Exact and Numerical Solution of the Fractional Sturm-Liouville Problem with Neumann Boundary Conditions.

In this paper, we study the fractional Sturm-Liouville problem with homogeneous Neumann boundary conditions. We transform the differential problem to an equivalent integral one on a suitable function space. Next, we discretize the integral fractional Sturm-Liouville problem and discuss the orthogonality of eigenvectors. Finally, we present the numerical results for the considered problem obtained by utilizing the midpoint rectangular rule.

HPV Infection Significantly Accelerates Glycogen Metabolism in Cervical Cells with Large Nuclei: Raman Microscopic Study with Subcellular Resolution.

Using Raman microscopy, we investigated epithelial cervical cells collected from 96 women with squamous cell carcinoma (SCC) or belonging to groups I, IIa, IIID-1 and IIID-2 according to Munich III classification (IIID-1 and IIID-2 corresponding to Bethesda LSIL and HSIL groups, respectively). All women were tested for human papillomavirus (HPV) infection using PCR. Subcellular resolution of Raman microscopy enabled to understand phenotypic differences in a heterogeneous population of cervical cells in the following groups: I/HPV-, IIa/HPV-, IIa/HPV-, LSIL/HPV-, LSIL/HPV+, HSIL/HPV-, HSIL/HPV+ and cancer cells (SCC/HPV+). We showed for the first time that the glycogen content in the cytoplasm decreased with the nucleus size of cervical cells in all studied groups apart from the cancer group. For the subpopulation of large-nucleus cells HPV infection resulted in considerable glycogen depletion compared to HPV negative cells in IIa, LSIL (for both statistical significance, ca. 45%) and HSIL (trend, 37%) groups. We hypothesize that accelerated glycogenolysis in large-nucleus cells may be associated with the increased protein metabolism for HPV positive cells. Our work underlines unique capabilities of Raman microscopy in single cell studies and demonstrate potential of Raman-based methods in HPV diagnostics.

Extremely Low Birth Weight Predisposes to Impaired Renal Health: A Pooled Analysis.

BACKGROUND: A number of studies examined the association between preterm delivery and kidney size and function later in life. However, the number of cases in published cohort studies is low. This study was aimed at performing a multicenter collaboration to pool data to obtain more accurate results to quantify the extent of renal impairment in former extremely low birth weight (ELBW; <1,000 g) children. METHODOLOGY: We performed a subject-level meta-analysis to pool data from Cracow (64 cases/34 controls) and Leuven (93 cases/87 controls). We assessed and analyzed cystatin C, estimated glomerular filtration rate (eGFR), ultrasound kidney length, and blood pressure (BP) in 11-year-old ELBW children compared with controls born at term. The prevalence of hypertension (HT) and prehypertension (preHT) in both groups was also analyzed. RESULTS: The study group comprised 157 former ELBW children (gestational age 23-33 weeks and birth weight 430-1,000 g) and 123 children born at term. Former ELBW children had lower mean eGFR (100.62 ± 16.53 vs. 111.89 ± 15.26 mL/min/1.73 m2; p < 0.001), smaller absolute kidney length (8.56 ± 0.78 vs. 9.008 ± 0.73 cm; <0.001), and higher systolic (111.8 ± 9.8 vs. 107.2 ± 9.07 mm Hg; p = 0.01) and diastolic (68.6 ± 6.8 vs. 66.3 ± 7.7 mm Hg; p = 0.03) BP. Smaller renal size in former ELBW children was positively associated with lower birth weight, shorter gestational age, and severity of perinatal complications (intraventricular hemorrhage, length of stay, mechanical ventilation, and oxygen therapy). CONCLUSION: ELBW is associated with lower eGFR and a high frequency of preHT and HT.

Usefulness of the most popular neurodevelopmental tests in preschool assessment of children born with very low birth weight.

