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Potential applications of therapeutic drug monitoring in treatment of neoplastic disease by antineoplastic agents.

Therapeutic drug monitoring of antineoplastic agents must be considered in terms of cytotoxicity to stem cells and toxicity of normal tissues. The dose-limiting toxicity which is myelosuppression is believed to be a necessary effect for maximal antitumour effect. The areas in which therapeutic drug monitoring can play a role include: predicting patients at risk of toxicity, e.g. high dose methotrexate therapy; low bioavailability, e.g. oral 6-mercaptopurine; identifying patients at risk of treatment failure, e.g. methotrexate clearance in acute lymphoblastic leukemia. Although therapeutic drug monitoring is not clinically useful at present, the potential role it can play is illustrated for cytosine arabinoside, adriamycin and cyclophosphamide. Two problems encountered with drug monitoring of antineoplastic disease are tumour heterogeneity manifested by clonal, nutritional and physical characteristics and the common practice of combination chemotherapy. These problems must be addressed in order to make therapeutic drug monitoring a practical tool in cancer chemotherapy. Future applications of therapeutic drug monitoring will require more studies to define therapeutic ranges for single agents, patient-to-patient variation, course-to-course variation and the effects of drug combinations on pharmacokinetic profiles of the drugs used.

Antineoplastic Agents↗

A therapeutic index that predicts the individual effects of prednisone in patients with cirrhosis.

Our aim was to construct an index that accurately predicts the degree of benefit or harm that prednisone therapy holds for patients with liver cirrhosis. The admission and survival data of 488 patients with cirrhosis who participated in a controlled clinical trial of prednisone in a dosage of 10-15 mg daily (251 patients) versus placebo (237 patients) and who were observed for up to 12 yr were analyzed using Cox's multiple regression model. Four variables each provided significant therapeutic information: antinuclear factor (p = 0.02) and large piecemeal necroses (p = 0.02) were associated with a beneficial effect, whereas ascites (p = 0.0004) and large regenerative nodules (p = 0.0007) were associated with a harmful effect of prednisone. From these four variables a therapeutic index was constructed. For a given patient the therapeutic index is a measure of how big the effect will be if prednisone is given. The gain in survival time obtained by administering prednisone according to the therapeutic index was estimated to be 349 yr, mainly confined to 217 patients with a significant positive (121) or negative (96) therapeutic index. The therapeutic index may prove useful for the optimal administration of prednisone treatment in new patients with cirrhosis.

Antibodies, Antinuclear↗

Major early complications from diagnostic and therapeutic ERCP: a prospective multicenter study.

BACKGROUND: There is a lack of multicenter prospective studies on complications of diagnostic and therapeutic endoscopic retrograde cholangiopancreatography (ERCP). METHODS: We studied 2769 consecutive patients undergoing ERCP at nine centers in the Triveneto region of Italy over a 2-year period. Six centers performed ERCP on less than 200 patients per year (small centers). General and ERCP-specific major complications were predefined. Data were collected at the time of ERCP, before discharge, and in cases of readmission within 30 days. ERCP was defined as therapeutic when endoscopic sphincterotomy (n = 1583), precut (n = 419), or drainage (n = 701) had been carried out, singularly or in combination. RESULTS: One hundred eleven major complications (4.0%) were recorded: moderate-severe pancreatitis 36 (1.3%), cholangitis 24 (0.87%), hemorrhage 21 (0.76%), duodenal perforation 16 (0.58%), others 14 (0.51%). Among 942 diagnostic ERCPs there were 13 major complications (1.38%) and 2 deaths (0.21%), whereas among 1827 therapeutic ERCPs there were 98 major complications (5.4%) and 9 deaths (0.49%). The difference in the incidence of complications between diagnostic and therapeutic ERCPs was statistically significant (p < 0.0001). Small center and precut were recognized as independent risk factors for overall major complications of therapeutic ERCP, whereas the following risk factors were identified in relation to specific complications: (1) pancreatitis: age less than 70 years, pancreatic duct opacification, and nondilated common bile duct; (2) cholangitis: small center, jaundice; (3) hemorrhage: small center; and (4) retroperitoneal duodenal perforation: precut, intramural injection of contrast medium, and Billroth II gastrectomy. CONCLUSIONS: Major complications are mostly associated with therapeutic procedures and low case volume. Present data support a policy of centralization of ERCP in referral centers. A more selected and safer use of precut may be expected to further limit the adverse events of ERCP.

