In situ optical sequencing and structure analysis of a trinucleotide repeat genome region by localization microscopy after specific COMBO-FISH nano-probing.

Trinucleotide repeat expansions (like (CGG)n) of chromatin in the genome of cell nuclei can cause neurological disorders such as for example the Fragile-X syndrome. Until now the mechanisms are not clearly understood as to how these expansions develop during cell proliferation. Therefore in situ investigations of chromatin structures on the nanoscale are required to better understand supra-molecular mechanisms on the single cell level. By super-resolution localization microscopy (Spectral Position Determination Microscopy; SPDM) in combination with nano-probing using COMBO-FISH (COMBinatorial Oligonucleotide FISH), novel insights into the nano-architecture of the genome will become possible. The native spatial structure of trinucleotide repeat expansion genome regions was analysed and optical sequencing of repetitive units was performed within 3D-conserved nuclei using SPDM after COMBO-FISH. We analysed a (CGG)n-expansion region inside the 5' untranslated region of the FMR1 gene. The number of CGG repeats for a full mutation causing the Fragile-X syndrome was found and also verified by Southern blot. The FMR1 promotor region was similarly condensed like a centromeric region whereas the arrangement of the probes labelling the expansion region seemed to indicate a loop-like nano-structure. These results for the first time demonstrate that in situ chromatin structure measurements on the nanoscale are feasible. Due to further methodological progress it will become possible to estimate the state of trinucleotide repeat mutations in detail and to determine the associated chromatin strand structural changes on the single cell level. In general, the application of the described approach to any genome region will lead to new insights into genome nano-architecture and open new avenues for understanding mechanisms and their relevance in the development of heredity diseases.

Analysis of Her2/neu membrane protein clusters in different types of breast cancer cells using localization microscopy.

The Her2/neu tyrosine kinase receptor is a member of the epidermal growth factor family. It plays an important role in tumour genesis of certain types of breast cancer and its overexpression correlates with distinct diagnostic and therapeutic decisions. Nevertheless, it is still under intense investigation to improve diagnostic outcome and therapy control. In this content, we applied spectral precision distance/position determination microscopy, a technique based on the general principles of localization microscopy in order to study tumour typical conformational changes of receptor clusters on cell membranes. We examined two different mamma carcinoma cell lines as well as cells of a breast biopsy of a healthy donor. The Her2/neu receptor sites were labelled by immunofluorescence using conventional fluorescent dyes (Alexa conjugated antibodies). The characterization of the Her2/neu distribution on plasma membrane sections of 176 different cells yielded a total amount of 20 637 clusters with a mean diameter of 67 nm. Statistical analysis on the single molecule level revealed differences in clustering of Her2/neu between all three different cell lines. We also showed that using spectral precision distance/position determination microscopy, a dual colour reconstruction of the 3D spatial arrangement of Her2/neu and Her3 is possible. This indicates that spectral precision distance/position determination microscopy could be used as an enhanced tool offering additional information of Her2/neu receptor status.

[Premature termination--a multifactoral phenomenon]. / Der Behandlungsabbruch--ein multifaktorielles Geschehen.

Twelve drop out cases in psychodynamic short term therapies according to the focus of cyclic maladaptive pattern by Strupp and Binder (CMP 1984) are reviewed regarding initial diagnostics, tape-recorded therapy sessions, and supervision records. Predictors of premature termination of therapy such as initially rated patients' therapy motivation, narcissistic personality traits, but also characteristics of interpersonal behavior of both patient and therapist are examined and discussed. Particularly important are the therapeutic alliance and narcissistic patients' difficulties to relate personally to their therapists instead of enacting projective dynamics of idealization and devaluation. Therapists frequently tend to respond to them by subconscious display of hostile countertransference. Four patterns of dynamics of therapy drop outs are described.

[The Dusseldorf short-term dynamic psychotherapy project (DKZP)]. / Das Düsseldorfer Kurzzeitpsychotherapie-Projekt (DKZP).