BACKGROUND: The aim of our study was multifaceted neurodevelopmental examination of children born prematurely with very low birth weight (VLBW) in order to evaluate the usefulness of popularly used tests. The second aim of the study was exploration of risk and protective factors of neurodevelopmental impairment. METHODS: Eighty-nine VLBW patients were evaluated at the age of 50 months. All children underwent anthropometric measurements and psychomotor tests: functional independence measure scale (WeeFIM), Gross Motor Function Measurement (GMFM), non-verbal psychometric evaluation (Leiter test), Developmental Test of Visual Perception (DTVP-2), temperament questionnaire (EAS-C) and children vocabulary test (TSD). RESULTS: Most severe deficits in ex-preterms' neurodevelopment were associated with verbal abilities, visual perception and temper abnormalities. WeeFIM, DTVP-2, Leiter and vocabulary tests' results correlated with each other. The lowest percent of children with deficits in WeeFIM test indicates, that it seems to be the most valuable tool for identification of the most seriously impaired children. Due to the highest percent of children with visual perception deficits, DTVP test seems to be good second choice in assessment of children born prematurely. In motor assessment GMFM appears to be more adequate than cerebral palsy (CP) diagnosis. Almost one fifth of VLBW did not reach 85% in Gross Motor Function Measurement, although only 9% of children had CP. CONCLUSIONS: Children born with VLBW had deficits in every part of psychometric evaluation. We believe that the most useful tests in assessment VLBW patients are WeeFIM, GMFM and DTVP. Children with severe prematurity complications could require more precise evaluation.

Comet assay is not useful to predict normal tissue response after radiochemotherapy in cervical and larynx cancer patients.

Normal tissues reactions after radiotherapy vary considerably even between patients receiving the same treatment. The ability to predict the differences in radiosensitivity before radiotherapy would have important implication. Patients with squamous cell carcinoma of the: (i) cervix (38 patients) and (ii) larynx (19 patients) were studied. Control group consisted of 9 healthy women. To assess individual radiosensitivity/chemoradiosensitivity alkaline version of comet assay was performed using isolated peripheral blood lymphocytes from cancer patients and healthy donors. The level of endogenous (0Gy), initial (immediately after 6Gy irradiation) and residual (after irradiation and 1h of repair) DNA damage was investigated. The mean value of endogenous damage was similar in control and cervical cancer (CCU) groups and significantly lower than in larynx cancer patients. Cancer patients showed slower DNA repair. For CCU and larynx patients, comet assay parameters were not helpful for unequivocal prediction of appearance of acute and late radiation reaction effects. Comet assay seems to be unable to predict normal tissue reaction after radiochemotherapy. Therefore, there is still need for developing predictive assays, however, due to complicated mechanism of chemoradiosensitivity, only assays assessing not one but many molecular pathways might gives us reliable score.

Somatic development and some indices of lipid metabolism in 11-year-old children born with extremely low birth weight (< 1000 g) (long-term cohort study).

