Royal jelly increased map-2 expression in hippocampal neurons of hypothyroid rats: an immunohistochemical study.

We investigated microtubule-associated protein 2 (MAP-2) immunoreactivity of hippocampal neurons and the potential role of royal jelly (RJ) in regulating MAP-2 during experimental hypothyroidism (HT). Thirty adult female Wistar albino rats were randomized into five groups: the control group was untreated, the sham control group was treated with 10 mg/kg 0.9% sterile saline injected intraperitoneally (i.p.), The RJ group was treated with 100 mg/kg RJ by oral gavage, the HT group was treated with 10 mg/kg propylthiouracil (PTU) i.p. to induce experimental hypothyroidism, and the HT + RJ group was treated with 10 mg/kg PTU i.p. + 100 mg/kg RJ by oral gavage. Oral and i.p. administrations were performed once daily for 20 days. Thyroid stimulating hormone (TSH) and free thyroxine (fT4) levels in the serum were measured biochemically, MAP-2 was measured immunohistochemically in the hippocampus and an immunohistochemical H score was calculated. MAP-2 immunoreactivity appeared in the cytoplasm of neuron cell bodies and dendrites in the hippocampal CA3 and CA1 regions in the control, sham control and RJ groups. MAP-2 immunoreactivity decreased significiantly in the HT group compared to control, sham control and RJ groups. Also, vascular dilation and swollen cells were observed following PTU administration. The degeneration that was observed in the HT group decreased by RJ administration. By contrast, MAP-2 immunoreactivity increased following administration of RJ. Experimental hypothyroidism reduced significiantly MAP-2 immunoreactivity in both the CA3 and CA1 regions and caused degeneration, including edema and vascular dilation, in the hippocampus. RJ increased MAP-2 expression and exhibited a therapeutic effect on the degenerative changes.

Treatment of an Esophageal Stricture in a 1-Month-Old Miniature Shetland Colt.

A 1-month-old Miniature Shetland colt was presented to the Burg Müggenhausen Equine Clinic. Primary complaints were regurgitation of milk, bilateral white nasal discharge, and weakness. Physical examination, endoscopy of the esophagus, and contrast radiography led to the diagnosis of an esophageal stricture and aspiration pneumonia. Surgical treatment by means of an esophagomyotomy was performed. The foal improved only temporarily and multiple sessions of endoscopic balloon dilation of the esophagus were performed afterwards. 12 months after the final treatment the foal was healthy and had no complaints regarding esophageal function.

The histopathological effects of levosimendan on liver injury induced by myocardial ischemia and reperfusion.

BACKGROUND: The aim of this study was to evaluate the histological and immunohistochemical effects of levosimendan on liver injury induced by myocardial ischemia and reperfusion (I/R) in a rat model. METHODS: Twenty-four male Wistar Albino rats were randomly divided into the four groups: Group C (Control, n = 6), Group I/R (n = 6), Group BI (I/R group treated with levosimendan before ischemia, n = 6), and Group AI (I/R group treated with levosimendan after ischemia, n = 6). Myocardial I/R was induced by ligation of the left anterior descending coronary artery for 30 min followed by two hours of reperfusion in I/R and I/R+Levosimendan groups. At the end of the study, liver tissue samples were obtained for histopathological and immunohistochemical examination. RESULTS: Masson Trichrome staining revealed significant hepatocyte degeneration and necrosis most marked in portal acinus Zone 3, especially around the central veins in Group I/R. Histopathological changes in Group AI were more similar to the changes in Group I/R. Milder hepatocellular degeneration was found in Group BI, when compared to groups I/R and AI. Immunohistochemical score was found to be significantly higher in Group I/R compared to groups C, BI and AI (p < 0.0001). The scores in groups BI and AI were found to be similar (p = 0.068). CONCLUSION: Levosimendan ameliorates liver injury induced by myocardial IR, especially when administered before induction of ischemia (Fig. 9, Ref. 37).

The effects of iloprost on lung injury induced by skeletal muscle ischemia-reperfusion.

