Maternal caloric restriction prior to pregnancy increases the body weight of the second-generation male offspring and shortens their longevity in rats.

Maternal undernutrition can affect offspring's physical status and various health parameters that might be transmittable across several generations. Many studies have focused on undernutrition throughout pregnancy, whereas maternal undernutrition prior to pregnancy is not sufficiently studied. The objective of our study was to explore the effects of food restriction prior to and during pregnancy on body weight and longevity of the second generation offspring. Adult female Wistar rats ("F0" generation) were 50% food restricted for one month prior to pregnancy (pre-pregnancy) or during pre-pregnancy and pregnancy. The third group was fed normally (control). The first generation offspring were normally fed until the 6(th) month of age to produce the second generation offspring; namely, the first-generation female rats were mated with male breeders from outside the experiment. The second generation offspring thus obtained were observed until natural death (up to 36 months). Compared to the controls, the second-generation male offspring whose "grandmothers (F0 females)" undernourished only during pre-pregnancy were significantly heavier from the 8(th) month of age, whereas no significant weight difference was found in the male offspring whose "grandmothers" were food-restricted during pre-pregnancy and pregnancy. Shorter lifespan was observed in the second-generation male offspring of "grandmothers" that were food-restricted either during pre-pregnancy or during pre-pregnancy and pregnancy. By contrast, no differences in body weight and lifespan were observed in all second-generation female offspring. In conclusion, maternal caloric restriction prior to pregnancy increases the body weight and shortens the longevity of the second-generation male offspring, indicating the sex-dependent transgenerational effect of maternal caloric restriction.

Nevomelanocytic atypia detection by in vivo reflectance confocal microscopy.

BACKGROUND AND OBJECTIVE: In vivo reflectance confocal microscopy (RCM) is a promising novel technology for non-invasive early diagnostics of cutaneous melanoma. However, the possibility to detect melanocytic atypia in nevi by means of in vivo RCM remains unknown. The aim of the study was to evaluate the significance of in vivo RCM features of melanocytic atypia for the diagnosis of melanocytic nevi, dysplastic nevi and cutaneous melanoma. MATERIALS AND METHODS: A total of 138 melanocytic skin lesions comprising 25 melanocytic nevi, 69 dysplastic nevi and 44 melanomas were analyzed by means of dermoscopy, in vivo RCM and routine histopathology. In vivo RCM images were analyzed for the arrangement of keratinocytes in epidermis, pagetoid cells and junctional melanocytic nests and correlated refractivity aspects of nests with histopathology. RESULTS: Separately and all together taken the in vivo RCM features of melanocytic atypia were significant in differential diagnosis of benign and malignant melanocytic skin lesions, though none of the features was significant in discriminating nevi without cytologic atypia of dysplastic nevi. In vivo RCM feature of dense cell clusters corresponded with melanin containing nevomelanocytes on histopathology though exact correspondence of non-homogeneous and atypical sparse cell clusters remained questionable. CONCLUSIONS: Nevus with histopathologically confirmed nevomelanocytic atypia (dysplastic nevus) could not be distinguished from nevus without atypia using analyzed in vivo RCM features of melanocytic atypia. More accurate diagnostics by means of in vivo RCM needs further investigation on reflectance of single and nested cutaneous melanocytes in benign and malignant skin lesions.

The effect of nanoparticles in rats during critical periods of pregnancy.

BACKGROUND AND OBJECTIVE. Nanotechnology works with substances at a nanometer scale, and it offers many solutions for biomedicine. Nanoparticles (NPs) have been shown as effective agents for imaging, drug delivery, pathogen detection, etc. However, to date, NP toxicity is poorly known. The aim of our study was to investigate the embryotoxicity and teratogenicity of quantum dots (QDs) at the different stages of rat embryogenesis. MATERIALS AND METHODS. Wistar rats were injected with CdSe/ZnS or CdTe QDs on the 6th, 13th, and 18th days of embryogenesis. Cyclophosphamide was chosen as a positive control of embryotoxicity. On the 21st day, the number of resorptions, weight, length, and external malformations of the embryos were estimated. Fluorescence spectroscopy and microscopy analysis were used to determine the accumulation of QDs in the tissues. RESULTS. Exposure to cyclophosphamide during the pregnancy decreased the embryonic weight and length when compared with the control group and produced numerous malformations. The effects depended on the stage of embryogenesis. Meanwhile, QDs did not cause any embryotoxic or teratogenic effects. However, CdTe QDs induced necrosis in the tissues of the peritoneal cavity. The necrotic tissues contained QDs with altered spectroscopic properties. Spectroscopic and microscopic tissue examination revealed that QDs accumulated in the placenta, but no penetration to the embryonic tissues was observed. CONCLUSIONS. QDs did not cause any direct embryotoxic or teratogenic effects, but they had adverse effects on the maternal organism. The observed QD effects and the long-term accumulation of QDs in the maternal organism may increase the risk of adverse effects on embryo development.

Transport of nanoparticles through the placental barrier.

