In vitro and in vivo evaluation of sandwich-like mesoporous silica nanoflakes as promising anticancer drug delivery system.

We present the new promising nanostructure- sandwich-like mesoporous silica nanoflakes synthesized on graphene oxide sheets core. In the first step biocompatibility of the nanoflakes with PEG and without functionalization in human fibroblast, melanoma and breast cancer cells was assessed. In order to define the cellular uptake in vitro and biodistribution in vivo the nanostructures were labelled with fluorescent dye. In the next step, the silica nanostructures were filled by the anticancer drug- methotrexate (MTX) and cytotoxicity of the complex in reference to MTX was evaluated. The WST-1 assay shows mild, but concentration dependent, cytotoxicity of the nanoflakes, most significant for the non-functionalized structures. PEG-modified silica nanoflakes didn't produce a disruption of cell membranes and lactate dehydrogenase (LDH) release. Cell imaging revealed efficient internalization of the silica nanoflakes in cells. Ex vivo organ imaging showed high accumulation of the nanostructures in lungs, bladder and gall bladder, whereas confocal imaging revealed wide nanoflake distribution in all tested tissues, especially at 1h and 4h post intravenous injection. Cytotoxicity of the nanoflake-MTX complex in reference to MTX showed similar cytotoxic potential against cancer cells. These findings may provide useful information for designing drug delivery systems, which may improve anticancer efficacy and decrease side effects.

Study on size effect of the silica nanospheres with solid core and mesoporous shell on cellular uptake.

The properties of mesoporous silica nanoparticles including large surface area, large pore volume, easy surface functionalization and control of structure and pore size has made them promising drug carriers. In this study, the effect of different diameters (50 nm, 70 nm, 90 nm, 110 nm and 140 nm) of silica nanospheres with a solid core and mesoporous shell (mSiO2/SiO2) on cellular internalization in mouse fibroblast cells (L929) was evaluated. The physical properties of the nanostructures were characterized with various methods, such as transmission electron microscopy with x-ray dispersion spectroscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy and zeta potential. In order to define the cellular uptake, the nanostructures were labelled with fluorescent dye Alexa647, and imaging and quantitative methods were applied: laser scanning confocal microscopy, flow cytometry and thermogravimetry. Our results indicate that cellular uptake of the studied nanospheres is size-dependent, and nanospheres of 90 nm in diameter showed the most efficient cell internalization. Thus, particle size is an important parameter that determines cellular uptake of nanoparticles and should be considered in designing drug delivery carriers.

Sandwich-like mesoporous silica flakes for anticancer drug transport-Synthesis, characterization and kinetics release study.

In this paper, we present the technology of synthesis, characterization and release kinetics of anticancer drug molecules from sandwich-like mesoporous silica nanoflakes. Mesoporous silica nanoflakes are a very attractive material due to their versatility, low cytotoxicity, large surface area, high pore volume and unique feature of containing parallel pores openon both sides. Nanosilica flakes were prepared through the formation of a mesoporous silica layer on a graphene oxide surface. After graphene oxide removal, the silica nanostructures were filled by an anticancer drug-methotrexate. Release kinetics studies were performed in different temperatures, imitating the conditions in living organisms. Release data was analyzed using the zero-order model, first-order model, Higuchi model and Korsmeyer-Peppas model. The optical properties of samples, and the kinetics of drug release from the nanostructure, were examined by UV-vis spectrophotometer. Data obtained from long term studies showed that the system can serve as an anticancer drug carrier system, since a significant amount of methotrexate was loaded to the material and released. The mechanism of MTX release from mesoporous silica nanoflakes appeared to be a parallel processes of diffusion through water-filled mesopores and degradation of the mSiO2 matrix. Physical and chemical characterization was undertaken by transmission electron microscopy (TEM) and X-ray dispersion spectroscopy (EDX). The specific surface area of the samples was measured through the adsorption of N2 isotherm, interpreted with the Brunauer-Emmett-Teller model (BET). TGA and UV-vis analyses were conducted in order to estimate the amount of the released drug.

Color Doppler assessment of blood flow in endometrial cancer.

