Mucoadhesion mechanism of chitosan and thiolated chitosan-poly(isobutyl cyanoacrylate) core-shell nanoparticles

The study is focused on the evaluation of the potential bioadhesive behaviour of chitosan and thiolated chitosan (chitosan-TBA)-coated poly(isobutyl cyanoacrylates) (PIBCA) nanoparticles. Nanoparticles were obtained by radical emulsion polymerisation with chitosan of different molecular weight and with different proportions of chitosan/chitosan-TBA. Mucoadhesion was ex vivo evaluated under static conditions by applying nanoparticle suspensions on rat intestinal mucosal surfaces and evaluating the amount of nanoparticles remaining attached to the mucosa after incubation. The analysis of the results obtained demonstrated that the presence of either chitosan or thiolated chitosan on the PIBCA nanoparticle surface clearly enhanced the mucoadhesion behaviour thanks to non-covalent interactions (ionic interaction and hydrogen bonds) with mucus chains. Both, the molecular weight of chitosan and the proportion of chitosan-TBA in the formulation influenced the nanoparticle hydrodynamic diameter and hence their transport through the mucus layer. Improved interpenetration ability with the mucus chain during the attachment process was suggested for the chitosan of high molecular weight, enhancing the bioadhesiveness of the system. The presence of thiol groups on the nanoparticle surface at high concentration (200 x 10(-6) micromol SH/cm2) increased the mucoadhesion capacity of nanoparticles by forming covalent bonds with the cysteine residues of the mucus glycoproteins.     

Natural history of human papillomavirus infection in non-vaccinated young males: low clearance probability in high-risk genotypes

In this study, we aimed to investigate the clearance of type-specific genital human papillomavirus (HPV) infection in heterosexual, non-HPV-vaccinated males whose female partners were positive to HPV DNA tests. All consecutive men attending the same sexually transmitted diseases (STD) centre between January 2005 and December 2006 were considered for this study. All subjects (n?=?1009) underwent a urologic visit and microbiological tests on first void, midstream urine and total ejaculate samples. One hundred and five patients were positive for HPV DNA (10.4 %; mean age: 34.8?±?5.8 years) and consented to clinical examination and molecular diagnostic assays for HPV detection scheduled every 6 months (median surveillance period of 53.2 months). HPV genotypes were classified as high risk, probable high risk and low risk. HPV-positive samples which did not hybridise with any of the type-specific probes were referred to as positive non-genotypeable. At enrollment, the distribution of HPV genotypes was as follows: high-risk HPV (n?=?37), probable high-risk HPV (n?=?6), low-risk HPV (n?=?23) and non-genotypeable HPV (n?=?39). A high HPV genotype concordance between stable sexual partners emerged (kappa?=?0.92; p?<?0.001). At the end of the study, 71/105 (67.6 %) subjects were negative for HPV (mean virus clearance time: 24.3 months). With regard to the HPV genotype, virus clearance was observed in 14/37 (37.8 %) high-risk HPV cases, 6/6 (100 %) probable high-risk HPV cases, 20/23 (86.9 %) low-risk HPV cases and 31/39 (79.5 %) non-genotypeable cases. The high-risk HPV genotypes showed the lowest rate and probability of viral clearance (p?<?0.001). In our series, high-risk HPV infections were more likely to persist over time when compared with other HPV genotypes.     

Lectin histochemistry on the dorsal epidermis of the Breton dog

Expression of sugar residues and the nature of oligosaccharide linkage during keratinocyte maturation in the epidermis of the Breton dog were studied with the use of lectin histochemistry. Thirteen lectins were used. Labelling was not observed with GSA I-B4, GSA II, UEA-I, and LTA. The cytoplasm of keratinocytes reacted with PNA, HPA, Con A, and WGA from the basal layer to the granular layer. PNA and Con A showed highest reactivity in the granular cell layer. The cell surface showed increased reactivity with PNA, HPA, and WGA with maturation of keratinocytes. KOH-neuraminidase treatment (KOH-Neu) increased PNA and RCA120 staining during keratinocyte differentiation thus indicating an increase in oligosaccharides terminating with sialic acid-Galbeta(1,3)GalNAc and sialic acid-Galbeta(1,4)GlcNAc, respectively. Labelling of the glycocalyx of basal and spinous keratinocytes with SNA and MAA revealed terminal Neu5acalpha(2,6)Gal/GalNAc and Neu5acalpha(2,3)Galbeta(1,4)GlcNAc. KOH-Neu-DBA showed oligosaccharides terminating with sialic acid-GalNAcalpha(1,3)GalNAc in the spinous and granular layers. A selective glycocalyx labelling of granular keratinocytes was observed with DBA and SBA. Reactions with MAA, PNA, DBA, RCA120, SBA, HPA, and WGA disappeared after the beta-elimination reaction. Our findings indicate that Breton dog epidermis contains more O-linked than N-linked oligosaccharides and confirm that different subpopulations of keratinocytes can be distinguished by lectin histochemistry.     

