The pharmacodynamic effects of combined administration of flibanserin and alcohol.

WHAT IS KNOWN AND OBJECTIVE: Flibanserin is a serotonin 5-HT1A agonist and 5-HT2A antagonist approved for the treatment of acquired, generalized hypoactive sexual desire disorder (HSDD) in premenopausal women. Because of the increased risk of hypotension- and syncope-related adverse events (AEs) observed with coadministration of flibanserin and alcohol, alcohol use is contraindicated. To provide a more comprehensive understanding of the interaction between flibanserin and alcohol, the results of a dedicated phase 1 alcohol-interaction study and a pooled analysis of phase 3 studies of premenopausal women with HSDD are presented. METHODS: In the phase 1 study, healthy participants (males [n=23] and females [n=2]) were randomly assigned to one of five sequence groups, which determined the order in which they were to receive flibanserin 100 mg or placebo, with or without ethanol 0.4 g/kg or 0.8 g/kg. Change from baseline in seated blood pressure, orthostatic vital signs, AEs and visual analogue scale sedation outcomes were examined. Blood samples were collected at baseline and for up to 4 hours after dosing to determine flibanserin area under the plasma concentration-time curve from 0 to 4 hours (AUC0-4 ). Pooled data from five phase 3 studies of patients receiving flibanserin 100 mg once daily (n=1543), or placebo (n=1905), were analysed. RESULTS: In the phase 1 study, the incidence of hypotension and syncope increased when flibanserin was coadministered with ethanol. Sedation increased 20% and 27% from baseline with flibanserin plus ethanol 0.4 g/kg and 0.8 g/kg, respectively, at 4 hours post-dose. In the pooled analysis of phase 3 studies, 58.2% and 63.6% of participants receiving flibanserin or placebo, respectively, reported baseline alcohol use. In patients receiving flibanserin, fatigue and dizziness occurred more frequently in patients with vs. without alcohol use. WHAT IS NEW AND CONCLUSION: Results from this study suggest that increased incidence of hypotension- and syncope-related events may result from a pharmacodynamic interaction between flibanserin and alcohol, although the clinical significance of these interactions in real-world populations remains unclear.

Tryptophan Metabolism Along the Kynurenine Pathway Downstream of Toll-like Receptor Stimulation in Peripheral Monocytes.

Tryptophan degradation along the kynurenine pathway is of central importance for the immune function. Toll-like receptors (TLRs), representing the first line of immune defence against pathogens, are expressed in various cell types. The most abundant expression is found on monocytes, macrophages and dendritic cells. The aim of this study was to investigate whether stimulation with different TLR ligands induces the kynurenine pathway in human peripheral monocytes. Cell supernatants were analysed using a liquid chromatography/mass spectrometry to measure kynurenine, kynurenic acid (KYNA), quinolinic acid (QUIN) and tryptophan. Stimulation of TLR-2, TLR-3, TLR-4, TLR-7/8 and TLR-9 was found to induce the production of kynurenine, but only stimulation of TLR-3 increased levels of further downstream metabolites, such as KYNA and QUIN. Stimulation of TLR-1, TLR-5 and TLR-6 did not induce the kynurenine pathway. Taken together, this study provides novel evidence demonstrating that TLR activation induces a pattern of downstream tryptophan degradation along the kynurenine pathway in monocytes. The results of this study may implicate that TLRs can be used as new drug targets for the regulation of aberrant tryptophan metabolism along this pathway, a potential therapeutic strategy that may be of importance in several disorders.

Repeated LPS Injection Induces Distinct Changes in the Kynurenine Pathway in Mice.