OBJECTIVE: To determine the location and intensity of angiogenesis as well as selected flow parameters by transvaginal color Doppler (TVCD) and to evaluate the relation of myometrial invasion, histological grading, lymph nodes, and omental and adnexal metastasis on blood flow characteristics in endometrial cancer. METHODS: Transvaginal colour Doppler and pulsed Doppler ultrasound were performed on 90 women with endometrial cancer. The degree of invasion as well as adnexal, omental, and pelvic lymph node metastasis was evaluated. Location of the blood vessels (peripheral, central, mixed) and vascular density as well as selected Doppler blood flow indices: PSV, RI of neoplastic infiltration was assessed. RESULTS: The median age of the 90 women was 63.3 +/- 12.3 years (range 32 to 86 years); of these 92.2% were postmenopausal. Cancer concerned only the endometrium (E), with superficial (S) and deep infiltration (D) established in 14.4%, 45.6% and 40%, respectively. The histological maturity was as follows: G1 - 17.6%, G2 - 66.7%, G3 - 16.6% of cases. Adnexal, omental and lymph node metastasis was found in 12.2%, 3.3% and 16.6%, respectively. Abnormal low impedance and high velocity flow (mean RI 0.38 +/- 0.09, PSV 20.45 +/- 9.6 cm/sec) were found in 88.9% of cases. In types E, S, D in 61.5%, 92.7% and 94.4%, respectively (p = 0.003). Differences in RI and PSV between groups with high and low vascular density were statistically significant (p = 0.005 and 0.001, respectively). In all cases peripheral and mixed vascularity were found more frequently (p < 0.05). A positive significant correlation between vascular density increase and surgicopathological stage of cancer was found more frequently (p < 0.005). There were significant differences in vascular density, Doppler blood flow indices and vascular location in each type of histological malignancy (p < 0.05). No significant differences in each flow parameter in hematogenous-adnexal/omental metastatic and non metastatic cases were found, whereas pelvic lymph node involvement and vascular density were shown to be statistically significant (p < 0.02). There were significant differences in vascular density in lymph-node positive cases whereas the remaining flow parameters did not differ. CONCLUSIONS: These results suggest that TVCD evaluation of endometrial cancer is a reliable method for assessing endometrial angiogenesis. Our results indicate that blood flow rates correspond with increased angiogenesis in endometrial cancers, and might potentially be used as a good prediction factor for tumor progression and metastasis in affected women. Preoperative ultrasound examination should be seen as an important tool in the establishment of individualized treatment programs for women with endometrial cancer.

The importance of fine needle aspiration biopsy and sonographic evaluation of parametria in cervical cancer.

OBJECTIVE: The aim of study was to estimate of the value of fine needle aspiration biopsy (FNAB) and transvaginal ultrasonography (TVS) in the preoperative assessment of the parametria in cervical cancer. We compared parametrial infiltration before and postoperatively by histopathology to verified and confirmed staging of disease. Correct staging qualification, especially evaluation of the parametrium, is very useful in choosing an adequate method of treatment, and thereby in patient survival. MATERIAL AND METHOD: 52 women (median age 56 years, range 33-85) with cervical cancer in Stage Ib and 49 (median age 51, range 36-71) in Stage II and III, were included in the study. Assessment of parametrial invasion before treatment was performed by fine needle aspiration biopsy (FNAB) with endovaginal ultrasound assistance. The sonographic evaluation of parametria was performed by Siemens Sonoline Versa Pro with a transvaginal 7 MHz mechanical transducer with a biopsy guide and 21-gauge needle. The probe was covered with a disposable latex sheath filled with ultrasound gel. The aspirated material was placed on a glass slide, fixed in 95% alcohol and submitted to cytologic evaluation. All of the patients with cervical cancer in Stage Ib underwent a Wertheim-Meigs hysterectomy. The preoperative findings were compared with data obtained by histopathology findings. Moreover, in the whole group of 101 patients a comparison of FNAB and sonography was performed. The sensitivity, specificity and diagnostic accuracy of this method were evaluated. RESULTS: Parametrial involvement assessed postoperatively by histopathology, in clinical Stage Ib cervical cancer was found in eight of 52 cases (15.4%). FNAB of parametrial involvement in the operated group was accurate in 14 of 18 (accuracy--83%, sensitivity--78%, specificity--84%, PPV--50%, NPV--95%). Sonographic assessment of parametrial involvement was correct in 12 of 18 cases (accuracy--58%, sensitivity--67%, specificity--56%, PPV--24%, NPV--89%). In the whole group of patients (operated and non-operated), sonographic evaluation of parametria verified by FNAB was correct in 104 of 202 cases (accuracy--78%, sensitivity--71%, specificity--86%, PPV--84%, NPV--74%). CONCLUSIONS: FNAB and TVS assessment of the parametria are very useful methods in confirmation of neoplastic infiltration. Correct preoperative diagnosis may improve staging, treatment and indirectly, survival of patients with cervical cancer.