Expression and association data strongly support JARID2 involvement in nonsyndromic cleft lip with or without cleft palate

Nonsyndromic cleft lip with or without cleft palate (CL/P) affects approximately 1 in 1,000 births. Genetic studies have provided evidence for the role of several genes and candidate loci in clefting; however, conflicting results have frequently been obtained and much have to be done to unravel the complex genetics of CL/P. In the present investigation we have focused on the candidate region in 6p23, a region that have been found linked to CL/P in several investigations, in the attempt to find out the susceptibility gene provisionally named OFC1. Gene expression experiments in mice embryo of positional candidate genes revealed that JARID2 was highly and specifically expressed in epithelial cells in merging palatal shelves. A family‐based linkage disequilibrium study confirmed the pivotal role of JARID2 in orofacial development and strongly supports a role for this gene in CL/P etiology (multiallelic haplotype test P=6×10^?5). Understanding the molecular role of JARID2 within facial development may offer additional information to further unravel the complex genetics of CL/P. Hum Mutat 31:1–7, 2010. © 2010 Wiley‐Liss, Inc.     

Sensory, motor somatic, and autonomic neurons projecting to the porcine cremaster muscle

The location of sensory, somatic, and autonomic neurons projecting to the pig cremaster muscle (CM) was studied by means of the retrograde neuronal tracer Fast Blue (FB) technique. FB was randomly injected in the left CM of four impuberal pigs and serial sections of sensory and autonomic ganglia and spinal cord were examined under a fluorescence microscope. Additionally, some indications about the number and size of labeled neurons were given. Sensory pseudounipolar somata were located ipsilaterally in the L2-L6 and S1-S2 dorsal root ganglia, their total number ranging between 125 and 194, their mean diameter between 24 and 89 microm. Somatic multipolar motoneurons were located ipsilaterally in the L2-L4 neuromeres of the spinal cord, their total number ranging between 53 and 169, their mean diameter between 29 and 53 microm. Autonomic multipolar paravertebral ganglia neurons were located ipsilaterally from L1 to S4 and contralaterally from L2 to S2. Their total number ranged from 2,015 to 3,067 and their mean diameter between 25 and 55 microm. The multipolar caudal mesenteric ganglia neurons were located bilaterally, their total number ranging between 14 and 1,408 and their diameter from 22 to 39 microm. In two subjects only, multipolar neurons were also found ipsilaterally in the microganglia of pelvic plexus (2 and 13 neurons). Their mean diameter ranged between 28 and 54 microm. Our study documented that the CM-projecting neurons were located at different neural levels, with a predominance in the autonomic ganglia.     

MR in the evaluation of new anterior cruciate ligament and tibial tunnel position: correlation with clinical and functional features

Purpose

This study aimed to evaluate correlations between the position of the tibial tunnel, its alignment with the ligament-screw system, presence of intratunnel fluid, position of the tibial tunnel with respect to the Blumensaat line and clinical knee stability in patients who underwent arthroscopic reconstruction of the anterior cruciate ligament (ACL), by using magnetic resonance (MR) imaging.

Materials and methods

Forty-eight patients (40 men, eight women; mean age, 31 years) underwent arthroscopic reconstruction of the ACL using double-strand semitendinosus and gracilis tendons. The new ACL was fixed to the tibial tunnel using Bio-Intrafix (Mitek). All patients underwent MR imaging 12 months after surgery and clinical evaluation at 6 and 12 months using the International Knee Documentation Committee (IKDC) scoring system. MR imaging and clinical features were correlated using the Mann-Whitney U test for continuous variables and Fisher??s exact test for categorical variables.

Results

Forty-one patients were clinically stable (groups A and B according to the IKDC test) and seven were unstable (group C). Mean values of tibial tunnel position in clinically unstable vs stable patients were, respectively, ?3.6 ±3.8 mm vs. ?2.8±3.8 mm in relation to the Blumensaat line (p=0.5712) and 77.3°±11.3 vs. 72.5°±5.5 necesas concerned the angle measured on the coronal view of the new ACL (p=0.3248); fluid was present in the tibial tunnel in 42.9% and 9.8% of cases, respectively (p=0.2104). MR imaging showed misalignment of ligament screw and tibial tunnel in 57.1% of patients in group C and in 12.2% in groups A and B (p=0.017).

Conclusions

Misalignment of the ligament-screw system and the tibial tunnel and the presence of fluid in the tibial tunnel appear to be directly correlated with clinical instability.     

