Morphologic variations in the pineal gland of the albino rat after a chronic alcoholisation process.

We studied the effect of alcohol on the pineal gland of 48 male Wistar rats. Animals were divided into control and experimental groups. The experimental group underwent a previous progressive alcoholisation period with ethanol diluted in water at a concentration of 40%. Animals were sacrificed at 3, 6, 9 and 12 months, and the ultrastructure, karyometric indices, and number of synaptic bodies in the pineal gland were analysed. The results showed progressive morphologic alterations in the ethanol-treated animals, which culminated in fatty degeneration of the pineal parenchyma after 6 months. The karyometric indices decreased in both the central and peripheral areas compared with the control group. Moreover, the seasonal rhythmicity observed in the controls disappeared in the experimental groups, whose number of different populations of synaptic bodies (synaptic ribbons and synaptic spherules) considerably lowered with inversion of their normal seasonal rhythm. These results support that chronic alcoholisation leads to fatty degeneration of the pineal parenchyma, and a considerable alteration in nuclear functional rhythms and synaptic bodies.

Importance of the Moskowitz artery in the laparoscopic medial approach to splenic flexure mobilization: a cadaveric study.

BACKGROUND: The medial approach in laparoscopic splenic flexure mobilization is based on the entrance to the lesser sac just above the ventral edge of the pancreas (VEOP). The artery of Moskowitz runs through the base of the mesocolon, just above the VEOP. The aim of this study was to assess the incidence of the artery of Moskowitz, its route and its distance from the VEOP. METHODS: We performed a cadaveric study on 27 human cadavers. The vascular arcades of the splenic flexure were dissected, the number of vascular arches, and the origin and localization of its terminal anastomosis were recorded. The splenic flexure avascular space (SFAS) was defined as the avascular zone in the mesocolon delimited by the VEOP, middle colic artery, ascending branch of the left colic artery and the vascular arch of the splenic flexure nearest to the VEOP and was quantified as the distance between the VEOP and the most proximal arch RESULTS: The artery of Drummond was identified in 100% of the cadavers. In 5 of 27 (18%) Riolan's arch was present, and in 3 of 27 (11%) the Moskowitz artery was found. The mean distance from the VEOP to the artery of Moskowitz was 0.3 cm (SD 0.04). This vascular arch travelled from the origin of the middle colic artery to the distal third of the ascending branch of the left colic artery. The SFAS was greater (p = 0.001) in cadavers that only presented the artery of Drummond (mean 6.8 cm; SD 1.25) than in those with Riolan's arch (mean 4.5 cm; SD 0.5) CONCLUSIONS: In the medial approach for laparoscopic mobilization of the splenic flexure, when only one of the arches is present, the avascular area is an extensive and secure territory. If the artery of Moskowitz is present, the area is nonexistent and this would contraindicate the approach due to risk of iatrogenic bleeding. A radiological preoperatory study could be essential for accurate and safe surgery in this area.

[Proposed difficult airway teaching methodology. Presentation of an interactive fresh frozen cadaver model]. / Propuesta de metodología docente en vía aérea difícil. Presentación de un modelo interactivo de cadáver fresco congelado.

BACKGROUND AND OBJECTIVE: The aim of this paper is to present a methodology based on the use of fresh-frozen cadavers for training in the management of the airway, and to evaluate the degree of satisfaction among learning physicians. MATERIAL AND METHODS: About 6 fresh-frozen cadavers and 14 workstations were prepared where participants were trained in the different skills needed for airway management. The details of preparation of the cadavers are described. The level of satisfaction of the participant was determined using a Likert rating scale of 5 points, at each of the 14 stations, as well as the overall assessment and clinical usefulness of the course. RESULTS: The mean overall evaluation of the course and its usefulness was 4.75 and 4.9, out of 5, respectively. All parts of the course were rated above 4 out of 5. The high level of satisfaction of the course remained homogeneous in the 2 editions analysed. The overall satisfaction of the course was not finally and uniquely determined by any of its particular parts. CONCLUSION: The fresh cadaver model for training physicians in techniques of airway management is a proposal satisfactory to the participant, and with a realism that approaches the live patient.

Efficacy of three drugs for protecting against gentamicin-induced hair cell and hearing losses.