Adolescent↗

Results of therapeutic penetrating keratoplasty.

PURPOSE: To determine the anatomic and visual results of therapeutic penetrating keratoplasty (PK) and its role in the management of corneal disease. METHODS: The authors reviewed the records of all of their patients who had undergone therapeutic PK over the past 9 years and evaluated each for the following criteria: cure of disease, graft clarity, and visual acuity. Patients were divided into seven categories: (1 and 2) bacterial and fungal keratitis, (3 and 4) herpetic keratitis, with and without inflammation, (5) acanthamoebic keratitis, (6) perforations due to keratoconjunctivitis sicca, and (7) other causes of perforation. RESULTS: In microbial keratitis, therapeutic PK eradicated the disease in all cases. Seventy-three percent of grafts for bacterial keratitis and 60% for fungal keratitis remained clear. A higher percentage of clarity was achieved when grafts were 9.0 mm or less. Seven patients with secondary endophthalmitis were cured with a surgical approach including therapeutic PK. In herpetic keratitis with active inflammation, only 36% of grafts remained clear, and inflammation recurred in 36%. All perforations due to post-herpetic persistent epithelial defects in "quiet" eyes were grafted successfully. In patients with severe keratoconjunctivitis sicca, eyes were anatomically stabilized in 83%, but all grafts failed because of complications from ocular surface disease. CONCLUSIONS: Therapeutic PK is valuable in the management of microbial keratitis that does not respond to antimicrobial therapy. Results are poorer for patients with herpetic keratitis, although selected patients respond to therapeutic PK when other methods of management have failed. Patients with perforations due to keratoconjunctivitis sicca have a uniformly poor prognosis for graft clarity.

Acanthamoeba Keratitis↗

Mechanism of action of serotonin selective reuptake inhibitors. Serotonin receptors and pathways mediate therapeutic effects and side effects.

Serotonin selective reuptake inhibitors (SSRIs) are currently among the most frequently prescribed therapeutic agents in all of medicine. Their therapeutic actions are diverse, ranging from efficacy in depression to obsessive-compulsive disorder, panic disorder, bulimia and other conditions as well. The plethora of biological substrates, receptors and pathways for serotonin are candidates to mediate not only the therapeutic actions of SSRIs, but also their side effects. Specifically, the immediate actions of SSRIs are mostly side effects, and may be mediated by the initiating actions of SSRIs, namely negative allosteric modulation of the serotonin transporter. A leading hypothesis to explain these immediate side effects is that serotonin is increased at specific serotonin receptor subtypes in discrete regions of the body where the relevant physiologic processes are regulated. Desensitization of post-synaptic receptors in these same discrete brain regions may explain the development of tolerance to these same side effects. The explanation for therapeutic effects characteristic of SSRIs may be found in delayed neurochemical adaptations. A leading hypothesis for this action is desensitization of somatodendritic serotonin 1A autoreceptors in the midbrain raphe. The hypothesis to explain why SSRIs have such diverse therapeutic actions is that somatodendritic 5HT1A autoreceptor desensitization increases serotonin in those critical brain regions and at those key serotonin receptor subtype(s) which may mediate the pathophysiologies of the various disorders. Understanding the topography of serotonin receptor subtypes in discrete anatomical pathways may enhance our understanding of both the therapeutic actions and side effects of these important pharmaceutical agents.

Autoreceptors↗

The effects of therapeutic vs. high-intensity ultrasound on the rabbit growth plate.

Six of 6-week-old NZ rabbits underwent ultrasound treatment using a therapeutic dose (0.5 W/cm(2)) and other six were treated with a higher dose (2.2 W/cm(2)) to the lateral aspect of the left knee joint for 20 min per day and a total of six weeks. The right knee joint served as a control. The goal of this study is to see if the therapeutic dose and high dose (approximately 45-fold therapeutic dose) will have toxic effects on the physis. Histological review appeared normal growth plate in the therapeutic group. In the high dosage group three of six cases displayed flattening of the distal femoral epiphysis and wedging of the proximal tibial plateau and indistinct growth plate lines. It is of interest to note that there are opening radiolucent area in the lateral aspect on the femoral metaphysis in five of six cases, where bone resorption has taken place. Histological results showed that there are disordered arrays of the cartilaginous cells in the proliferative zone. The height of the lateral physis in the high dose group is not only greater than that in the therapeutic dose (1083.8 vs. 500.3 micro m), but also greater than that in their contralateral control (530.7 micro m) (P<0.05). This short-term study demonstrates that high dose ultrasound has profound pathologic effects in growing bone. Therapeutic doses of ultrasound do not have an adverse effect on bone growth in the short-term follow-up.

