The impact of postmastectomy and regional nodal radiation after neoadjuvant chemotherapy for clinically lymph node-positive breast cancer: a National Cancer Database (NCDB) analysis.

BACKGROUND: Following neoadjuvant chemotherapy (NAC), the optimal strategies for postmastectomy radiotherapy (PMRT) and regional nodal irradiation (RNI) after breast-conserving surgery (BCS) are controversial. In this analysis, we evaluate the impact of these radiotherapy (RT) approaches for women with clinically node-positive breast cancer treated with NAC in the National Cancer Database (NCDB). PATIENTS AND METHODS: Women with cT1-3 cN1 M0 breast cancer treated with NAC were divided into four cohorts by surgery [Mastectomy (Mast) versus BCS] and post-chemotherapy pathologic nodal status (ypN0 versus ypN+). Overall survival (OS) was estimated using the Kaplan-Meier method and RT approaches were analyzed using the log-rank test, multivariate Cox models, and propensity score-matched analyses. RESULTS: From 2003 to 2011, 15 315 cases were identified including 3040 Mast-ypN0, 7243 Mast-ypN+, 2070 BCS-ypN0, and 2962 BCS-ypN+ patients. On univariate analysis, PMRT was associated with improved OS for both Mast-ypN0 (P = 0.019) and Mast-ypN+ (P < 0.001) patients. On multivariate analyses adjusted for factors including age, comorbidity score, cT stage, in-breast pathologic complete response, axillary surgery, ypN stage, estrogen receptor status and hormone therapy, PMRT remained independently associated with improved OS among Mast-ypN0 [hazard ratio (HR) = 0.729, 95% confidence interval (CI) 0.566-0.939, P = 0.015] and Mast-ypN+ patients (HR = 0.772, 95% CI 0.689-0.866, P < 0.001). No differences in OS were observed with the addition of RNI to breast RT for BCS-ypN0 or BCS-ypN+ patients. Propensity score-matched analyses demonstrated identical patterns of significance. On subset analysis, OS was improved with PMRT in each pathologic nodal subgroup (ypN0, ypN1, and ypN2-3) (all P < 0.05). CONCLUSIONS: In the largest reported analysis of RT for cN1 patients treated with NAC, PMRT was associated with improved OS for all pathologic nodal subgroups. No OS differences were observed with the addition of RNI to breast RT.

Improved survival with dose-escalated radiotherapy in stage III non-small-cell lung cancer: analysis of the National Cancer Database.

BACKGROUND: Concurrent chemoradiation is the standard of care in non-operable stage III non-small-cell lung cancer (NSCLC). Data have suggested a benefit of dose escalation; however, results from the randomized dose-escalation trial RTOG 0617 revealed a lower survival rate with high-dose radiation. To evaluate the impact of dose escalation on overall survival (OS) in stage III NSCLC treated with chemoradiotherapy outside the controlled setting of a randomized trial, we carried out an observational, population-based investigation of the National Cancer Database (NCDB). PATIENTS AND METHODS: A total of 33 566 patients with stage III NSCLC treated with chemoradiation from 2004 to 2012 and radiation doses between 59.4 and 85 Gy were included. The primary end point was OS, with median survival calculated via Kaplan-Meier. Univariate, multivariable and propensity-score matching analyses were carried out. RESULTS: Patients were stratified by dose with median OS of: 18.8, 19.8 and 21.6 months for cohorts receiving 59.4-60, 61-69 and ≥70 Gy, respectively (P < 0.001). Granular dose analyses were carried out demonstrating increased OS with increasing radiation dose: median survival of 18.8, 21.1, 22.0 and 21.0 months for 59.4-60, 66, 70 and ≥71 Gy, respectively. While 66, 70 and ≥71 Gy resulted in increased OS in comparison with 59.4-60 Gy, no significant difference in OS was observed when comparing 66 with ≥71 Gy (P = 0.38). CONCLUSIONS: Dose escalation above 60 Gy was associated with improved OS in this cohort of stage III NSCLC patients treated with chemoradiotherapy. A plateau of benefit was observed, with no additional improvement in OS with increased dose (≥71 Gy) compared with 66-70 Gy. With evidence suggesting worse OS and quality of life with increased dose, these data support investigation of the role of intermediate-dose radiation, and in the absence of randomized evidence, may be leveraged to justify utilization of intermediate-dose radiation.