The immune system has been recognized as a potential contributor to psychiatric disorders. In animals, lipopolysaccharide (LPS) is used to induce inflammation and behaviors analogous to some of the symptoms in these disorders. Recent data indicate that the kynurenine pathway contributes to LPS-induced aberrant behaviors. However, data are inconclusive regarding optimal LPS dose and treatment strategy. Here, we therefore aimed to evaluate the effects of single versus repeated administration of LPS on the kynurenine pathway. Adult C57BL6 mice were given 0.83 mg/kg LPS as a single or a repeated injection (LPS + LPS) and sacrificed after 24, 48, 72, or 120 h. Mice receiving LPS + LPS had significantly elevated brain kynurenine levels at 24 and 48 h, and elevated serum kynurenine at 24, 48 and 72 h. Brain kynurenic acid and quinolinic acid were significantly increased at 24 and 48 h in mice receiving LPS + LPS, whereas serum kynurenic acid levels were significantly decreased at 24 h. The increase of brain kynurenic acid by LPS + LPS was likely unrelated to the higher total dose as a separate group of mice receiving 1.66 mg/kg LPS as single injection 24 h prior to sacrifice did not show increased brain kynurenic acid. Serum quinolinic acid levels were not affected by LPS + LPS compared to vehicle. Animals given repeated injections of LPS showed a more robust induction of the kynurenine pathway in contrast to animals receiving a single injection. These results may be valuable in light of data showing the importance of the kynurenine pathway in psychiatric disorders.

Susceptibility of Lenticels within the Stem Depression of Tomato Fruit to Bacterial Soft Rot.

The postharvest development of bacterial soft rot in tomato fruit caused by Pectobacterium carotovorum is herein linked with inoculation of lenticel-like apertures located around the stem attachment. Stem scars misted with aqueous cell suspensions of P. carotovorum (1 × 106 CFU/ml) or briefly (5 s) immersed in the suspension were likely to become infected during subsequent storage, with a disease incidence exceeding 70% within 7 days at 24°C. Water soaking was initially observed beneath the fruit surface at the juncture of radial walls of endocarp tissues and a corky ring around the stem attachment. If fruit were swirled for 1 min in chlorine at 150 ppm, pH 6.5, within 5 s after inoculation, lesions did not occur. However, if the chlorine treatment was delayed by 120 s, lesions occasionally developed. A 60-min delay usually assured that decay would occur before fruit ripened (within 5 days). Delaying the chlorine wash by 18 h, as would occur if commercial harvests were held overnight prior to packing operations, led to the same incidence of bacterial soft rot as observed among control fruit. Inoculation of fruit with attached stems did not lead to as much decay during storage compared with stemless fruit, suggesting that an intact calyx physically reduced lenticel exposure to external aqueous cell suspensions. Tomato fruit exposed to uncontrolled free moisture during or shortly after harvest have a high potential for postharvest decays and the likely site for infection development is the area surrounding the stem attachment.

A randomized, placebo-controlled, double-blind study of sapropterin to treat ADHD symptoms and executive function impairment in children and adults with sapropterin-responsive phenylketonuria.

Symptoms of attention deficit-hyperactivity disorder (ADHD), particularly inattention, and impairments in executive functioning have been reported in early and continuously treated children, adolescents, and adults with phenylketonuria (PKU). In addition, higher blood phenylalanine (Phe) levels have been correlated with the presence of ADHD symptoms and executive functioning impairment. The placebo-controlled PKU ASCEND study evaluated the effects of sapropterin therapy on PKU-associated symptoms of ADHD and executive and global functioning in individuals who had a therapeutic blood Phe response to sapropterin therapy. The presence of ADHD inattentive symptoms and executive functioning deficits was confirmed in this large cohort of 206 children and adults with PKU, of whom 118 responded to sapropterin therapy. In the 38 individuals with sapropterin-responsive PKU and ADHD symptoms at baseline, sapropterin therapy resulted in a significant improvement in ADHD inattentive symptoms in the first 4 weeks of treatment, and improvements were maintained throughout the 26 weeks of treatment. Sapropterin was well-tolerated with a favorable safety profile. The improvements in ADHD inattentive symptoms and aspects of executive functioning in response to sapropterin therapy noted in a large cohort of individuals with PKU indicate that these symptoms are potentially reversible when blood Phe levels are reduced.

