AMSA injection using the computer-assisted injection system was more successful than the conventional syringe technique, the rather modest to low success rates, slow onset, and declining duration of pulpal anesthesia over 60 minutes would not ensure predictable pulpal anesthesia from the second premolar to the central incisor. (+info)