Disparities in disease presentation in the four screenable cancers according to health insurance status.

OBJECTIVES: Current guidelines support the use of screening for early detection in breast, prostate, colorectal and cervical cancer. The purpose of this study was to evaluate whether insurance status predicts for more advanced disease in these four currently screened cancers. STUDY DESIGN: The Surveillance, Epidemiology, and End Results (SEER) database was queried for breast, prostate, colorectal and cervix in patients aged 18-64 years. The database was queried from 2007 to 2011, with 425,614 patients with known insurance status included. METHODS: Multinomial logistic regression was used to evaluate insurance status and cancer presentation. RESULTS: Under multivariate analysis for breast cancer, uninsured patients more often had invasive disease (odds ratio [OR]: 1.55), T- (OR: 2.00), N- (OR: 1.59) stage, and metastatic disease (OR: 3.48), and were more often high-grade (OR: 1.21). For prostate cancer, uninsured patients again presented more commonly with higher T-stage (OR: 1.45), nodal (OR: 2.90) and metastatic (OR: 4.98) disease, in addition to higher prostate-specific antigen (OR: 2.85) and Gleason score (OR: 1.65). Colorectal cancer had similar findings with uninsured individuals presenting with more invasive disease (OR: 1.78), higher T (OR: 1.86), N (OR: 1.22), and M (OR: 1.58) stage, in addition to higher carcinoembryonic antigen levels (OR: 1.66). Similar results were seen for cervical cancer with uninsured having higher T (OR: 2.03), N (OR: 1.21), and M (OR: 1.45) stage. CONCLUSION: In the four cancers detected by screening exams, those without health insurance present with more advanced disease, with higher stage and grade, and more elevated tumour markers.

Effects of hydrostatic pressure on mouse sperm.

OBJECTIVES: The objective of this study was to investigate the abnormalities in sperm after exposure to hydrostatic pressure. BACKGROUND: Hydrostatic pressure acting on the cells is one of the fundamental environmental mechanical forces. Disorders of relationship between the cells and this mechanical force, such as when pressure varies beyond physiological limits, can lead to disease or pathological states. Sperm exposed to different range of hydrostatic pressure within male reproductive system and after entering the female reproductive system. METHODS: Sexually mature male NMRI mice, 8-12 weeks-old were sperm donors. Sperms were separated from the caudal epididymis and maintained in Ham's F-10 culture medium supplemented with 10 % FBS and divided into control and treatments. Sperm suspensions in the treatments were placed within pressure chamber and were subjected to increased hydrostatic pressure of 25, 50 and 100 mmHg (treatment I, II and III) above atmospheric pressure for 2 and 4 h. Sperm viability, motility, morphology, DNA integrity and fertilizing ability were assessed and compared with control. RESULTS: Results showed that hydrostatic pressure dependent on ranges and time manner reduced sperm quality due to adverse effect on viability, motility , morphology, DNA integrity and fertilizing ability in all of treatments, especially after 4h (p<0.05). CONCLUSION: Our data revealed hydrostatic pressure reduces sperm quality as a consequence of adverse effects on sperm parameters and may cause male infertility or subfertility (Tab. 5, Ref. 5).

Possible role of lncRNAs in amelioration of Parkinson's disease symptoms by transplantation of dopaminergic cells.

