Effect of the method of conception and embryo transfer procedure on mid-gestation placenta and fetal development in an IVF mouse model.

BACKGROUND: Abnormal placentation is a potential mechanism to explain the increased incidence of low birthweight observed after IVF. This study evaluates, in a mouse model, whether the method of conception and embryo transfer affect placentation and fetal development. METHODS: IVF blastocysts (CF1 x B6D2F1/J) were cultured in Whitten's medium (IVF(WM), n = 55) or K modified simplex optimized medium with amino acids (IVF(KAA), n = 56). Embryos were transferred to the uteri of pseudo-pregnant recipients. Two control groups were created: unmanipulated embryos produced by natural mating (in vivo group, n = 64) and embryos produced by natural mating that were flushed from uterus and immediately transferred to pseudo-pregnant recipients (flushed blastocysts, FB group, n = 57). At gestation age 12.5 days, implantation sites were collected and fixed; fetuses and placentas were weighed and their developmental stage (DS) evaluated. Placental areas and vascular volume fractions were calculated; parametric statistics were applied as appropriate. RESULTS: IVF fetuses showed a modest but significant delay in development compared with FB mice (P < 0.05). In addition, IVF conceptuses were consistently smaller than FB (P < 0.05). Importantly, these differences persisted when analyzing fetuses of similar DS. The placenta/fetus ratio was larger in the IVF group (IVF(WM) 0.95; IVF(KAA) = 0.90) than the FB group (0.72) (P < 0.05 for all comparisons). Gross morphology of the placenta and ratio labyrinth/fetal area were equivalent in the IVF and FB groups, as were percentage of fetal blood vessels, maternal blood spaces and trophoblastic components. CONCLUSIONS: In vitro embryo culture affects fetal and placental development; this could explain the lower birthweight in IVF offspring.

Mammary implants: laboratory simulation of recreational diving conditions.

To ascertain whether mammary implants are prone to changes in conformation or structure if they are submitted to recreational dives, eight mammary implants were submitted to 40 simulated dives to imitate an average recreational diving schedule. Matching implants were used as a control group. Photographs were taken before and after completion of the protocol. All implants were observed for changes in volume and checked for integrity. Variations in density were evaluated using a Tc scan. No changes in volume occurred after each dive. None of the implants showed ruptures, and Tc scanning failed to reveal any differences in density between tested and control implants. Cohesive-gel implants submitted to the simulated dives showed some morphological alterations. This study indicates that the mammary implants tested could be implanted in a sports diver, but raises concern about whether the increased exposure to stress could negatively affect their durability.

[The role of hysteroscopy in the follow up of post-mastectomy patients undergoing adjuvant therapy with Tamoxifen]. / Ruolo dell'isteroscopia nel follow-up di pazienti mastectomizzate sottoposte a terapia adiuvante con tamoxifene (TAM).

The authors report on up to date knowledge of the risk of endometrial carcinoma women operated on mastectomy for breast carcinoma and treated with TAM. Starting from their own clinical and scientific experience, the authors follow a group of such patients with a strict monitoring, to ascertain the eventual comparison of dysplastic and neoplastic endometrial pathologies. The group numbers 18 patients and the aim of the study is to evaluate the importance of hysteroscopy as a diagnostic approach for this iatrogenic pathology. The authors affirm the validity of this partially invasive diagnostic method that has to be integrated with clinical and laboratory parameters that are justified by the cost/benefit ratio always favourable for the diagnosis of a neoplastic pathology.

The role of the administration time of prophylactic antibiotic therapy in colorectal cancer surgery: a review of 6,069 patients from 36 randomized clinical trials.

The aim of the present report was to establish the effectiveness of different prophylactic antibiotic regimens and administration times in colorectal cancer surgery. Six thousand and sixty nine patients from 36 selected randomized clinical trials, published between 1980 and 1989, were reviewed. The occurrence of septic events, isolated bacterial strains, fever and postoperative hospitalization times were also analyzed. The therapeutic schedules that included the perioperative administration of antibiotics provided better results that those that did not (p. less than .0001 for infections both specifically related and unrelated to colorectal surgery). The number of postoperative administrations did not affect the clinical results, even if the predominant choice was to give more than one administration of antibiotics. A factorial design demonstrated that prolonging the perioperative administrations up to the postoperative period provided statistically significant benefits (p less than .0001) only with regard to the risk of infections that were not specifically related to colorectal surgery.

The role of natural killer cells in murine cytomegalovirus eye infection.

