Electromechanical cell lysis using a portable audio device: enabling challenging sample preparation at the point-of-care.

Audio sources are ubiquitously available on portable electronic devices, including cell phones. Here we demonstrate lysis of Mycobacterium marinum and Staphylococcus epidermidis bacteria utilizing a portable audio device coupled with a simple and inexpensive electromagnetic coil. The resulting alternating magnetic field rotates a magnet in a tube with the sample and glass beads, lysing the cells and enabling sample preparation for these bacteria anywhere there is a cell phone, mp3 player, laptop, or other device with a headphone jack.

Identification of the complement regulatory proteins CD46, CD55, and CD59 in human fallopian tube, endometrium, and cervical mucosa and secretion.

PROBLEM: Complement lytic activity has been demonstrated, and a potential for its activation is present in human cervical and tubal secretions and in the endometrium. This necessitates the presence of regulatory mechanisms for protection of the sperm and the implanting allogeneic conceptus in the female genital tract. Complement regulatory proteins demonstrated on sperm and in seminal fluid have been attributed such a role. It is however likely that additional protection is required for a successful conception and implantation to take place. This lead us to investigate the distribution of the complement regulatory factors in cervical mucus and mucosa, uterine endometrium, and fallopian tube. METHOD: Endometrium and cervical mucosa were obtained from patients undergoing hysterectomy for benign conditions, and specimens were selected from different stages of the menstrual cycle. Fallopian tubes were obtained from patients submitted for sterilization, while cervical mucus was aspirated from volunteers undergoing gynecological examination. Immunohistochemistry was performed on all tissue samples, using monoclonal antibodies to membrane cofactor protein (MCP), decay accelerating factor (DAF), CD59 and complement receptor 1 (CR1). Western blot analysis was performed on cervical mucus under nonreducing conditions. RESULTS: MCP, DAF, and CD59 were found to be expressed in human endometrium and fallopian tube. No variation in expression was detected throughout the menstrual cycle. CR1 was not expressed. Soluble forms of DAF and CD59 were found to be present in cervical mucus. CONCLUSION: The complement regulatory proteins MCP, DAF, and CD59 are expressed throughout the female genital tract, and may thus play an important role in protecting the traversing sperm and implanting blastocyst from complement mediated damage.

The localization and concentration of copper in the fallopian tube in women with or without an intrauterine contraceptive device.

The localization and concentration of copper in cryostat sections of human fallopian tubes from women using an intrauterine contraceptive device (IUCD) and controls was determined by light microscopic- and atomic absorption spectrophotometric techniques. The copper was visually accumulated in the epithelium and the copper concentration in the tubal tissue was increased in the IUCD user group. We also measured the concentration of copper and ceruloplasmin in serum, but there was no statistical difference between the two groups studied. Our results indicate that the IUCD has effects beyond the uterine cavity. The accumulation of copper may be associated with earlier observations of the morphological changes and infiltration of inflammatory cells observed in the fallopian tube in IUCD users.

PIP: During laparoscopic sterilization at a hospital in Bergen, Norway, surgeons removed a 1 cm segment of the macroscopic isthmic portion of the fallopian tube from 40 healthy, nonpregnant women. They used light microscopic and atomic absorption spectrophometric techniques to localize and measure the concentration of copper in cryostat sections of the fallopian tubes of 20 women who had used a copper releasing IUD for more than six months and of 20 other women who used neither an IUD nor an hormonal contraceptive. Copper was visually present in the epithelium, but weakly so, in few of the biopsies from the non-IUD users. In IUD users, it accumulated throughout the epithelium and dwindled in the lamina propria. Its mean concentration in the fallopian tube was 58% higher in IUD users than non-IUD users (128.3 vs. 81.4 nmol/mg protein; p = 0.003). IUD use did not affect protein levels. The IUD affected neither the concentration of copper nor of the copper-transporting protein, ceruloplasmin, in serum. These findings suggest that the accumulation of copper in the fallopian tube in IUD users may effect morphological changes and infiltration of inflammatory cells in the fallopian tube.

In situ characterization of leukocytes in the fallopian tube in women with or without an intrauterine contraceptive device.