Long non-coding RNAs (lncRNAs) are biomarkers for diagnosis and treatment of Parkinson's disease (PD). Since dopaminergic cell transplantation is a clinical method to treat PD, this study investigated the effects of dopaminergic cell therapy on the expression of some lncRNAs and genes related to PD. In this study, Twenty-eight rats were randomly assigned to four experimental groups. The control group (Sal group) received saline injections. The Par group was a PD rat model with 6-hydroxydopamine (6-OHDA) injection in right striatum (ST). PD animals were transplanted by undifferentiated P19 stem cells (Par-E group), and P19-derived dopaminergic cells (Par-N group). Cell transplant effects were evaluated using behavioral tests (cylinder, open field, and rotarod tests), and histological methods (H&E and Nissl staining, and immunohistochemistry). Moreover, the expression of lncRNAs MALAT1, MEG3, and SNHG1, alongside specific neuronal (synaptophysin) and dopaminergic (tyrosine hydroxylase) markers was evaluated by qRT-PCR. Behavioral and histopathological examinations revealed that cell transplantation partially compensated dopaminergic cell degeneration in ST and substantia nigra (SN) of PD rats. The expression of MALAT1, SNHG1, and MEG3 was decreased in the ST of the Par group, while MEG3 and SNHG1 gene expression was increased in PBMC relative to the Sal group. In PBMC of the Par-N group, all three lncRNAs showed a reduction in their expression. Conversely, MALAT1 and SNHG1 expression was increased in ST tissue, while MEG3 gene expression was decreased compared to the Sal group. In conclusion, dopaminergic cell transplantation could change the lncRNAs expression. Furthermore, it partially improves symptoms in PD rats.

Clinical significance of mild inferolateral wall ischemia of the left ventricle on 99mTc-MIBI myocardial perfusion single photon emission computed tomography (SPECT).

INTRODUCTION: Mild ischemia in the inferolateral wall on myocardial perfusion imaging is seen frequently in practice. The aim of this study is to assess the importance of the above issue on myocardial perfusion SPECT with coronary angiography. PATIENTS AND METHODS: All patients enrolled in this study exhibited mild ischemia of the inferolateral wall on myocardial single photon emission computed tomography (SPECT) with 99mTc-MIBI, using the 20 left ventricular segments model. Each patient completed a questionnaire, including type of chest pain, risk factors, and previous examinations, and all cases were followed up for one year. Luminal stenosis of >50% was classified as significant stenosis on coronary angiography. A p value < 0.05 was considered statistically significant. RESULTS: During investigation, 105 cases had mild ischemia on myocardial perfusion imaging (MPI) of which 36 subjects (22 male and 14 female) underwent coronary angiography. The mean age was 56.62±10.23 years old (age range: 36-73 years). The inferolateral wall was compared to the left circumflex (LCX) territory. Nineteen out of 36 (52.7%) cases had stenosis in the LCX. Twenty-three of 105 (21.90%) underwent revascularization during the one year follow up. In multiple logistic regressions, with LCX stenosis on angiography as the dependent variable, only abnormal MPI was independently associated significantly. CONCLUSIONS: The findings of the study may indicate that even a mild perfusion defect in the inferolateral wall should be carefully managed, especially in high-risk subjects for coronary artery disease.

Complete improvement in a patient with multiple irreversible defects of the left ventricle on 99m technetium-sestamibi SPECT after percutaneous coronary intervention.

99mTc-sestamibi has been investigated as a potential viability marker; initial studies have shown good concordance between 201Tl and 99mTc-sestamibi activities in both viable and nonviable myocardium. However, assessment of myocardial viability by 99mTc-sestamibi remains controversial for tissue recovery after revascularization. Here, we present a patient with several regions of severely diminished and irreversible (defect persisting in both early and delay images of each set scanning) defects on initial scan which were dissolved completely on the follow up scan after an intervention. In a 75 year-old Asian woman with acute myocardial infarction who received thrombolytic therapy and subjected to percutaneous coronary angiography (PCI) on day 28 after acute myocardial infarction(MI), resting 99mTc-sestamibi SPECT was applied on day 4 (initial scan) and 138 (follow up scan) after acute MI at 30 and 180 min after injection of tracer (740 MBq); Two-dimensional echocardiography was carried out at the same time. On the initial image set, there was irreversible defects in the apex, anteroapical, inferoapical, anteroseptal, septal and also anterior walls, while the follow up image was normal in all regions.The angiography intervention showed just significant stenosis on left anterior descending (LAD) vessel (95%). This may highlight the failure of 99mTc-sestamibi as a marker of myocardial viability and also mandate further validating of the procedure with follow up scan or other modalities for myocardial viability investigation.

The investigation of ABO and Rh blood groups distribution in patients with endometriosis needs new project design.

