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Biomedical subjects

G Rispoli

Publications and source records attributed to G Rispoli.

At least 19 recordsLinked to original sources

One-trocar appendectomy.

BACKGROUND: Laparoscopic appendectomy is a feasible and a safe alternative to open appendectomy. Several laparoscopic procedures have been described that use one or more trocars. We report our experience with the treatment of acute appendicitis using a laparoscopy-assisted technique by means of only one transumbilical trocar. METHODS: From February 1996 to February 1999 we performed 65 laparoscopic appendectomies. In the procedures, a 10-mm operative telescope was used, with a 450-mm atraumatic grasper introduced through the operative channel. After the intraabdominal laparoscopic dissection, the appendix was exteriorized through the umbilical trocar. The appendectomy was performed outside the abdomen as in the open procedure. The procedure was completed using only one trocar in 55 patients (84.6%). Regarding the other 10 cases (15.3%), in 5 we used more than one trocar and in 5 conversion to open surgery was needed. RESULTS: The average operating time in our series was 25 min and the median time to discharge was 2 days. There were six (11%) postoperative complications (three serous umbilical secretion and three umbilical hematomas). CONCLUSIONS: Our results suggest that this technique, which combines the advantages of both the open and the laparoscopic procedures, is a valid alternative for the treatment of acute appendicitis. However, this procedure cannot always be completed using only one trocar, as happened in 10 cases in our series.

Adolescent↗

No evidence for calcium electrogenic exchanger in frog semicircular canal hair cells.

We investigated the possibility that, in hair cells mechanically isolated from frog semicircular canals, Ca2+ extrusion occurs via a Na+ : Ca2+ (cardiac type) or a Na+ : Ca2+,K+ (retinal type) exchanger. Cells concurrently imaged during whole-cell patch-clamp recordings using the Ca2+ sensitive fluorescent dye Oregon Green 488 BAPTA-1 (100 micro m) showed no voltage dependence of Ca2+ clearance dynamics following a Ca2+ load through voltage-gated Ca2+ channels. Reverse exchange was probed in hair cells dialyzed with a Ca2+- and K+-free solution, containing a Na+ concentration that saturates the exchanger, after zeroing the contribution to the whole-cell current from Ca2+ and K+ conductances. In these conditions, no reverse exchange current was detected upon switching from a Ca2+-free external solution to a solution containing concentrations of Ca2+ alone, or Ca2+ + K+ that saturated the exchanger. By contrast, the same experimental protocol elicited peak exchange currents exceeding 100 pA in gecko rod photoreceptors, used as positive controls. In both cell types, we also probed the forward mode of the exchanger by rapidly increasing the intracellular Ca2+ concentration using flash photolysis of two novel caged Ca2+ complexes, calcium 2,2'-([1-(2-nitrophenyl)ethane-1,2-diyl]bis(oxy))bis(acetate) and calcium 2,2'-([1-(4,5-dimethoxy-2-nitrophenyl)ethane-1,2-diyl]bis(oxy)) bis(acetate), in the presence of internal K+ and external Na+. No currents were evoked by UV-triggered Ca2+ jumps in hair cells, whereas exchanger conformational currents up to 400 pA, followed by saturating forward exchange currents up to 40 pA, were recorded in rod photoreceptors subjected to the same experimental conditions. We conclude that no functional electrogenic exchanger is present in this hair cell population, which leaves the abundant plasma membrane Ca2+-ATPases as the primary contributors to Ca2+ extrusion.

Animals↗

Cholecystoenteric fistula (CF) is not a contraindication for laparoscopic surgery.