BACKGROUND AND PURPOSE Exposure to an ototoxic level of an aminoglycoside can result in hearing loss. In this we study investigated the otoprotective efficacy of dexamethasone (DXM), melatonin (MLT) and tacrolimus (TCR) in gentamicin (GM)-treated animals and cultures. EXPERIMENTAL APPROACH Wistar rats were divided into controls (treated with saline); exposed to GM only (GM); and three GM-exposed groups treated with either DXM, MLT or TCR. Auditory function and cochlear surface preparations were studied. In vitro studies of oxidative stress, pro-inflammatory cytokine mRNA levels, the MAPK pathway and caspase-3 activation were performed in organ of Corti explants from 3-day-old rats. KEY RESULTS DXM, MLT and TCR decreased levels of reactive oxygen species in GM-exposed explants. The mRNA levels of TNF-α, IL-1ß and TNF-receptor type 1 were significantly reduced in GM + DXM and GM + MLT groups. Phospho-p38 MAPK levels decreased in GM + MLT and GM + TCR groups, while JNK phosphorylation was reduced in GM + DXM and GM + MLT groups. Caspase-3 activation decreased in GM + DXM, GM + MLT and GM + TCR groups. These results were consistent with in vivo results. Local treatment of GM-exposed rat cochleae with either DXM, MLT or TCR preserved auditory function and prevented auditory hair cell loss. CONCLUSIONS AND IMPLICATIONS In organ of Corti explants, GM increased oxidative stress and initiated an inflammatory response that led to the activation of MAPKs and apoptosis of hair cells. The three compounds tested demonstrated otoprotective properties that could be beneficial in the treatment of ototoxicity-induced hearing loss.

Review of the anatomic concepts in relation to the retrorectal space and endopelvic fascia: Waldeyer's fascia and the rectosacral fascia.

OBJECTIVE: A precise anatomical study of the fascias within the retrorectal space is reported, analyzing and clarifying the anatomical concepts previously employed to describe Waldeyer's and the rectosacral fascia. METHOD: The pelvis was dissected in 15 cadavers (10 males and five females). All specimens were divided in the median sagittal plane including the middle axis of the anal canal, to allow a correct visualization of and access to the retrorectal space. RESULTS: The retrorectal space was limited anteriorly by the rectum and posterior mesorectum covered by a fine visceral fascia, and posteriorly by the sacrum covered by the parietal presacral fascia. The rectosacral fascia divided the retrorectal space into inferior and superior portions in 80% of the male and 100% of the female specimens. It originated from the presacral parietal fascia at the level of S2 in 15%, S3 in 38% and S4 in 46% of specimens. In all cases it passed caudally to join the rectal visceral fascia 3-5 cm above the anorectal junction. As described by Waldeyer, the floor of the retrorectal space is formed by the fusion of the presacral parietal fascia and the rectal visceral fascia and lies above the levator ani muscle at the level of the anorectal junction. CONCLUSION: The rectosacral fascia divides the retrorectal space into inferior and superior portions. This must be differentiated from Waldeyer's description of the fascia lying in the inferior limit of the retrorectal space, formed by the fusion of the rectal visceral and parietal fascias.

Corneal endothelial cell density decreases with age in emmetropic eyes.

PURPOSE: To analyze the corneal endothelial cell density in healthy adult emmetropic subjects. METHODS: We analyzed the corneal endothelial cell density of a group made up of 225 emmetropic subjects (n=225). As age-matched control groups we analyzed two other groups, one made up of myopic subjects (n=209) and the other made up of hyperopic subjects (n=203). We recorded the mean of three consecutive measurements of the corneal endothelial cell density using the Topcon SP-2000P non-contact specular microscope (Topcon Corp., Tokyo, Japan). RESULTS: The mean age was 38.6+/-11.8 years, 40.7+/-12.2 years, and 39.2+/-10.5 years for emmetropic, myopic and hyperopic subjects respectively (p=0.994). No significant differences (p=0.920) in endothelial cell density values were found between emmetropic (2985+/-245 cells/mm2), myopic (2936+/-258 cells/mm2) and hyperopic eyes (2946+/-253 cells/mm2). Lower corneal endothelial cell density values were found in older emmetropic (p<0.001), myopic (p<0.001), and hyperopic subjects (p<0.001). A significant correlation between endothelial cell density and age was found in emmetropic (r=-0.958; p<0.001), myopic (r= -0.954; p<0.001) and hyperopic subjects (r= -0.948; p<0.001). CONCLUSIONS: In healthy emmetropic subjects there is a reduction in corneal endothelial cell density with age although there are no differences in corneal endothelial cell density values between emmetropic, myopic and hyperopic subjects.