The value of ultrasonography in preoperative assessment of selected prognostic factors in endometrial cancer.

OBJECTIVE: To determine the efficiency of transabdominal and transvaginal ultrasonography (TAS and TVS) in the assessment of myometrial invasion, cervical involvement, pelvic lymph nodes, adnexal and omental metastases (preoperative staging) of endometrial cancer. METHODS: Transabdominal and transvaginal 2D, ultrasound were performed on 90 women to classify myometrial invasion, cervical involvement, pelvic lymph nodes and adnexal metastases in endometrial cancer. According to this 13 type E (invasion involving the endometrium), 41 type S (superficial, of less than 50% of myometrial infiltration), 36 type D (deep infiltration) and 22 cervical involvement were identified. There were 15 G1, 60 G2 and 15 G3 cases. Adnexal, omental and lymph-node metastases were found in 11, two and 15 cases, respectively. Endometrial cancer was diagnosed on the basis of dilatation and curettage. The degree of invasion was evaluated preoperatively. Ultrasonographic findings were compared to surgical staging and histopathology of the surgical specimen. RESULTS: The median age of the 90 women was 63.3 +/- 12.3 years (range 32 to 86 years). The median thickness of malignant endometrium was 19.5 +/- 9.6 (range 7 to 54 mm). In type E the median thickness was 11.76 +/- 4.2, in type S 17.3 +/- 7.6, in type D 24.8 +/- 9.8 and in cases with cervical involvement 23.2 +/- 11 mm. Myometrial invasion evaluated by TVS was accurate in 76 of 90 cases (accuracy 84.4%). In type E sensitivity was 92.3%, specificity 87.0%, positive and negative predictive value, respectively, 63.1% and 98.6%. In type S these values were respectively: 78.0%, 93.9%, 91.4%, 80.0% and in type D--88.9%, 92.6%, 88.9% and 100.0%. Tumor extension to the cervix was properly assessed in 19 of 22 women in which it was present (sensitivity 86.4%, specificity 85.3%, positive predictive value 85.5%, negative predictive value 95.1%). Adnexal metastasis was correctly diagnosed in 8 of 11 cases in which it was present (sensitivity 72.7%, specificity 97.5%, positive predictive value 80%, negative predictive value 96.3%), and lymph-node metastasis in only 5 of 15 cases (sensitivity 33.3%, specificity 100%, positive predictive value 100%, negative predictive value 88.2%). CONCLUSION: These results suggest that 2D TAS and TVS evaluation of endometrial cancer are reliable methods for preoperative assessment of selected prognostic factors, e.g. myometrial invasion, cervical involvement and adnexal metastases. However in assessing lymph-node metastases, TVS with its low sensitivity, did not provide additional information. Preoperative ultrasound examination should be speculated as an important tool in the establishment of different surgical choices which can be made after a correct pretreatment prognosis.

The effectiveness of hysteroscopic polypectomy in cases of female infertility.

The authors, based on their own material, evaluated the value of hysteroscopic treatment of endometrial polyps in a group of infertile women. Polypectomy was performed in 25 patients using either endoscopic microscissors or electric loop. Follow-up hysteroscopy was performed after two months as an integral part of the treatment. All the patients were observed for 12 months. The primary intrauterine investigation was complete in all patients without regard to kind of instrument which was confirmed by second-look hysteroscopy. About 80% of the patients who underwent surgery conceived. Restoration of reproductive ability did not depend on the size of the removed lesion.

Hysteroscopy in cases of cervical polyps.

To assess the usefulness of hysteroscopy in cases of cervical polyps, we studied 78 patients with a lesion found during a routine gynecological examination. All women were referred for diagnostic hysteroscopy followed by endoscopic polypectomy. In 83.3% of the cases intrauterine investigation confirmed primary diagnosis - a polyp peduncle in the cervical canal. Sixty-five of those lesions were the only pathologic condition, and six (7.7%) were associated with an endometrial polyp. In the remaining 16.7% of examined patients, primary polyps identified as cervical polyps appeared to be endometrial. All women admitted to the study were successfully treated by hysteroscopy. Our results suggest that endoscopic evaluation of the uterine cavity in women with cervical polyps can clarify the initial diagnosis. Hysteroscopy allows not only a precise visualisation of the polyp peduncle but also gives the possibility to identify and treat concurrent asymptomatic intrauterine pathological conditions.

