Influence of the glass packing on the contamination of pharmaceutical products by aluminium. Part III: Interaction container-chemicals during the heating for sterilisation

The interaction of chemicals with the container materials during heating for sterilisation was investigated, storing the components of parenteral nutrition solutions individually in sealed glass ampoules and in contact with a rubber stopper, and heating the system at 121 degrees C for 30 min. Subsequently, the aluminium content of the solutions was measured by atomic absorption spectrometry (AAS). The assay was also carried out with acids, alkalis and some complexing agents for Al. The containers were decomposed and also assayed for aluminium. 30 different commercial solutions for parenteral nutrition, stored either in glass or in plastic containers, were assayed measuring the aluminium present in the solutions and in the container materials. The results of all investigated container materials revealed an aluminium content of 1.57% Al in glass, 0.05% in plastic and 4.54% in rubber. The sterilisation procedure showed that even pure water was able to extract Al from glass and rubber, 22.5 +/- 13.3 microg/L and 79.4 +/- 22.7 microg/L respectively, while from plastic the aluminium leached was insignificant. The Al released from glass ampoules laid between 20 microg/L for leucine, ornithine and lysine solutions and 1500 microg/L for solutions of basic phosphates and bicarbonate; from rubber stoppers it reached levels over 500 microg/L for cysteine, aspartic acid, glutamic acid and cystine solutions. Ion-exchange properties and influence of pH can explain the interaction of glass with some chemicals (salts, acids and alkalis), but only an affinity for aluminium could explain the action of some amino acids and other chemicals, as albumin and heparin, on glass and rubber, considering the aluminium release. Experiments with complexing agents for Al allowed to conclude that the higher the stability constant of the complex, the higher the Al release from the container material.     

Fungal growth inside saline-filled implants and the role of injection ports in fungal translocation: in vitro study

Infection is a serious complication of breast augmentation and tissue expansion with inflatable devices. Several reports have shown that fungi may be able to survive, colonize, and even cause infection in saline-filled devices. The mechanism of how they penetrate, spread, and colonize inside the inflatable implants is not exactly understood. The authors assessed both the expander membrane and the port in terms of leakage and penetration of Candida albicans and Aspergillus niger in an in vitro model. Thirty saline-filled expanders connected to the injection port were placed in sterile containers filled with tryptic soy broth culture medium to simulate the clinical situation in phases I and II. Intact and multipunctured ports were used in the first and second phases of the study, respectively. Either the container or the implant was inoculated with one of these fungi, and six implants in containers without fungal inoculation served as controls. As a third phase, intraluminal survival of fungi was investigated in saline-filled containers (n = 12) in 21 days. The silicone membrane, with its intact connecting tube and port, was impermeable to these fungi, whereas both fungi were able to diffuse inside-out or outside-in through the punctured ports. C. albicans did not survive beyond 18 days in saline, whereas A. niger continued to multiply at day 21. Chemical analyses of the implant fluids revealed that the contents of the culture medium diffused into the implants in phases I and II. The data show that an intact silicone membrane is impermeable to fungi, and punctured ports allow translocation of fungi into the implants. Fungi can grow and reproduce in a saline-only environment, and their survival periods differ among the species. Furthermore, their survival may be enhanced by the influx of substances through the implant shell.     

A simple technic for repeated collection of blood samples from mice.

A device for repeated collection of small blood samples from mice was constructed from a plastic syringe. Blood was collected into a 3.33 lambda capillary tube. Bleeding was stopped by a hemostat made from a rubber stopper. This technic allows easy collection of approximately 20 serial samples within an 8-hr period.     