Comparison of Adding Treatment of PTSD During and After Shelter Stay to Standard Care in Residents of Battered Women's Shelters: Results of a Randomized Clinical Trial

This study explored the acceptability, feasibility, and initial efficacy of an expanded version of a PTSD treatment developed for residents of battered women's shelters, Helping to Overcome PTSD through Empowerment (HOPE) in women who received standard shelter services (SSSs). A Phase I randomized clinical trial comparing HOPE + SSSs (n = 30) to SSSs (n = 30) was conducted. Primary outcome measures included the Clinician‐Administered PTSD Scale (Blake et al., 1995) and the Revised Conflict Tactic Scales (Straus, Hamby, Boney‐McCoy, & Sugarman, 1996 ). Participants were followed at 1‐week, and 3‐ and 6‐months posttreatment. Only 2 women dropped out of HOPE + SSS treatment. Latent growth curve analyses found significant treatment effects for PTSD from intimate partner violence (IPV) (β = ?.007, p = .021), but not for future IPV (β = .002, p = .709) across follow‐up points. Significant effects were also found for secondary outcomes of depression severity (β = ?.006, p = .052), empowerment (β = .155, p = .022), and resource gain (β = .158, p = .036). Additionally, more women in HOPE + SSSs were employed at 3‐ and 6‐month follow‐up compared to those in SSSs only. Results showed the acceptability and feasibility of adding IPV‐related treatment to standard services. They also suggested that HOPE may be a promising treatment for residents of battered women's shelters. Further research with a larger sample, utilizing more diverse shelter settings and a more rigorous control condition, is needed to confirm these findings.     

Lumbosacral epidural lipomatosis: MRI grading

Lumbosacral epidural lipomatosis (LEL) is characterized by excessive deposition of epidural fat (EF). The purpose of our retrospective study was to quantify normal and pathologic amounts of EF in order to develop a reproducible MRI grading of LEL. In this study of 2528 patients (1095 men and 1433 women; age range 18–84 years, mean age 47.3 years) we performed a retrospective analysis of MRI exams. We obtained four linear measurements at the axial plane parallel and tangent to the superior end plate of S1 vertebral body: antero-posterior diameter of dural sac (A-Pd DuS), A-Pd of EF, located ventrally and dorsally to the DuS, and A-Pd of the spinal canal (Spi C). We calculated (a) DuS/EF index and (b) EF/Spi C index. We developed the following MRI grading of LEL: normal, grade 0: DuS/EF index ≥1.5, EF/Spi C index ≤40%; LEL grade I: DuS/EF index 1.49–1, EF/Spi C index 41–50% (mild EF overgrowth); LEL grade II: DuS/EF index 0.99–0.34, EF/Spi C index 51–74% (moderate EF overgrowth); LEL grade III: DuS/EF index ≤0.33, EF/Spi C index ≥75% (severe EF overgrowth). The MRI exams were evaluated independently by three readers. Intra- and interobserver reliabilities were obtained by calculating Kappa statistics. The MRI grading showed the following distribution: grade 0, 2003 patients (79.2%); LEL grade I, 308 patients (12.2%); LEL grade II, 165 patients (6.5%); and LEL grade III, 52 patients (2.1%). The kappa coefficients for intra- and interobserver agreement in a four-grade classification system were substantial to excellent: intraobserver, kappa range 0.79 [95% confidence interval (CI), 0.65–0.93] to 0.82 (95% CI, 0.70–0.95); interobserver, kappa range 0.76 (95% CI, 0.62–0.91) to 0.85 (95% CI, 0.73–0.97). In LEL grade I, there were no symptomatic cases due to fat hypertrophy. LEL grade II was symptomatic in only 24 cases (14.5%). In LEL grade III, all cases were symptomatic. A subgroup of 22 patients (42.3%) showed other substantial spinal pathologies (e.g., disk herniation). By means of simple reproducible measurements and indexes MRI grading enables a distinction between mild, moderate, and severe EF hypertrophy. Kappa statistics indicate that LEL can be reliably classified into a four-grade system by experienced observers.     

In vivo MRI of cancer cell fate at the single-cell level in a mouse model of breast cancer metastasis to the brain.

Metastasis (the spread of cancer from a primary tumor to secondary organs) is responsible for most cancer deaths. The ability to follow the fate of a population of tumor cells over time in an experimental animal would provide a powerful new way to monitor the metastatic process. Here we describe a magnetic resonance imaging (MRI) technique that permits the tracking of breast cancer cells in a mouse model of brain metastasis at the single-cell level. Cancer cells that were injected into the left ventricle of the mouse heart and then delivered to the brain were detectable on MR images. This allowed the visualization of the initial delivery and distribution of cells, as well as the growth of tumors from a subset of these cells within the whole intact brain volume. The ability to follow the metastatic process from the single-cell stage through metastatic growth, and to quantify and monitor the presence of solitary undivided cells will facilitate progress in understanding the mechanisms of brain metastasis and tumor dormancy, and the development of therapeutics to treat this disease.