AIM: To continue the prospective follow-up cohort study on the somatic development and lipid metabolic parameters of 11-year-old-children born with extremely low birth weight and to compare them with the results obtained in the study of 7-year-old children from the same group. MATERIAL AND METHODS: 204 newborns with birth weight ≤1000 g were born in the Malopolska voivodship between 1.09.2002 and 31.08.2004. From this group 115 (56%) children died during infancy and 89 (44%) infants survived. At the age of 7 years 81 (91%) of the children from this group were examined. At the age of 11 years investigations were carried out in 62 (75%) of the children, while 19 (26%) were lost to follow-up. All the children underwent anthropometric measurements. Moreover, the lipid profile (serum total cholesterol, triglycerides, HDL-cholesterol, LDL cholesterol) was evaluated. The control group consisted of 36 children born at term chosen randomly from the general population and matched with regard to age and sex. RESULTS: Children born with extremely low birth weight were generally smaller than their peers. At 7 years, they were shorter (113.75 cm(-0.72) vs.124.52 cm(0.53)), lighter (19.47 kg(-1.12)vs.25.23 kg(0.39)), had a smaller head circumference (49.81 cm(-2.19) vs.52.5 cm(-0.377)), waist circumference (50.14 cm(-0.83) vs.55.45 cm (0.34)), mid-upper arm circumference (17.51 cm vs. 19.29 cm), skinfold thickness (0.76cm(-0.817) vs.0.92cm (-0.19)) and body mass index (14.5 kg/m2 (-0.99)vs.16.16 kg/m2 (0.12)) expressed both as absolute values and z-score values compared to the control group. At 11 years old, the height (141.7 cm(-0.368) vs.146.26 cm(0.65)), weight (33.88 kg (-0.59)vs.40.45 kg(0.66)), head circumference (51.37 cm(-2.05)vs.54.02 cm(-0.33)), waist circumference (61.7 cm (0.26) vs.67.84 cm(1.06)), mid-upper arm circumference (20.95 cm vs. 22.85 cm), skinfold thickness (1.17 cm(-0.25)vs.1.68 cm(0.78)) and body mass index (16.74 kg/m2 (-0.62) vs.18.72 kg/m2 (0.36)) expressed both as absolute values and z-score values were still lower in children born with extremely low birthweight than in the control group. However, their gains over the time period between 7 and 11 years were comparable to their born-at-term peers in all the measured anthropometric parameters. There were no statistically evident differences in the indices of lipid metabolism. CONCLUSIONS: Preterm children with extremely low birth weight (<1000 g) are at an increased risk of growth failure. Once they reach teenage years they are shorter and lighter than their age- and sexmatched born-at-term peers. They also have smaller heads. In our study we did not find statistically evident differences between the investigated and control group in lipid indices. There is a need for longitudinal studies to observe somatic, mental and metabolic development in order to organize multidisciplinary holistic medical care for them.

Ocular outcomes in 4-year old prematurely born children.

Purpose: The aim of the study was to assess functional and structural ophthalmologic outcomes in 4-year old very low birth weight children (VLBW). Material and Methods: A group of 82 VLBW children including: (1) children without retinopathy of prematurity ­ group O (n = 30), (2) children with retinopathy of prematurity with no indications for laser coagulation- group 1 (n = 20 ), (3) children with retinopathy of prematurity treated with laser coagulation ­ group 2 (n = 32) were enrolled. Functional (visual acuity, visual evoked potentials, stereopsis, color vision test) and structural (anterior eye segment examination and fundoscopy) outcome, cycloplegic refraction, intraocular pressure and angle of squint were examined in all children. Developmental Test of Visual Perception was also assessed. Results: Very good visual acuity was presented in 56 (68.3%) patients, good visual acuity in 11 (13.4%) children, visual acuity between 0.4 and 0.2 in 13 (15.9%) and unfavorable function (equal or less than 0.1) was observed in 2 (2.4%) children. Twenty-three patients (28.1%) were myopic, 57 patients (69.5%) were hyperopic. Astigmatism > 1D occurred in 49 (59.8%) patients. Anisometropia larger than 2 D occurred in 7 patients (8.7%). There was no statistical difference between frequency of mentioned above complications between the groups. Abnormal VEP results were more common (14 children -54%) in group 2 as compared to other groups (group 0 - 7 children -25%, group 1 ­ 4 children -24%; p = 0.044). Moreover, the lower percentage of group 2 children presented stereopsis vision (46.9%, group 0 ­ 93.3%, group 1 ­ 90%; p<0.05). Children with retinopathy of prematurity treated with laser coagulation had significantly lower Developmental Test of Visual Perceptions scores. In group 1, in 1 child naevus pigmentosus of the eyelid was observed, in group 2 in one child eyeball atrophy in the right eye and aphakia in the left eye were diagnosed. In 1 child in group 0 optic disc drusen were observed, in 2 children optic discs were pale. In 2 children in group 2 retinal detachment stage 5 was diagnosed. Conclusion: An important factor affecting final ophthalomolgical assessment in prematurely born children is retinopathy of prematurity treated with laser coagulaion. Children with retinopathy of prematurity without laser coagulation had examinations results comparable wih children without retinopathy of prematurity. Diode laser photocoagulation is an effective method of treatment fot active stages of retinopathy of prematurity.