Loss of vacuum in rubber stoppered vials stored in a liquid nitrogen vapor phase freezer

A total of 1885 rubber stoppered 3-ml vials (NS-33 and NS-51 glass (expansion sizes), sealed under vacuum, were subjected to the ultracold temperatures of a liquid nitrogen vapor phase (LNVP) freezer for 16.5 hr and tested for vacuum loss after equilibration back to 25 °C. The presence of a vacuum in each vial was checked before and after freezing. Results with two lots of vials per glass expansion size showed positive or no negative pressure rather than a vacuum in 46.25% (NS-33), 98.25% (NS-33), 64.72% (NS-51), and 99.38% (NS-51) of the vials. Apparently, the fit or slight misfit of the stopper into the vial opening and the hardness of the butyl rubber stopper at ultracold temperatures enhance the chances of leakage around the stopper area under extremely cold temperatures. Storage of rubber stoppered vials containing lyophilized products sealed under vacuum in LNVP freezers is not advisable.     

Multiplication in liquid medium of Treponema sp. isolated from intestinal contents of swine

Treponema hyodysenteriae and Treponema innocens were multiplied by a simple culture method in liquid medium. TSB medium was prepared by the PRAS method in plasma bottles containing glass beads. Spirochaetes were injected through the rubber stopper and the bottles were incubated while revolving round their axes. The most abundant growth of spirochaetes in rotary culture was observed after 72 h incubation at 40 degrees C. whereas the highest number of viable cells in stationary culture was observed after 120 h. However, in the latter case the number of cells was lower than introduced at inoculation. Growth of the bacteria was stimulated by equine serum and 5% addition of rumen fluid. Optimal growth temperature was 40 degrees C.     

影响生物制品冻干粉针剂水分的探讨

探讨生物制品冻干粉针剂样品放置一时间后残余水分增高的原因。进行了水分测定,真空度检测,二甲硅油和丁基橡胶药用瓶塞干燥失重的检测。冻干后每只丁基橡胶药用瓶塞平均含水分0.00224g。结果表明丁基橡胶药用瓶塞灭苗,干燥和冻干过程中去除水分不彻底是引起样品中水分升高的直接原因。     

Exogenous bacterial contamination of donor blood.

The Canadian Red Cross blood transfusion service has followed a set protocol for phlebotomy and collection of a unit of blood. Recent requirements for automated testing have necessitated that a second tube of blood be obtained from the blood line following collection of the unit. Evaluation of the techniques used, however, has indicated the possibility of bacterial contamination from the skin of donors, from insertion of the needle through an unsterile rubber stopper, and through backflow from a nonsterile vacuum tube. To test these possibilities swabs were taken from skin and stoppers of vacuum tubes. Further, vacuum tubes were deliberately contaminated with Escherichia coli. The normal sampling procedure, which involves stripping the donor line to refill and mix the blood, was then followed. This resulted in contamination of the segments and even the blood bag. These findings led to modification of the standard bleeding technique, whereby stripping was eliminated and sterile vacuum tubes were to be used at all times.     

Spectrométrie de masse et hémocultures: un nouvel outil de diagnostic immédiat

Delay in identification of microorganisms hamper the prompt initiation of adequate empirical antibiotic therapy of bacteremia. MALDI-TOF-MS allows identification of microorganisms directly from colonies within minutes. Identification of microorganisms responsible for bacteremia can be made directly from blood culture bottles. Many different protocols have been published, all based on the separation of microorganisms from erythrocytes and blood culture broth that alter the quality of spectra. Correct identifications were obtained for most of the procedures used in at least 80% of cases, within 20 to 60 min once the blood culture is detected positive. MALDI-TOF-MS, by far the fastest identification technique, offers a significant breakthrough in the management of bacteremia.     

Cytologic diagnosis by transthoracic fine needle sampling without aspiration

The cytologic findings of transthoracic fine needle sampling without aspiration (fine needle capillary [FNC] sampling) are reported. Eleven patients were examined by FNC sampling while four were examined by the classic fine needle aspiration (FNA) method. In contrast to FNA sampling, FNC sampling produced less patient trauma and admixture of the sample with blood, while giving a better perception of the tumor and its consistency. The quality of the samples obtained by the FNC technique was equal to that of the FNA samples. The results demonstrate that fine needle sampling without aspiration may be used in the study of deep-seated as well as of superficial organs.     