BACKGROUND: Cholecystoenteric fistula (CF) is a rare complication of cholelithiasis. The aim of this study was to evaluate the safety and risk of complications when the laparoscopic approach is applied in patients with CF. METHODS: A questionnaire was mailed to all surgeons with experience of >100 cholecystectomies working in Naples, Italy, and the neighboring area. RESULTS: Between February 1990 and May 1999, 34 patients presented with cholecystoenteric fistula (0.2% of >15,000 laparoscopic cholecystectomies performed in the same period). These patients were allocated into two groups: the LT group (those who underwent laparotomic conversion after the diagnosis of CF), which consisted of 20 patients, four men and 16 women, with a mean age of 66.5 +/- 9.3 years (range, 46-85) and the LS group (laparoscopically treated patients), which consisted of 14 patients, three men and 11 women, with a mean age of 65.6 +/- 8.8 years (range, 51-74). They types of CF observed were as follows: in the former group of patients, cholecystoduodenal fistulas (n = 11, 55%), cholecystocolic fistulas (n = 5, 25%), cholecystojejunal fistulas (n = 3, 15%), and cholecystogastric fistulas (n = 1, 5%); in the latter group, cholecystoduodenal fistulas (n = 8, 5.1%), and cholecystocolic fistulas (n = 4, 28.6) and cholecystojejunal fistulas (n = 2, 14.3%). Stapler closure of CF was done in four LT patients and three LS patients with cholecystoduodenal fistula; it was also done in three LT patients and three LS patients with cholecystocolic fistula. Hand-sutured fistulectomy was performed in six LT patients and three LS patients with cholecystoduodenal fistula, in two LT patients with cholecystocolic fistula, and in all patients with cholecystojejunal or cholecystogastric fistula. There were no deaths or intraoperative complications in either group. One patient in the LT group developed a bronchopneumonia postoperatively. Postoperative hospital stay was significantly longer in LT patients-17 +/- 4 vs 3+/-1 days (p < 0.001). CONCLUSION: Cholecystoenteric fistula is an occasional intraoperative finding during laparoscopic cholecystectomy. The results of this study, which are based on the collective experiences of 19 surgeons, illustrate the growing success of the laparoscopic approach to this condition, including a decreasing rate of conversion to open surgery over the last 3 years.

Aged↗

Regional distribution of calcium currents in frog semicircular canal hair cells.

In the present work we studied the regional expression of voltage-dependent Ca channels in hair cells from the frog semicircular canals, employing whole-cell patch-clamp on isolated and in situ hair cells. Although Ca channels are thought to play a major role in afferent transmission, up to now no data were available regarding their distribution in vestibular organs. The problem appears of interest, especially in the light of recent results showing the presence of multiple Ca current components in semicircular canal hair cells. Our data suggest the presence, in all regions of the crista ampullaris, of two classes of cells, one displaying an inactivating Ca current (R1) and one lacking it. In the former cells, Ca current amplitude decreased from the central to the peripheral zone (the maximal currents being observed in the intermediate zone). Only L-type and R2 current components displayed regional differences in expression, whereas the size and properties of R1, although variable among cells, were not regionalized. However, in cells lacking R1, Ca current amplitudes were similar regardless of cell shape and location. The possible contributions of this Ca current distribution to afferent discharge properties are discussed.

Animals↗

Calcium-activated potassium current clamps the dark potential of vertebrate rods.

Vertebrate photoreceptors respond to light with a graded hyperpolarization from a membrane potential in the dark of approximately -35 mV. The present work investigates the physiological role of the Ca2+-activated K+ current in the photovoltage generation in mechanically isolated rods from salamander retina. Membrane current or voltage in isolated rods was recorded from light- and dark-adapted rods under voltage- or current-clamp conditions, respectively. In light-adapted rods of the salamander, selective blockade of Ca2+-activated K+ channels by means of charybdotoxin depolarized the plasma membrane of current-clamped rods by approximately 30 mV, from a resting potential of approximately -35 mV. A similar depolarization was observed if external Ca2+ (1 mM) was substituted with Ba2+ or Sr2+. Under control conditions, the injection of currents of increasing amplitude (up to -100 pA, to mimic the current entering the rod outer segment) could not depolarize the membrane potential beyond a saturating value of approximately -20 mV. However, in the presence of charybdotoxin, rods depolarized up to +20 mV. In experiments with dark-adapted current-clamped rods, charybdotoxin perfusion lead to transient depolarizations up to 0 mV and steady-state depolarizations of approximately 5 mV above the dark resting potential. Finally, the recovery phase of the voltage response to a flash of light in the presence of charybdotoxin showed a transient overshoot of the membrane potential. It was concluded that Ca2+-activated K+ current is necessary for clamping the rod photovoltage to values close to the dark potential, thus allowing faithful single photon detection and correct synaptic transmission.

Ambystoma mexicanum↗

Dynamics of intracellular calcium in hair cells isolated from the semicircular canal of the frog.