We carefully studied all the three published papers in your journal as "ABO and Rh Blood group distribution in patients with endometriosis" and "Associations of ABO blood groups with various gynecologic diseases" and would like to express our point of view about them.

The association of rate pressure product (RPP) and myocardial perfusion imaging (MPI) findings: a preliminary study.

INTRODUCTION: The product of heart rate and systolic blood pressure, termed as rate-pressure product (RPP), is a very reliable indicator of myocardial oxygen demand and is widely used clinically. There have been previous attempts to describe the relationship between RPP and the onset of pain in angina pectoris. The current study aimed to evaluate the association between RPP results and scan findings. MATERIALS AND METHODS: In total, 497 patients with suspected coronary artery disease (CAD) underwent gated, single-photon emission computed tomography (SPECT) imaging with dipyridamole, exercise, or dobutamine stress, and were included in this study. Baseline and maximum heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), and electrocardiogram (ECG) results were recorded. The rate-pressure product (RPP) was calculated as the product of heart rate and systolic arterial pressure for both baseline and maximum measures. The difference between the RPP max and the basal RPP is known as the RPP reserve. Researchers also obtained semi-quantitative analyses of myocardial perfusion imaging (MPI), using gated software, demographic information, risk factors of CAD, and pretest likelihoods of CAD using nomograms. RESULT: Four hundred and ninety-seven cases, including 426 patients with dipyridamole stress, 59 with exercise stress, and 12 with dobutamine stress, underwent myocardial perfusion imaging. Scan results were positive in 194 (45.5%) and negative in 232 (54.5%) patients with dipyridamole stress. In patients with exercise stress, the scan was positive in 24 (40.7%) cases and negative in 35 (59.3%) cases. In dobutamine stressed patients, the scan was positive in 6 (50%) cases and negative in the 6 remaining cases. Dipyridamole stress resulted in a significant difference between HR at rest and at maximum (28.95 ± 24.53, p-value<0.0001), between systolic BP at rest and maximum (6.75 ± 12.50, p-value<0.0001) and between diastolic BP at rest and maximum (1.45 ± 5.80; p-value<0.0001). There was a significant correlation between sum stress scores (SSS) and reserved RPP (r= -0.12, p-value<0.001) which, in dipyridamole patients, was r=-0.18, p-value=0.0001). In addition, there was a significant association between reserved RPP and risk of CAD (p-value<0.001). In the patients with dipyridamole stress, the ejection fraction (EF) change (odds ratio =0.92; 95% CI: 0.86-0.98; p=0.01), reserve RPP (odds ratio =1.00; 95% CI: 1.00-1.00; p=0.04), risk of CAD (odds ratio =5.80; 95% CI: 3.21-10.50; p<0.0001) and age (odds ratio =0.94; 95% CI: 0.89-0.98; p=0.01) were associated significantly with MPI results, using multiple logistic regressions. CONCLUSION: The study demonstrated that RPP is associated with MPI findings using gated SPECT imaging with dipyridamole stress. However, to confirm this preliminary result, further studies are mandatory.

Size-dependent molecular interaction of nontraditional 2D antibiotics withStaphylococcus aureus.

The application of nanomaterials for their antibacterial properties is the subject of many studies due to antibiotic resistance of pathogen bacteria and the necessity of omitting them from food and water resources. Graphene oxide (GO) is one of the most popular candidates for antibacterial application. However, the optimum condition for such an effect is not yet clear for practical purposes. To shed light on how GO and bacteria interaction depends on size, a wide range of GO flake sizes from hundreds of µm2going down to nano-scale as low as 10 N m2was produced. In anin-vitrosystematic study to inhibitStaphylococcus aureusgrowth, the correlation between GO flake size, thickness, functional group density, and antibacterial activity was investigated. The GO suspension with the average size of 0.05 µm2, in the order of the size of the bacteria itself, had the best bacteriostatic effect onS. aureuswith the minimum inhibitory concentration value of 8 µg ml-1, well within the acceptable range for practical use. The bacteriostatic effect was measured to be a 76.2% reduction of the colony count over 2 h of incubation and the mechanism of action was the wrapping and isolation of cells from the growth environment. Furthermore,in-vivoanimal studies revealed that 16 µg ml-1of the optimum GO has efficient antibacterial performance against the methicillin-resistant strains of the bacteria with an enhanced wound healing rate and tensiometrial parameters which is important for realized targets.