Animals↗

T-cell design: optimizing the therapeutic potential of autoreactive T cells by genetic modification.

Autoreactive CD4+ Th2 cells have been shown to be therapeutic in the treatment of EAE. However, their full therapeutic potential has yet to be realized. Genetic modification of autoreactive Th2 T cells may provide the means for delivering therapeutic transgene factors to autoimmune inflammatory lesions. Optimum therapeutic effects may be achieved by designing Th2 T cells in such a way that expression of transgene factors is regulated by antigen-inducible IL4, IL5 or IL10 transgene promoters. The innate antiinflammatory effects of the native autoreactive Th2 T cell may be enhanced by incorporating transgene regenerative growth factors in the T-cell design. Such factors may include remyelination growth factors (PDGF-A, bFGF, and IGF-I) that complement each other by acting predominantly at different stages in the development of mature myelinating oligodendrocytes. Moreover, in light of recent findings indicating extensive axonal damage during MS, neuroprotective transgene factors may prove to be therapeutic when delivered to EAE lesions by autoreactive Th2 T cells. Thus, optimum therapeutic effects may require multiple transfers of autoreactive Th2 T cells producing several distinct complementary transgene factors. In addition, the pathogenicity of epitope spreading and the inherent instability of self recognition during EAE may require serial transfer of genetically modified T cells reacting to multiple self determinants.

Animals↗

Are therapeutic relationships in psychiatry explained by patients' symptoms? Factors influencing patient ratings.

OBJECTIVE: To investigate the therapeutic relationship and how it is influenced by sociodemographic and clinical factors. METHOD: This study analysed self-ratings of the therapeutic relationship in 90 first-admitted, 72 long-term hospitalised and 41 out-patients with schizophrenia along with 249 alcoholic and 42 depressive in-patients and their association with sociodemographic and clinical variables. In all the patients, the therapeutic relationship was assessed using a simple scale based on three items. RESULTS: The therapeutic relationship differed significantly across groups. It was rated most positively by alcoholic patients and least positively by long-term hospitalised schizophrenia patients. Increased observer-rated psychopathology was significantly associated with a poorer therapeutic relationship in all groups except the hospitalised schizophrenia patients. In this group, increased self-rated symptoms were associated with a poorer relationship. In multiple regression analyses, 3-28% of the relationship variance was explained by psychopathology. CONCLUSIONS; Patient ratings of the therapeutic relationship were partially explained by psychopathology, leaving the greater part of the variance to be explained by factors other than sociodemographic and clinical characteristics.

Adult↗

Predictors of barriers to treatment and therapeutic change in outpatient therapy for antisocial children and their families.

This study examined the role of parent psychopathology and quality of life in predicting barriers to participation in outpatient treatment and therapeutic change among clinically referred children. Children (N = 169) referred for oppositional, aggressive, and antisocial behavior and their families participated. The major findings were that (1) higher levels of parent psychopathology and lower levels of quality of life predicted the subsequent emergence of perceived barriers to treatment in the parents and therapeutic changes among the children, (2) these effects were not explained by socioeconomic disadvantage or severity of child dysfunction, (3) perceived barriers and therapeutic changes were related and this relation was not explained by other family and child predictors, (4) as the level of perceived barriers increased among families, the amount of therapeutic change and the proportion of children who made a marked change decreased, and (5) parent perception of the relevance and demandingness of treatment were salient dimensions contributing to the relation between perceived barriers and therapeutic change. The conceptual and applied implications of relating barriers to treatment and therapeutic change are discussed.

Adolescent↗

Absence of bacterial colonization of the airways after therapeutic rigid bronchoscopy without stenting.