Changes in cytosolic free Ca(2+) concentration ([Ca(2+)]i) were monitored optically in hair cells mechanically isolated from frog semicircular canals using the membrane-impermeant form of the Ca(2+)-selective dye Oregon Green 488 BAPTA-1 (OG, 100 microM). Cells stimulated by depolarization under whole-cell voltage clamp conditions revealed Ca(2+) entry at selected sites (hotspots) located mostly in the lower (synaptic) half of the cell body. [Ca(2+)]i at individual hotspots rose with a time constant tau1 approximately 70 ms and decayed with a bi-exponential time-course (tau2 approximately 160, tau3 approximately 2500 ms) following a 160 ms depolarization to -20 mV. With repeated stimulation [Ca(2+)]i underwent independent amplitude changes at distinct hotspots, suggesting that the underlying Ca(2+) channel clusters can be regulated differentially by intracellular signalling pathways. Block by nifedipine indicated that the L-type Ca(2+)channels are distributed at different densities in distinct hotspots. No diffusion barrier other than the nuclear region was found in the cytosol, so that, during a prolonged depolarization (lasting up to 1s), Ca(2+) was able to reach the cell apical ciliated pole. The effective Ca(2+) diffusion constant, measured from the progression of Ca(2+) wavefronts in the cytosol, was approximately 57 microm(2)/s. Our results indicate that in these hair cells, buffered diffusion of Ca(2+) proceeds evenly from the source point to the cell interior and is dominated by the diffusion constant of the endogenous mobile buffers.

Animals↗

[Human castration: historical notes].

Human castration has been performed from early times for different reasons: to punish and revenge, to display one's religious fanaticism, to protect or to control women, for eunuchs' trade, for therapeutical purposes. In early modern times men were castrated to obtain sopranos voices, or for eugenic or racial reasons. Nowadays chemical castration is used as a therapeutic treatment or as a way to punish rape and other criminal behaviours. Castration is surgical or chemical act that may obviously cause serious physical and psychological consequences.

History, Ancient↗

[ARS mingendi cum instrumentis, probes: candelette, minugie, catheters].

The Museum of History of Medicine of the University of Rome "La Sapienza" has a rich collection of probes and metal catheters, some of which in surgical kits. Tracing the history of catheters, the authors follow the technical evolution in the field of urology techniques, also highlighting the intelligent craft of artisans, surgeons, barbers - and the industrial effort to ameliorate techniques. The use and the crafting of these instruments are to be related to new knowledge in the fields of urethral, vesical and prostatic occlusion. Italian contribution of the field is stressed, highlighting the original experience of Michele Troja, who invented and diffused metal catheters covered in caouthouc.

Andrology↗

Ca2+-dependent kinetics of hair cell Ca2+ currents resolved with the use of cesium BAPTA.

Hair cells in the frog semicircular canal, studied by the whole-cell patch-clamp technique, display three distinct Ca2+ currents: two non-inactivating components (L type and R type, the latter termed R2 in the following) and a second R type current (termed R1), which runs down first and inactivates in a Ca2+-dependent fashion. Since intracellular EGTA, up to 5 mM, did not display major effects on such inactivation, we used increasing amounts of BAPTA in the patch pipette, to control [Ca2+]i more efficiently and investigate whether modifications in [Ca2+]i at the cytoplasmic side of the channel affect the inactivation of the RI component and in general the gating of all channel types. The results here reported show that (1) K+ currents heavily contaminate recordings obtained using high concentrations of BAPTA in its commercially available K+ salt form; (2) BAPTA Cs+ salt can be satisfactorily employed to obtain reliable recordings; (3) the kinetics of channel gating and R1-channel inactivation are indeed markedly affected by effectively buffering [Ca2+]i.

Animals↗

Removal of a foreign body from the distal colon using a combined laparoscopic and endoanal approach: report of a case.

PURPOSE: We describe the case of a of 53-year-old homosexual male, from whom a cigar container (22 cm in length and 3.5 cm in diameter), which was introduced into the rectum and migrated upwards to the distal colon, was successfully extracted by combining laparoscopic and anal approaches. METHODS: The foreign body was mobilized laparoscopically, pushed down the rectum, and then extracted transanally with the aid of a dilator anoscope. CONCLUSIONS: Combining the laparoscopic and anal approaches in selected cases of colorectal foreign bodies is useful.

Colon↗

Calcium currents in hair cells isolated from semicircular canals of the frog.

L-type and R-type Ca(2+) currents were detected in frog semicircular canal hair cells. The former was noninactivating and nifedipine-sensitive (5 microM); the latter, partially inactivated, was resistant to omega-conotoxin GVIA (5 microM), omega-conotoxin MVIIC (5 microM), and omega-agatoxin IVA (0.4 microM), but was sensitive to mibefradil (10 microM). Both currents were sensitive to Ni(2+) and Cd(2+) (>10 microM). In some cells the L-type current amplitude increased almost twofold upon repetitive stimulation, whereas the R-type current remained unaffected. Eventually, run-down occurred for both currents, but was prevented by the protease inhibitor calpastatin. The R-type current peak component ran down first, without changing its plateau, suggesting that two channel types generate the R-type current. This peak component appeared at -40 mV, reached a maximal value at -30 mV, and became undetectable for voltages > or =0 mV, suggestive of a novel transient current: its inactivation was indeed reversibly removed when Ba(2+) was the charge carrier. The L-type current and the R-type current plateau were appreciable at -60 mV and peaked at -20 mV: the former current did not reverse for voltages up to +60 mV, the latter reversed between +30 and +60 mV due to an outward Cs(+) current flowing through the same Ca(2+) channel. The physiological role of these currents on hair cell function is discussed.