PURPOSE: The aim of this study was to investigate the effects of iloprost (I) on lung injury as a remote organ following skeletal muscle ischemia-reperfusion injury in a rat model. MATERIALS AND METHODS: Twenty-four Wistar Albino rats were randomized into four groups (n = 6). Laparotomy was performed in all groups under general anesthesia. Only laparotomy was applied in Group S (Sham). Ischemia reperfusion group (Group I/R) underwent ischemia and reperfusion performed by clamping and declamping of the infrarenal abdominal aorta for 120 minutes. Group iloprost (Group I) received intravenous infusion of iloprost 0.5 ng/kg/min, without ischemia and reperfusion. Group I/R/I received intravenous infusion of iloprost 0.5 ng/kg/min immediately after 2 hours of ischemia. At the end of the study, lung tissue was obtained for determining total oxidant status (TOS) and total antioxidant status (TAS) levels, histochemical and immunohistochemical determination. RESULTS: Diffuse lymphocyte infiltration was detected in immunohistochemical examination of lung tissue in Group I/R. The connective tissue around bronchi, bronchioles and vessel walls was found to be increased. Although minimal local lymphocyte infiltration was detected in some fields in Group I/R/I, the overall tissue was found to be similar to Group S. iNOS expression was significantly higher in Group I/R, when compared with Group S and significantly lower in Group I/R/I compared to Group I/R.TOS levels were significantly higher in Group I/R, when compared with groups S and I (p = 0.028, p = 0.016, respectively) and significantly lower in group I/R/I, when compared with Group I/R (p = 0.048). TAS levels were significantly higher in Group I/R, when compared with groups S, I (p = 0.014, p = 0.027, respectively) and significantly lower in Group I/R/I, when compared with Group I/R (p = 0.032). CONCLUSION: These results indicate that administration of iloprost may have protective effects against ischemia reperfusion injury (Fig. 8, Tab. 1, Ref. 30)

The protective effects of dexmedetomidine on liver injury-induced myocardial ischemia reperfusion.

PURPOSE: The aim of this study was to evaluate the effect of dexmedetomidine (100 µg/kg-ip) on liver injury-induced myocardial ischemia and reperfusion (IR) in rats. MATERIALS AND METHODS: Twenty-four Wistar Albino rats were separated into four groups. There were four experimental groups (Group C (Control; n = 6), Group IR (ischemia-reperfusion, n = 6), Group D (Dexmedetomidine; n = 6) that underwent left thoracotomy and received ip dexmedetomidine without IR administered via 100 µg/kg ip route 30 minutes before ligating the left coronary artery, and Group IR-D (IR-Dexmedetomidine; n = 6). A small plastic snare was threaded through the ligature and placed in contact with the heart. To produce IR, a branch of the left coronary artery was occluded for 30 min followed by two hours of reperfusion. However, after the above procedure, the coronary artery was not occluded or reperfused in the control rats. At the end of the study, liver tissue was obtained for histochemical and immunohistochemical determination.Some part of tissue samples were stained with Masson-trichrome for the evaluation of ultrastructural changes and inducible nitric oxide synthase (iNOS) expression was evaluated in other part of samples for immunohistochemical examination. RESULTS: Histopathological changes were detected in Group IR when compared with Group C. iNOS expression was found to be increased and stronger particularly in the vascular wall, perisinusoidal space and hepatocytes around vena centralis in this group compared to the control group. Perivascular oedema was detected to be decreased in Group IR-D compared to Group IR. It was also observed that the impairment in the radial arrangement of hepatocytes significantly recovered in Group IR-D. The immunoreactivity was found to be significantly decreased in the assessment of iNOS expression in the same group when compared with Group IR. CONCLUSION: Administration of dexmedetomidine ameliorates liver injury induced by myocardial ischemia and reperfusion (Fig. 8, Ref. 33).

Immunohistochemical and ultrastructural changes in rat fat tissue related to the local hCG injection.