一项小鼠采血新技术——颌下静脉丛采血法

目的介绍小鼠颌下静脉丛采血方法。方法选用2月龄昆明小鼠,固定小鼠后,将采血注射针头刺入颌下静脉丛血管取血。结果单人操作约1 min内可完成小鼠颌下静脉丛采血,采血量达到0.3-0.5 mL。结论此方法无需麻醉、创伤小、采血量大并可重复采血,是一种简便、安全、快速、采血量多的采血方法。     

A microbial sampler for freshwaters

A freshwater sampler using five sterile evacuated glass tubes is described. Water enters when a rubber stopper is mechanically removed from the end of a sterile hypodermic needle inserted into each tube. Plate counts of bacterial colonies were compared with those obtained with other samplers.     

A practical device for pinpoint delivery of molecules into multiple neurons in culture

We have developed a device for pinpoint delivery of chemicals, proteins, and nucleic acids into cultured cells. The principle underlying the technique is the flow of molecules from the culture medium into cells through a rupture in the plasma membrane made by a needle puncture. DNA transfection is achieved by stabbing the needle tip into the nucleus. The CellBee device can be attached to any inverted microscope, and molecular delivery can be coupled with conventional live cell imaging. Because the position of the needle relative to the targeted cultured cells is computer-controlled, efficient delivery of molecules such as rhodamine into as many as 100 HeLa cells can be completed in 10 min. Moreover, specific target cells within a single dish can be transfected with multiple DNA constructs by simple changes of culture medium containing different plasmids. In addition, the nano-sized needle tip enables gentle molecular delivery, minimizing cell damage. This method permits DNA transfection into specific hippocampal neurons without disturbing neuronal circuitry established in culture.     

长白山阔叶红松林凋落物组成及其季节动态

为了解群落尺度上凋落物组成及其时空变化,对长白山阔叶红松林25 hm² 样地内2008年度收集的凋落物进行统计分析.结果表明: 一年间,150个收集器内收集的凋落叶分属35种树木,占样地内胸径≥1 cm树种数(52种)的67.3%;凋落物总量为29.39 kg,折合3918.4 kg·hm^-2,其中,阔叶、杂物、针叶和枝条凋落量分别占凋落物总量的61.7%、18.0%、11.7%和 8.6%;紫椴、水曲柳、蒙古栎、色木槭和春榆5个树种的叶凋落量占阔叶总凋落量的83.8%;不同树种的凋落量季节差异很大,61.9%的凋落物产生于9月13日至10月10日.其中,红松和紫椴叶凋落高峰出现在9月13-26日,蒙古栎、春榆和色木槭叶凋落高峰出现在9月27日-10月10日.收集器间凋落物量差异较大,其中68个收集器的年凋落量在150～200 g,1个收集器大于500 g;单个收集器全年最多可收集到18个树种的凋落叶,凋落叶种数为12种的收集器最多(32个).叶凋落量与样地内该树种的胸高断面积总和成正比.样地内凋落叶的分布存在明显的空间异质性,且收集器内凋落物的收集量与其所处的位置有关.     

The determination of acetaldehyde in blological samples by head-space gas chromatography

A method for the determination of acetaldehyde (AcH) in biological samples by head-space gas chromatography is presented. Human venous blood (antecubital), rat blood (heart-punctured) rat liver (freeze-clamped), and rat and mouse brain (freeze-clamped) were used as the biological samples. The method involves two steps, in the first of which the samples are deproteinized with perchloric acid (PCA). Rat blood can, alternatively, be hemolyzed with water. In the second step, the deproteinized supernatant or hemolyzed blood is pipetted into a serum bottle, which is sealed with a rubber stopper and brought to 65°C in a sampling turntable. Head-space samples are then automatically taken for GLC analysis. Ethanol causes a nonenzymatic formation of AcH in the PCA supernatants of liver and brain, which can be inhibited by the use of thiourea. This reaction is minor in the blood supernatants and cannot be inhibited there by thiourea. The method described measures the total AcH content without regard to any binding. Some of the AcH in rat blood was shown to be bound.     