[Transanal ultrasonography for diagnosis of obstetric anal sphincter trauma]. / Mozliwosci ultrasonografii przezodbytniczej w diagnostyce uszkodzen polozniczych zwieraczy odbytu.

Fecal incontinence affects up to 2% of the general population. The major cause of it is related to the birth trauma of the anal sphincters. In this paper the possibilities of transanal endosonography in the assessment of the anal canal muscles defects after deliveries were presented. Anal canal sonographic anatomy together with the technique of the examination with the use of rotating endoprobe were presented. Transanal rotating endoprobe was compared with exoanal probes for visualization of the anal canal. Ultrasonic images of internal and external anal sphincters defects were shown with specific patterns of defects allowing differentiation between morphological and neurological damages of the sphincters. Anal endosonography was compared with clinical examination, electromyography and manometry in the assessment of the integrity of the anal sphincters.

Preoperative discrimination between malignant and benign adnexal masses with transvaginal ultrasonography and colour blood flow imaging.

BACKGROUND: Ovarian cancer is one of the causes of death in women, and in about 70% of cases is recognized only in advanced stages. This study was undertaken to evaluate distinctive values of transvaginal and color Doppler ultrasonography in differentiating malignant and benign adnexal masses through analysis of ultrasonic morphological features of malignancy and estimation of location and intensification of angiogenesis as well as values of resistance of flow in examined masses. PATIENTS AND METHODS: 329 women with malignant and benign adnexal masses underwent ultrasonographic and colour Doppler examination 1-5 days before surgery (laparotomy, laparoscopy) thus allowing histological verification of diagnosis. The ultrasonographic structure was assessed using a morphological scoring system devised by Sassone, Jain and Benacerraf. Regions showing vasculature, especially within septae and solid parts of tumours were examined by means of transvaginal colour Doppler. Location and intensification of angiogenesis as well as resistance index (RI) were investigated. Sensitivity, specificity, PPV and NPV of both techniques were assessed. Statistical analysis of obtained data were based on the Student's t test; p < 0.05 level was considered significant. RESULTS: Postoperatively 255 (77.5%) benign and 74 (22.5%) malignant tumours were seen. In the group of benign masses the average age of women was 42.6+/-12.3 and in the malignant it was 53.1+/-12.6 (p<0.0001). The transverse dimension of benign lesions was 77.2+/-19, whereas for malignant it was 107.0+/-31 (p<0.0001). Benign tumours in 63.0% were cystic, in 26.0% mixed cystic-solid and in 11.0% solid echostructures while in malignant they were respectively, 6.8%, 56.8% and 36.4% (p<0.0001). Doppler flow within the tumour was 74.5% in benign and 98.6% in malignant masses (p<0.0001). In benign lesions homogenous superficial or peripheral vasculature was visualized, and in the majority of cases (82.7%) it was of medium intensification. However in malignant central, peripheral or mixed vascularisation. in the majority intensified character was found. Average value of the resistance index in all benign masses amounted to 0.77+/-0.14, however in malignant it was 0.39+/-0.07 (p<0.0001). CONCLUSIONS: We contend that complete ultrasonographic estimation of ovarian neoplasms outside the qualification of structural details should include Doppler analysis of vasculature parameters. Most important is the qualification of resistance of flow, and location and intensification of vascularisation in examined masses which permit the differentiation of malignant and benign lesions. Preoperatively recognizing malignant processes with colour Doppler ultrasonography shows higher accuracy, specificity and PPV.

Transvaginal colour flow imaging in assessment of ovarian tumor neovascularization.

Localization and intensity of angiogenesis in benign and malignant ovarian neoplasms recorded by color Doppler transvaginal ultrasound has been assessed. Increased intratumoral angiogenesis has been affirmed in 60% of malignant and only in 9% of benign lesions.

Transvaginal Doppler ultrasound with colour flow imaging in benign and malignant ovarian lesions.

