Biomedical subjects
J C Pérez
Publications and source records attributed to J C Pérez.
[Therapeutic guide to primary care based on the evidence, 2nd edition: an honest and practical work].
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Telomeric associations in cigarette smokers exposed to low levels of X-rays.
Telomeric association (TA), i.e. fusion of chromosomes by their telomeres, predisposes a cell to genetic instability. Because of this we investigated the effect of X-rays exposure and cigarette smoking on the frequency of TA in peripheral blood lymphocytes of exposed individuals, in order to determine if TA can be a chromosomal marker in populations exposed to these carcinogens and if there is an synergistic effect between both agents. We found that the exposed groups show a greater percentage of TA when compared with the control group (P<0.001). However, although the percentage of metaphases with TA in the group with combined exposure (12.6%) was greater than in the others exposed groups (P<0.05), this value was less than the sum of the two individual effects (15.1%). Our results suggest that probably there is not an additive or synergistic effect between X-rays and smoking, and that TA may be a useful cytogenetic marker for evaluating populations exposed to mutagens.
Partial characterization of a basic protein from Crotalus molossus molossus (northern blacktail rattlesnake) venom and production of a monoclonal antibody.
The venom of Crotalus molossus molossus (blacktailed rattlesnake) is very basic compared to that of other Crotalinae venoms. Unlike other Crotalinae venoms that are separated by anion exchange chromatography, C. m. molossus venom must be fractionated by cation exchange chromatography. Electrophoretic titration (ET) was used to predict the isoelectric point (pI) and optimal conditions for isolation. The specific hemorrhagic activity for C. m. molossus venom was 7.5 mm/microg, making it one of the most hemorrhagic of Crotalinae venoms. Basic hemorrhagic and fibrinolytic proteins from the venom of C. m. molossus venom were further fractionated by cation exchange chromatography. A basic fibrinolytic/hemorrhagic protein (CMM4) was isolated. CMM4 has a molecular weight between 23 and 26 kDa and a pI of approximately 11.3. SDS electrophoresis revealed one band and ET curve revealed 3 bands with very similar surface charges at all pH. CMM4 did not activate plasminogen when tested with a Chrom Z-PLG assay. The proteins in CMM4 had similar N-terminal amino acid sequences to each other (D-Q-Q-N-L-P-Q-(S/A/R)-Y-(V/R/I)-E-L-V-V-V-A-D-H-R-L-F-M-K-Y-K-S-D-L- N-T). The differences in these proteins are in positions 8 and 10. CMM4 may contain isoforms that differ by minor sequence variations at their amino-termini. The amino acid sequences of CMM4 were very similar to other fibrinolytic and hemorrhagic metalloproteinases isolated from venoms of the genera Crotalus. The specific hemorrhagic activity of CMM4 decreased as the specific fibrinolytic activity increased. A monoclonal antibody (CMM1b) was produced against C. m. molossus venom that neutralized the hemorrhagic activity of some of its fractions. CMM1b also reacted with 11 of 29 venom samples tested via ELISA.
An internet database of crotaline venom found in the United States.
Many snake venoms have been shown to be complex mixtures of pharmacologically important molecules, some of which have potential therapeutic value in the treatment of clot-induced ischemia, cancer and other human disorders. The literature contains many references on how venom and/or venom components are being used in medicine. Within the United States, there are 44 subspecies of poisonous snakes. Despite this rather vast diversity, 90% of the venom-related biomedical research conducted on native snakes found in the United States has been done on a limited number of the more common species. Since the venoms from most of the native species are not available or characterized, their composition and potential usefulness in medicine and applied biomedical research has not been explored. The Natural Toxins Research Center (NTRC) at Texas A&M University-Kingsville has developed a serpentarium that presently houses a population of over 250 snakes composed of 11 species and 20 subspecies. These snakes are cataloged on the Internet database along with their geographical location data, proteolytic activities, high performance liquid chromatography (HPLC) and electrophoretic titration (ET) profiles. Many of these snake venoms have never been characterized and few locale-specific differences within a species have been examined. These venoms can be queried through an on-line search routine. The database will be a useful starting point for anyone interested in isolating fibrinolytic enzymes, specific toxins, hemorrhagins, or other pharmacologically active proteins from snake venoms.
