Endothelial cell apoptosis is accelerated by inorganic iron and heat via an oxygen radical dependent mechanism

BACKGROUND: Iron participates in diverse pathologic processes by way of the Fenton reaction, which catalyzes the formation of reactive oxygen species (ROS). To test the hypothesis that this reaction accelerates apoptosis, we used human umbilical vein endothelial cells (HUVECs) as surrogates for the microvasculature in vivo. METHODS: HUVECs were loaded with Fe [III](ferric chloride and ferric ammonium citrate) with 8-hydroxyquinoline as carrier and were then challenged with two stimuli of the heat shock response, authentic heat or sodium arsenite. Iron dependence was tested with two chelators, membrane-impermeable deferoxamine and membrane-permeable o-phenanthroline. The role of ROS was assessed with superoxide dismutase, catalase, and the reporter compound dichlorofluorescein diacetate. The mechanism of cell death was assessed with three complementary techniques, Annexin V/propidium iodide labeling, the TUNEL stain, and electron microscopy. RESULTS: Iron-loaded HUVECs executed apoptosis after a heat shock stimulus. Iron-catalyzed formation of ROS appeared to be a critical mechanism, because both chelation of iron and enzymatic detoxification of ROS attenuated this apoptosis. CONCLUSIONS: Inorganic iron, in concert with chemical and physical inducers of the heat shock response, may trigger apoptosis. The accumulation of iron in injured tissue may thereby predispose to accelerated apoptosis and account, in part, for poor wound healing and organ failure.     

Modulation of the action of gonadotrophin surge-attenuating factor by gonadotrophin-releasing hormone

Gonadotrophin surge-attenuating factor (GnSAF) is a putative non-steroidal ovarian factor which attenuates the luteinizing hormone (LH) surge in superovulated women through the reduction of the pituitary response to gonadotrophin-releasing hormone (GnRH). The mechanism of action of GnSAF on gonadotrophin secretion was further studied by investigating six normally ovulating women in two cycles--a spontaneous and a follicle-stimulating hormone (FSH)-treated cycle. The response of the pituitary to five consecutive pulses of GnRH was investigated in late follicular phase (follicle size 15 mm) of both cycles. GnRH pulses, 10 micrograms each, were injected i.v. every 2 h and LH was measured in blood samples taken before and 30, 60 and 120 min after each pulse. FSH was injected daily at the fixed dose of 225 IU starting on cycle day 2. Peak values of LH increment occurred 30 min after each pulse. However, maximal LH increment occurred in both cycles after the second GnRH dose. In the FSH cycles the response of LH to the first three pulses was significantly attenuated compared with the spontaneous cycles, while the response to the fourth and fifth pulses was similar in the two cycles. In both cycles, LH increment 30 min post GnRH (net increase above the previous value) was similar after the fourth and fifth pulses. Serum concentrations of oestradiol and immunoreactive inhibin, although higher in the FSH cycles, remained stable throughout the GnRH experimental period in both cycles. These results demonstrate that multiple submaximal doses of GnRH can override the attenuating effect of GnSAF on LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the lymph nodes of rats under the influence of natural sorbents in damaging exposure to carbophos

The experiment was performed in male Wistar rats to study the influence of natural sorbents (Tyumen diatomites and Novosibirsk kudurites) administered with food at dose 6% of its dry weight on lymph nodes. The data obtained demonstrated that addition of natural sorbents to food does not exert pathological influence on lymph nodes of experimental animals. Sorbent diet affects follicular and paracortical lymph node structures, changing their sizes and cytological composition; natural sorbents increase immune potential which is realized after the impairing action.     

Characterization of clinical isolates of Streptococcus agalactiae by random amplified polymorphic DNA using degenerate oligonucleotides

Epidemiological studies of Streptococcus agalactiae strains have been limited by the lack of sensitive and discriminatory methods for comparing clinical isolates. Serotyping, albeit a widely used methodology, has been shown to possess low capability to distinguish between epidemiologically related and unrelated isolates. We have employed here a random amplification of polymorphic DNA (RAPD) assay, using degenerate oligonucleotides as primers, to characterize S. agalactiae isolates from related or unrelated clinical samples. Epidemiologically-related isolates (mother-infant pairs) showed identical profiles by this methodology. On the contrary, 12 epidemiologically-unrelated isolates (classified into 5 different serotypes) resulted in 11 distinct RAPD patterns. This suggests that the proposed modified RAPD assay provides a highly discriminatory tool for the analysis of genomic diversity among isolates from pathogenic organisms.     

Auditory assessment in a protocol of concurrent carboplatin and irradiation use in fractionated therapy

Possible ototoxicity related with a hyperfractionated concurrent chemo-radiation schedule was studied in 36 patients with head and neck cancer. The schedule consisted of two daily fractions, each consisting of 5 mg/m2 of carboplatin plus 115 cGy. Therapy was given 5 days a week for 7 weeks up to a total dose of 700 mg carboplatin and 8050 cGy. Tonal audiograms and superliminal tests were carried out before treatment, at conclusion, six months post-treatment, and every year thereafter. After a mean follow-up of 18 months (maximum 30 months), no treatment-related sensorineural hearing loss was observed. However, recruitment disappeared in post-treatment superliminal tests in 37% of patients who presented sensorineural deafness with recruitment, although it reappeared in audiograms recorded six months later. We attributed this to transient demyelinization. After treatment, a previous tympanosclerosis reactivated in 1 case and effusive otitis media occurred in 8 patients. Previous effusive otitis media disappeared in 1 patient (nasopharyngeal tumor).     

Mutations in filamin 1 prevent migration of cerebral cortical neurons in human periventricular heterotopia

Long-range, directed migration is particularly dramatic in the cerebral cortex, where postmitotic neurons generated deep in the brain migrate to form layers with distinct form and function. In the X-linked dominant human disorder periventricular heterotopia (PH), many neurons fail to migrate and persist as nodules lining the ventricular surface. Females with PH present with epilepsy and other signs, including patent ductus arteriosus and coagulopathy, while hemizygous males die embryonically. We have identified the PH gene as filamin 1 (FLN1), which encodes an actin-cross-linking phosphoprotein that transduces ligand-receptor binding into actin reorganization, and which is required for locomotion of many cell types. FLN1 shows previously unrecognized, high-level expression in the developing cortex, is required for neuronal migration to the cortex, and is essential for embryogenesis.     

Stability and throughput optimization of multichannel slotted non-persistent CSMA protocol

This paper describes two protocols suitable for a multichannel, multiaccess slotted non-persistent CSMA environment for infinite population, analyses their stability and examines their throughput optimization. The common sensing multichannel slotted non-persistent CSMA (CSCSMA) protocol is an extension of the slotted single channel non-persistent CSMA protocol with an appropriate policy for the selection of the channel in which a station (re)transmits. This policy restricts the control information among the stations, and consequently reduces the cost of the station interface. Pakes's Lemma criteria are applied to define sufficient conditions for ergodicity of the Markov chain, which describes the evolution of the busy station population, and to specify regions in which the multichannel system is stable. Control parameters are the retransmission probabilities. Optimization rules are derived which show that the optimal retransmission probabilities may be expressed as a function of the number of busy stations. The separate sensing multichannel slotted non-persistent CSMA protocol (SSCSMA), using a different policy, distinguishes the channels into two groups: the retransmission channel group devoted in collision resolution; and the transmission channel group for the access of free stations. Stability regions are defined using the results for CSCSMA, and rules for optimal allocation of channels among the two groups are derived for improvement of system performance.