[Development of diagnosis-related groups in different surgical disciplines]. / Entwicklung der Fallpauschalenvergütung in verschiedenen chirurgischen Fachgebieten.

BACKGROUND: Since the introduction of Diagnosis-Re-lat-ed Groups (DRGs) in Germany, the variables of remuneration have continuously changed. Subjectively, reimbursement by DRG has a negative connotation among all specialities. We analysed the development of reimbursement and length of stay in different surgical specialties. MATERIAL AND METHODS: By grouping the top-10-diagnoses and therapies in hand surgery, trauma surgery, general surgery as well as cardiothoracic and vascular surgery between 2004 and 2010, DRGs were obtained, compared and the data deduced. RESULTS: While the lower threshold of length of stay remained almost the same, mean value and upper threshold became shorter in most of the top-10-diagnoses. During the observation period, total reimbursement increased by 30 % in hand surgery, 20 % in general surgery and 17 % in cardiothoracic and vascular surgery, while in trauma surgery it decreased by 1 %. This corresponds to mean annual growth rates of 4.47 %, 3.08 %, 2.68 % and - 0.15 %, respectively. No correlation was found between the 4 disciplines and macro-economic parameters. CONCLUSION: Reductions of mean and upper thresholds of length of stay are present in all surgical disciplines. Total reimbursements developed partially in a contradictory manner. Negative growth involves the danger that hospital investments cannot be realised, especially in the presence of high personnel costs.

Reimbursement for tissue reconstruction by pedicled and free flaps across five European countries. The importance of autologous breast reconstruction.

AIM: Case payment mechanisms have become the principle means of remunerating hospitals in many countries. We analysed the reimbursement for different types of autologous tissue transfer in five European countries. METHODS: We looked at common surgical options for breast reconstruction and flaps at other body regions. The principle diagnosis was systematically modified and processed with national grouper software to identify the relevant Diagnosis-Related Groups. RESULTS: The mean difference in payment was 4509 € in breast reconstruction versus only 2599 € in other locations. According to the underlying diagnosis for reconstruction, procedures after resection of malignant breast cancer showed higher reimbursement (mean 8319 €) than of other body parts (mean 6454 €). Sweden had the highest mean reimbursement (9589 €) followed by Austria (8032 €), Germany (7259 €), Italy (6667 €) and the UK (6037 €). Austria, Italy and the UK showed significant differences of reimbursement between pedicled flaps of the breast and other parts of the body. CONCLUSION: International data for the benchmarking and refinement of a national compensation system can be a useful instrument in identifying ways of improving each system. Across a spectrum of European countries, reimbursement for the reconstruction of the breast and other body parts was analysed and characteristics were identified. As rationalisation of healthcare becomes widespread in European countries, the need for individualised reimbursement which correlates accordingly is becoming ever more important.

Psychotherapy as an epigenetic 'drug': psychiatric therapeutics target symptoms linked to malfunctioning brain circuits with psychotherapy as well as with drugs.

WHAT IS KNOWN AND OBJECTIVE: Psychotherapy has traditionally competed with psychopharmacology. As drugs have become the more dominant treatment in psychiatry and primary care, this approach is increasingly criticized as limited in scope, lacking in robust outcomes and too heavily influenced by the pharmaceutical industry. Our objective is to show that recent advances in neurobiology are clarifying that learning and environmental experiences, such as psychotherapy, change brain circuits as do drugs. The leading notion of how therapeutic effects occur in psychiatric disorders is that they happen when symptoms are reduced by improving the efficiency of information processing in hypothetically malfunctioning brain circuits. COMMENT: With this formulation of psychiatric symptoms and their relief, it is not surprising that both psychotherapy and psychopharmacology can be clinically effective for treating psychiatric disorders, or indeed that combining them can be therapeutically synergistic. Psychotherapy, including a new spinoff of cognitive behavioural therapy called trial-based therapy, like many other forms of learning, can hypothetically induce epigenetic changes in brain circuits that can enhance the efficiency of information processing in malfunctioning neurons to improve symptoms in psychiatric disorders, just like drugs. WHAT IS NEW AND CONCLUSION: Psychotherapies can be conceptualized as epigenetic 'drugs', or at least as therapeutic agents that act epigenetically in a manner similar or complementary to drugs. These findings are leading to a paradigm shift in psychiatry such that psychotherapy is experiencing a come-back as various standardized, brief, goal-directed psychotherapies are being integrated with drug treatment of psychiatric disorders by psychopharmacologists who have traditionally relied on a drugs-only approach.