Use of oxygen-permeable silicone rubber pouches for growing mass cultures of bacteria

Growth of most bacteria often involves the use of expensive incubated shaker systems. In this report, oxygen-permeable silicone rubber pouches, with oxygen permeability over 100 times higher than other polymers, were employed for growing bacterial cultures. With little, if any, agitation oxygen-permeable silicone rubber pouches produced bacterial growth rates equivalent to growth rates obtained in shaker flasks. The silicone rubber pouch described has a glass cuvette integrated into its design that permits readings of bacterial density without opening the pouch. One can sterilize and store powdered bacterial culture medium in silicone rubber pouches ; therefore, bacterial cultures can be initiated by simply adding water and bacteria.     

A simple modification of the Baermann method for diagnosis of strongyloidiasis

The diagnosis of Strongyloides stercoralis infections is routinely made by microscopic observation of larvae in stool samples, a low sensitivity method, or by other, most effective methods, such as the Baermann or agar culture plate methods. We propose in this paper a practical modification of Baermann method. One hundred and six stool samples from alcoholic patients were analyzed using the direct smear test, agar culture plate method, the standard Baermann method, and its proposed modification. For this modification the funnel used in the original version of the method is substituted by a test tube with a rubber stopper, perforated to allow insertion of a pipette tip. The tube with a fecal suspension is inverted over another tube containing 6 ml of saline solution and incubated at 37 degrees C for at least 2 h. The saline solution from the second tube is centrifuged and the pellet is observed microscopically. Larva of S. stercoralis were detected in six samples (5.7%) by the two versions of the Baermann method. Five samples were positive using the agar culture plate method, and only in two samples the larva were observed using direct microscopic observation of fecal smears. Cysts of Endolimax nana and Entamoeba histolytica/dyspar were also detected in the modification of Baermann method. Data obtained by the modified Baermann method suggest that this methodology may helps concentrate larvae of S. stercoralis as efficiently as the original method.     

育苗容器规格对橡胶树籽苗芽接苗生长的影响

采用不同规格控根育苗容器对橡胶树籽苗芽接苗进行悬空和近地培养,观测其生长情况。结果表明,悬空培养对橡胶树籽苗芽接苗的纵向伸长(株高、主根长度)影响较为显著。育苗容器的高度一致时,体积越大的育苗容器,根系伸展空间越多,苗木地上部分长势越好;体积一致时,高度越高的育苗容器,苗木主根长度越长,但对籽苗地上部分及主根粗度影响较小。     

Effect of climate and type of storage container on aflatoxin production in corn and its associated risks to wildlife species

The effects of grain storage containers on aflatoxin production, and the relationship between the level of aflatoxin and the number and weight of fluorescing kernels were determined in corn (Zea maize) stored in controlled climate regimes. Two hundred and forty 100-g samples were held up to 3 mos using four types of storage containers placed in four climates. Storage containers included corn placed in metal cans, paper bags, plastic bags, and paper bags placed in plastic bags. Climates were constant during the duration of the project and included a combination of temperatures and humidities. Temperatures were 29-32 C and 14-18 C; relative humidities were 85-88% and 35-40%. In addition, corn was exposed to environmental conditions conductive for aflatoxin production and 100 g samples were randomly collected, examined under ultraviolet light for fluorescence, and then quantified for aflatoxin levels. Corn samples tested negative for aflatoxin at the beginning of the project. Main (i.e., container, climate, and month) and interactive effects were not observed. Mean levels of aflatoxin ranged from 0 to 151 microg/kg. Aflatoxin was produced regardless of type of storage container, time of storage, and climatic conditions; however, only 8% of the samples produced aflatoxin levels that exceeded 50 microg/kg. Fluorescing corn ranged from 0 to 19 kernels per sample, while aflatoxin levels ranged from 0 to 1,375 microg/kg for the same samples. No relationships were found between the number and weight of fluorescing kernels of corn and aflatoxin levels. The black light test yielded a false negative rate of 23% when in fact the aflatoxin concentrations exceeded 50 microg/kg. Therefore, quantifying fluorescing grain under UV light should not be considered a feasible alternative for aflatoxin testing of grain intended for wildlife.     

