Cell culture model predicts human disease: Altered expression of junction proteins and matrix metalloproteinase's in cervical dysplasia

ABSTRACT: BACKGROUND: Cervical cancer is necessarily caused by human papillomaviruses, which encode three oncogenes manifesting their functions by interfering with a number of cellular proteins and pathways: the E5, E6, and E7 proteins. We have earlier found in our microarray studies that the E5 oncogene crucially affects the expression of cellular genes involved in adhesion and motility of epithelial cells. METHODS: In order to biologically validate our previous experimental findings we performed immunohistochemical staining of a representative set of tissue samples from different grades of high-risk human papillomavirus associated cervical disease as well as normal squamous and columnar cervical epithelium. Three-dimensional collagen raft cultures established from E5-expressing and control epithelial cells were also examined. The expression of p16, matrix metalloproteinase (MMP) -7, MMP-16, cytokeratin (CK) 8/18, laminin, E-cadherin and beta-catenin was studied. RESULTS: In agreement with our previous microarray studies, we found intense staining for E-cadherin and beta-catenin in adherens junctions even in high-grade cervical lesions. Staining for MMP-16 was increased in severe disease as well. No significant change in staining for MMP-7 and cytokeratin 8/18 along with the grade of cervical squamous epithelial disease was observed. CONCLUSIONS: Here we have confirmed, using tissue material from human papillomavirus associated lesions, some of the cellular gene expression modifications that we earlier reported in an experimental system studying specifically the E5 oncogene of papillomaviruses. These findings were partially surprising in the context of cervical carcinogenesis and emphasize that the complexity of carcinogenesis is not yet fully understood. Microarray approaches provide a wide overwiev of gene expression in experimental settings, which may provide biologically valid biomarkers in disease diagnostics, prognosis, and follow-up.     

Physical functioning, health and survival: a ten-year follow-up study

BACKGROUND AND AIMS: Cutbacks in human resources are putting home care personnel under increasing pressure in their work. Home health care personnel need to know how they can prevent undesirable conditions and support older people towards successful aging. The aim of this ten-year follow-up study was to identify factors associated with increased mortality among 70-year-old subjects living in the community. METHODS: Data were collected in 1991 by a postal questionnaire sent to all residents of Turku, Finland, born in 1920. A physical examination was also conducted. Ten years later, in 2001, the mortality rate of this population was determined. The data were examined statistically. RESULTS: Many health-related factors, such as smoking, poor subjective health, and mobility, were related to an increased level of mortality. In addition, several diseases (e.g., diabetes, angina pectoris, cancer) at age 70 were associated with increased mortality over ten years. Difficulties in daily activities at age 70 also increased the risk of mortality. CONCLUSIONS: The findings offer useful clues for planning health care services and preventive interventions provided by home health care personnel. Home health care personnel should give special attention to older people who feel ill or very ill.     

Hereditary gelsolin amyloidosis mimicking Sj?gren’s syndrome

Hereditary gelsolin amyloidosis (AGel amyloidosis) belongs to the wide group of amyloidotic diseases, which comprise various hereditary but also sporadic forms, such as inflammation-associated AA amyloidosis, primary or myeloma-associated AL amyloidosis and common Alzheimer's disease and type II diabetes-associated local amyloidoses. AGel amyloidosis caused by a gelsolin G654A gene mutation is autosomally dominantly inherited and presents typically in the 30s with progressive corneal lattice dystrophy, followed by cutis laxa and cranial polyneuropathy. Here, we present a case of sicca syndrome, originally diagnosed as primary Sjögren's syndrome (SS) but later found to represent an initial disease manifestation of AGel amyloidosis, not recognised earlier. This case emphasises both the importance of specific amyloid stainings and comprehensive salivary gland histopathology as well as family history in SS differential diagnostics.     

Fronto-cerebellar systems are associated with infant motor and adult executive functions in healthy adults but not in schizophrenia

Delineating longitudinal relationships between early developmental markers, adult cognitive function, and adult brain structure could clarify the pathogenesis of neurodevelopmental disorders such as schizophrenia. We aimed to identify brain structural correlates of infant motor development (IMD) and adult executive function in nonpsychotic adults and to test for abnormal associations between these measures in people with schizophrenia. Representative samples of nonpsychotic adults (n = 93) and people with schizophrenia (n = 49) were drawn from the Northern Finland 1966 general population birth cohort. IMD was prospectively assessed at age 1 year; executive function testing and MRI were completed at age 33-35 years. We found that earlier motor development in infancy was correlated with superior executive function in nonpsychotic subjects. Earlier motor development was also normally associated with increased gray matter density in adult premotor cortex, striatum, and cerebellum and increased white matter density in frontal and parietal lobes. Adult executive function was normally associated with increased gray matter density in a fronto-cerebellar system that partially overlapped, but was not identical to, the gray matter regions normally associated with IMD. People with schizophrenia had relatively delayed IMD and impaired adult executive function in adulthood. Furthermore, they demonstrated no normative associations between fronto-cerebellar structure, IMD, or executive function. We conclude that frontal cortico-cerebellar systems correlated with adult executive function are anatomically related to systems associated with normal infant motor development. Disruption of this anatomical system may underlie both the early developmental and adult cognitive abnormalities in schizophrenia.     