What do web-use skill differences imply for online health information searches?

BACKGROUND: Online health information is of variable and often low scientific quality. In particular, elderly less-educated populations are said to struggle in accessing quality online information (digital divide). Little is known about (1) how their online behavior differs from that of younger, more-educated, and more-frequent Web users, and (2) how the older population may be supported in accessing good-quality online health information. OBJECTIVE: To specify the digital divide between skilled and less-skilled Web users, we assessed qualitative differences in technical skills, cognitive strategies, and attitudes toward online health information. Based on these findings, we identified educational and technological interventions to help Web users find and access good-quality online health information. METHODS: We asked 22 native German-speaking adults to search for health information online. The skilled cohort consisted of 10 participants who were younger than 30 years of age, had a higher level of education, and were more experienced using the Web than 12 participants in the less-skilled cohort, who were at least 50 years of age. We observed online health information searches to specify differences in technical skills and analyzed concurrent verbal protocols to identify health information seekers' cognitive strategies and attitudes. RESULTS: Our main findings relate to (1) attitudes: health information seekers in both cohorts doubted the quality of information retrieved online; among poorly skilled seekers, this was mainly because they doubted their skills to navigate vast amounts of information; once a website was accessed, quality concerns disappeared in both cohorts, (2) technical skills: skilled Web users effectively filtered information according to search intentions and data sources; less-skilled users were easily distracted by unrelated information, and (3) cognitive strategies: skilled Web users searched to inform themselves; less-skilled users searched to confirm their health-related opinions such as "vaccinations are harmful." Independent of Web-use skills, most participants stopped a search once they had found the first piece of evidence satisfying search intentions, rather than according to quality criteria. CONCLUSIONS: Findings related to Web-use skills differences suggest two classes of interventions to facilitate access to good-quality online health information. Challenges related to findings (1) and (2) should be remedied by improving people's basic Web-use skills. In particular, Web users should be taught how to avoid information overload by generating specific search terms and to avoid low-quality information by requesting results from trusted websites only. Problems related to finding (3) may be remedied by visually labeling search engine results according to quality criteria.

[100 years after Kienböck's description: review of the etiology of Kienböck's disease from a historical perspective]. / 100 Jahre nach Kienböck: Literaturübersicht über die Ursachenforschung der Mondbeinnekrosen.

Kienböck's disease (KD) leads to collapse of the lunate bone with severe consequences for the wrist function which for some patients may result in occupational invalidity. The many synonyms of KD (aseptic necrosis or avascular necrosis) insinuate that the true etiopathology remains poorly understood. This reviews aims at exploring the level of evidence which brought forward the different hypotheses on the origin of KD. The widespread theories about the origin were formed about 100 years ago but a specific therapy is still not within reach. Although the cause of the disease remains essentially unknown it is officially recognized as an occupational disease in Germany. Empirical attempts to explain the etiopathology are based on compression of the lunate, impaired vascularity through vibration exposition, fracture and dislocation of the lunate from the radiolunate fossa. The level of evidence urges a cautious interpretation of currently discussed hypotheses on the etiology of KD.

Role of antibiotic treatment after surgical debridement of superficial hand infections in 180 patients.