Percutaneous tracheostomy: comparison of Ciaglia and Griggs techniques.
BACKGROUND: Although the standard tracheostomy described in 1909 by Jackson has been extensively used in critical patients, a more simple procedure that can be performed at the bedside is needed. Since 1957 several different types of percutaneous tracheostomy technique have been described. The purpose of the present study was to compare two bedside percutaneous tracheostomy techniques: percutaneous dilatational tracheostomy (PDT) and the guidewire dilating forceps (GWDF). MATERIALS AND METHODS: A prospective study in two medical/surgical intensive care units (ICUs) was carried out. Sixty-three critically ill patients who required endotracheal intubation for longer than 15 days were consecutively selected to undergo PDT (25 patients) or GWDF (38 patients) technique. Intraoperative and postoperative complications were recorded. RESULTS: Age (mean +/- standard error) was 63 +/- 1.1 years. The patients had been mechanically ventilated for an average of 19.8 +/- 1.2 days. The GWDF technique was significantly faster than PDT technique (P = 0.02). Fifteen complications occurred in 10 out of 63 (15%) patients. They were as follows: tracheal tear (one patient in each group; in one case this was due to false passage); transient hypotension (one patient in the PDT group and two patients in the GWDF group); atelectasis (one patient in the PDT group); and haemorrhage (one patient in the PDT group and three patients in the GWDF group). In both patients with tracheal tear, reduced arterial oxygen saturation (SaO2) with concomitant subcutaneous emphysema ensued. CONCLUSION: We found no statistical differences between complications with both techniques. The surgical time required for the GWDF technique was less than that for PDT.
The DeltaF508 mutation in Ecuador, South America.
There are few reports about the incidence of the DeltaF508 mutation in Latin American countries. We show the study of the DeltaF508 mutation and the seven most common "European" mutations in 10 Ecuadorian CF affecteds. The incidence of DeltaF508 mutation found was 25% and none of the other seven was detected in our population, which indicates that at least 60% of the mutations in the studied population are different from most common in Europe. Similar data have been reported in other Amerindian populations, therefore it is suggested that Cystic Fibrosis in Ecuador-and other Amerindian countries in Latin America-have a different ethiology than that of Caucasian populations.
Natural protease inhibitors to hemorrhagins in snake venoms and their potential use in medicine.
Snake venoms are complex mixtures of many toxins and enzymes which effectively immobilize prey without a struggle and assist in digestion. Certain animals have a remarkable resistance to envenomation of snakes. Naturally occurring factors that neutralize snake venoms have been found in the sera of most snakes and a few warm-blooded animals. These antihemorrhagic and antineurotoxic factors have been purified from snake and mammalian sera. The antihemorrhagins are not immunoglobulins since they have different physical and chemical characteristics. The natural immunity to hemorrhagins is the result of tissue inhibitors of metalloproteinases (TIMP) found in animal sera of resistant animals. Most animals have matrix metalloproteinases (MMP) and TIMP that are implicated in a wide variety of normal physiological processes and pathological conditions. MMP in animals have many biological functions in embryogenesis, morphogenesis and tissue remodeling. MMP activities are precisely regulated by endogenous TIMP. Disruption of the balance between MMP and TIMP causes various diseases such as arthritis, periodontal diseases, diabetes, ophthalmologic conditions, neoplasia, metabolic bone disease, atherosclerosis and orthopedic conditions. Resistant animals that have a high titer of TIMP would have a survival advantage when bitten by poisonous snakes. Snake venoms are abundant and stable sources of MMP which are medically important. The venom MMP which cause unregulated destruction of tissue have sequences which have some degree of homology with mammalian MMP which control normal biological processes. Resistant animals are important sources of TIMP which can be used to study metalloproteinase related diseases. For these reasons the MMP in snakes and TIMP in resistant animal are excellent candidates for developing new drug therapies.
The antihemorrhagic factor of the Mexican ground squirrel, (Spermophilus mexicanus).