Scintigraphic parameters with emphasis on perfusion appraisal in rest 99mTc-sestamibi SPECT in the recovery of myocardial function after thrombolytic therapy in patients with ST elevation myocardial infarction (STEMI).

INTRODUCTION: This study was performed to determine the clinical application of rest 99mTc-sestamibi in the assessment of viability and functional improvement of the left ventricle (LV) myocardium in the post-thrombolytic therapy of acute myocardial infarction (AMI). MATERIAL AND METHODS: In 37 patients with AMI who received thrombolytic therapy, 2-dimensional (2D) echocardiography, as well as the resting redistribution of 99mTc-sestamibi, was investigated, both within 1 week and 3-5 months after AMI. The predictive capacity of the perfusion percentage for myocardial function recovery was evaluated. Also, the capacities of the possible variables in the prediction of recovery of myocardial function resulting from a change in LV ejection fraction (EF) were evaluated using stepwise multiple regression analysis. RESULTS: Thirty-seven patients (30 men and 7 women; mean age: 58±14 years) with AMI were enrolled in the study. Redistribution was observed in 35 and 50 segments of the initial and follow-up scans, respectively. In addition, 146 segments with reverse redistribution (RR), both in the initial scan (118 segments) and the follow-up scan (86 segments), were also observed. An apparent difference in wall motion scores was seen between the initial and follow-up echocardiographs (p<0.001). Furthermore, using the optimal cut-off point of perfusion percentage in each image set, sensitivity as well as specificity and likelihood ratio (LR) for the improvement of regional wall motion after 3-5 months were defined. CONCLUSION: These data showed that redistribution and reverse redistribution of 99mTc-sestamibi post thrombolytic therapy can be used as a marker of viability to predict the recovery of segmental wall motion abnormality (stunning), as well as the improvement of segmental perfusion uptake. This study also demonstrates that the resting 99mTc-sestamibi SPECT can be used for an approximate assessment of LV function status and can predict the recovery of jeopardized myocardium function after thrombolytic therapy.

Systemic infection with Candida albicans in breast tumor bearing mice: Cytokines dysregulation and induction of regulatory T cells.

OBJECTIVE: The effect of candidemia on immunologic parameters in breast tumor bearing patients is not well studied. Here, we hypothesised that candidemia in the tumor background may change the outcome of immunologic parameters and tumor condition. METHOD: Mice were divided into four groups, including normal, tumor, Candida infected (only Candidiasis) and tumor/Candidiasis groups. Tumor changes were recorded daily after tumor transplantation and induction of candidemia. Splenocytes of mice were harvested, cultured, and stimulated with PHA; afterwards, IL-4, IL-10, IFN-γ, TNF-α and TGF-ß cytokines were assessed using ELISA kits. We also evaluated the population of CD4+CD25+Foxp3+ regulatory T cells in the tumor infiltrated and splenocytes. RESULTS: The results showed that infection with C. albicans decreased the IFN-γ/IL-4 ratio in tumor/candidiasis and candidiasis groups versus their non-infected controls. IL-10, TGF-ß and TNF-α levels increased in the candidiasis group. In addition, Candidemia led to an increase in the Treg population in tumor microenvironment and splenocytes of experimental groups compared with non-infected controls. Finally, candidemia increased tumor growth of tumor/Candidiasis group compared with the tumor group. CONCLUSION: It seems that systemic infection with C. albicans could not only induce regulatory T cells but also result in dysregulation of cytokine network and thereby facilitate tumor growth.

Patterns of lymph node sampling and the impact of lymph node density in favorable histology Wilms tumor: An analysis of the national cancer database.