OBJECTIVES: Recently, it has been observed that weight loss is accelerated by human chorionic gonadotropin (hCG) hormone preparation used for hypothalamic dysfunction in obesity treatment in both sexes. hCG is also used for in vitro fertilization and in treatment of hypogonadotropic hypogonadism. Our aim was to observe the ultrastructural changes caused by local injections of hCG made for purpose of weight loss and to present them to inform those receiving such therapy. MATERIALS AND METHODS: In our study, 10 obese female, 10 male obese, 10 non-obese female and 10 non-obese male rats were used. In each group, single dose of subcutaneous hCG injection has been applied to 7 rats for 5 weeks in 5 days of the week, and placebo has been applied to the remaining 3 rats. Following the injection, the tissues were evaluated morphologically, immunohistochemically and ultrastructurally. RESULTS: Leptin immunoreactivity was similar in all groups. When the adipose tissue samples were examined under electron microscope, they were observed to exhibit normal structure with organelles located around the nuclei and nucleoli, and no distinctive features were found among the groups. CONCLUSIONS: Administering hCG in addition to diet had no advantage on weight reduction in rats.

Dose-dependent immunohistochemical and ultrastructural changes after oral methylphenidate administration in rat heart tissue.

Methylphenidate, more commonly known as Ritalin, is a piperidine derivative and is the drug most often used to treat attention deficit/hyperactivity disorder, one of the most common behavioural disorders of children and young adults. Our aims were to investigate dose-dependent immunohistochemical D2 expression and ultrastructural changes of the rat heart tissue, and to demonstrate possible toxicity of the long-term and high dose use of the methylphenidate. In this study, 27 female pre-pubertal Wistar albino rats, divided into three different dose groups (5, 10 and 20 mg/kg) and their control groups, were used. They were treated orally with methylphenidate dissolved in saline solution for 5 days/week during 3 months. At the end of the third month, after perfusion fixation, left ventricle of cardiac tissue was removed. Paraffin, semi-thin and thin sections were collected and immunohistochemical, terminal deoxynucleotidyl transferase-mediated Dig-dUTP nick end labelling assay and ultrastructural studies were performed. In conclusion, we believe that Ritalin is dose-related affecting dopaminergic system to increase heart rhythm and contraction. Thus, this drug may cause degenerative ultrastructural changes in mitochondrial path.

Age-related immunohistochemical and ultrastructural changes in rat oculomotor nerve.

During ageing process, multiple changes occur on nervous tissue composed of cells and extracellular matrix. Changes on nervous tissues are usually known as degenerative changes on axon structure and connective tissue covering the nerve such as a decrease in the number of fibre or general structural changes. For this purpose, we have studied age-dependent ultrastructural changes in the rat oculomotor nerve with electron microscopy and also demonstrated collagen structure of the neural sheaths with immunohistochemical techniques. This study was conducted in Gazi University Faculty of Medicine, Department of Anatomy with a total of nine Wistar albino rats. We observed strong collagen type I immunoreactivity in endoneurium and slight to moderate reactivity in fibroblast cytoplasm in 3-month- and 12-month-old groups and mild reactivity in 24-month-old group. Collagen type IV immunoreactivity was stronger in endoneurium and perineurium in the 3-month- and 12-month-old groups compared with collagen type I and fibroblast cytoplasm showed a very strong reactivity. On the other hand, in the 24-month-old group, there was slight reactivity in endoneurium and a strong reactivity in perineurium. NGF staining showed moderate to strong reactivity on Schwann cells of the 3-month-old group. The immunoreactivity decreased in the 12-month- and 24-month-old groups. In the 3-month-old rat group, Schwann cell cytoplasm, mitochondrial structure and neurofilaments were normal. In the 12-month-old group, there were no changes in organelle distribution, mitochondrial structure and neurofilaments, but there was an increase in the connective tissue. An inconsiderable number of degenerated myelinated nerves were observed. We detected an important decrease in the collagen type I immunoreactivity, which could suggest that the endoneurium, perineurium and epineurium are less resistant to the age-related collagen loss and that the peripheral nerve is protected by a weaker barrier in the old group. The collagen type IV immunoreactivity was significantly decreased with age. NGF synthesis decreases with age because of Schwann cell structural degeneration or for different reasons. Thus, this could explain the diminished capacity of regeneration and damage of the myelination of the peripheral nerve.

The effects of dexfenfluramine administration on brain serotonin immunoreactivity and lipid peroxidation in mice.