The Mexican ground squirrel (Spermophilus mexicanus) has a natural resistance to western diamondback rattlesnake venom (Crotalus atrox). The LD50 for the Mexican ground squirrel is 53 mg/kg body weight, which is 13 times higher than that of BALB/c mice. An antihemorrhagic factor from serum of the Mexican ground squirrel was isolated using Sephadex G-200 gel filtration, ion exchange A-50, G-75 gel filtration and HPLC DEAE 5PW ion exchange chromatography. The purified factor neutralized proteolytic and hemorrhagic activity of crude C. atrox venom. The results of this research suggest that the antihemorrhagic factor in the serum of the Mexican ground squirrel is not an antibody and neutralizes hemorrhagic activity of C. atrox venom.
Screening for fibrinolytic activity in eight Viperid venoms.
Snake venoms contain direct-acting fibrinolytic metalloproteinases (MMP) that could have important applications in medicine. Fibrinolytic enzymes isolated from venom can induce in vitro clot lysis by directly acting on a fibrin clot. The most ideal fibrinolytic enzyme would have high affinity for clots, dissolve clots directly without causing hemorrhage, and would not be neutralized in vivo by endogenous metalloproteinase inhibitors. The purpose of this study was to compare DEAE/HPLC venom profiles from Viperid snakes and identify fractions that contain fibrinolytic activity with no hemorrhagic activity and are not neutralized by animal sera. The sera selected were from four (Virginia opossum, Gray woodrat, Mexican ground squirrel, and Hispid cottonrat) animals known to neutralize hemorrhagic activity in snake venoms. Nineteen fractions from the Viperid venoms had fibrinolytic activity. Agkistrodon venom fractions contained the highest specific fibrinolytic activities. A. piscivorus leucostoma fraction 4 contained a high specific fibrinolytic activity, no hemorrhagic activity, and the fibrinolytic activity was not neutralized by the proteinase inhibitors of the four animal sera. A. contortrix laticinctus fraction 1 also had a high specific fibrinolytic activity and no hemorrhagic activity. However, the fibrinolytic activity was neutralized by Didelphis virginiana (Virginia opossum) serum.
The detection of hemorrhagic proteins in snake venoms using monoclonal antibodies against Virginia opossum (Didelphis virginiana) serum.
Most snakes and a few warm-blooded animals have a resistance to snake venoms because of naturally occurring antihemorrhagins found in their sera. The antihemorrhagins in serum of Virginia opossum (Didelphis virginiana) neutralize hemorrhagic activity by binding to hemorrhagins in snake venoms. The binding characteristic of antihemorrhagins in D. virginiana serum was used to develop a five-step western blot. The detection of hemorrhagic proteins were measured indirectly with antihemorrhagins in Virginia opossum serum and with DV-2LD#2, a monoclonal antibody specific for Virginia opossum antihemorrhagins. Snake venoms were separated by native-PAGE, transferred to a Millipore Immobilon-P membrane and then incubated with crude Virginia opossum serum. The hemorrhagins in snake venom bind to antihemorrhagins in Virginia opossum serum which react with DV-2LD#2 a monoclonal antibody that is specific for Virginia opossum antihemorrhagins. DV-2LD#2 monoclonal antibody inhibits antihemorrhagic activity in Virginia opossum serum when mixed in equal amounts. The inhibition of antihemorrhagins by DV-2LD#2 monoclonal antibody suggests specificity. DV-2LD#2 monoclonal antibody does not recognize antihemorrhagins in gray woodrat (Neotoma micropus) serum. The five-step western blot reveals two well-defined bands which represent hemorrhagins found in Western diamondback rattlesnake (Crotalus atrox) venom. Venoms from 15 different snake species were examined to determine the usefulness of the five-step western blot. Other hemorrhagic venoms (Great Basin rattlesnake (C. viridis lutosus), Prairie rattlesnake (C. viridis viridis), Tancitaran dusky rattlesnake (C. pusillus), Northern Mojave rattlesnake (C. scutulatus scutulatus type B) and Northern Pacific rattlesnake (C. v. oreganus)) had one single band in the five-step western blot. DV-2LD#2 did not bind to the non-hemorrhagic venoms and reacted with 50% of the hemorrhagic venoms used in this study. The monoclonal antibody, CAH, reacted with all the hemorrhagic venoms except for the venom of the King cobra (Ophiophagus hannah) and did not react with the non-hemorrhagic venoms. The hemorrhagic binding site of CAH monoclonal antibody and the antihemorrhagin in Virginia opossum are different binding sites. The five-step western blot will be a very useful assay for determining hemorrhagic activity without using live animals.