Following airway stenting, bacterial colonization of the airways with potentially pathogenic micro-organisms occurs within 4 weeks after treatment in the majority of patients. The objective of this study was to prospectively investigate whether nonstenting therapeutic rigid bronchoscopy (using laser, cryotherapy, mechanical dilatation or debridement) is followed by airway colonization or infection. Protected specimen brush sampling of the central airways and quantitative culture were performed immediately prior to, and 4 weeks after nonstenting therapeutic rigid bronchoscopy in 20 consecutive patients with central airway lesions. Prior to therapeutic bronchoscopy, airway colonization/infection was present in nine of 20 (45%) patients. In these nine patients, 10 different potential pathogens were identified: Streptococcus pneumoniae (four cases), Pseudomonas aeruginosa (three), Haemophilus influenzae (two), and Serratia marcescens (one). Eight of these nine patients had a history of postobstructive infections, of which three were currently being treated with antibiotics. Four weeks following therapeutic bronchoscopy, airway colonization/infection was present in five of 20 (25%) patients, each of whom had airway colonization/infection prior to bronchoscopy. In three of these five patients, the same organisms were found 4 weeks after bronchoscopy as at baseline bronchoscopy. In two of five patients new organisms were identified: one case of Streptococcus viridans and one case of Haemophilus parainfluenzae, both considered to be nonpathogens. In four of nine patients with airway colonization/infection prior to bronchoscopy, the airways were clear of micro-organisms after the procedure. The authors conclude that: 1) nonstenting therapeutic rigid bronchoscopy is not complicated by airway colonization or infection by new potential pathogens; and 2) therapeutic rigid bronchoscopy led to clearing of airway colonization/infection in almost half of the patients studied.

Adult↗

A reinforcement-based therapeutic workplace for the treatment of drug abuse: three-year abstinence outcomes.

Long-term Therapeutic Workplace effects were evaluated in heroin- and cocaine-dependent, unemployed, treatment-resistant young mothers. Participants were paid to work or to train in the Therapeutic Workplace but had to provide drug-free urine samples to gain daily access. Participants (N = 40) were randomly assigned to a Therapeutic Workplace or usual care control group. Therapeutic Workplace participants could work for 3 years. Relative to controls, Therapeutic Workplace participants increased cocaine (28% vs. 54% negative; p = .04) and opiate (37% vs. 60% negative; p = .05) abstinence on the basis of monthly urine samples collected until 3 years after intake. The Therapeutic Workplace can be an effective long-term treatment of cocaine and heroin addiction in poor and chronically unemployed young mothers.

Adolescent↗

Precision diagnostic and therapeutic interventions in rare genetic neurodevelopmental disorders.

Neurodevelopmental disorders (NDDs) include a broad spectrum of phenotypes spanning from intellectual disability (ID) to developmental delay (DD) and autism spectrum disorder (ASD). As neurodevelopmental phenotypes are a common presenting feature of an underlying genetic condition, professional medical organizations recommend genetic testing for all individuals with a NDD. When testing is pursued, identified genetic differences can lead to personalized clinical management with early diagnosis supporting the development of surveillance and intervention for co-occurring adverse health outcomes. Despite this, barriers to testing have prevented individuals from receiving a genetics referral and testing. Current therapeutic modalities including small molecule drugs, gene therapies, and antisense oligonucleotide therapies have emerged and shown promise in preclinical trials with therapeutic drugs gaining FDA approval. However, translational challenges are extensive, especially for identifying biomarkers of drug effects in the CNS. In this review, we discuss diagnostic approaches and clinical utility of genetic testing for rare genetic neurodevelopmental disorders, emerging development of individualized therapies, and progress for current therapeutics in addition to challenges with clinical translation and delivery. We will highlight opportunities for early diagnosis and treatment that are steadily gaining ground in favor of optimizing long-term health outcomes and improving quality of life for neurodiverse individuals. IMPACT: The path from genomics to therapeutics for neurodevelopmental disorders continues to present multiple opportunities and challenges. While emerging genome-wide sequencing and gene editing technologies deliver increased diagnostic yields and alternatives to life-long small molecule therapies, clinical translation has been challenging due to inherent cost and genetic heterogeneity. Limited access to genetic testing despite practice guidelines remains a barrier towards precision therapeutics for rare neurodevelopmental disorders, while pre-clinical investigations face obstacles when translating to human subjects. This review will summarize the impact of existing successes in diagnosis and therapeutics for neurodevelopmental disorders while highlighting ongoing challenges and areas of future opportunities.

Humans↗

A therapeutic trial of growth hormone in hypopituitary adults and its influence upon continued prescription by general practitioners.