Considering the rising prevalence of antimicrobial resistance and the lack of recommendations on antibiotic treatment, the present study evaluated the necessity of local and systemic antibiotic therapy in addition to surgical debridement in superficial hand infections. Superficial hand infections were defined as not involving tendons, joints or bone. Data were analyzed for 180 patients, assigned to three study groups according to postoperative antimicrobial treatment. Patients in group I were treated with both systemic and local antibiotics; in group II antimicrobial therapy was limited to local antibiotics in the form of subcutaneous gentamicin bead chains; group III did not receive any antibiotic treatment after surgical debridement. Patients were followed up at two weeks and at three months. Immobilization time and length of stay were longer in group I, but there were no significant differences between the groups in terms of revision rate or recovery of hand function assessed by measurement of finger mobility, grip strength and the Disabilities of the Arm, Shoulder and Hand score at follow-up. Antibiotic treatment of hand infections should be considered carefully and reserved for specific indications: e.g., severe infections and selected patients such as those with diabetes or immunocompromising diseases.

[Comparison of hand surgery in the German and Italian DRG systems]. / Vergleich der Handchirurgie im deutschen und italienischen DRG-System.

Diagnosis-related groups (DRGs) are a patient classification system grouping related types of patients treated to the resources they consumed. In this analysis we compared the Italian and the German DRG systems regarding hand surgery with an emphasis on reimbursement of clinical cases. The 15 most common hand surgical diagnoses and their corresponding operative treatment in our clinic in 2009 were processed using a DRG grouper. The underlying data were transferred to the Italian system. Thus, the length of stay and the reimbursement of both countries could be obtained and compared. The latter was adjusted and corrected by the purchasing power of each country. The mean of the upper threshold of length of stay was 10 days in the German as well as the Italian system whereas the median was 2 times higher in Italy (6 vs. 12 days). Fifteen out of 19 cases showed higher reimbursement in Germany. The case mix index (CMI) of 0.917 in Germany represents a mean payment of 2,676 € per case. In Italy the hypothetical CMI of 0.635 resulted in a mean reimbursement of 1,853 € per case. The biggest difference in remuneration could be found for replantation of multiple fingers. For this service the German health-care system pays 12,320 € more than the Italian. Total proceeds of the top 15 diagnoses applying the number of cases treated in our clinic revealed 1.7 million € in the German and 1.2 million € in the Italian DRG system. Considering the purchasing power utilizing consumer prize parities, the difference of reimbursement between the countries decreased to 300,000 €. There is no mean length of stay per DRG in Italy, only the upper threshold of length of stay is determined. In most cases the latter is higher in Italy compared to Germany. The consumption of resources for finger replantation is not adequately represented in the Italian DRG system compared to finger amputation. Reimbursement of inpatient care is influenced by multiple factors not being subject to the free market economy. For this reason only descriptive comparison is feasible.

[Primary treatment of complicated flexor tendon injuries of the hand]. / Primärversorgung komplizierter Beugesehnenverletzungen an der Hand.

Complicated flexor tendon injuries are classified into lacerations, avulsions, ruptures, and defects. They are often a challenge for hand surgeons and frequently they present unsatisfactory functional results postoperatively. Lacerations and avulsions are usually treated by pull-out sutures and suture anchors. In ruptures, the causality should be sought. Tendon-linking, transposition and tenodesis/arthrodesis are the domain of rheumatoid arthritis. The primary transplantation of tendons is rarely indicated, ideally in non-contaminated flexor tendon defects in zones III-V with an uninjured surrounding soft tissue situation. Postoperative rehabilitation programs are very the same as in normal flexor tendon injuries.

Re-targeted adenovirus vectors with dual specificity; binding specificities conferred by two different Affibody molecules in the fiber.