Longitudinal assessment of renal size and function in extremely low birth weight children at 7 and 11 years of age.

BACKGROUND: There are a lack of studies describing a longitudinal association between preterm delivery and renal complications later in life. We assessed renal size and function in preterm infants born with extremely low birth weight (ELBW) during 4 years of follow-up, comparing these parameters to age-matched children born full term (term controls). METHODS: The results of selected renal laboratory tests [levels of cystatin C, creatinine, blood urea nitrogen (BUN)] and of renal ultrasound evaluations were compared between the ELBW group and the term control group at age 7 and 11 years. RESULTS: The study population consisted of 64 children born with ELBW (ELBW children) who had been recruited at birth and 36 children born at term (term children) who took part in both follow-up assessments. Renal ultrasound examination revealed a significantly smaller renal volume in the 7- and 11-year-old ELBW children compared to the term controls [right kidney volume: 50.8 vs. 61.2 ml/m(2), respectively, at 7 years (p <0.01) and 51.4 vs. 58.2 ml/m(2), respectively, at 11 years (p <0.01); left kidney volume: 51.4 vs. 60.3 ml/m(2), respectively, at 7 years (p <0.01) and 55.2 vs. 60.7 ml/m(2), respectively, at 11 years (p = 0.02)]. Renal function in ELBW children was also affected. Serum cystatin C levels were significantly higher in ELBW children than in the controls at 7 years of age, and this difference remained statistically significant at 11 years of age [0.63 vs. 0.59 mg/l, respectively, at 7 years (p = 0.02) and 0.72 vs. 0.61 mg/l, respectively, at 11 years (p = 0.01)]. Six ELBW children also had elevated cystatin C levels (0.97-1.11 mg/l) at 11 years of age. Cystatin C levels were within normal range in the ELBW children at age 7 years and in term children in both follow-up studies. BUN levels were higher in ELBW children at the age of 11 years (4.49 vs. 4.15 mmol/l; p = 0.028). CONCLUSION: Continued follow-up of these patients will reveal whether the observed worsening in renal function will persist into adulthood.

Comparison of effectiveness of treatment of patients with sporadic and germline BRCA1-related ovarian cancer.

OBJECTIVES: Numerous reports suggest that the clinical course of ovarian cancer (OC) in BRCA, including BRCA1, mutation carriers (BRCA1-OC) is different than in patients with sporadic ovarian cancer (SOC). Most of the authors indicate more fa-vourable treatment results in patients with BRCA1-OC. The aim of the study was to compare the effectiveness of treatment of patients with advanced-stage (FIGO III/IV) SOC and BRCA1-OC. MATERIAL AND METHODS: Between 2004 and 2009, 957 OC patients were treated in Cracow Branch of Cancer Center, M. Sklodowska-Curie Memorial Institute, Poland. Germline BRCA1 mutation was found in 66 patients. To compare the effective-ness of treatment, the group of 47 advanced-stage BRCA1-OC patients was matched with the group of 47 advanced-stage SOC patients. Pairs of patients were matched in terms of the most important prognostic factors, i.e. stages according to FIGO, primary cytoreduction extent, tumour histologic subtype and grade, as well as year of diagnosis and treatment. RESULTS: The 5-year overall survival rate was 42.9% for BRCA1-OC patients and 34.3% for SOC patients (p = 0.354). Mean time to progression was 22.7 and 14.5 months for BRCA1-OC and SOC group, respectively (p = 0.05). Complete response to pri-mary surgery and first line chemotherapy was obtained in 42.5% and 37.9% of cases, respectively; the difference, however, did not reach the statistical significance. CONCLUSIONS: Results of combined treatment in the group of BRCA1-related OC patients seem to be better than in the group of sporadic ovarian cancer patients.