OBJECTIVES: Adult GH deficiency (GHD) is associated with profound alterations in body composition, lipid profiles and quality of life which frequently improve after GH therapy. However, the beneficial effects of treatment are not derived by all and consequently some scepticism persists with regard to the use of GH therapy in adults. We assessed whether a 3-month therapeutic assessment with GH therapy could be used to determine which GHD adults should be treated over the longer term. We also assessed the continued prescription of GH by general practitioners (GPs) following the initial therapeutic assessment. DESIGN: A three month open therapeutic trial of GH in GHD adults. Patients were treated with GH at an initial dose of 0.01 iU/kg/d, increased after 1 month to 0.015 iU/kg/d for males and 0.02 iU/kg/d for females. After completion of the three months the continued prescription of GH by the GPs was assessed. PATIENTS: All adult GHD patients were considered for GH therapy. Thirty-nine GHD adults wanted GH therapy (group 1) and their baseline characteristics such as age, duration of GHD, and IGF-1 concentration were compared with 24 subjects who declined to receive GH (group 2). MEASUREMENTS: Measurements of body composition using bioelectrical impedance analysis, lipids and quality of life measured using a dedicated questionnaire were made before and after GH therapy. The response of the general practitioners to continued GH therapy after the initial therapeutic assessment was also noted. RESULTS: Compared with subjects who declined GH therapy (group 2), subjects of group 1 were younger (46.4 +/- 14.4 vs. 54.2 +/- 15.7 years, P < 0.05) and had lower peak GH responses to provocative testing (1.4 +/- 2.1 vs. 2.9 +/- 2.7 mU/l, P < 0.001), though there were no differences between IGF-1 concentration (11.7 +/- 6.2 vs. 14. 2 +/- 7.9 nmol/l). Following three months of GH therapy, there were significant improvements in all measured parameters including increased free fat mass (50.2 vs. 52.4 kg, P < 0.005) and total body water (37 vs. 38.7 l, P < 0.005), reduced fat mass (31.6 vs. 29.8 kg, P < 0.005), reduced AGHDA score (7 vs. 4, P < 0.001) and reduced cholesterol (6.3 vs. 5.8 mmol/l, P < 0.001), LDL (4 vs. 3.33 mmol/l, P < 0.001) and cholesterol/HDL ratio (5.57 vs. 4.67, P < 0.001). IGF-1 concentrations were significantly increased following treatment (12 vs. 32.4 nmol/l). Six subjects decided to discontinue GH therapy, 2 before the end of the study due to potential drug-related side-effects and 4 subjects derived no benefit from treatment. Despite the demonstrable benefits of treatment for the remaining 33 GHD adults, 6 GPs refused to continue to prescribe GH therapy for reasons of lack of familiarity with the drug or advice from their health authority. CONCLUSION: Patients who wanted GH therapy were usually younger and more severely GHD than counterparts who elect not to be treated. However, a therapeutic trial of GH therapy is required to distinguish those subjects who derive benefit from treatment. We have shown that three months of low dose GH therapy is a sufficient period to elicit significant beneficial responses in quality of life, body composition parameters and lipids for the majority of patients and appears to be a sufficient period for patients to decide whether they want longer term therapy. The initial therapeutic trial also provides the objective evidence for the general practitioners to decide upon the continued prescription of therapy. Despite the positive evidence provided by this study, a small minority of general practitioners still refuse to prescribe GH therapy.

Adult↗

Therapeutic action of ghrelin in a mouse model of colitis.

BACKGROUND & AIMS: Ghrelin is a novel growth hormone-releasing peptide with potential endogenous anti-inflammatory activities ameliorating some pathologic inflammatory conditions. Crohn's disease is a chronic debilitating disease characterized by severe T helper cell (Th)1-driven inflammation of the colon. The aim of this study was to investigate the therapeutic effect of ghrelin in a murine model of colitis. METHODS: We examined the anti-inflammatory action of ghrelin in the colitis induced by intracolonic administration of trinitrobenzene sulfonic acid. Diverse clinical signs of the disease were evaluated, including weight loss, diarrhea, colitis, and histopathology. We also investigated the mechanisms involved in the potential therapeutic effect of ghrelin, such as inflammatory cytokines and chemokines, Th1-type response, and regulatory factors. RESULTS: Ghrelin ameliorated significantly the clinical and histopathologic severity of the trinitrobenzene sulfonic acid-induced colitis; abrogating body weight loss, diarrhea, and inflammation; and increasing survival. The therapeutic effect was associated with down-regulation of both inflammatory and Th1-driven autoimmune response through the regulation of a wide spectrum of inflammatory mediators. In addition, a partial involvement of interluekin-10/transforming growth factor-beta1-secreting regulatory T cells in this therapeutic effect was demonstrated. Importantly, the ghrelin treatment was therapeutically effective in established colitis and avoided the recurrence of the disease. CONCLUSIONS: Our data demonstrate novel anti-inflammatory actions for ghrelin in the gastrointestinal tract, ie, the capacity to deactivate the intestinal inflammatory response and to restore mucosal immune tolerance at multiple levels. Consequently, ghrelin administration represents a novel possible therapeutic approach for the treatment of Crohn's disease and other Th1-mediated inflammatory diseases, such as rheumatoid arthritis and multiple sclerosis.