Obesity continues to be an increasing health problem in worldwide and antiobesity drugs have commonly been used by obese patients. During the use of anorectic drugs, the antioxidant defense may be affected, especially by reactive oxygen species. It was decided to investigate the effects of dexfenfluramine on body weight, daily food intake, brain thiobarbituric acid-reactive substances (TBARS), glutathione (GSH) and nitric oxide (NO) levels, and 5-HT immunoreactivity. Mice were divided into two groups each containing 8 Swiss Albino adult (6 months) mice. Group 1, untreated, was used as a control; group 2 was treated with dexfenfluramine 0.4 mg/kg per day intraperitoneally for 7 days. Brain TBARS and GSH levels were assayed spectrophotometrically. The stable end-products of NO, nitrite and nitrate, were analyzed spectrophotometrically. Brain tissue 5-HT immunoreactivity was observed using an immunohistochemical method. There were significant decreases in body weight in the dexfenfluramine group (p < 0.05). Although brain GSH and NO(x) levels decreased significantly, brain TBARS levels increased in the dexfenfluramine group (p < 0.05). Brain 5-HT immunoreactivity also increased in the dexfenfluramine-treated group compared to control. In conclusion, our findings show that dexfenfluramine is effective in achieving weight loss and also increases lipid peroxidation in mouse brain.

Visualization of sperm by immunohistochemistry: application of alternative counterstains.

The aim of this study was to evaluate the application of hematoxylin, safranin, light green and picric acid as counterstain in sperm immunohistochemistry. This is important to visualize the best staining procedure and to determine the advantage of picric acid as a counterstain in some situations. Picric acid used for counterstaining in the immunohistochemical procedure gives the best image of reaction on sperms when DAB was used as a chromogen.

Effects of dopexamine on rat cardiorenal functions during lipopolysaccharide-induced experimental sepsis.

We aimed to test the protective effect of dopexamine on renal function and systemic haemodynamics in rats with induced sepsis. Female Sprague-Dawley rats were randomized into three equal groups: group 1 (control, received 3% creatinine throughout the experiment); group 2 (sepsis, received 3% creatinine and Escherichia coli lipopolysaccharide [LPS] endotoxin [8 mg/kg per h]); and group 3 (sepsis plus dopexamine, received 3% creatinine, E. coli LPS and dopexamine [1 microgram/kg per min]). Time-adjusted heart rate, systolic, diastolic and mean arterial pressures, urine volume and glomerular filtration rate (from creatinine clearance) were recorded. After bacterial infusion heart rate increased and mean arterial pressure decreased; the fall in mean arterial pressure was less pronounced with dopexamine (group 3) than without (group 2). Dopexamine also induced significant and moderate increases in urine volume and heart rate, respectively. We concluded that dopexamine has some positive inotropic-chronotropic effects and induces favourable responses in renal function.

Atrophic and regenerative changes in rabbit mimic muscles after lidocaine and bupivacaine application.

Destruction and denervation atrophy in skeletal muscles caused by the injection of local anaesthetics was investigated by injecting lidocaine or bupivacaine around the rabbit facial nerve to produce facial paralysis. Animals were then sacrificed at 2, 4, 6, and 8 weeks post-injection, and changes in mimic muscle tissue were assessed at each stage by light microscopy and electron microscopy. Atrophic changes were observed at 2-6 weeks after injection, and regeneration started at 6-8 weeks. Compared to bupivacaine, lidocaine caused more dramatic atrophic changes and was associated with slower muscle regeneration.

Expression of growth factors in fetal human olfactory mucosa during development.

The olfactory epithelium is one of the unique sites in the mammalian nervous system at which there is continual neurogenesis. Constant turnover of primary sensory neurons in the periphery results in remodeling of neuronal circuits and synapses in the olfactory bulb throughout life. Most of the specific mechanisms and factors that control this process are still unknown. Recent studies suggest that growth factors and their receptors may play a significant role in the development and continuous regeneration of olfactory neurons. In this study, standard immunohistochemical methods were used to detect the presence of epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta in the human olfactory epithelium of different developmental stages (7 to 30 weeks) in formalin-fixed tissue specimens. Immunoreactivity to epidermal growth factor-receptor was seen in basal cells, supporting cells, neurons, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm. Similarly transforming growth factor-alpha immunoreactivity was recognized in basal cells, supporting cells, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm but not in neurons. Nerve growth factor-beta detection was restricted on olfactory nerve cells, dentritic knob and basal cell layer. So, this study confirms the development of human olfactory mucosa and shows epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta proteins are reliable markers for developing olfactory epithelium.

