Recombinant human thyrotropins of the twenty-first century

In recent years, many new recombinant protein therapeutics have been developed and tested in clinical trials [1]. Current and future clinical uses of recombinant human thyroid-stimulating hormone (rhTSH; Thyrogen, Genzyme) in thyroid diseases are discussed in the review published in this issue of Expert Opinion on Pharmacotherapy [2]. As Thyrogen is a wild-type rhTSH produced in Chinese hamster ovary cells, it has relatively low affinity to the human TSH receptor. Such low affinity and weak intrinsic bioactivity of rhTSH, compared to the bovine or rodent TSH, may help to explain the results of several studies indicating limited clinical efficacy of Thyrogen. TSH analogues with largely increased receptor affinity, potency and efficacy, are expected to provide not only more effective than currently used diagnostic methods, but should also serve as indispensable second-generation thyrotropins for the diagnosis and treatment of thyroid carcinomas with a largely limited number of TSH receptors.     

An attempt to use Gore-Tex surgical membrane in lumbar disc surgery

BACKGROUND: Peridural fibrosis developing after lumbar discectomy may be responsible for as much as 20% of all Failed Back Surgery Syndrome. A variety of biological and non-biological materials have been used as a barrier to invasion of fibrous tissue into the vertebral canal. AIM: The purpose of this study was to evaluate the use of expanded polytetrafluoroethylene (ePTFE) surgical membrane (Gore-Tex membrane) to inhibit peridural fibrosis and reduce FBSS symptoms after lumbar discectomy. MATERIAL AND METHODS: In a prospective study we compared postoperative results in 20 patients who had an ePTFE membrane implanted during lumbar discectomy with the results in 20 patients in whom no material was implanted. The outcomes were evaluated using a questionnaire on activities of daily living according to the Low Back Outcome Score, pain grading scale -- Visual Analog Scale, assessment of Lasegue sign and MRI 18-24 months after the operation for all patients. RESULTS: The authors found no evident positive clinical and radiological effects of using ePTFE surgical membrane during lumbar discectomy. CONCLUSIONS: 1. It is impossible to prove that ePTFE membrane used during lumbar discectomy essentially prevents postoperative peridural scar formation. 2. The use of ePTFE membrane does not improve the outcome of the surgical treatment of lumbar disc herniation.     

Spontaneous spinal cord herniation: a rare cause of the Brown-Sequard syndrome

Recent reports have shown that spontaneous spinal cord herniation in the thoracic segment of the spine may be one of the causes of the progressive spinal cord lesion. Although it was described for the first time by Wortzman in 1974, it was only in early 1990s that a growing number of publications on single cases of the condition started to be observed. In the relevant literature collected by us we found reports on 53 patients altogether. In a number of cases herniation was diagnosed only intraoperatively in spite of complex radiological diagnostics. The analysis of relevant literature shows that spontaneous or idiopathic spinal cord herniation has a very typical clinical picture and most cases share a number of features, such as the clinical condition, age and sex of the patient as well as the level and location of the entity and its radiological picture. We conclude that spontaneous herniation of the spinal cord may be not as rare as previously thought and that it should be always taken into account in differential diagnostics of progressive myelopathy in the thoracic segment in middle-aged patients, especially females.     

New score indicating placental vascular resistance

BACKGROUND: Umbilical artery Doppler velocimetry is a routine method for fetal surveillance in high-risk pregnancy. Uterine artery Doppler seems to give comparable information, but it can be difficult to interpret as there are two arteries, which might show notching and/or increased pulsatility index (PI) as signs of increased vascular impedance. Combining the information on vascular resistance on both sides in a new score might simplify and improve evaluation of placental circulation. METHODS: Uterine and umbilical artery Doppler velocimetry was evaluated in 633 high-risk pregnancies. The managing clinician was informed only about the umbilical artery flow. The umbilical artery flow spectrum was semiquantitatively divided into four blood flow classes (BFC), expressing signs of increasing vascular resistance. The uterine artery Doppler flow spectrum was divided into five uterine artery scores (UAS), taking into account presence/absence of notching and/or increase in PI. By adding UAS to BFC, a new placental score (PLS) was constructed with values ranging from 0 to 7, indicating general placental vascular resistance. The scores were related to three outcome variables: small-for-gestational age (SGA), premature delivery (<37 weeks), and cesarean section. RESULTS: All three score systems showed a significant relationship between signs of increasing vascular resistance and outcome. The new PLS showed the best association to adverse outcomes, with optimal cut-off at values exceeding score 3. CONCLUSION: Doppler velocimetry on both sides of the placenta showed a strong relationship to an adverse outcome of pregnancy. The new PLS showed a better relationship to adverse perinatal outcome than the BFC and the UAS. The PLS can simplify evaluation of uteroplacental and fetoplacental Doppler velocimetry.     

Risk of lymph node metastases in patients with ovarian cancer

AIM: The aim of this study was the assessment of incidence of the lymph node spread in patients with ovarian cancer. Additionally, some of clinical and histopathology factors, as well as patients age were analyzed in relation with lymph nodes metastases. MATERIAL AND METHOD: Based on 112 operations performed in patients with ovarian cancer FIGO stage I-IV, analysis of pelvic and paraaortic lymph node metastasis was carried out. In this group only in 70 patients paraaortic lymph nodes were removed. The rest of patients underwent pelvic lymphadenectomy only because of poor general condition or very intensive cytoreductive surgery. Statistical analysis was provided using unvaried regression test and Pearson test. RESULTS: In early stages of ovarian cancer (I and II) the percent of patients with involved lymph nodes was 17.4 and in advanced stages 37.9. Strong correlation between involvement of pelvic and paraaortic lymph nodes was seen. The most frequent localization of lymph node metastases was the site around intercrossing of left renal vein and aorta. It should be stressed that in 8 cases isolated paraaortic metastases were seen. Risk factors of lymph node metastases were clinical stage, tumor grade and age of patients. Clear cell carcinoma and mixed carcinoma had also prognostic significance. CONCLUSION: This analysis proved that incidence of lymph node metastases was high even in early stage, and therefore lymphadenectomy should be an integral part of standard surgical procedures in patients with ovarian cancer.     