Analysis of Variance↗

[Therapeutic goals in psychological medicine--state of research and development of a standardized self-rating instrument].

From the patient's as well as from the therapist's point of view goals of therapy undoubtedly play an important role in all fields of psychological medicine (psychiatry, psychotherapy, psychosomatic medicine). On the other hand, there is a substantial lack of systematic i.e. empirical studies on this issue. Giving an overview on the available scientific literature we start by characterising therapeutic goals as a secondary psychological construction. Therapeutic goals and their multiple associations and different aspects of psychotherapy are discussed. However, a variety of definitions of therapeutic goals, often influenced by the therapeutic orientation of the authors, make it difficult to integrate concepts and to compare findings. Hence, we focus on those results which were found in relation to general concepts, i.e. beyond specific theoretical approaches. Furthermore, we discuss the pros and cons of individual measures of therapeutic goals compared with standardised self-rating instruments. Finally, we underline the lack of studies on therapeutic goals in the psychiatric sphere and report on some results of our own pilot study on the Personal Goals Questionnaire in psychiatric inpatients.

Humans↗

[Clinical relevance of computed tomography under emergency conditions--diagnostic accuracy, therapeutic consequences].

PURPOSE: To evaluate the diagnostic accuracy and therapeutic consequences of computed tomography performed on an emergency basis in a primary care hospital. MATERIAL AND METHODS: In 418 patients, 463 computed tomographies (thorax, abdomen, pelvis, spine, aorta, neck and extremities) were performed with-in 12 months, providing 999 diagnoses. The computed tomography diagnoses were retrospectively evaluated and correlated to surgery and discharge diagnoses. Therapeutical consequences were analyzed and allocated to a time period < 36 h (urgent) and > or = 36 - 72 h (elective). Average age was 49 (1 - 94) years (41% female and 59% male). Discharge diagnosis was defined as gold standard, provided that it was supported by clinical, blood chemical, diagnostic and possible surgical data. RESULTS: In 176 of 999 diagnoses (18%), the diagnoses were classified as "non-correlatable". Of the 823 correlated diagnoses, 431 were true positive, 14 false positive, 66 false negative and 312 true negative. Sensitivity, specificity and diagnostic accuracy of computed tomography was 87, 96 and 90%. Computed tomography had therapeutic consequences (surgery, drainage, puncture, reposition, thrombolytic therapy, chemotherapy, bronchoscopy, endoscopy, percutaneous transluminal angioplasty, coiling etc.) in 57% and no direct therapeutic interventions in 43%. Computed tomography excluded the suspected diagnosis in 36% and resulted in a conservative therapeutic regiment in 7%. Surgery was performed on 134 of the 418 patients (32%) who underwent computed tomography, with the surgery urgent in 71 (17%) and elective in 63 (15%) of the 418 patients. CONCLUSION: The high diagnostic accuracy and consecutive therapeutic consequences of computed tomography in establishing a precise, timely and reproducible diagnosis confirm its important role as primary diagnostic method on an emergency basis.

Adolescent↗

Therapeutic window of serum haloperidol concentration in acute schizophrenia and schizoaffective disorder.