Nanoparticles (NP) are organic or inorganic substances, the size of which ranges from 1 to 100 nm, and they possess specific properties which are different from those of the bulk materials in the macroscopic scale. In a recent decade, NP were widely applied in biomedicine as potential probes for imaging, drug-delivery systems and regenerative medicine. However, rapid development of nanotechnologies and their applications in clinical research have raised concerns about the adverse effects of NP on human health and environment. In the present review, special attention is paid to the fetal exposure to NP during the period of pregnancy. The ability to control the beneficial effects of NP and to avoid toxicity during treatment requires comprehensive knowledge about the distribution of NP in maternal body and possible penetration through the maternal-fetal barrier that might impair the embryogenesis. The initial in vivo and ex vivo studies imply that NP are able to cross the placental barrier, but the passage to the fetus depends on the size and the surface coating of NP as well as on the experimental model. The toxicity assays indicate that NP might induce adverse physiological effects and impede embryogenesis. The molecular transport mechanisms which are responsible for the transport of nanomaterials across the placental barrier are still poorly understood, and there is a high need for further studies in order to resolve the NP distribution patterns in the organism and to control the beneficial effects of NP applications during pregnancy without impeding the embryogenesis.

The effect of ß-carotene against adriamycin toxicity on the embryo formation.

UNLABELLED: Adriamycin is an anthracycline antibiotic widely used for the treatment of many types of cancer. The cytotoxic effect of Adriamycin occurs by a free radical-mediated mechanism. Thus, to prevent or reduce the toxic effect of Adriamycin, it is possible to use it in combination with antioxidants. The aim of this study was to evaluate a potential effect of ß-carotene against Adriamycin-induced toxicity on the embryo formation. MATERIALS AND METHODS: Pregnant rats were treated with Adriamycin, ß-carotene, and their combination during the critical stages of embryogenesis. The first group was control group. Adriamycin was administered on day 9 (group 2a) and day 12 (group 2b) of gestation by a single intraperitoneal injection at a dose of 5 mg/kg. ß-Carotene was given at a dosage of 0.6 mg/(kg·day) from day 6 to 10 or from day 9 (group 3a) to 13 (group 3b) of gestation 5 times per os; in the case of their combination, ß-carotene was given per os 3 times before Adriamycin injection, one time simultaneously with Adriamycin and one time after its injection (groups 4a and 4b). Animals were euthanized on day 21 of gestation. Embryo resorptions and alive fetuses were counted, weighed, and measured. The embryos of each litter were examined macroscopically after the Buen solution fixation for the embryo defects. In order to render the skeleton visible, the soft tissues were macerated using caustic soda, stained with alizarin red, and cleared with glycerin. RESULTS: Adriamycin induced embryotoxicity; the combination of Adriamycin and ß-carotene decreased the number of Adriamycin-induced embryo resorptions about two times. A gavage with Adriamycin alone decreased fetal body weights (P<0.05), while giving it in combination, the fetal body weight was similar to that in the control group. Adriamycin induced the retardation of skeletogenesis and external fetal malformations (microphthalmia, hydrocephaly, anencephaly, and others). After an exposure to ß-carotene, external malformations (diaphragmatic hernia) of embryos were found only occasionally. ß-Carotene in combination with Adriamycin produced no positive effect on Adriamycin-induced skeletodysgenesis or external malformations. CONCLUSIONS: Antioxidant ß-carotene in combination with Adriamycin slightly reduced the Adriamycin-induced embryotoxicity, but produced no positive effect on Adriamycin-induced skeleto-dysgenesis or external malformations.

Effect of Biological Environment on Polyester Surgical Suture's Physical Properties: An Experimental Study in Rats.

Objective The following study aims to analyze the alteration of nonabsorbable polyester surgical suture physical properties after in vivo incubation. Methods A comparative study of braided nonabsorbable polyester 2/0 (U.S.P) sutures was performed. The control in vitro group and three experimental in vivo subgroups, composed of ten sutures in each, were created. All 30 experimental sutures were implanted into a total of 15 laboratory rats subcutaneous tissue (two sutures in each rat) and removed after seven, eight and nine weeks, respectively. Further, they were attached to the mechanical testing device and affected with a mechanical force, increasing the load by 0,1 N/s until complete breakage. Tensile strength (TS), failure displacement (FD), failure strain (FS) and failure stress (FST) were measured at the point of failure and compared to the same parameters of the control group. Results No statistically significant difference was found in the physical parameters of the samples between the experimental and control groups (TS [p = 0.358], FD [p = 0.258], FS [p = 0.258] FST [p = 0.358]). A statistically significant difference was found in the failure load between sutures that break on the knot site (KS) and the rest of the samples: significantly less force was needed to break the suture on the KS. Moreover, most of the breaks on the KS occurred in sutures that were incubated for the longest period of nine weeks (n = 4). An anomaly of partial failure (PF) was noticed. Sutures with PF elongated significantly more compared to the sutures that did not undergo PF in the control and in experimental groups (p = 0,044; p = 0,017; p = 0,016; p = 0,013). Conclusion The biological environment had no radical aftereffects to the suture's physical properties. In vivo exposure may cause the suture to break on the KS more frequently and may lead to PF, when a few sutures composed of fiber fail. Sutures that undergo PF tend to elongate further.