Animals↗

[Surgical therapy of varicocele. Technical note on the inguinal approach].

BACKGROUND: There are several therapeutic proposals for idiopathic varicocele treatment, but all of them present a certain number of relapses. The technique proposed consists, through an inguinal approach, in a systematic ligation and section of all veins potentially responsible of a venous reflux. METHODS: During a period from 1990-1995 a total of 73 patients were surgically treated for idiopathic varicocele with this technique. All patients underwent preoperative spermiogram, hormonal assay and Doppler velocimetry. Through an inguinal approach, under selective spinal anaesthesia, a ligature and section of the internal spermatic vein at the internal inguinal ring is performed, followed by searching, ligature and section, of other veins responsible of venous reflux (external spermatic vein, anastomosis with sapheno-femoral veins, arteriovenous micro-anastomosis, pubic veins). The patient is discharged within twenty-four hours. RESULTS: An external dilated spermatic vein was discovered in 30.1%, pubic veins in 49.3%, anastomosis with safeno-femoral veins in 10.9% and arteriovenous micro-anastomosis in 43.8%. There were no mortality or serious postoperative morbidity; only in five cases there were complications consistent in two orchitis and three hydrocele. No relapse at follow-up (12-36 months), performed with palpation and a Doppler study, was found, while there was a spermiogram improvement in 61.6% of cases. CONCLUSIONS: This surgical approach is a safe and radical treatment without any relapse, of all types of varicocele.

Adolescent↗

Calcium regulation of phototransduction in vertebrate rod outer segments.

The biochemical events underlying the phototransduction cascade in retinal photoreceptors of vertebrates are now well established, on the basis of a wealth of electrophysiological and biochemical evidence. In this review the Ca2+ regulation of the enzymes that generates the photoreceptor light response is analyzed, as well as the Ca2+ transport across the plasma membrane. Most of the results discussed in the following were collected from electrophysiological experiments.

Animals↗

Electrophysiological characterization of ionic transport by the retinal exchanger expressed in human embryonic kidney cells.

The retinal Na+:Ca2+, K+exchanger cDNA was transiently expressed in human embryonic kidney (HEK 293) cells by transfection with plasmid DNA. The correct targeting of the expressed protein to the plasma membrane was confirmed by immunocytochemistry. The reverse exchange offrent (Ca2+ imported per Na+ extruded) was measured in whole-cell voltage-clamp experiments after intracellular perfusion with Na+ (Na+i, 128 mM) and extracellular perfusion with Ca2+ (Ca2o+, 1 mM) and Ko+ (20 mM). As expected, the exchange current was suppressed by removing Ca2o+. Surprisingly, however, it was also abolished by increasing Na+o to almost abolish the Na+ gradient, and it was almost unaffected by the removal of Ko+. Apparently, then, at variance with the exchanger in the rod outer segment, the retinal exchanger expressed in 293 cells acts essentially as a Na+:Ca2+ exchanger and does not require K+ for its electrogenic activity.

Calcium↗

Expression of an active Na+/Ca2+ exchanger isoform lacking the six C-terminal transmembrane segments.

The short isoform of the Na+/Ca2+ exchanger (67 kDa) that is produced by alternative splicing during the expression of the 6 kb canine exchanger cDNA in 293 cells was separately expressed in the same system. The protein consisted of the five N-terminal transmembrane segments and of a large portion of the main hydrophilic loop, but lacked the six C-terminal hydrophobic segments of the regular protein (108 kDa). Very high RNA levels were found after transient cell transfection with plasmid DNA encoding this truncated isoform. The RNA processing, the translation and targeting of the resulting protein to the plasma membrane appeared to be less efficient than those of the 108-kDa polypeptide produced in the same system. The Na(+)-dependent Ca(2+)-uptake activity of 293 cells expressing the short isoform was measured by an isotopic rapid filtration method, whereas the current associated with Ca2+ extrusion was measured in electrophysiological patch-clamp experiments. The results showed that the expressed isoform functioned in the typical reverse and forward Na+/Ca2+ exchange modes. In both the electrophysiological and the isotopic measurements the activity of the short isoform was 6-7-fold lower than that of the 108-kDa protein expressed in the same system. However, lower amounts of the short isoform reached the plasma membrane: its specific activity could thus be significantly higher. Possibly, the short isoform could form a dimer in which a second 67 kDa polypeptide replaces the C-terminal part of the 108-kDa protein.