X-radiation actions on the neuromuscular transmission.
The effects of high doses of X-radiation (100 kilorads) on the neuromuscular transmission of isolated sciatic nerve-sartorius muscle preparations of the frog, as evaluated by bioelectrical characteristics, were explored. Intracellular microelectrode recordings after X-irradiation showed that the resting, excitation and action potentials of nerve fibers approaching the synaptic terminal region of the motor end-plate became lessened, and also presented a slower velocity of impulse propagation, earlier than that observed in muscle cells. After forty minutes following the irradiation period, the neuromuscular transmission became blocked, although muscle fibers still responded to direct electrical stimulation. Records taken at the motor end-plate region of muscle cells, demonstrated the presence of postsynaptic miniature end-plate potentials (m.e.p.p.'s), the sequence of which fits closely into a random Poisson distribution. X-irradiation elicited an increase of the rate of m.e.p.p.'s and induced membrane changes over fine terminal nerve branches, leading into a failure to initiate and propagate action potentials. Only as time progressed, this nerve bioelectrical impairment was accompanied by a similar one in muscle cells, associated to the inability to develop contractile tension. The increase of m.e.p.p.'s frequency due to depolarization by a high K+ concentration, of presynaptic nerve membranes of control and irradiated preparations, was reversed by a high Mg+--Ca2+--free media. However, a concentration of Mg2+, which normally reduced quantal release of acetylcholine (ACh), without altering presynaptic nerve membrane potentials, failed to modify the spontaneous basal frequency of m.e.p.p.'s, both in irradiated and control preparations. The findings of the present study suggest that the presynaptic ACh synthesis, storage, and availability for ACh liberation are not early affected by X-rays, i.e. at a time when transmission from nerve to muscle had already failed. On the contrary, the most precocius membrane changes following X-irradiation, seem related to the mechanisms underlying nerve resting, excitation, and action potentials. The smallest presynaptic nerve terminal fibers close to the motor end-plate, are the most radiosensitive portion of the myoneural junction. Indeed, their bioelectric characteristics are early affected by X-rays, resulting in a failure to generate and propagate nerve impulses which lead to a blockade of neuromuscular transmission.
Programmed stimulator for threshold of action potential.
A programmed digital stimulator is described, in which the pulse generator can be regulated in frequency and duration. The output pulses amplitude increases automatically by increments under 5% of the previous step, sequence which allows a much better approximation to the threshold value of action potential. A photographic camera is synchronized for recording. The power supplies, pulse duration generator, binary counter, converter, pulse control, and detector circuits are given in detail.
Electronic system for the obtention of phase plane curves for action potentials.
This system is designed for the amplification of the bioelectrical signal, obtention of its derivative in respect to time, automatic brightness control for both beams of the oscilloscope, and stability. The system includes a central measuring section which is a modified Tektronix oscilloscope type 565. Automatic control of brightness as a function of sweeping speed allows photographic recordings.
Automatic microelectrode compensator (AMC).
A solid-state system designed for compensation of asymmetry potentials between microelectrodes, for bioelectric measurements, is described. Advantages over the manual compensator are the following: synchronic compensation of electrodes, application to automatized systems against aging effects in electordes, high speed, and elimination of general muisances of manual circuits. Besides, the system allows to measure digitally the polarization difference and resting membrane potential with precision of +/- 1 mV.
Automatic silver-plater of electrodes. Reference silver-silver chloride electrodes.
The silver-plater developed for the preparation of silver-silver chloride electrodes attains constancy in electrochemical characteristics, circulating current during exactly identical periods with inverted polarity.
Denervation effects on choline depolarization of muscle membrane.
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[On analeptics: Remeflin and its clinical application].
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