Quantum dots mediated embryotoxicity via placental damage.

The increasing use of nanoparticles in consumer products raises the concerns of their safety. This study investigated the biological effects of quantum dots (QD) exposure to rats during pregnancy. CdTe QD were injected on the 13th gestation day. Morphological features of 121 fetuses and histological analysis of placentas were performed on the 20th gestation day. The results showed that QD exhibit dose dependent embryotoxicity: survival rates of fetuses were 97% (5mg/kg dose), 86% (10mg/kg dose) and 43% (20mg/kg dose). QD exposure also resulted in the reduction of fetal body length and mass, disturbed ossification of limbs and caused placental tissue damage. QD exhibit no teratogenic effects at the applied doses. It is hypothesized that embryogenesis was impeded due to the placental damage rather than QD penetration and accumulation in the fetuses. To conclude, mothers should be protected from QD exposure during pregnancy.

[Accumulation of photosensitizer in rat embryos (a spectroscopic study)]. / Fotovaisto pasiskirstymas ziurkes ir embriono audiniuose (spektroskopinis tyrimas).

UNLABELLED: The aim of the present study was to investigate the accumulation of photosensitizer Photofrin II in the different organs as well as the placenta and embryos of pregnant rats and to determine during which stage of embryogenesis the photosensitizer is accumulated the most effectively. MATERIALS AND METHODS: The experiments were carried out on 25 fetuses from 10 Wistar rats (weight 160-240 g). Female rats were mated with male rats in the evening. Vaginal smears were collected from each female rat next morning and were examined by microscope in order to determine the presence of sperm. The day when sperm was detected in the vagina was considered to be pregnancy day 0. Photofrin II (a dose of 5 mg/kg) was administered intravenously to pregnant rats on days 7, 14, 16, 18 and 20 of embryogenesis. Rats were euthanized 24 hours after intravenous injection of Photofrin II and the following organs were taken: brain, spleen, liver, kidneys, lungs, uterus, placenta, and embryos. The accumulation of the photosensitizer was observed in the samples prepared from these parts of body. Fluorescence measurements ex vivo were performed with an S2000-FI fluorescence spectrometer (Ocean Optics Inc., Florida, USA) by exciting the samples with a blue light emitting diode (lambda=400 nm). RESULTS: A comparative study of fluorescence spectra on days 7, 14, 16, 18 and 20 of embryogenesis showed that the most intense accumulation of Photofrin II in the embryo was on day 7, while on the other days of embryogenesis the accumulation of Photofrin II increased obviously in the uterus and placenta. CONCLUSIONS: The obtained data show that the accumulation of Photofrin II in the embryo depended on the stage of embryogenesis as well as on permeability of the placental barrier. Further photodynamic therapy studies are necessary to determine the total effect of Photofrin II on the embryo.

[Elbow joint formation in grafts of the limbs lacking the anterior or posterior necrotic zones]. / Alkunes sanario sklaida persodintose galuniu uzuomazgose, stokojanciose priekines arba uzpakalines nekrozes zonu.

UNLABELLED: The purpose of this experimental work is to examine the joint formation in limb bud grafts lacking the anterior or posterior necrotic zones where programmed cell death (apoptosis) occurs. METHODS: The embyonic chick limbs were transplanted onto the chorioallantoic membrane. The limbs of experimental group were transplanted without the prospective anterior or posterior necrotic zones. Transplants containing all limb bud material served as the control group. RESULTS: Most experimental grafts showed wide or complete fussions of the bones forming the elbow joint in the early developmental stages. The regression of the elbow joint in the control grafts was gradually progressive: from the narrow cartilaginous bridges between articular surfaces in early stages to extensive fusion of bone primodia in late developmental stages. CONCLUSIONS: The elimination of the anterior or posterior necrotic zones suppressed apoptosis in the "opaque patch" and caused the fussion of the bones in elbow joint.

[The influence of azathioprine on the osteogenesis of the limbs]. / Azatioprino itaka galuniu osteogenezei.

BACKGROUND: The purpose of this experimental work was to examine the osteogenesis of rats limbs under the influence of immunosuppressant azathioprine. MATERIAL AND METHODS: Two doses of azathioprine (50mg/kg and 80 mg/kg) were administered intraperitoneally to experimental and control pregnant rats once during the period from the 1(st) to the 14(th) day of embryogenesis. The size of the centers of ossification was measured in the limbs fixed in ethanol, cleared and stained with alizarin red. RESULTS: The analysis of the experimental data revealed that osteogenesis was sensitive to the influence of azathioprine administered to embryos from the 4(th) to the 13(th) day of embryogenesis. The centers of ossification in embryos treated with azathioprine on the 10 and 12(th) days were most damaged: shorter, deformed or absent. CONCLUSIONS: The limb osteogenesis was delayed and damaged by azathioprin in the period of active organogenesis. The bigger the doze of azathioprine the more osteogenesis was delayed. The most sensitive was the skeleton of the autopod.