Prevalence of respiratory tract infections, allergies and assessment of humoral immunity within the Malopolska region's cohort of 11- year old children born with extremely low birth weight in comparison with to their term born peers.

BACKGROUND: Children born with extremely low birth weight (ELBW) have more respiratory tract complications during childhood. Little is known about respiratory and allergy problems in ELBW children at the threshold of adolescence. MATERIALS AND METHODS: A follow-up study was conducted at the age of 11 among ELBW children (n=65) and age-matched controls (n=36). The primary outcomes in the study were the occurrence of respiratory and allergy problems and the rate of hospitalization due to respiratory complications at the age of 11 years, assessed with a questionnaire. Secondary outcome variables were serum levels of immunoglobulin classes. RESULTS: ELBW children had more respiratory tract infections (31 vs.11%, p = 0.03), but less allergies (3 vs. 22%, p < 0.01) compared with controls and had lower level of serum tIgE (geometric mean: 46.5 vs. 89.3 kU/l, p = 0.02). The risk factors for the occurrence of respiratory tract disorders in the ELBW group were: low gestational age, need for surfactant therapy and length of ventilatory support in the neonatal period. CONCLUSIONS: ELBW children have more frequent respiratory tract complications, but fewer allergies at the age of 11 years compared with children born at term. Lower respiratory tract problems decrease in ELBW children with age. Respiratory tract infections are not connected with deficiency in humoral immunity.

Nephrectomy or nephron-sparing surgery - how to decide?

AIM OF THE STUDY: Radical nephrectomy in the treatment of renal cell carcinoma (RCC) remains the gold standard, but nephron-sparing surgery (NSS) is still increasing in importance. The main goal of this study was to compare the diagnostic accuracy of ultrasound and multi-detector computed tomography in RCC staging and its influence on deciding about further patient treatment. MATERIAL AND METHODS: 87 patients (age range 27-90 years; median 61.5) underwent ultrasound (US) scan and contrast-enhanced computed tomography (CE-CT) of the abdomen and pelvis. 28 patients were qualified for NSS. The remaining group of patients underwent nephrectomy. RESULTS: NSS was performed more frequently among patients with lesions in the lower pole of the kidney and there was no infiltration to the calyx and renal pelvis. Radical nephrectomy (RN) was pursued in cases with lesions in the central or upper pole. Lesion diameter in patients qualified for NSS was smaller than in patients qualified for radical nephrectomy. CONCLUSIONS: Determining the relationship between tumour and adjacent structures is not a simple matter. According to our study, 50% of CT results differ from histopathology assessment. Tumour diameter determined in CT examination is larger than in ultrasound and histopathological measurements while US scanning tends to underestimate tumour size in relation to histopathological assessment.

Radiotherapy-induced lumbosacral plexopathy in a patient with cervical cancer: a case report and literature review.

Radiotherapy-induced lumbosacral plexopathy in cervical cancer treatment is a very rare, but extremely serious complication. The clinical course is associated with severe bilateral lower leg pain, reduced sensation, different degrees of weakness, paresis or paralysis, and sometimes also urinary or fecal incontinence. Patient quality of life becomes significantly deteriorated. Escalating neurological disorders may make self-sufficient functioning impossible. Neurological symptoms, most often irreversible, may develop at different times after irradiation, even after more than 30 years. We present a case of neurological toxicity in a patient successfully treated for cervical cancer with pelvis and para-aortic lymph node irradiation and weekly cisplatin. Neurological symptoms developed a few weeks after completion of external irradiation, were gradually escalating and resulted in complete immobilization of the woman. We underline the significance of long-term, systematic physiotherapy and pharmacological therapy which has resulted in significant improvement of motion efficiency. The literature review concerns the questions of frequency, clinical course and mechanisms of radiation-induced plexopathy.