INTRODUCTION: There is controversy about the role of lymph node (LN) sampling or dissection in the management of favorable histology (FH) Wilms tumor (WT), specifically how it performed and how it may impact survival. OBJECTIVE: The objective of this study was to analyze factors affecting LN sampling patterns and the impact of LN yield and density (number of positive LNs/LNs examined) on overall survival (OS) in patients with advanced-stage favorable histology Wilms tumor (FHWT). METHODS: The National Cancer Database (NCDB) was queried for patients with FHWT during 2004-2013. Demographic, clinical and OS data were abstracted for those who underwent surgical resection. Poisson regression was performed to analyze how factors influenced LN yield. Patients with positive LNs had LN density calculated and were further analyzed. RESULTS: A total of 2340 patients met criteria, with a median age at diagnosis of 3 years (range 0-78 years). The median number of LNs examined was three (range 0-87). Lymph node yield was affected by age, race, insurance, tumor size, laterality, advanced stage, LN positivity, and institutional volume. A total of 390 (16.6%) patients had LN-positive disease. Median LN density for these LN-positive patients was 0.38 (range 0.02-1) (Summary Figure). Estimated 5-year OS was significantly improved for those with LN density ≤0.38 vs. >0.38 (94% vs. 84.6%, P = 0.012). In this population, on multivariate analysis, age and LN density were significant predictors of OS. DISCUSSION: It is difficult to compile large numbers of cases in rare diseases like WT, and fortunately a large administrative database such as the NCDB can serve as a great resource. However, administrative data come with inherent limitations such as missing data and inability to account for a variety of factors that may influence LN yield and/or OS (specimen designation, pathologist experience, surgeon experience/volume, institutional Children's Oncology Group (COG) association, etc.). In this specific disease, the American Joint Committee on Cancer staging (captured by the NCDB) is different than the COG WT staging system that is used clinically, and the NCDB does not capture oncologic outcomes beyond OS. CONCLUSIONS: In a review of the NCDB, various factors associated with LN yield and observed LN density were identified to be significantly associated with OS in patients with LN-positive FHWT. This reinforces the need for adequate LN sampling at the time of WT surgery, to maximize surgical disease control. It was proposed that LN density as a metric may allow for improved risk-stratification, and possibly allow for therapeutic reduction in a sub-set of patients with low LN density.

Pseudomonas aeruginosa infections are prevented in cystic fibrosis patients by avian antibodies binding Pseudomonas aeruginosa flagellin.

Pseudomonas aeruginosa (PA) is the main cause of morbidity and mortality in cystic fibrosis (CF) patients. CF patients with chronic PA infections have a more rapid deterioration of their lung function and the bacteria become impossible to eradicate from the lungs. Antibiotic resistance among PA strains in CF patients is steadily increasing. Specific chicken (IgY) antibodies against PA have been shown to have potential to prevent PA infections in CF. Anti-Pseudomonas IgY reduces PA adhesion to epithelia, but the mechanism has not been fully elucidated. To gain further insight into the prophylactic effect of these antibodies, the immunoreactivity was investigated by 2D electrophoresis of PA strains, immunoblotting and MALDI-TOF-MS. To confirm the identity of the proteins, the tryptic peptides were analyzed by MALDI-TOF-MS to accurately measure their monoisotopic masses as well as determine their amino acid sequences. In order to facilitate fragmentation of the peptides they were N-terminally or C-terminally labeled. Several strains were investigated and anti-Pseudomonas IgY was immunoreactive against all of these strains, which strengthens its potential as a prophylactic treatment against PA. Flagellin was identified as the major antigen. Flagellin is the main protein of the flagella and is crucial for establishing infections in hosts as well as being involved in PA chemotaxis, motility, adhesion and inflammation. Furthermore, secreted flagellin elicits an inflammatory response. In conclusion, anti-Pseudomonas IgY binds flagellin, which may prevent PA infections in CF patients by hindering host invasion.

Is mucin a determinant of peritoneal dissemination of gastrointestinal cancer? Analysis of mucin depletion in two preclinical models.