PURPOSE: To study the role of natural killer cells in the dissemination of virus to the eye and virus growth as well as the production of lesions in the eye during primary infection of murine cytomegalovirus. METHODS: Virus activity and lesion production by murine cytomegalovirus in various organs of natural killer cell-depleted BALB/c strain of inbred mice were compared with those of immunocompetent normal BALB/c mice. RESULTS: In mice injected intraperitoneally with murine cytomegalovirus, virus could be isolated from 50% of eyes in the natural killer cell-depleted group whereas no virus was detected from any eye in the control group. In natural killer cell-depleted mice with positive virus isolation from eyes, no murine cytomegalovirus was isolated from either optic nerves or trigeminal ganglia whereas virus was detected from blood lymphocytes, macrophages, granulocytes, and plasma. After intravitreal injection of murine cytomegalovirus, virus titers in eyes of natural killer cell-depleted mice were significantly higher than those in the control group. Derangement of retinal cell layer and inflammatory cells as well as cytomegalic cells in iris and ciliary body were noted in natural killer cell-depleted group, whereas no such changes were observed in the eyes of the control mice. CONCLUSION: Natural killer cell depletion enhances the dissemination of murine cytomegalovirus to the eye through the hematogenous route, and increases virus multiplication as well as lesion production in the eye.

Can evoked phonomyography be used to recognize fast and slow muscle in man?

The present study is aimed at ascertaining if muscle sound might be used as a detector of the contractile properties of individual human muscles "in vivo". In order to test this hypothesis, Soleus muscle (slow) and Vastus Lateralis Femoris muscle (fast) were investigated in three healthy subjects during electrically elicited contractions. Evoked phonomyograms were obtained from isometric single twitch contractions using a microphonic apparatus. Time and frequency domain analysis were performed. Evoked phonomyogram rising time values obtained from the two muscles are significantly different (p less than .01) and this difference is clearly due to their different mechanical properties. The power spectrum of all signals was obtained by means of harmonic analysis routine and mean frequency thus obtained was taken into account. Power spectrum values are approximately 1.5 times greater in fast muscle than in slow muscle (p less than .01). These findings lead us to the conclusion that evoked phonomyography can be considered a useful technique for the assessment of mechanical properties of individual human muscles.

Protective effect of anti-glycoprotein D antibody on herpetic chorioretinitis in newborn rabbits.

Subcutaneous injection of type 2 herpes simplex virus (HSV-2) (10(3) PFU) to newborn rabbits produced severe skin lesions and wide dissemination of the virus to various organs including the eye. Ocular lesions were characterized by retinal folds and choroiditis. HSV could be isolated from mononuclear cells (MNCs) of infected animal blood. Newborn rabbits treated with monoclonal antibody (MAb) against glycoprotein D (gD) of HSV on days 0, 2 and 4 postinfection had little or no skin lesions (0.0-2.3 mm) compared to controls (2.8-13.0 mm). In addition, the MAb treatment significantly suppressed dissemination of the virus to the eye (0% in MAb-treated vs 83% in control) and other organs and reduced the rate of chorioretinitis (0% in MAb-treated vs 50% in control). The treatment of HSV-infected MNCs with MAb resulted in 91-100% reduction in infectivity of the cells. The results suggest that anti-gD MAb protects newborn rabbits from HSV-2 eye infection by neutralizing the virus in skin and inactivating HSV-infected MNCs in blood.

In vivo reactivation of latent murine cytomegalovirus in the eye by immunosuppressive treatment.

Intravitreal inoculation of 10(3) pfu of murine cytomegalovirus (MCMV) to 3-week-old BALB/c mice resulted in virus isolation from eye homogenates for 2 weeks and from co-cultured specimens of the same eye up to 5 weeks after inoculation, indicating that MCMV in the eye became latent 2 weeks after the virus inoculation. Immunosuppressive treatment with daily intramuscular injections of cyclosporine (40 micrograms/g) and cortisone acetate (125 micrograms/g) 9 weeks after intravitreal MCMV inoculation resulted in isolation of infectious virus from ten of 44 eye homogenates (10 of 22 mice) during a 3-week period, indicating in vivo reactivation of latent ocular MCMV. No virus was isolated from eye homogenates of similarly infected mice with sham immunosuppression (daily intramuscular saline injections), nor was any virus isolated from uninfected eyes with immunosuppressive treatment. Three weeks of daily cyclosporine and cortisone injections depleted L3T4+ cells to 6.0%, Lyt-2+ cells to 20% and anti-MCMV antibody to 10% of untreated mice. The results suggest that the eye can serve as a site of latent MCMV that can be reactivated by immunosuppressive means.