Histological evaluation of sections from the human fallopian tube revealed an inflammatory reaction in 21 of 31 women using an intrauterine contraceptive device (IUCD) and in four of 29 controls (non-IUCD users). The inflammatory cells were mainly localized at the epithelium-lamina propria interface and at the center of the mucosal folds. The immunohistochemical study revealed leukocytes (CD45+), T lymphocytes (CD3+), T helper cells (CD4+), T suppressor/cytotoxic cells (CD8+), B lymphocytes (CD22+, CD19+), granulocytes, monocytes and null cells (CD11b+) mainly localized at the lamina propria in both groups. T lymphocytes were the predominant cell type, and the ratio between T helper and T suppressor/cytotoxic cells was fairly close to one both in IUCD-users and controls. B lymphocytes were the least frequent cell type identified. In IUCD users, the numbers of the different leukocytes were increased. In both groups, IgA-, IgG- and IgM-positive cells were demonstrated and were predominantly located at the lamina propria of the mucosal folds. The IgA-positive cells dominated in both groups, whereas IgG- and IgM-positive cells were less frequent. Cell positive for IgA, IgG or IgM were significantly increased in number in the IUCD users. The data confirm the presence of an immune system in the normal human fallopian tube and indicate that the IUCD can induce a prominent recruitment of inflammatory cells, with a tubal inflammation as the result. The IUCD may disturb the immunological function of the fallopian tube and its rôle in fertilization.

PIP: In Norway, a 1.5 cm segment of the isthmic portion of the fallopian tube was removed for immunohistochemical and histological analyses from 60 healthy nonpregnant women. Researchers compared data on the 31 women who had used a copper IUD for 2-10 years with data on the remaining 29 women who did not use any IUD for the last year to examine the localization, number, and subtypes of leukocytes in the tube. An inflammatory reaction was present in 68% of the IUD users compared to only 14% of controls (p = 0.005). A chronic inflammatory reaction was the predominant type in both groups (52% for IUD users and 8% in controls). The inflammatory cells were largely found in the epithelium-lamina propria interface and in the center of the mucosal folds. In both groups, leukocytes (CD45+), T lymphocytes (CD3+), T helper cells (CD4+), T suppressor/cytotoxic cells (CD8+), B lymphocytes (CD22+, CD19+), granulocytes, monocytes, and null cells (CD11b+) were chiefly located at the lamina propria. CD3+ cells were the primary cell type in both groups. The ratio of CD4+ and CD8+ was close to one in both groups. CD22+ and CD19+ were relatively rare in both groups. The number of the different leukocytes was significantly greater among IUD users than the controls (e.g., total leukocytes, 1127 vs. 609; p 0.0001). The lamina propria of the mucosal folds housed most IgA, IgG, and IgM positive cells. IgA positive cells were the most frequent cells in both groups. The number of cells positive for IgA, IgG, or IgM were significantly higher in the IUD user group than the control group (e.g., total Ig in endosalpinx, 188 vs. 50; p 0.0001). These findings show that the IUD can incite inflammatory cells, resulting in inflammation of the fallopian tube. Thus, the IUD may interfere with the immunological function of the tube and the tube's role in fertilization.

Altered ciliary substructure in the endosalpinx in women using an IUCD.

Fallopian tube segments were removed from 20 women undergoing sterilization by laparoscopy or laparotomy. Ten of these patients used an intrauterine contraceptive device (IUCD). The other 10 had used neither IUCD nor oral contraceptives and served as controls. The ciliary ultrastructure was examined by light- and transmission electron microscopy. The IUCD users had a significantly reduced ciliary length and less well oriented cilia, as compared with the control group. Also, the proportion of cilia with a ciliary crown structure was significantly smaller in IUCD users than in the non-users. The mechanism that may cause these alterations and their putative consequences are discussed.

Fertility following ectopic pregnancy with special reference to previous use of an intra-uterine contraceptive device (IUCD).

A group of 304 women operated on for ectopic pregnancy were followed up with respect to subsequent reproductive performance. The group was distributed into current users, earlier users and non-users of an intra-uterine contraceptive device (IUCD). Pelvic inflammatory disease (PID) and infertility were more common among non IUCD users. Reproductive performance was statistically significantly better in ever-users of an IUCD than in never-users. The proportions of women wishing to become pregnant and who later gave birth to a live baby in the three groups were 69.2%, 61.4% and 38.0% respectively.