Time-limited psychodynamic therapies in the outpatient setting are offered for a broad spectrum of psychogenic disorders. Meta-analyses prove their effectiveness mainly for neurotic disorders. We are interested in the suitability of the here applied CMP/SASB model (Tress et al. 1996) of psychodynamic short-term therapy for patients with psychosomatic and severe personality disorders. We extensively present the concept of Cyclic Mal-adaptive Pattern (CMP) in its clinically relevant parts as the core of psychogenic pathological developments. Particularly the introject, as the patient's attitude towards him- or herself, is of great importance for therapeutic change and its follow-up development. Within the DKZP (Duesseldorf Short-Term Psychotherapy Project), 36 male and female therapists, mostly interns at the local university clinic, treated 82 patients (m: 23/f: 59) with personality disorders (n = 26) and psychosomatic disorders (n = 31) as research groups and, in comparison, neurotic patients (n = 25) in a naturalistic design. 68 treatments have been completed so far, 11 patients have dropped out, the remaining 3 are still in therapy. Relevant research instruments are the Beeinträchtigungsschwere Score (BSS, an impairment score), Global Assessment of Functioning Scale (GAF), Cyclic Maladaptive Pattern (CMP), Symptom Check List (SCL-90-R), and Structural Analysis of Social Behavior (SASB) including process and content ratings, as well as the Intrex questionnaire. The treatments last 25 sessions. We carried out follow-up examinations after 6 months, 1 year, 2, and 5 years. Effect sizes for the whole sample by BSS and GAF are remarkably high, with further increase at follow-up interviews. Psychosomatic patients came off best, but personality disorders in comparison to neurotic patients profited surprisingly well. Effect sizes in self-rating (SCL and Intrex) were less high. However, in self-rating the changes of social functions are not sufficiently addressed. As a result, our form of focal therapy is well suited not only for neuroses, but also for psychosomatic disorders and personality disorders. An enlargement of the indication spectrum for this form of psychodynamic therapy to include these disorders is well founded and promising.

Five-year survival rates of melanoma patients treated by diet therapy after the manner of Gerson: a retrospective review.

OBJECTIVE: Compare 5-year melanoma survival rates to rates in medical literature. DESIGN: Retrospective. SETTING: Hospital in Tijuana, Mexico. PATIENTS: White adult patients (N = 153) with superficial spreading and nodular melanoma, aged 25-72 years. INTERVENTION: Gerson's diet therapy: lactovegetarian; low sodium, fat and (temporarily) protein; high potassium, fluid, and nutrients (hourly raw vegetable/fruit juices). Metabolism increased by thyroid; calorie supply limited to 2600-3200 calories per day. Coffee enemas as needed for pain and appetite. MAIN OUTCOME MEASURE: 5-year survival rates by stage at admission. RESULTS: Of 14 patients with stages I and II (localized) melanoma, 100% survived for 5 years, compared with 79% of 15,798 reported by Balch. Of 17 with stage IIIA (regionally metastasized) melanoma, 82% were alive at 5 years, in contrast to 39% of 103 from Fachklinik Hornheide. Of 33 with combined stages IIIA + IIIB (regionally metastasized) melanoma, 70% lived 5 years, compared with 41% of 134 from Fachklinik Hornheide. We propose a new stage division: IVA (distant lymph, skin, and subcutaneous tissue metastases), and IVB (visceral metastases). Of 18 with stage IVA melanoma, 39% were alive at 5 years, compared with only 6% of 194 from the Eastern Cooperative Oncology Group. Survival impact was not assessed for stage IVB. Male and female survival rates were identical for stages I-IIIB, but stage IVA women had a strong survival advantage. CONCLUSIONS: The 5-year survival rates reported here are considerably higher than those reported elsewhere. Stage IIIA/B males had exceptionally high survival rates compared with those reported by other centers.

[Relation between research and practice in analytic psychotherapy]. / Zum Verhältnis von Forschung und Praxis in der analytischen Psychotherapie.

The technique of psychoanalytic treatment as an objective, operative-technological theory inevitably experiences a subjective refraction on analyst's personality and his or her own cognitive and interactional styles influencing therapeutic efficiency. Systematic clinical trials of treatment data integrating motions from psychoanalytic, social and cognitive science, reconcile the gap between research and practice, between basic science and application. It will be discussed how empirical research can delineate therapists' cognitive and perceptual skills and the process of transformation from declarative to procedural knowledge, further elaborated in reciprocal connection with practice. The models of Structural Analysis of Social Behavior (SASB) and Cyclic Maladaptive Pattern (CMP) are especially considered. We will also discuss the consequences for training in psychoanalysis.