[Cardiovascular effects of isometric contraction of the masticatory muscles in humans]. / Effetti cardiovascolari della contrazione isometrica dei muscoli masticatori nell'uomo.

The cardiovascular responses to static exercise do not include, until now, studies about masticatory muscles. In this work the cardiocirculatory effects of the isometric contraction of masticatory muscles were compared with those evoked by the handgrip exercise. Non invasive and, in one case, invasive (aortic catheterism) monitoring of arterial blood pressure was performed. The findings show that mean blood pressure and especially heart rate do not increase during static exercise of masticatory muscles as reported in the literature for other muscular groups: it is suggested that the peculiar architecture of masticatory muscles may explain the absence of significant cardiovascular response to their isometric contraction.

Induction of Fc and C3b receptors on rabbit corneal cells by herpes simplex virus.

The expression of receptors for Fc portion (FcR) of immunoglobulin G (IgG) and for a C3b component of complement (C3bR) by herpes simplex virus (HSV) was studied in primary cultures of rabbit corneal cells. Monolayer cultures of epithelial, stromal and endothelial cells of the rabbit cornea were infected with three strains each of type 1 (HSV-1) and type 2 HSV (HSV-2). Rosette methods were used to detect receptors by means of sheep erythrocytes sensitized with rabbit IgG for FcR and C3b-coated sheep erythrocytes for C3bR. The FcR were expressed regularly on epithelial, stromal and endothelial cells by all three strains of both HSV-1 and HSV-2. The C3bR, however, were expressed only by HSV-1 on epithelial and stromal cells. Little or no C3bR activities could be detected on endothelial cells infected with any strain of HSV-1 or HSV-2. The FcR and C3bR expressed on corneal cells were induced by HSV and were blocked by monoclonal antibody to HSV-1 glycoprotein E(gE) or glycoprotein C(gC) respectively, confirming findings of other investigators that gE acts as FcR and gC as C3bR.

Ocular toxoplasmosis in immunosuppressed nonhuman primates.

To investigate the role of cellular immunodeficiency in recurrent toxoplasmic retinochoroiditis, six Cynomolgus monkeys (Macaca fascicularis) with healed toxoplasmic lesions of the retina were immunosuppressed by total lymphoid irradiation. Three months prior to irradiation 30,000 Toxoplasma gondii organisms of the Beverley strain had been inoculated onto the macula of eye in each monkey via a pars plana approach. Toxoplasmic retinochoroiditis developed in each animal, and lesions were allowed to heal without treatment. During total lymphoid irradiation animals received 2000 centigrays (cGy) over a 7-week period. Irradiation resulted in an immediate drop in total lymphocyte counts and decreased ability to stimulate lymphocytes by phytohemagglutinin. Weekly ophthalmoscopic examinations following irradiation failed to show evidence of recurrent ocular disease despite persistent immunodeficiency. Four months after irradiation live organisms were reinoculated onto the nasal retina of the same eye in each animal. Retinochoroidal lesions identical to those seen in primary disease developed in five of six animals. Toxoplasma organisms therefore were able to proliferate in ocular tissue following the administration of immunosuppressive therapy. This study fails to support the hypothesis that cellular immunodeficiency alone will initiate recurrent toxoplasmic retinochoroiditis. Results suggest that reactivation of disease from encysted organisms involves factors other than suppression of Toxoplasma proliferation. If reactivation occurs by other mechanisms, however, cellular immunodeficiency then may allow development of extensive disease.

Protective effect of passive immunization on herpetic retinitis of newborn rabbits.

We studied the protective effects of passive immunization with virus specific antibody in newborn rabbits inoculated subcutaneously with type 2 herpes simplex virus (HSV-2). Newborn rabbits given anti-HSV-2 antibody intraperitoneally (IP) on days 0, 2 and 4 post infection had smaller herpetic skin lesions and reduced mortality when compared to controls. In addition, the IP treatment using this schedule reduced virus growth in the skin lesions and virus dissemination, so that it decreased the frequency of herpetic retinitis. When the IP antibody administration was started at 24 hours post virus inoculation, according to the schedule days 1, 3 and 5, there was less protection; larger skin lesions, higher mortality, and greater evidence of virus dissemination. Also HSV-infected mononuclear cells (MNCs) treated with anti-HSV serum resulted in a significant reduction in the number of infected MNCs. The results of these studies suggest that anti-HSV-2 antibody contributes to protection against HSV-2 infection of skin as well as eyes, probably by inactivation of the virus locally at the skin inoculation site, and by combating the hematogenous spread of HSV-infected MNCs as well as free virus to various organs including the eye.