Irradiation-induced bone sarcoma in a patient treated for cervix cancer 28 years earlier.

AIM OF THE STUDY: To present a case of a patient with cervical carcinoma in stage IIA who was diagnosed with pelvic bone sarcoma 28 years after radiotherapy. CASE PRESENTATION: A 37-year-old woman with IIA cervix cancer was treated with external beam irradiation and brachytherapy. The patient had undergone conventionally fractionated external beam irradiation using the "box" technique, with the total dose of 50 Gy and brachytherapy with radium applicators (intrauterine tube and fornix applicator) with the dose of 60 Gy calculated at point A. After treatment she was followed up for 2 years. Twenty-six years later, inoperable pelvic bone sarcoma was diagnosed within the irradiated field. The clinical course was aggressive and rapid progression during chemotherapy was observed. CONCLUSIONS: For patients receiving radiotherapy, long-term careful follow-up is mandatory due to second cancer risk. In the case of any suspicious symptoms, such patients need proper diagnosis to detect any disease as early as possible.

Assessment of long-term renal complications in extremely low birth weight children.

We assessed the long-term renal complications in a regional cohort of extremely low birth weight (ELBW) children born in 2002-2004. The study group, comprising 78 children born as ELBW infants (88% of the available cohort), was evaluated with measurement of serum cystatin C, urinary albumin excretion, renal ultrasound, and 24-h ambulatory blood pressure measurements. The control group included 38 children born full-term selected from one general practice in the district. Study patients were evaluated at a mean age of 6.7 years, and had a median birthweight of 890 g (25th-75th percentile: 760-950 g) and a median gestational age of 27 weeks (25th-75th percentile: 26-29 weeks). Mean serum cystatin C levels were significantly higher (0.64 vs. 0.59 mg/l; p = 0.01) in the ELBW group. Hypertension was diagnosed in 8/78 ELBW and 2/38 of the control children (p = 0.5). Microalbuminuria (>20 mg/g of creatinine) was detected only in five ELBW children (p = 0.17). The mean renal volume was significantly lower in the ELBW group (absolute kidney volume 81 ml vs. 113 ml; p < 0.001, relative kidney volume 85 vs. 97%; p < 0.001). Abnormally small kidneys (<2/3 of predicted size) were detected in 19 ELBW and four control children (p = 0.08). Multivariate logistic regression revealed that the only independent risk factor for renal complications was weight gained during neonatal hospitalization (odds ratio: 0.67; 95% confidence interval: 0.39-0.94). Serum cystatin C and kidney volume are significantly lower in school-age ELBW children. It is important to include systematic renal evaluation in the follow-up programs of ELBW infants.

Insulin-like growth factor-1 (IGF-1) serum concentration among 7-year-old extremely low birth weight children--an indicator of growth problems.

BACKGROUND: The study aim was to compare somatic development, body composition, insulin growth factor-1 (IGF-1) serum concentration and serum lipid profile between small-for-gestational-age (SGA) and appropriate-for-gestational-age (AGA) 7-year-old extremely low birth weight (ELBW) children and full-term (FT) controls. MATERIALS AND METHODS: Fifty-seven AGA ELBW children [mean birth weight (BW) 850 g and mean gestational age (GA) 26.4 weeks] and 24 SGA ELBW children (mean BW 833 g, mean GA 29.5 weeks) were evaluated. The control group included 37 age- and sex-matched FT children from one outpatient center. RESULTS: Nine AGA and four SGA were found to have short stature defined as height < 3rd percentile for chronological age (p = 0.9). In the AGA cohort, the results of weight, head circumference, triceps skin fold and body mass index (BMI) measurements were significantly reduced in the short-stature subgroup. IGF-1 serum concentrations differed significantly between short-stature and normal-stature subgroups (121 vs. 193 ng/mL; p = 0.02). In the SGA cohort, weight and serum concentrations of total cholesterol (5.03 vs. 4.26 mmol/L; p = 0.04), LDL cholesterol (3.24 vs. 2.38 mmol/L; p = 0.01) and IGF-1 (113 vs. 211 ng/mL; p = 0.01) differed significantly between the short-stature and normal-stature subgroups. CONCLUSIONS: Short stature at the age of 7 years was diagnosed in a similar percentage of AGA and SGA former ELBW infants. In both cohorts short-stature children have significantly lower weight and serum IGF-1 levels. IGF-1 seems to be involved in prolonged growth restriction among ELBW infants, regardless of whether they were AGA or SGA.