Vectors based on Adenovirus type 5 (Ad5) are among the most common vectors in cancer gene therapy trials to date. However, for increased efficiency and safety, Ad5 should be de-targeted from its native receptors and re-targeted to a tumor antigen. We have described earlier an Ad5 vector genetically re-targeted to the tumor antigen HER2/neu by a dimeric version of the Affibody molecule ZH inserted in the HI-loop of the fiber knob of a coxsackie and adenovirus receptor-binding ablated fiber. This virus showed almost wild-type growth characteristics and infected cells through HER2/neu. Here we generate vectors with double specificity by incorporating two different Affibody molecules, ZH (HER2/neu-binding) and ZT (Taq polymerase-binding), at different positions relative to one another in the HI-loop. Receptor-binding studies together with viral production and gene transfer assays showed that the recombinant fiber with ZT in the first position and ZH in the second position (ZTZH) bound to both its targets, whereas surprisingly, the fiber with ZHZT was devoid of binding to HER2/neu. Hence, it is possible to construct a recombinant adenovirus with dual specificity after evaluating the best position for each ligand in the fiber knob.

HIV-1 rev depolymerizes microtubules to form stable bilayered rings.

We describe a novel interaction between HIV-1 Rev and microtubules (MTs) that results in the formation of bilayered rings that are 44-49 nm in external diameter, 3.4-4.2 MD (megadaltons) in mass, and have 28-, 30-, or 32-fold symmetry. Ring formation is not sensitive to taxol, colchicine, or microtubule-associated proteins, but requires Mg(2+) and is inhibited by maytansine. The interaction involves the NH(2)-terminal domain of Rev and the face of tubulin exposed on the exterior of the MTs. The NH(2)-terminal half of Rev has unexpected sequence similarity to the tubulin-binding portion of the catalytic/motor domains of the microtubule-destabilizing Kin I kinesins. We propose a model wherein binding of Rev dimers to MTs at their ends causes segments of two neighboring protofilaments to peel off and close into rings, circumferentially containing 14, 15, or 16 tubulin heterodimers, with Rev bound on the inside. Rev has a strong inhibitory effect on aster formation in Xenopus egg extracts, demonstrating that it can interact with tubulin in the presence of normal levels of cellular constituents. These results suggest that Rev may interact with MTs to induce their destabilization, a proposition consistent with the previously described disruption of MTs after HIV-1 infection.

NMR structure of a specific DNA complex of Zn-containing DNA binding domain of GATA-1.

The three-dimensional solution structure of a complex between the DNA binding domain of the chicken erythroid transcription factor GATA-1 and its cognate DNA site has been determined with multidimensional heteronuclear magnetic resonance spectroscopy. The DNA binding domain consists of a core which contains a zinc coordinated by four cysteines and a carboxyl-terminal tail. The core is composed of two irregular antiparallel beta sheets and an alpha helix, followed by a long loop that leads into the carboxyl-terminal tail. The amino-terminal part of the core, including the helix, is similar in structure, although not in sequence, to the amino-terminal zinc module of the glucocorticoid receptor DNA binding domain. In the other regions, the structures of these two DNA binding domains are entirely different. The DNA target site in contact with the protein spans eight base pairs. The helix and the loop connecting the two antiparallel beta sheets interact with the major groove of the DNA. The carboxyl-terminal tail, which is an essential determinant of specific binding, wraps around into the minor groove. The complex resembles a hand holding a rope with the palm and fingers representing the protein core and the thumb, the carboxyl-terminal tail. The specific interactions between GATA-1 and DNA in the major groove are mainly hydrophobic in nature, which accounts for the preponderance of thymines in the target site. A large number of interactions are observed with the phosphate backbone.

Which comes first: atypical antipsychotic treatment or cardiometabolic risk?

OBJECTIVE: To provide an overview for practicing clinicians on the pharmacological basis of cardiometabolic risk induced by antipsychotic drugs in patients with serious mental illness, to propose hypotheses to explain these risks and to give tips for managing cardiometabolic risk during antipsychotic treatment. METHOD: A MEDLINE search using terms for atypical antipsychotics (including individual drug names), metabolic, cardiovascular, weight gain and insulin resistance, cross-referenced with schizophrenia was performed on articles published between 1990 and May 2008. RESULTS: Strong evidence exists for significant cardiometabolic risk differences among several antipsychotic agents. Histamine H1 and serotonin 5HT2C antagonism are associated with risk of weight gain, but receptor targets for dyslipidemia and insulin resistance have not yet been identified. Convincing data indicate that hypertriglyceridemia and insulin resistance may occur in the absence of weight gain with certain antipsychotics. CONCLUSION: Although lifestyle and genetics may contribute independent risks of cardiometabolic dysfunction in schizophrenia and other serious mental illness, antipsychotic treatment also represents an important contributor to risk of cardiometabolic dysfunction, particularly for certain drugs and for vulnerable patients. Mental health professionals must learn to recognize the clinical signposts indicating antipsychotic-related cardiometabolic problems to forestall progression to type II diabetes, cardiovascular events and premature death.