Intraobserver reproducibility of retinal nerve fiber layer measurements using scanning laser polarimetry and optical coherence tomography in normal and ocular hypertensive subjects.

PURPOSE: To evaluate quantitatively the intraobserver reproducibility of measurements of the retinal nerve fiber layer (RNFL) in healthy subjects and an ocular hypertensive population using two nerve fiber analyzers. METHODS: Sixty eyes of normal (n=30) and ocular hypertensive subjects (n=30) were consecutively recruited for this study and underwent a complete ophthalmologic examination and achromatic automated perimetry. RNFL were measured using scanning laser polarimeter (GDx-VCC) and optical coherence tomography (OCT Model 3000). Reproducibility of the RNFL measurements obtained with both nerve fiber analyzers were compared using the coefficient of variation. RESULTS: In both groups the authors found fair correlations between the two methods in all ratio and thickness parameters. The mean coefficient of variation for measurement of the variables ranged from 2.24% to 13.12% for GDx-VCC, and from 5.01% to 9.24% for OCT Model 3000. The authors could not detect any significant differences between healthy and ocular hypertensive eyes, although in normal eyes the correlations improved slightly. Nevertheless, the test-retest correlation was slightly better for GDx-VCC than for OCT Model 3000 (5.55% and 7.11%, respectively). CONCLUSIONS: Retinal mapping software of both nerve fiber analyzers allows reproducible measurement of RNFL in both healthy subjects and ocular hypertensive eyes, and shows fair correlations and good intraobserver reproducibility. However, in our study, GDx showed a better test-retest correlation.

NGF and TGF-beta mRNA expression during pregnancy in a rat corneal wound healing model.

BACKGROUND: Growth factors seem to play a major role in corneal wound healing and TGF-beta seems to be associated with abnormal healing after corneal surgical procedures. Few studies have analysed the role of NGF and TGF-beta on corneal wound healing during pregnancy. The aim of the present study was to create an animal model to evaluate the expression of NGF and TGF-betas during corneal wound healing in two groups: control and pregnant rats. METHODS: Corneal mRNA for NGF and the three isoforms of TGF-beta were analysed by RT-PCR, in a time-course experiment on different days after epithelial wounding (2, 7, 14 days) in pregnant and control groups RESULTS: The results show high corneal mRNA expression for NGF and TGF-beta1 without any variation throughout the healing process or pregnancy evolution. However, we detected a different expression of corneal mRNAs for TGF-beta2 and TGF-beta3 in the control group. This data was not detected in the pregnant group. DISCUSSION: Our results suggested that pregnancy could have a relevant role on TGF-beta2 and TGF-beta3 mRNA expression during the corneal wound healing process. Additional research should be performed to corroborate these findings.

Differences in corneal anatomy in a pair of monozygotic twins due to continuous contact lens wear.

PURPOSE: To study the differences in the corneal anatomy in a pair of 31-year-old monozygotic female twins, one of whom has been a contact lens wearer for the past 15 years and the other is a nonwearer. METHODS: We analyzed the corneal thickness and the corneal endothelial cell density of the monozygotic twins and of two age-matched control groups, one made up of non-contact lens wearers (35 eyes of 35 patients; mean age, 29.32 +/- 2.83 years) and the other made up of soft contact lens wearers (30 eyes of 30 patients; mean age, 33.08 +/- 3.72 years; mean duration of contact lens use, 10.62 +/- 4.32 years). Five corneal thickness measurements were carried out with the Orbscan Topography System II. Three consecutive measurements of the corneal endothelial cell density were carried out with the Topcon SP-2000P noncontact specular microscope. RESULTS: Lower central corneal endothelial cell densities were found in both eyes of the monozygotic contact lens-wearing twin. The greater corneal thickness was shown at superonasal cornea except in the right cornea of the monozygotic contact lens twin. Moreover, significant differences in corneal thickness values were found between the monozygotic twins. CONCLUSIONS: The differences between the monozygotic twins seem to confirm that daily use of soft contact lenses can modify normal corneal anatomy.

Influence of light/dark, seasonal and lunar cycles on the nuclear size of the pinealocytes of the rat.