Evaluation of bacteremia in a pediatric intensive care unit: epidemiology, microbiology, sources sites and risk factors

Bacteremia is a common cause of morbidity and mortality in children treated in pediatric intensive care unit (PICU). We have investigated the causative agents of bacteremia in our PICU over a one-year period, to determine mortality associated with such infection and identify the dependent predictors for morbidity and mortality. From 1 January till 31 December 2006, 479 patients were admitted in the PICU and 379 blood culture samples were taken. Samples were incubated in the BACTEC 9050 System, and isolates identified by routine microbiological methods. A pair of samples taken for aerobic and anaerobic culture were statistically regarded as one sample. Data collected from the medical records of each patient were recorded onto standardized collections sheets and included demographic information, predisposing conditions, source(s) of infection, important clinical and laboratory parameters at the time of infection, and microbiological data. Based on these data, positive blood cultures were classified as either contaminants or true bacteremias. During a year period, 117 episodes of bacteremia were documented in 72 patients. The most frequent isolates were the coagulase-negative staphylococci 32.2% (39), followed by Candida spp. 30.5% (36). The mean white blood cell count (WBC) on the day of bacteremia was 15.2 x 10(9)/L (range 0.1-48.0 x 10(9)/L), and 3.3% of episodes occurred in neutropenic (WBC count < 1 x 10(9)/L) children. The mean temperature on the day of infection was 38.2 +/- 1.1 degrees C (range, 34-41 degrees C). Some newborns 23% (n = 5) had a significantly lower mean temperature (p < 0.02) and lower mean WBC count (p < 0.05) than older children. Hemodynamic instability was noted in 11% of bacteremic episodes. Among all bacteremias, intravascular catheters were implicated in 22.6%, pneumonia in 20.4%, genitourinary tract in 14.2%, surgical wounds in 11.7% and, gastrointestinal tract in 9.8%. Seven patients died because of sepsis. Early diagnosis, prompt blood culture reports, followed by appropriate antibiotic treatment is essential in reducing mortality in such patients. Short hospital stay and restricted use of invasive devices should be the aims to reduce the risk of bacteremia during the stay in the PICU.     

Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry improves appropriateness of antibiotic treatment of bacteremia

Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows the identification of microorganisms directly from positive blood culture broths. Use of the MALDI-TOF MS for rapid identification of microorganisms from blood culture broths can reduce the turnaround time to identification and may lead to earlier appropriate treatment of bacteremia. During February and April 2010, direct MALDI-TOF MS was routinely performed on all positive blood cultures. During December 2009 and March 2010 no direct MALDI-TOF MS was used. Information on antibiotic therapy was collected from the hospital and intensive care units' information systems from all positive blood cultures during the study period. In total, 253 episodes of bacteremia were included of which 89 during the intervention period and 164 during the control period. Direct performance of MALDI-TOF MS on positive blood culture broths reduced the time till species identification by 28.8-h and was associated with an 11.3% increase in the proportion of patients receiving appropriate antibiotic treatment 24 hours after blood culture positivity (64.0% in the control period versus 75.3% in the intervention period (p0.01)). Routine implementation of this technique increased the proportion of patients on adequate antimicrobial treatment within 24 hours.