Role of lymphocyte and antibody in the pathogenesis of immune-mediated herpetic uveitis.

We investigated the role of various immune components in the pathogenesis of immune-mediated experimental herpetic uveitis. Inbred III/J strain of rabbits were sensitized with an intravitreal injection of 10(3) PFU of type 1 herpes simplex virus (HSV), and sensitized cervical lymph node (LN) cells were obtained on postinfection day 12. Intravitreal injection to the normal III/J rabbit eye of HSV antigen with either sensitized LN cells or anti-HSV serum failed to induce uveitis, whereas intravitreal injection of HSV-antigen with both sensitized LN cells and anti-HSV serum produced severe uveitis within six hours. The combination of sensitized LN cells, HSV-antigen and normal rabbit serum, or that of normal LN cells, HSV antigen and anti-HSV serum, did not induce uveitis. Further studies using B lymphocyte and T lymphocyte fractions from sensitized LN showed that only the combination of sensitized T lymphocytes, HSV antigen and anti-HSV serum regularly produced uveitis following intravitreal injection. These results indicate that the interaction of HSV antigen, sensitized T lymphocytes and anti-HSV antibody may play a role in the pathogenesis of immune-mediated herpetic uveitis.

Suppression of secondary herpes simplex uveitis by cyclosporine.

The authors studied the effect of an immunosuppressive agent, cyclosporine (CyA), on experimental secondary herpes simplex (HS) uveitis. Secondary HS uveitis was induced in a rabbit eye that had recovered from primary HS uveitis by challenging it with an intravitreal injection of herpes simplex virus (HSV) antigen. Daily intramuscular injections of CyA (25 mg/kg body weight) for 7 days prior to the intravitreal challenge with HSV antigen significantly suppressed the induction of secondary HS uveitis, but daily injections of CyA after the challenge with HSV antigen was ineffective. Intravitreal injections of CyA (5 mg) 7 days and 3 days prior to the HSV challenge were less effective, but the combined treatment with seven daily intramuscular CyA and two intravitreal CyA injections prior to the HSV challenge was most effective in the prevention of the uveitis. The daily intramuscular treatment with CyA resulted in a marked reduction of cell-mediated immunity while leaving the level of circulating HSV specific antibody high. No reactivation of latent HSV was detected in trigeminal and superior cervical ganglia of CyA-treated rabbits.

The presence of cytotoxic autoantibody to lacrimal gland cells in NZB/W mice.

New Zealand Black and White F1 hybrid mice (NZB/W mice) spontaneously develop an autoimmune disease which provides us with a suitable animal model for Sjögren's syndrome. With increasing age, these mice develop foci of mononuclear cell infiltration in the lacrimal and salivary glands, which closely resemble the lesions seen in patients with Sjögren's syndrome. We studied the cell-mediated and antibody-mediated immune responses of NZB/W mice to lacrimal gland cells. Lacrimal gland acinar cells were isolated from 2-month-old NZB/W or BALB/c mice for the target of 51Cr-release assay. There was no statistically significant difference in the spleen cell-mediated cytotoxicity to lacrimal gland cells among NZB/W mice of different ages (2, 5, and 8 months old). With increasing age, on the other hand, we found a statistically significant increase in the titers of autoantibodies to lacrimal gland cells in NZB/W mice, while aged BALB/c mice did not develop such antibodies. Fractionation of pooled positive sera by gel filtration revealed that this cytotoxic activity was mostly recovered in the IgM fraction. The tissue absorption study showed that these antibodies cross-reacted with salivary gland and kidney.

Different susceptibilities of skin to type 1 and type 2 herpes simplex viruses in newborn rabbits.

Skin infections with type 1 herpes simplex virus (HSV-1) were compared with skin infections with type 2 virus (HSV-2). Five strains each of HSV-1 and HSV-2 were tested by injecting 10(3) 50% tissue culture infective doses of each strain subcutaneously into 1-day-old New Zealand white rabbits. All five strains of HSV- 2 produced severe skin lesions that resulted in wide dissemination of the infection to many organs, paralysis of the hind legs, and finally death. The virus could be isolated frequently from skin lesions, from various organs (liver, lungs, adrenal glands, brain, and eyes), and from circulating leukocytes and plasma. In contrast, all five strains of HSV-1 failed to produce significant skin lesions or dissemination of virus, only half of the skin lesions yielded HSV, and no virus could be isolated from the blood. These results indicate that HSV-1 dose not grow well in the skin of newborn rabbits and fails to disseminate, whereas HSV-2 is dermatotropic and disseminates readily to many organs by hematogenous routes.