Dual Switch in Lipid Metabolism in Cervical Epithelial Cells during Dysplasia Development Observed Using Raman Microscopy and Molecular Methods.

Cellular lipid metabolism is significantly transformed during oncogenesis. To assess how dysplasia development influences lipid cellular metabolisms and what is the molecular background behind it, cervical epithelial cells of 63 patients assigned to seven groups (based on the cytological examination and HPVhr test results) were studied using a multimethodological approach including Raman microscopy and molecular methods. The consistent picture obtained studying the lipid content, cell inflammation, SREBF1 gene methylation (hence SREBP1 inhibition) and level of mitochondrial DNA copies (indirectly the number of mitochondria) showed that changes in lipid metabolism were multidirectional. Cells from patients classified as mildly dysplastic (LSIL) exhibited a unique behavior (the highest level of inflammation and SREBF1 methylation, the lowest lipid content and mitochondrial DNA). On the contrary, cells from severe dysplastic (HSIL) and cancer (SCC) groups showed the opposite characteristics including the lowest SREBF1 gene methylation as well as the highest level of mitochondrial DNA and lipid cellular concentration (for HSIL/HPVhr+ and SCC groups). Following dysplastic progression, the lipid content decreases significantly (compared to the control) for mildly abnormal cells, but then increases for HSIL/HPVhr+ and SCC groups. This intriguing dual switch in lipid metabolism (reflected also in other studied parameters) on the way from normal to squamous carcinoma cells is of potential diagnostic interest.

Sequence variation analysis of the E1 and E2 genes of human papillomavirus type 16 in cervical lesions in women from the south of Poland.

The E1 and E2 genes of the human papillomavirus encode the so-called early proteins, their sequences are conserved, and regulatory functions are associated with the viral oncoproteins. The purpose of this study is to determine the HPV16 E1 and E2 mutations appearing in the female population of southern Poland, depending on the severity of cervical pathological changes. We also take into account the number of E1 and E2 mutations detected in the E6 gene variant (350G or 350T). This publication is one of the first in the Central and Eastern Europe to deal with this topic. We identified 4 mutations in the E1 gene and 24 mutations in the E2 gene that have not been described so far. In three cases of squamous cell carcinoma a C3409T mutation occurred, which is widely described as oncogenic. This mutation lies in the 3243-3539 area of the E2 hinge region. Statistical analyses show a possible relationship of mutations in this area with oncogenesis. The discovered dependencies may be important in the context of oncogenesis, however, a study with a larger group of patients is needed in order to confirm this view.

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.

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.

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.

Regional lung ventilation pattern in preschool children with bronchopulmonary dysplasia is modified by bronchodilator response.

BACKGROUND: Bronchopulmonary dysplasia (BPD) remains a significant long-term complication of prematurity. A standardized method of pulmonary function testing is still not available in preschool children with BPD. We investigated the feasibility of Electrical Impedance Segmentography (EIS) monitoring in this group and the impact of bronchodilator response (BDR) to salbutamol on the pattern of lung ventilation. METHODS: We conducted a follow-up study of 4-year-old premature children who had been treated in the tertiary NICU. The cohort was divided into two groups based on the presence of BPD. EIS monitoring was performed before and 15 min after the administration of 400 µg of salbutamol (pMDI with spacer) in all subjects during spontaneous tidal breathing in upright position. Data were expressed as median segmental impedance amplitude differences and segmental ventilation inhomogeneity index (II) changes. RESULTS: We included 51 children in our analysis: 33 with BPD (median birth weight-840 g; median gestational age-27 weeks) and 18 without BPD (1,290 g; 30 weeks, respectively). There was a significant increase in median segmental impedance amplitude after salbutamol in gravity non-dependent segments in children with BPD: upper left (UL): 462 versus 534 AU; (P = 0.003); upper right (UR): 481 versus 595 AU (P < 0.001) and II in these segments: UL: 0.046 versus 0.078 (P = 0.003) UR: 0.049 versus 0.064 (P = 0.006). There were no changes in the lower segments. There were no changes in ventilation pattern in children without BPD. CONCLUSION: BDR to salbutamol increases breath amplitude in gravity non-dependent segments of the lungs during spontaneous tidal breathing in preschool children with BPD. Pediatr Pulmonol. 2017;52:353-359. © 2016 Wiley Periodicals, Inc.

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.

HPV16 E6 polymorphism and physical state of viral genome in relation to the risk of cervical cancer in women from the south of Poland.

The aim of this study was to analyse the correlation between HPV16 E6 variants and the physical status of viral genome (integrated, mixed, episomal) among patients with cervical cancer (n=40) and low-grade squamous intraepithelial lesions - LSIL (n=40). The study was performed on 80 HPV16 positive samples. HPV16 E6 variants were identified using PCR and DNA sequencing. Nucleotide sequences of E6 were compared with the prototype sequence (EUR-350T). The physical state of HPV DNA was determined as the ratio of E2/E6 copy number per cell. Twelve different intratypic variants were identified as belonging to European (in 77 samples) and North-American 1 (in 3 samples) sublineages. The most prevalent non-synonymous variant was EUR-350G, which occurred with similar frequency in cervical cancer and LSIL. The frequencies of additional mutations in variants with EUR-350T or EUR-350G sequences differed significantly. For the first time, missense mutations G122A, C153T and G188A were discovered in EUR-350G variant. The integrated viral genome was predominant in women with cervical cancer. The EUR-350T prototype and EUR-350G without additional mutations variants were prevalent in cervical cancer samples with the HPV16 characterized by integrated DNA. In summary, European variants of HPV16 E6 dominated in both cancer and LSIL group. The presence of EUR-350G favoured the occurrence of additional nucleotide changes. We showed that nucleotide changes occur significantly more often in the mixed form of viral DNA and in LSIL group and that the variants without additional mutations may promote integration of HPV16 genome.