Morphological and physiological studies suggest a possible division of the pineal parenchyma into an external or "cortical" and another central or "medullar" layer. We have studied the possible influence of the light/dark, seasonal and lunar cycles on the nuclear size of the pinealocytes of the rat in both the hypothetical "cortical" and "medullar" layers. Forty male Wistar rats were used. Experiment was carried out in two seasons, winter and spring, two lunar phases, full moon and new moon, and the two circadian phases, photophase and scotophase. The nuclear volume of the pinealocytes, calculated from the Jacobj's formula, was the karyometric parameter used as measurement of the nuclear size. Main results showed that nuclear volume of the cortical pinealocytes was greater than that of the medullar pinealocytes only during the photophases of winter new-moon days and spring full moon days, whereas in all the remaining situations, the greater nuclear sizes were found in the pinealocytes of the medullar layer. These results support the existence of independent morphological variations of the pinealocyte in the central and peripheral zones of the pineal gland.

Influence of light/dark, seasonal and lunar cycles on serum melatonin levels and synaptic bodies number of the pineal gland of the rat.

Synaptic bodies (SB) are ultrastructural organelles observed in the pinealocytes of mammals. According to its shape, they have been classified into synaptic ribbons (SR), synaptic spherules (SS), and intermediate synaptic bodies (ISB). They have been related to the melatonin regulation and production mechanisms of the pineal gland. Circadian and circannual fluctuations of both melatonin and SB have been reported. The possibility that other external factors, apart from light-dark or seasonal cycles, might influence pineal function has been suggested. We studied the evolution of the number of SB and serum melatonin levels not only during light-dark and seasonal phases but also during lunar cycles. Forty male wistar rats were used. Experiment was first carried out in winter and repeated identically in spring. Each season, one group of animals was killed during the new-moon days and a second group during the full-moon days: half of both groups in the photophase and the other half in the scotophase. The number of SB was measured at electron microscopic level whereas serum melatonin levels were determined by radioimmunoassay techniques. Main results showed that SR number and serum melatonin levels were higher during scotophases, winter and full-moon days. The SS only showed a light predominance during winter, whereas predominance of the ISB was found only during the scotophases. These results support the influence of the photophasic factors on the SR and ISB variations. In the case of the SS the influence of the lunar cycles is always dependent on the other factors. Finally, the serum level of melatonin is clearly influenced by the photophasic rhythms and the seasonal periods but not by the lunar cycles.

The medial sural artery perforator free flap.

The medial sural artery supplies the medial gastrocnemius muscle and sends perforating branches to the skin. The possible use of these musculocutaneous perforators as the source of a perforator-based free flap was investigated in cadavers. Ten legs were dissected, and the topography of significant perforating musculocutaneous vessels on both the medial and the lateral gastrocnemius muscles was recorded. A mean of 2.2 perforators (range, 1 to 4) was noted over the medial gastrocnemius muscle, whereas in only 20 percent of the specimens was a perforator of moderate size noted over the lateral gastrocnemius muscle. The perforating vessels from the medial sural artery clustered about 9 to 18 cm from the popliteal crease. When two perforators were present (the most frequent case), the perforators were located at a mean of 11.8 cm (range, 8.5 to 15 cm) and 17 cm (range, 15 to 19 cm) from the popliteal crease. A series of six successful clinical cases is reported, including five free flaps and one pedicled flap for ipsilateral lower-leg and foot reconstruction. The dissection is somewhat tedious, but the vascular pedicle can be considerably long and of suitable caliber. Donor-site morbidity was minimal because the muscle was not included in the flap. Although the present series is short, it seems that the medial sural artery perforator flap can be a useful flap for free and pedicled transfer in lower-limb reconstruction.

Inner ear lesion alters acoustically induced c-Fos expression in the rat auditory rhomboencephalic brainstem.

The pattern of c-Fos expression was mapped in the adult rat's brain following unilateral cochlear lesions. In normal and cochlear lesioned rats, c-Fos expression was induced with sound stimuli. Acoustic stimulation consisted of pulses of four tones. An additional control group consisted of non-stimulated rats. In the cochlear nuclei (CN), c-Fos activation was scarce in isolated rats and increased strongly following sound stimulation. Following unilateral cochlear lesion, acoustically driven expression was decreased in all CN in both the lesioned and the untreated sides. The ventromedial periolivary nucleus and the rostral periolivary nucleus showed c-Fos activation in isolated conditions and were strongly activated following sound stimulation. The rest of the superior olivary complex showed no c-Fos activation in isolated rats and a weak activation following sound stimulation. Following unilateral cochlear lesions, acoustically driven expression was decreased in some, but not all superior olivary nuclei in both the lesioned and the untreated sides. In the lateral lemniscus complex, c-Fos activation was scarce in isolated rats and increased strongly after stimulation. Following unilateral cochlear lesion, acoustically driven expression decreased bilaterally in all nuclei. We have found that unilateral inner ear lesions lead to bilateral impairment of the capability of acoustic pathway neurons, to being c-Fos-activated following sound stimulation.