Neurocognitive test profiles of extremely low birth weight five-year-old children differ according to neuromotor status

The neurocognitive outcome of children born with extremely low birth weight (ELBW) is highly variable due to the complexity of morbidity. So far, no study has compared comprehensive neuropsychological test profiles in groups with different neuromotor status. In a national cohort of ELBW children neuropsychological test profiles were assessed in 4 groups defined according to a neurological examination at 5 years of age: normal neuromotor status (N = 56), motor coordination problems (N = 32), multiple subtle neuromotor signs including both motor coordination problems and deviant reflexes (N = 20), and spastic diplegia (N = 12). The neurocognitive assessment included a test of intelligence, the Wechsler Primary and Preschool Scale of Intelligence-Revised (WPPSI-R) and 14 subtests of attention and executive functions, verbal functions, manual motor functions, visuoconstructional functions and verbal learning (NEPSY). The children with normal neuromotor status performed within the average range; children with motor coordination problems had widespread impairment; and children with spastic diplegia and children with multiple minor neuromotor signs had uneven test profiles with stronger verbal results but weaknesses in attention and executive functions, and in manual motor and visuoconstructional tasks. In conclusion, very preterm children with neuromotor signs, including motor coordination problems, are at risk for neurocognitive impairment, in spite of average intelligence. More impaired children have more irregular test profiles. Follow-up and neuropsychological assessment of very preterm children with minor neuromotor signs are therefore indicated.     

Feedback in patient counselling training--pharmacy students' opinions

OBJECTIVE: The aims were to analyse Finnish pharmacy students' opinions of feedback given to them in patient counselling training during their internship period, and to evaluate the guidelines used. METHODS: The study was based on reports written by 136 students, analysed qualitatively. To examine the significance of the guidelines and the perceived quality of the feedback, students were categorised into groups. Correlations were assessed by frequencies and cross-tabulations. RESULTS: A statistically significant correlation was found (p<0.001) between the perceived quality of the feedback and the use of the guidelines. The relations between the perceived usefulness of the guidelines and the use of the guidelines in patient counselling exercises were also statistically significant (p=0.034). CONCLUSIONS: The results indicate that the perceived quality of the feedback was indeed associated with the use of the guidelines. They not only offer a training method focusing on essential communication skills in patient counselling, but are also a useful tool for developing counselling practices in general. PRACTICAL IMPLICATIONS: Since mentors are probably unfamiliar with the guidelines, students should be trained to explain their use. To ensure that mentors are competent in analysing communication skills, further training for them is essential.     

Neuroanatomical mapping of juvenile rat brain regions with prominent basal signal in [(35)S]GTPgammaS autoradiography

[(35)S]GTPgammaS autoradiography represents a powerful functional approach to detect receptor-dependent G(i/o) protein activity in anatomically defined brain structures. Inherent to this technique, however, is the notable basal signal evident in several brain regions in the absence of receptor stimulation by exogenously added agonist. In the rat brain, much of this basal labelling derives from tonic activation of adenosine A(1) and lysophosphatidic acid LPA(1) receptors in the gray and white matter regions, respectively. Despite the elimination of the two receptor activities, prominent basal [(35)S]GTPgammaS labelling is still evident in discrete brain structures, possibly reflecting regional enrichment of G(i/o) and/or constitutive receptor activity or the presence of still unknown endogenous ligands activating their orphan receptors. Here, the anatomical distribution of the enhanced basal signal was systematically mapped in brain sections of 4-week-old male Wistar rats. Regions with prominent basal [(35)S]GTPgammaS labelling represented neuroanatomically distinct structures, in particular various thalamic and hypothalamic nuclei. For instance, the paraventricular thalamic nucleus, the bed nucleus of the stria terminalis and the subfornical organ were highly labelled, as were the periaqueductal gray and the nucleus of the solitary tract. Pre-treatment with N-ethylmaleimide (NEM), an alkylating agent preventing all known receptor-driven G protein activity in cryostat sections markedly decreased the basal binding in all examined regions. In preliminary screening, selective antagonists for various brain-enriched G(i/o)-coupled receptors failed to suppress the basal signal in any of the studied regions.     

A technique for simultaneous needle insertion in prostate seed implantation

The purpose was to develop a fast needle insertion system to shorten the implantation time and to restrain prostate swelling during the implantation, thus reducing the seed setup error. The basic idea is to insert all the needles simultaneously using ultrasound guidance. The developed system consists of two similar templates that are connected. All the needles are set and locked to a moveable rear template according to the dose plan. The needle pack is then pushed into position, the lock released and seeds implanted needle by needle. A test and training phantom was also built.     

S100B proteins in febrile seizures

S100B protein concentrations correlate with the severity and outcome of brain damage after brain injuries, and have been shown to be markers of blood-brain barrier damage. In children elevated S100B values are seen as a marker of damage to astrocytes even after mild head injuries. S100B proteins may also give an indication of an ongoing pathological process in the brain with respect to febrile seizures (FS) and the likelihood of their recurrence. To evaluate this, we measured S100B protein concentrations in serum and cerebrospinal fluid from 103 children after their first FS. 33 children with acute infection without FS served as controls for the serum concentrations. In the FS patients the mean S100B concentration in the cerebrospinal fluid samples was 0.21 μg/L and that in the serum samples 0.12 μg/L. The mean serum concentration in the controls was 0.11 μg/L (difference 0.01 μg/L, 95% confidence interval -0.02 to 0.04 μg/L, P = 0.46). There was a correlation between age and serum S100B concentration (r = -0.28, P = 0.008) in children under four years, but S100B concentrations did not predict the clinical severity of the FS nor their recurrence. There was no correlation between time of arrival at the hospital after FS and S100B concentration in serum (r = -0.130, P = 0.28) or in cerebrospinal fluid samples (r=-0.091, P = 0.52). Our findings indicate that FS does not cause significant blood-brain barrier openings, and increase the evidence that these seizures are relatively harmless for the developing brain.