Although several studies have been performed on the serum level-therapeutic effect relationship of neuroleptic drugs, the application of therapeutic drug-monitoring of neuroleptics is still a matter of controversy. Until now, haloperidol provided the most promising results. For this reason, an investigation of the dependence of clinical improvement on haloperidol serum levels was conducted in an acute psychiatric ward (Landeskrankenhaus Bernburg). In an open clinical trial haloperidol serum levels were measured in 57 acute schizophrenic patients for at least three weeks and correlated with clinical outcome (percent change of Brief Psychiatric Rating Scale, BPRS). A bisigmoidal model was used for data analysis. After three weeks of treatment, the major result proved to be a significant relationship between haloperidol serum level and therapeutic effect with pseudo-r2=0.316 and p=0.0031. Clinical improvement is enhanced by increasing haloperidol concentration up to about 10 ng/ml. It attains a maximum at about 10 ng/ml and decreases at haloperidol serum levels in a range of 10 ng/ml to 50 ng/ml. A simulation of this dependence of clinical improvement on serum levels, mediated by the variable dose design, can be excluded because of the results of a retrospective analysis of dosing behavior. Further evidence is thus provided for the dependence of therapeutic effect on the serum haloperidol concentration in acute schizophrenia. For practical application the position of a therapeutic window can be defined by a lower threshold level of about 5 ng/ml and an upper threshold of about 17 ng/ml. However, a maximal therapeutic effect is assured at 10 ng/ml. This should be the target value in serum level-guided dose adjustments.

Adult↗

Improved herpes simplex virus type 1 amplicon vectors for proportional coexpression of positron emission tomography marker and therapeutic genes.

For the development of efficient and safe gene therapy protocols for clinical application it is desirable to determine the tissue dose of vector-mediated therapeutic gene expression noninvasively in vivo. The herpes simplex virus type 1 thymidine kinase gene (HSV-1-tk) has been shown to function as a marker gene for the direct noninvasive in vivo localization of thymidine kinase (TK) expression by positron emission tomography (PET). Using bicistronic or multicistronic gene-expressing cassettes with tk as the PET marker gene, the quantitative analysis of tk gene expression may indirectly indicate the distribution and the level of expression of linked and proportionally coexpressed genes. Here, we describe the construction and functional evaluation of HSV-1 amplicon vectors mediating proportional coexpression of HSV-1-tk as PET marker gene and the enhanced green fluorescent protein gene (gfp) as proof of principle and cell culture marker gene and the Escherichia coli cytosine deaminase (cd) as therapeutic gene. Several double-/triple-gene constructs expressing HSV-1-tk, gfp, and E. coli cd were engineered based on gene fusion or the use of an internal ribosome entry site (IRES). Functional analysis in cell culture (green fluorescent protein [GFP] fluorescence and sensitivity to the prodrugs ganciclovir [GCV] and 5-fluorocytosine [5-FC]) and Western blots were carried out after infection of proliferating rat 9L gliosarcoma and human Gli36 glioma cells with helper virus-free packaged HSV-1 amplicon vectors. To study the ability of PET to differentiate various levels of tk expression noninvasively in vivo, retrovirally transduced and selected populations of rat F98 and human Gli36dEGFR glioma cells with defined levels of proportionally coexpressed tk and gfp genes were grown as subcutaneous tumors in nude rats and nude mice, and tk imaging by PET was performed. To study HSV-1 amplicon vector-mediated gene coexpression in vivo, HSV-1 amplicon vectors bearing coexpression constructs were injected (4 x 10(7) to 1 x 10(8) transducing units) into subcutaneously growing Gli36dEGFR gliomas in nude animals, and tk imaging was performed 24 hr later. All vector constructs mediated GFP expression and sensitized 9L and Gli36 cells toward GCV- and 5-FC-mediated cell killing in a drug dose-dependent manner, respectively. The levels of gene expression varied depending on the location of the genes within the constructs indicating the influence of the IRES on the level of expression of the second gene. Moreover, functional proportional coexpression of the PET marker gene HSV-1-tk and the linked therapeutic E. coli cd gene was observed. In selected tumor cell populations, subtle IRES-dependent differences of tk gene expression could be noninvasively distinguished by PET with good correlation between quantitative assays for IRES-dependent attenuated GFP and TK expression in culture and in vivo. After infection of subcutaneously growing gliomas with HSV-1 amplicon vectors, various levels of TK expression were found ranging from 0.011-0.062 percentage injected dose per gram (%ID/g). These values were 4.0- to 5.7-fold lower than positive control tumor cells. TK expression could be imaged by PET in vivo even with the tk gene located at the weak position downstream from the IRES. In conclusion, these HSV-1 amplicon vectors carrying HSV-1-tk as PET marker gene and any linked therapeutic gene will serve an indirect noninvasive assessment of the distribution of therapeutic gene expression by PET. Monitoring the correlation between primary transduction and therapeutic efficiency of a given vector is highly desirable for the development of safe and efficient gene therapy and vector application protocols in clinical applications.

Animals↗