Flow cytometric "rare event analysis": a standardized approach to the analysis of donor cell chimerism

Lack of standardization of methods and detection limits contributes to the controversial results regarding microchimerism after organ transplantation and has prompted the development of a standardized, reproducible, "easy-to-use" flow cytometric method for this type of "rare event analysis".EDTA-blood of healthy, HLA-typed donors was stained simultaneously with FITC- and biotin-labeled HLA-class I antibodies (One Lambda) as well as Cy5-PE-labeled CD45 (Medac, Hamburg) according to a standard protocol and analysed on a Coulter EPICS XL Flow cytometer (FCM).An absolute range of positivity (mean MESF+/- 1 STD) was determined for 22 HLA-specific antibodies. The range of positivity ranged between 5000 and 20,000 MESF (anti-A23, 24(9) FITC) and 40,000-140,000 (anti-Bw6 FITC). The frequency of nonspecific positive signals using nonstained cells, isotype-controls and irrelevant HLA antibodies was between 0.01% and about 0.5%, in some samples up to 1.4%, with an MESF between 8000 and 150,000, thus interfering clearly with the defined positive range of most antibodies tested. Using an "HLA antibody cocktail", combining FITC- and PE-labeled antibodies for different HLA specificities and thereby creating an internal control, the identification of donor cells was improved but was only rarely applicable.Due to the lack of highly reactive antibodies, FCM analysis was not suitable for the reliable identification of very low numbers of donor hematopoetic cells despite the theoretical advantages of flow cytometric detection of rare events. The single parameter approach was hampered by a significant frequency of nonspecific positive signals, which were easily mistaken as specific (true) positive signals, whereas the multiparameter approach could only be used in selected cases.     

Prion diseases: a dual view of the prion hypothesis as seen from a distance

We review the historical background and principles of the prion theory in its current shape. We showed that most of data may be still interpreted dually according to the protein only hypothesis and according to the theory in which additional component is necessary to comprise the infectivity. The enormous impact of structural biological studies is also stressed.     

Influence of vigabatrin, a novel antiepileptic drug, on the anticonvulsant activity of conventional antiepileptics in pentetrazole-induced seizures in mice

Vigabatrin is a novel antiepileptic drug, which increases GABA levels by irreversible inhibition of GABA-aminotransferase. The aim of this study was to evaluate the effects of vigabatrin on the anticonvulsant activity of valproate, ethosuximide and clonazepam against pentetrazole-induced seizures in mice. In addition, the effects of antiepileptic drugs alone or in combination with vigabatrin were studied on motor performance and long-term memory. Chemical seizures were induced by subcutaneous injection of pentetrazole at its CD(97) and defined as a clonus of the whole body with an accompanying loss of righting reflex, lasting for over 3 s. Vigabatrin inhibited the clonic pentetrazole-induced seizures and ED(30) of the drug was 879 mg/kg. Vigabatrin (at the subthreshold dose of 250 mg/kg) potentiated the protective activity of ethosuximide, reducing its ED(30) from 142 to 95 mg/kg against clonic seizures induced by pentetrazole, but simultaneously elevated its plasma level. The protective activity of valproate and clonazepam remained almost unchanged. However, vigabatrin (250 mg/kg) decreased TD(30) (50% toxic dose - corresponding to the impairment of motor coordination in 50% of the animals) of ethosuximide and clonazepam from 549 and 3.84 to 460 and 1.1 mg/kg, respectively, in the chimney test. Vigabatrin (250 mg/kg) did not influence TD(30) value of valproate in this test. Vigabatrin (at the dose of 250 mg/kg) did not impair long-term memory in combination with antiepileptics. Potentiation of the ethosuximide's protective activity was apparently due to a pharmacokinetic interaction. Consequently, no pharmacodynamic interactions between vigabatrin and the studied conventional antiepileptic drugs were evident.     

Loss of melanin from human RPE with aging: possible role of melanin photooxidation

The pigment melanin, which is believed to play a photoprotective role, was quantified here in human RPE cells from donors of different age. Electron spin resonance (ESR) spectroscopy was shown to provide a quantitative measure of melanin and was used as a non-destructive measure of melanin content. Results indicated an age-related melanin loss in RPE cells, with melanin content diminishing 2.5-fold between the first and the ninth decade of life. To determine whether photo-oxidation may contribute to age-related changes in RPE melanin, RPE in human eyecups, isolated human and bovine RPE cells, purified melanin granules, or synthetic dopa melanin were irradiated with various wavelengths and intensities of visible light. Samples were analysed for changes in melanin content by ESR spectroscopy, and by absorption and emission spectrophotometry. The concentration of hydrogen peroxide was measured in some samples, and some human eyecups were examined by transmission electron microscopy. Irradiation of RPE in eyecups with intense visible light was found to produce a time-dependent photobleaching of melanosomes that was accompanied by the formation of hydrogen peroxide. Photobleaching of isolated RPE melanosomes and synthetic dopa melanin resulted in enhanced melanin fluorescence, as previously shown for melanin from aged donors by others, and significantly reduced ESR signal intensity, resembling the changes in melanin with aging observed here. We conclude that the content of melanin in RPE cells undergoes an age-related change to which photo-oxidation may contribute. This observation raises the question of whether age-related changes in melanin reduce the photoprotective role of the pigment in aging RPE cells.