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.

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.

Physical state of human papillomavirus type 16 in cervical intraepithelial lesions and cancers determined by two different quantitative real-time PCR methods.

The aim of this study was to analyse the correlation between a new multiplex qPCR assay and a reference qPCR assay for assessment of the human papillomavirus (HPV16) load and the viral genome status. The study was performed on 100 HPV16 positive samples containing premalignant lesions and carcinomas. HPV16 E2 and E6 gene loads were assessed by two PCR methods. The load of E2 and E6 was normalized to the cell number by qPCR targeting the RNase P open reading frame. The physical state of the viral genome was determined as a ratio of E2/E6 copies number per cell. Among 100 samples analysed, there were no statistically significant differences in the E2 and E6 viral load evaluated by multiplex qPCR and qPCR, the correlation coefficients were 0.98 and 0.97, respectively. There were 19% of samples with the integrated, 73% with mixed and 8% with episomal state of viral genome detected by multiplex qPCR and 17%, 79%, 4%, respectively, found by qPCR. Prevalence of integrated and episomal forms estimated by multiplex qPCR was higher than the one obtained by qPCR (Chi2, p < 0.0001), but in samples with premalignant and malignant diagnoses no significant differences were demonstrated regardless of the methods used. Sensitivity and specificity of multiplex qPCR were 93.7% and 100% as compared with qPCR, the positive predictive value was 100%. In summary, the multiplex qPCR assay in respect of HPV16 load and the frequency of viral genome status was shown to be a sensitive and specific reference method. Simultaneous estimation of E2 and E6 genes in one reaction tube reduces the cost of testing.

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.

The prevalence and risk factors of allergic and respiratory symptoms in a regional cohort of extremely low birth weight children (<1000 g).

BACKGROUND: Children who were <1000 g (ELBW extremely low birth weight) at birth more frequently present with wheezing which is the most common reason that pediatric consultation is sought. Therefore asthma is diagnosed very often. However is the asthma that is diagnosed in ELBW subjects atopic in origin, or is there a different etiology? AIM: To determine if ELBW infants are at higher risk for the development of allergic and respiratory symptoms and to establish if there were any specific risk factors for these symptoms. METHODS: 81 children born with a mean birthweight of 845 g (91% of available cohort) were evaluated at the mean age 6.7 years. The control group included 40 full-term children. The children were examined for clinical signs of allergy, and were subjected to the following tests: serum total IgE, skin prick tests (SPT), exhaled nitric oxide measurement (FeNO) and spirometry. RESULTS: ELBW children had wheezing episodes more often (64% vs. 25%; OR (odds ratio): 5.38; 95% CI (confidence interval): 2.14-13.8) and were diagnosed more frequently with asthma (32% vs. 7.5%; OR: 5.83, 95% CI: 1.52-26) than their term born peers. The most important risk factors for wheezing persistence were hospitalization and wheezing episodes in first 24 months of life. Mean serum tIgE level (geometric mean: 32+/-4 vs. 56+/-4 kU/L; p=0.002) was higher and the number of children with positive results of tIgE level (12% vs. 32%; p=0.02) were more frequent in the control group. Children from the control group also more frequently had SPT, however this data was not statistically significant (11% vs. 24%; p=0.09). All of the ELBW had normal FeNO level (<=20 ppb), but 5 children from the control group had abnormal results (p=0.02). There was no difference between the groups in the occurrence of allergic symptoms. CONCLUSION: ELBW children have more frequent respiratory, but not allergic problems at the age of 6-7 years compared to children born at term. The need for rehospitalization in the first 2 years of life, was a more important risk factor of future respiratory problems at the age of 7 than perinatal factors, the diagnosis of bronchopulmonary dysplasia or allergy.

Comparison of the sensitivity and specificity of real-time PCR and in situ hybridization in HPV16 and 18 detection in archival cervical cancer specimens.

The aim of this study was to analyze the correlation between real-time PCR (RT-PCR) treated as a reference method and in situ hybridization with tyramide amplification system (ISH-TSA) in the detection of HPV16 and 18 infection and the assessment of viral genome status. The study was performed on cervical cancer biopsies fixed in 10% neutral buffered formalin and embedded in paraffin obtained from 85 women. TaqMan-based 5'exonuclease RT-PCR with type-specific primers was used to assess HPV16 and 18 infections and genome status. Viral infection and genome status was also assessed by ISH-TSA. RT-PCR revealed 76 (89.4%), and ISH-TSA 81 (95.3%) cancers with HPV16 and 18 infections. The ISH-TSA sensitivity and specificity were: 96.1% and 11.1% compared to RT-PCR. The difference between these techniques in HPV detection was significant (p = 0.000). Among 76 HPV16/18 positive cancers in RT-PCR, there were 30 (39.5%) with integrated and 46 (60.5%) with mixed viral genome form. According to ISH-TSA, there were 39 (51.3%) samples with integrated and 37 with mixed form (48.7%). The sensitivity and specificity of ISH-TSA in genome status assessment were 70.0% and 60.9%, respectively. The difference between RT-PCR and ISH-TSA in genome state detection was not statistically significant (p = 0.391). These results suggest that ISH-TSA shows insufficient specificity in HPV detection for use in clinical practice. However, this assay could be applied for viral genome status assessment.

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.

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.

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.

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.