Characteristics of immune-competent amoebocytes non-invasively retrieved from populations of the sentinel earthworm Lumbricus rubellus (Annelida; Oligochaeta; Lumbricidae) inhabiting metal polluted field soils.

Lumbricus rubellus is a cosmopolitan earthworm devoid of riboflavin-storing eleocytes; its immune competent coelomocytes are predominantly amoebocytes. Our aim was to determine whether amoebocyte cytometrics in L. rubellus are robust biomarkers for innate immunological responses to environmental pollutants. Investigations were conducted on populations inhabiting three unpolluted and five metalliferous (mainly Pb+Zn+Cd) habitats in the UK and Poland. Inter-population differences in worm mass and amoebocyte numbers did not consistently reflect soil or tissue metal concentrations. Flow cytometry indicated that autofluorescence of the amoebocytes differs between cells from the unpolluted and metal-polluted worms, and pinocytosis of neutral red by amoebocytes was lower (especially at 15 versus 60 min incubation) in worms from the polluted Poland site compared with the reference population. To conclude, amoebocyte cytometrics and functionality are potentially useful for environmental diagnostics; deployment is contingent on better understanding potential confounders.

Clinico-morphological parameters affecting survival of patients with advanced cervical cancer.

The aim of the study was to investigate the prognostic significance of selected clinico-morphological parameters including Ki-67 antigen expression and microvessel density. The data of 122 patients with squamous cell carcinoma, FIGO stages IB-IIIB and treated with radiochemotherapy and brachytherapy were studied. Significant prognostic factors for disease-free survival in univariate analysis were the FIGO stage and the presence of atypical mitoses in carcinoma cells. Multivariate Cox analysis confirmed prognostic significance of the FIGO stage and Ki-67 expression with regard to disease-free survival. With regard to overall survival, the most important prognostic factor was Ki-67 antigen expression. The data concerning the pretreatment status of these parameters may be helpful in clinical practice.

Effects of nickel, zinc, and lead-contaminated soil on burrowing rate and coelomocytes of the earthworm, Allolobophora chlorotica.

We have shown previously that stubby worms Allolobophora chlorotica are sensitive to environmental stress, including metal-polluted soil. In order to discern the mechanisms of this sensitivity, adult (clitellate) Al. chlorotica were exposed in the laboratory to soil samples soaked with water (control) or Ni (1 and 2 mg/kg), Zn (1.25 and 2.5 g/kg) or Pb (5 and 10 g/kg) chlorides. Worms avoided contact with metal contaminants by prolonging burrowing time in metal-soaked samples, especially in the case of lead. Higher concentrations of the investigated metals were lethal for worms. During a 3 week exposure to lower metal concentrations, nickel and lead readily accumulated in the bodies of worms while zinc was efficiently regulated. However, body weights and numbers of non-invasively retrieved free coelomocytes (consisting of amoebocytes and riboflavin-loaded eleocytes) were significantly lower only in zinc-exposed worms. We assume that zinc regulation in worm bodies is more energy-demanding than nickel or lead bioaccumulation, thus this might be responsible for inhibition of the body gain and diminution of immunocompetent cells in zinc-exposed earthworms. Alternatively, missing free coelomocytes may actually be involved in Zn trafficking and removal through nephridia and/or in the formation of multicellular brown bodies, since metal can unbalance host/bacteria relationships.