The chemical architecture of the rat's periaqueductal gray based on acetylcholinesterase histochemistry: a quantitative and qualitative study.

The chemoarchitecture of the periaqueductal gray has been extensively studied, based on acetylcholinesterase reaction and comparing it to other chemical markers. We have divided the periaqueductal gray into four main longitudinal columns, namely dorsomedial, dorsolateral, lateral and ventrolateral. We also identified the dorsal midline column, the supraoculomotor cap and the juxta-aqueductal ring. The acetylcholinesterase gave rise to a strong reaction in the outer half of the lateral column, the outer half of the dorsomedial column, the supraoculomotor cap and the ventral half of the juxta-aqueductal ring. This labeling was in part complementary to that of the NADPH diaphorase and allowed the lateral column to be differentiated from the ventrolateral column. However, the inner half of both lateral and ventrolateral columns displayed the same chemical properties including acetylcholinesterase, tyrosine hydroxilase and serotonin. Thus, from the chemical view, these inner halves should be considered as one different region. Finally, the juxta-aqueductal ring was composed of two clearly different halves, i.e. dorsal and ventral. The dorsal half did not show any clear differences from the above columns and was negative for acetylcholinesterase, NADPH diaphorase and tyrosine hydroxilase, while the ventral half was clearly different from the lateral and ventrolateral columns and displayed a positive reaction to all those chemical markers. From these results, we strongly suggest the use of acetylcholinesterase histochemistry as a tool for accurate parcellation of the periaqueductal gray.

Circadian and seasonal changes of synaptic bodies in different parts of the rabbit pineal gland.

In the mammalian pineal gland, synaptic bodies (SBs) are poorly understood organelles. Previous studies in rabbits have shown that the organelles are rather heterogeneous in shape, are few in number during the day and increase in number at night. No studies are currently available on seasonal changes in this species and it is unknown whether the biological rhythms are identical in the proximal, intermediate and distal parts of the elongated pineal. To this end, a study was made of 84 rabbits kept under natural lighting conditions to examine numerical variations of the different types of SBs in the proximal, intermediate and distal regions of pineal glands procured at different timepoints of a 24-hour cycle and in each of the four annual seasons. In the present study, rod-like, sphere-like, ovoid, rectangular and triangular SB profiles were distinguished; the first two types being the most abundant. In addition to the well-known circadian changes, with low numbers of SB profiles during the day and high numbers at night, we found pronounced season-related differences as well as differences related to pineal regions. In autumn and winter, nighttime SR profile numbers were significantly higher than in spring and summer. With respect to regional differences it was found that the amplitude of the circadian rhythm increased in a proximo-distal direction in the gland. In autumn the strongly enhanced nocturnal increase was restricted to the distal region of the gland, whereas in winter it was seen in both the distal and the intermediate regions. The regional differences are probably related to the fact that the postganglionic sympathetic fibres, which regulate pineal function, enter the gland distally and proceed rostrally to the proximal region. Taken together, the results show that day- and nightlength are structurally coded in the pineal gland by means of SB numbers. Provided the SBs of the mammalian pineal gland are involved in synaptic processes, the results suggest that synaptic processes are enhanced at night as well as in autumn and winter.

Chronic ethanol feeding induces cellular antioxidants decrease and oxidative stress in rat peripheral nerves. Effect of S-adenosyl-L-methionine and N-acetyl-L-cysteine.

Chronic ethanol feeding promotes oxidative stress in rat peripheral nerve. Malondialdehyde, a lipid peroxidation product, content increases in sciatic nerves of rats fed an ethanol-containing diet, when compared with pair-fed animals. Moreover, glutathione content and glutathione peroxidase activity in this same tissue decrease in ethanol-fed vs. pair-fed rats. S-Adenosyl-L-methionine and N-acetyl-L-cysteine, both with possible therapeutic action on alcoholism, were tested in this animal model. Only N-acetyl-L-cysteine was able to normalize malondialdehyde content and to restore glutathione content and glutathione peroxidase activity, to values not significantly different from those of sciatic nerves from pair-fed animals. The reasons for the different effect of both substances tested is also discussed.