[The course of pregnancy and teratogenicity of antiepileptic agents in 66 patients with epilepsy]. / Untersuchungen zum Schwangerschaftsverlauf und zur Teratogenität der Antiepileptika bei 66 Epilepsie-Patientinnen.

We report on 79 pregnancies in 66 female outpatients with epilepsy. An increase of seizure frequency was significantly more frequent in complex partial seizures than in grand mal seizures and in absences. The reason for these disparities are not clear. In most patients a raised frequency of seizures during pregnancy decreased again after delivery. Carbamazepine was the antiepileptic drug prescribed most frequently followed by valproic acid. The course of the blood levels of carbamazepine and valproic acid was nonuniform during pregnancy. Total concentrations of carbamazepine in cord blood were on average 84.5% of those in maternal blood (n = 22). Valproic acid blood levels were on average 183% of those in maternal blood (n = 15). It is still unclear whether these differences are clinically relevant. During the last weeks of pregnancy we found an increase of the free fraction of carbamazepine and valproic acid. Simultaneously the total protein concentration decreased. Until now these findings are without clinical relevance. The course of labor did not differ from normal population concerning the ratios of spontaneous labor, cesarean section and delivery by forceps. Miscarriage and perinatal mortality were 2.7% each and outnumbered the risk in the general population. In 42.8% of the neonates one to three perinatal complications were observed. The ratio of perinatal complications is not different between patients with monotherapy and combined therapy respectively. There was a tendency to lower values of length, weight and head circumference in the male neonates but not in the female neonates. The risk of minor malformations was 26%, the risk of major malformations was 14% (including one case of suspected malformation) without a discernible correlation with a specific antiepileptic drug.

[The effect of posterior chamber lenses or YAG laser capsulotomy on sensitivity to glare and twilight vision in comparison to phakic control groups (corrected)]. / Der Einfluss von Hinterkammerlinsen oder YAG-Laser-Kapsulotomie auf Blendempfindlichkeit und Dämmerungssehen im Vergleich zu phaken Kontroll gruppen [corrected]

Patients who had undergone extracapsular cataract extraction with implantation of a posterior chamber intraocular lens, and normal subjects of corresponding age and vision were examined with the mesoptometer and nyktometer to compare their twilight vision and glare sensitivity. All the results were poorer in patients with intraocular lenses. Statistically, some of the differences in the results of examinations performed in the two groups were highly significant. As the controls were strictly matched in age and visual acuity, these results confirm other studies with similar objectives. Additionally, some patients with posterior chamber lenses were examined who had also undergone YAG laser capsulotomy for secondary cataract. Although the number of cases was too small for statistical evaluation, it appeared that in this group twilight vision and glare sensitivity could not be improved in the same way as daylight visual acuity by means of this treatment. These findings indicate that since intraocular lenses are increasingly being implanted in younger patients, special attention should also be paid to scotopic vision and glare sensitivity during examinations for drivers' licenses, particularly in cases of good daylight vision. Special examinations, e.g., with the mesoptometer or nyktometer, should be carried out in all such cases. Increased glare sensitivity in eyes with lens implants should also be taken into consideration with regard to the increasing importance of computer and TV screens.

Investigations on the intraindividual constancy of the ratio of carbamazepine to carbamazepine-10,11-epoxide in man.

The constancy of the ratio of carbamazepine (5H-dibenzo[b,f]azepine-5-carboxamide) to carbamazepine-10,11-epoxide was investigated in epileptic patients receiving carbamazepine (Tegretal) monotherapy. We observed a significant interindividual difference, but not between the times in a single patient. Together with the similarly evaluated concordance coefficient, this indicates a certain intraindividual constancy of the ratio between carbamazepine and its epoxide. The carbamazepine:carbamazepine epoxide ratio was dose-dependent, while a sex difference could not be demonstrated.