[Differentiation of an integrated and episomal HPV-16 DNA using real-time PCR in cervical specimens of women diagnosed with intraepithelial lesions and invasive cervical cancer]. / Róznicowanie postaci zintegrowanej i episomalnej DNA HPV-16 metoda real-time PCR w wydzielinie szyjki macicy kobiet z rozpoznan sródnablonkowa neoplazja i rakiem szyjki macicy.

UNLABELLED: Persistent high-risk HPV infection, especially HPV-16, is considered to be an important step in the process of cervical carcinogenesis. Integration of viral DNA into the host genome through the destruction of HPV E2 sequences, increases the expression of viral proteins E6 and E7 and their participation in the transformation of cervical cancer. OBJECTIVE: The aim of this study was to apply real-time PCR (RT-PCR) to assess the prevalence of integrated and episomal HPV-16 DNA and determine viral DNA load in women with cervical intraepithelial lesions and invasive cervical cancer MATERIAL AND METHODS: A total of 84 women infected with HPV-16, including 44 with LSIL, 7 with HSIL and 33 with invasive cervical cancer participated in the study Cervical specimens were collected using the cytobrush. The presence of a sequence of E2 and E6 HPV-16 and human gene RNasy P was detected by quantitative RT-PCR. The viral load presented as the form of the virus genome copy numbers per 1,000 cells. RESULTS: The integrated form of HPV-16 genome was found in 97% of women with cervical cancer. In women with LSIL and HSIL mixed form (simultaneous occurrence of an integrated and episomal form) of the viral genome (84% and 57%, respectively) prevailed. The frequency of the integrated HPV-16 DNA increased with progression of dysplastic lesions of the cervix (p<0.001). Statistically significant differences in average number of copies of the virus in women with LSIL and HSIL compared to patients with cancer (p<0.001) were observed. The highest viral load was detected in women demonstrating an integrated HPV-16 DNA. CONCLUSIONS: Quantitative analysis of the sequence of E2 and E6 HPV-16 tested by RT-PCR can be used to determine the degree of integration of the viral genome and quantitative evaluation of viral load in clinical material. It can also serve as an additional parameter defining risk of progression of transformation in the cervix.

Multimodal longitudinal respiratory function assessment in very low birth weight 7-year-old children.

PURPOSE: Preterm birth is associated with adverse pulmonary outcomes. We aimed to evaluate respiratory morbidities and lung function of very low birth weight (VLBW) Polish children followed up at the age of 7 years old, and to compare with electrical impedance segmentography (EIS) results recorded at 4 years of age. MATERIALS AND METHODS: VLBW children were compared with term controls using impulse oscillometry and spirometry. Perinatal data and current respiratory morbidities were analyzed and pulmonary function test results were compared with previous EIS results. RESULTS: We included 40 VLBW children and 30 controls in the analysis. Elevated total airway resistance and forced expiratory volume in the first second below the lower limit of normal were more prevalent in VLBW children compared with term controls (15 vs 0%; 18 vs 0%). A positive bronchodilator response was more common in VLBW children (R5 Hz: 46 vs 13.3%; R5-20 â€‹Hz: 65 vs 36.7%). Children with bronchopulmonary dysplasia (BPD) had higher total airway resistance (R5 Hz/R5 Hz pred: 1.35 vs 0.95; p â€‹< â€‹0.001), large airway resistance (R20 Hz/R20 Hz pred: 0.89 vs 0.66; p â€‹= â€‹0.001), small airway resistance (R5-20 â€‹Hz: 0.57 vs 0.34 â€‹kPa â€‹L-1 â€‹s-1; p â€‹= â€‹0.009), than controls. Strong correlation between BDR in EIS and R5 Hz/R5 Hz pred was observed in children with BPD (r â€‹= â€‹0.7). CONCLUSION: VLBW school-aged children with BPD presented with substantial respiratory morbidity and persistent reduction of lung function, affecting small and large airways and lung parenchyma. EIS may be an alternative tool for lung function assessment in children with BPD.