PIP: Pregnancy outcomes of 304 women operated for ectopic pregnancy from 1979-1983 in Bergen Norway were analyzed with regard to prior use of IUDs. It is commonly known that wearing an IUD increases the risk of ectopic pregnancy, and that ectopic pregnancy rates are increasing. The women were classified into those who never used an IUD, those ever using an IUD and those currently using an IUD. The never user group were slightly younger and of lower mean parity. During the study, the percentages of the groups who delivered live babies were 32.5% (69.2% corrected for contraception) among current users, 48.2% (61.14% corrected) among past users and 31% (38%) among never users of IUDs. Combining all pregnancies, miscarriages and legal abortions, the figures were 84.6%, 75% and 42.3%. Thus, fertility after ectopic pregnancy was significantly (p0.0001) better in IUD users than in nonusers. This held for both primigravidae and multigravidae. There was no difference in rates of subsequent ectopic pregnancy. Examining medical records, pelvic inflammatory disease (PID) and infertility were more common in non-IUD users and ex-IUD users. Apparently, the consequences for future fertility or PID were more severe in the group never using an IUD.

Mast cells in the tubal wall in women using an intrauterine contraceptive device.

The number of mast cells in the tubal wall of 33 healthy nonpregnant women, 17 of whom had an intrauterine contraceptive device (IUCD), was investigated. Light microscopy showed that both the muscularis externa and the lamina propria of the tubal wall contained more mast cells in the 17 IUCD users than in the 16 non-users (control group). In both patient groups the mast cell concentration was higher in the muscularis externa than in the lamina propria. Most mast cells of the muscularis externa were more closely related to smooth muscle cells than to blood vessels. The increased number of mast cells in IUCD users may be a factor in the pathogenesis of pelvic inflammatory disease and the ectopic pregnancies that occur in IUCD users.

Morphological changes in tubal mucosa associated with the use of intrauterine contraceptive devices.

We examined the endosalpinx of 28 healthy non-pregnant women, 12 of whom had an intrauterine contraceptive device (IUCD) and 16 of whom had neither an IUCD nor used oral contraceptives. Measurements made on light- and scanning electron micrographs showed that the percentage area covered by ciliated cells was approximately 40% in non-IUCD users and only 20% in IUCD users.

PIP: Given that IUD users have a high risk of ecotopic pregnancy and since most of these pregnancies are tubal, a study was undertaken in Norway to determine if there were any differences in the tubal epithelium of IUD users and women who never used IUDs. The proportion of the epithelium surface covered by ciliated cells was the major focus of the investigation. 1 cm segments from the isthmic section of the oviduct were removed from 12 IUD users and 16 nonusers while they were undergoing sterilization procedures. The women were all healthy and nonpregnant. Both groups of women were similar in terms of average age and parity. The IUDs used by the women included 5 copper Ts, 4 Multiloads, 1 Lippes Loop, and 2 IUDs of unknown type. All of the users had worn the devices for at least 6 months. The specimens of epithelium were immersed in a fixative and stained. The proportion of the epithetial surface covered by ciliated cells was measured using both light and scanning electron micrographs. The results obtained from the 2 methods of measuring differed somewhat for individual specimens; however, the overall results obtained from both methods were similar. Light microscropy measurements indicated that the proportion of epithetial surface covered by ciliated cells was 39.9% for nonuser and 21.2% for IUD users. Respective results obtained by scanning electron microscopy were 38.8% and 20.0%. The results were obtained from women in different menstrual cycle phases. A comparison of specimens by menstrual cycle phase revealed that the proportion of surface covered by ciliated cells was not affected by hormonal changes during the menstrual cycle. IUD users had a slightly higher intraepithelial leucocyte count than the control group. Whether or not reduced ciliation in IUD users plays a role in tubal pregnancy is not known; however, egg transport might be slowed down when there are fewer ciliated cells. Reduced ciliation might also affect sperm migration which in turn might enhance the risk of abnormal embryos. Most tubal pregnancies are characterized by abnormal embryos.