Alternative Splicing↗

Transport of K+ by Na(+)-Ca2+, K+ exchanger in isolated rods of lizard retina.

Transport of K+ by the photoreceptor Na(+)-Ca2+, K+ exchanger was investigated in isolated rod outer segments (OS) by recording membrane current under whole-cell voltage-clamp conditions. Known amounts of K+ were imported in the OS through the Ca(2+)-activated K+ channels while perfusing with high extracellular concentration of K+, [K+]o. These channels were detected in the recordings from the OS, which probably retained a small portion of the rest of the cell. The activation of forward exchange (Na+ imported per Ca2+ and K+ extruded) by intracellular K+, Ki+, was described by first-order kinetics with a Michaelis constant, Kapp(Ki+), of about 2 mM and a maximal current, Imax, of about -60 pA. [Na+]i larger than 100 mM had little effect on Kapp(Ki+) and Imax, indicating that Nai+ did not compete with Ki+ for exchange sites under physiological conditions, and that Na+ release at the exchanger intracellular side was not a rate-limiting step for the exchange process. Exchanger stoichiometry resulted in one K+ ion extruded per one positive charge imported. Exchange current was detected only if Ca2+ and K+ were present on the same membrane side, and Na+ was simultaneously present on the opposite side. Nonelectrogenic modes of ion exchange were tested taking advantage of the hindered diffusion found for Cai2+ and Ki+. Experiments were carried out so that the occurrence of a putative nonelectrogenic ion exchange, supposedly induced by the preapplication of certain extracellular ion(s), would have resulted in the transient presence of both Cai2+ and Ki+. The lack of electrogenic forward exchange in a subsequent switch to high Nao+, excluded the presence of previous nonelectrogenic transport.

Animals↗

Visual transduction in dialysed detached rod outer segments from lizard retina.

1. Properties of a new preparation for studying the physiology and biochemistry of phototransduction in retinal rods are described. Whole-cell voltage clamp was used to record the generation, maintenance and light-sensitivity of dark current in rod outer segments that had been isolated from the rest of the receptor cell by detachment at the connecting cilium. 2. Detached outer segments dialysed with standard internal solution supplemented with physiological amounts of ATP (5 mM) and GTP (1 mM) developed a standing inward dark current that was the sum of three components: approximately 91% light-sensitive current, approximately 6% Na(+)-Ca2+,K+ exchange current and approximately 3% leakage current. Light-sensitive dark current (mean amplitude approximately -63 pA) was suppressed transiently by brief flashes in an intensity-dependent manner. Light responses had the same kinetics, sensitivity and intensity-response relationship as those recorded from intact rods. 3. Dialysed outer segments differed from intact rods in that intense flashes evoked saturating responses that recovered incompletely to a plateau of reduced dark current caused by incomplete inactivation of the transduction cascade. Light sensitivity was reduced for a short time following an intense flash and then recovered despite persistent reduction of dark current. This suggests that there is no fixed relationship between dark current amplitude and light sensitivity. 4. Light-sensitive dark current faded rapidly when outer segments were not supplied with nucleotides. Outer segments dialysed with solution that contained cyclic GMP, but no ATP or GTP, supported dark current at a level that increased with [cyclic GMP]. When basal phosphodiesterase (PDE) activity is inhibited, 8 microM cyclic GMP supports a dark current of approximately 70 pA. 5. Light sensitivity decreased during recordings made with solution that contained only cyclic GMP, consistent with the inhibition of G protein activation by loss of GTP. After thorough nucleoside triphosphate depletion, however, intense illumination evoked a transient increase rather than a decrease in dark current, i.e. an inverted light response. This result suggests that isomerized rhodopsin may generate a signal that causes either inhibition of basal PDE activity or release of bound cyclic GMP. 6. Sustained Na(+)-Ca2+,K+ exchange current was recorded during steady illumination when Ca2+, but not when Mg2+, was added to the dialysis solution. Exchange current increased with the amount of added Ca2+ and saturated at approximately 18 pA when the dialysis solution contained > or = 10 mM Ca2+.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine Triphosphate↗