The metabolic syndrome and schizophrenia.

OBJECTIVE: To summarize the accumulated data on metabolic syndrome prevalence in patients with schizophrenia, examine evidence for a biological contribution of the mental illness to metabolic risk and review novel options available for management of prediabetic states. METHOD: A Medline search using metabolic syndrome, insulin resistance and insulin sensitivity cross-referenced with schizophrenia was performed on articles published between 1990 and May 2008. RESULTS: Recent evidence indicates that schizophrenia increases predisposition towards metabolic dysfunction independent of environmental exposure. Both fasting and non-fasting triglycerides have emerged as important indicators of cardiometabolic risk, while metformin, thiazolidinediones and GLP-1 modulators may prove promising tools for managing insulin resistance. CONCLUSION: Because of lifestyle, disease and medication effects, schizophrenia patients have significant risk for cardiometabolic disease. Routine monitoring, preferential use of metabolically neutral antipsychotics and lifestyle education are critical to minimizing risk, with a possible role for antidiabetic medications for management of insulin resistant states that do not respond to other treatment strategies.

Comparison of CO(2) laser and 4.0 MHz radiosurgery for making incisions in the skin and muscles of green iguanas (Iguana iguana).

Four green iguanas scheduled for euthanasia were used to compare the extent of collateral tissue damage associated with CO(2) laser and 4.0 MHz radiosurgery. The iguanas were anaesthetised and a series of three skin and three muscle incisions was made by 4.0 MHz radiosurgery (0.18 mm wire electrode, 25 W, cut mode) and CO(2) laser (0.3 mm ceramic tip, 15 W focused beam super-pulse mode), and three incisions were made with a scalpel blade as controls. Following euthanasia, a total of 60 skin and 36 muscle sections were evaluated histologically. Radiosurgery and the laser both produced bloodless incisions, but radiosurgery caused significantly less collateral tissue damage in the skin (307 [97] v 386 [108] microm) and the muscle (18 [7] v 91 [15] microm).

In vivo and in vitro uptake of 111In, delivered with the affibody molecule (ZEGFR:955)2, in EGFR expressing tumour cells.

The epidermal growth factor receptor, EGFR, is overexpressed in many carcinomas. Targeting this receptor with radionuclides is important for imaging and therapy applications in nuclear medicine. We investigated the in vitro and in vivo properties of a new high affinity EGFR binding affibody molecule, (ZEGFR:955)2, when conjugated with CHX-A''-DTPA and labelled with 111In. The binding time patterns and retention studies were performed using cultured squamous carcinoma A431 cells that overexpress EGFR. In the in vivo studies, female BALB/c nu/nu mice carrying tumours from xenografted A431 cells were used. The in vitro studies showed EGFR specific binding, high uptake and good retention of 111In when delivered as [111In](ZEGFR:955)2. The retention after 72 h of incubation was 38.0+/-1.15% of the initial level. The biodistribution study showed a tumour specific 111In uptake of 3.8+/-1.4% of injected dose per gram tumour tissue 4 h post-injection. The tumour to blood ratio was 9.1 and the tumours could easily be visualized with a gamma camera at this time-point. 111In delivered with [111In](ZEGFR:955)2 gave an EGFR specific uptake and the results indicated that the (ZEGFR:955)2 affibody molecule is a candidate for radionuclide-based tumour imaging. Potential therapy applications are discussed.