BACKGROUND: Mucinous gastrointestinal cancers may indicate a higher propensity for widespread peritoneal seeding than their non-mucinous counterparts. We hypothesized that mucin content of gastrointestinal cancer cells and tumors is an indicator of cell viability and a determinant of the peritoneal tumor burden and tested our hypothesis in relevant experimental models. METHODS: MKN45 and LS174T models of human gastrointestinal cancer were treated with known mucin-depleting agents in vitro and in vivo, their mucin production was evaluated with Western blot immunohistochemistry, PAS staining and ELISA, and its correlation with cell viability and peritoneal tumor burden was analyzed. RESULTS: A relationship was found between the viability of cancer cells and their mucin levels in vitro. In agreement, when treated animal models were categorized into low- and high-burden groups (based on the weight and number of the peritoneal nodules), tumoral mucin levels were found to be significantly higher in the latter group. CONCLUSIONS: Tumoral mucin is apparently among the factors that dictate the pattern and extent of the peritoneal spread of gastrointestinal cancer, where it allows for enhanced dissemination and redistribution. If further tested and validated, our hypothesis could lay the basis for the development of novel mucin-targeted strategies.

Remote cerebellar hemorrhage.

Remote cerebellar hemorrhage (RCH) is a rare but benign, self-limited complication of supratentorial craniotomies that, to the best of our knowledge, has not been described in the imaging literature. RCH can be an unexpected finding on routine postoperative imaging studies and should not be mistaken for more ominous causes of bleeding such as coagulopathy, hemorrhagic infarction, or cortical vein occlusion. Cerebellar hemorrhage in the typical setting can be identified as RCH and does not require more extensive or invasive evaluation.

Strength of conjugate binding to plasmid DNA affects degradation rate and expression level in vivo.

In vitro assays have demonstrated the capability of poly-L-lysine to protect plasmid DNA from serum nucleases and cellular lysates. Our purpose was to evaluate the stability and potency of poly-L-lysine-DNA polyplexes after intravenous injection into mice. Polyplexes consisted of 32P-radiolabeled plasmid DNA complexed with poly-L-lysine at specified charge ratios. Variations in conjugate hydrophobicity and levels of modification with polyethylene glycol were investigated. Our results show that, in contrast to in vitro studies, the systemically administered polyplexes exhibited marked DNA degradation in the vascular compartment within 5 min. Substitution of poly-L-lysine epsilon-amino sites with polyethylene glycol or hydrocarbon chains resulted in faster degradation even when complexed at higher charge (+/-) ratios. Use of excess cationic charge in the polyplexes (+/- 2.5) diminished degradation rates only slightly. An analysis was made of the strength of the poly-L-lysine:DNA interaction by competition with poly-aspartic acid. Polyplexes with the strongest binding between conjugate and DNA in the competition assay were also the most stable in blood. However, tighter binding was not enough to fully protect the polyplex in vivo and polyplex DNA was substantially degraded within 10 min. Increased polyplex stability did not correlate with improved in vivo transfection efficiency.

Stabilization of poly-L-lysine/DNA polyplexes for in vivo gene delivery to the liver.

We are developing a self-assembling non-viral in vivo gene delivery vehicle based on poly-l-lysine and plasmid DNA. We have characterized poly-l-lysines of different chain lengths for DNA condensation and strength of DNA binding. Poly-l-lysine chains >20 residues bound DNA efficiently in physiological saline, while shorter chains did not. Attachment of asialoorosomucoid to PLL increased the PLL chain length required for efficient DNA binding in saline and for efficient DNA condensation. By electron microscopy, poly-l-lysine/DNA polyplexes appeared as toroids 25-50 nm in diameter or rods 40-80 nm long; conjugation of asialoorosomucoid to the polylysine component increased the size of resulting polyplexes to 50-90 nm. In water, poly-l-lysine and asialoorosomucoid-PLL polyplexes have effective diameters of 46 and 87.6 nm, respectively. Polyplexes containing only poly-l-lysine and DNA aggregated in physiological saline at all charge ratios and aggregated at neutral charge ratios in water. Attachment of asialoorosomucoid lessened, but did not eliminate, the aggregation of PLL polyplexes, and did not result in efficient delivery of polyplexes to hepatocytes. Conjugation of polyethylene glycol to poly-l-lysine sterically stabilized resulting polyplexes at neutral charge ratios by shielding the surfaces. For efficient in vivo gene delivery, polyplexes will need to be sterically stabilized to prevent aggregation and interaction with serum components.