Since early detection of ovarian cancer is difficult, most cases are diagnosed at advanced stages. The imaging diagnosis is based on pattern classification and is limited with respect to the precise determination of malignancy. High-frequency transvaginal sonography improves the ability to detect malignant ovarian tumors over that of transabdominal route, however, the predictive values are unsatisfactory because of inability to distinguish between malignant and benign tumors that have similar morphologic characteristics. The introduction of transvaginal colour flow imaging has allowed detection of low-resistance intratumoral blood vessels, characteristic of malignant tumors, and visually reflected the state of blood flow of an ovarian tumor. These two ultrasonographic methods were used for diagnosis of ovarian tumors in 65 women treated in our Department of Obstetrics and Gynecology. Waveforms of the parenchymal tumor arteries or tumor surface arteries were compared using value of the resistance index (RI). The sensitivity, specificity and accuracy of the preoperative RI in detecting malignant ovarian tumors were 100%, 94%, 95.4% respectively. The sensitivity, specificity and accuracy of preoperative suspicious sonographic findings in detecting malignant ovarian tumors were 100%, 61% and 71%. Positive and negative predictive values of colour flow imaging were 85% and 100%, whereas for grey-scale transvaginal ultrasonography they were 46% and 100% respectively. The findings of this study suggest that transvaginal colour Doppler is a method which is superior to the other methods for preoperative evaluation of ovarian malignancy.

[Analysis of the course of pregnancies and labor in girls under 18 years of age]. / Analiza przebiegu ciazy i porodu u pacjentek przed 18 rokiem zycia.

Authors analysed the course of pregnancy and labor at 141 under 18 aged girls, between 1981-1990. The rise of number of labors at teen-agers was observed in the last 5 years. This tendency should pay our attention on this group of patients.

PIP: During 1981-90, there were 141 births to women under the age of 18 from among 22,037 deliveries at the II Department of the Obstetrical and Gynecological Clinic of the Academy of Medicine in Warsaw. The control group consisted of 141 consecutive primiparas aged 19-30. Among the 22,037 births, the underage women accounted for 0.64%. 10 women were under 16 years of age, and two patients each were 13 and 14 years of age, respectively, while 23 were aged 16, and 108 were aged 17. The percentage of pregnant adolescents increased from 0.13% in 1982 to 0.84% in 1986 and to 1.24% in 1990. There were 47 (33.67%) pregnancy complications among adolescents and 55 (39%) among adults. The threat of premature birth occurred in 11 adolescents (7.0%) vs. 13 adults (9.22%); premature discharge of amniotic fluid in 21 adolescents (14.89%) and 25 adults (17.73%); EPH gestosis in 4 adolescents (2.84%) and 5 adults (3.55%); infection of the urinary tract in 7 adolescents (4.96%) and 10 adults (7.10%). Normal delivery occurred in 124 (87.94%) adolescents and in 112 (79.4%) adults. Cesarean sections were performed in 11 (7.8%) adolescents and 24 (17.1%) adult primiparas. There were 26 (18.4%) instances of delivery complications in adolescents and 66 (46.7%) in adults. Incomplete delivery occurred in 21 (14.9%) adolescents vs. 23 (16.3%) adults. Significantly more (39) cases of rupture of the uterine cervix occurred in adults (27.6%) than in adolescents (4 instances or 2.8%). The duration of labor ranged from 2 hours to 12.5 hours in the group of adolescents with an average of 7.1 hours compared to from 2 hours to 25 hours with an average of 7.9 hours in the control group. The average values in the first minute of life amounted to 8.85 marks on the Apgar score in the case group compared to 9.36 scores in controls. The average birth weight of neonates was 3190 g in the case group compared to 3284 g in the control group.

[Ectopic spleen in pelvis minor--a case report]. / Sledziona ektopowa zlokalizowana w miednicy mniejszej--opis przypadku.

A case of ectopic spleen diagnosed as a left adnexal tumor in a 19 year old female was presented. Considering the rareness and lack of typical symptoms, diagnosis of this pathology still is very difficult, and establish most often intraoperatively. In every case of ascertainment irregular mass in minor pelvis one should differentiate it among: colonic diverticulosis, wandering kidney, coproliths, colon and mesentery tumors and ectopic spleen. We consider that ultrasonography plays the special role in these cases. In every case of stating the pathologic structure in minor pelvis it seems advisable not to content only with estimation of genital organs, but also locate such as: kidneys, liver, spleen. Such a procedure allows to state preoperative diagnosis more precisely.