Long-term degeneration and regeneration of the rabbit facial nerve blocked with conventional lidocaine and bupivacaine solutions.

The alterations due to the long-term degeneration and regeneration of the rabbit facial nerve were investigated. The facial nerve was blocked with conventional lidocaine and bupivacaine solutions. The rabbits were sacrificed and facial nerves were removed 2, 4, 6 and 8 weeks after the administration of normal saline, lidocaine and bupivacaine. The samples were then processed for electron microscopic observation. Degenerations were seen 2-4 weeks after the injection of normal saline, lidocaine and bupivacaine. In the normal saline group the regeneration process was observed after 6 weeks, whereas in the lidocaine and bupivacaine groups this process was first noticed after 8 weeks. In the lidocaine and bupivacaine groups, regeneration was not completed at the end of 8 weeks; however, it was faster in the bupivacaine group. Bupivacaine is therefore suggested as the drug of choice for neural blockade as it caused less damage and a faster regeneration.

Branching patterns of rabbit oculomotor and trochlear nerves demonstrated by Sihler's stain technique.

A modified Sihler's stain technique was used to visualize the branching patterns of oculomotor and trochlear nerves. The levator palpebrae, superior rectus, inferior rectus, medial rectus, inferior oblique, superior oblique and tensor trochlea muscles were isolated from the eyes of normal rabbits and processed using modified Sihler's technique. The distributions and terminal ramifications of the oculomotor and trochlear nerves were observed. Two distinct divisions and terminal branches of the oculomotor nerve were shown in detail together with the trochlear nerve distribution. The application of Sihler's technique enables researchers to trace nerve branching within relatively transparent muscles, whereas the nerve fibers are counterstained and clearly visible. This technique could be useful for detailed studies of the motor control of extraocular muscles.

Combined inguinal paravascular (3-in-1) and sciatic nerve blocks for lower limb surgery.

BACKGROUND AND OBJECTIVES: The effects of increasing the maximum recommended dose of lidocaine 1% plus epinephrine from 500-650 mg in combined 3-in-1/sciatic nerve block were investigated in 45 adult patients scheduled for surgery on the lower limb. The goal of the study was to show primarily, whether pneumatic tourniquet pain could be reduced, whether toxic plasma levels of local anesthetic were reached, and whether any other clinical side effects occurred. RESULTS: The results indicated that the incidence of tourniquet pain was significantly reduced and, most important, that there were no noteworthy differences in venous lidocaine plasma levels between the two groups investigated. CONCLUSIONS: There was a significant decrease in time required for complete neural blockade after administration of the 650 mg lidocaine dose, and little or no difference in the duration of postoperative analgesia.

[Combined sciatic 3-in-1 block. Application in lower limb orthopedic surgery]. / Kombinierter Ischiadicus 3-in-1-Block. Anwendung bei orthopädischen Operationen im Unterschenkelbereich.

The effects of increasing the maximum prescribed volume of 50 ml lidocaine 1% to 65 ml in a combined sciatic 3-in-1 block were investigated in 25 adult patients. The goal of the study was (1) to show possible increase in the success rate and (2) to determine if toxic plasma levels of local anaesthetic would be reached. Further more, we wanted to find out if there were any side-effects. The patients were divided into three groups according to body weight: group I, 50-69 kg (n = 7); group II, 70-80 kg (n = 9); group III, over 80 kg (n = 9). Besides clinical observation, the plasma levels of local-anaesthetic were determined over a 90-min period. RESULTS. Following the neural blockade, the obturator nerve was the first to be rendered insensitive after 4.8 +/- 3.6 min, followed by the saphenous (7.2 +/- 4.4 min) and femoral nerves, and finally by the lateral femoral cutaneous nerve after 12.5 +/- 5.3 min. Around the knees, the motor blockade was complete after 11.9 + 4.3 min and around the hips 11.2 +/- 4.6 min. Intraoperatively, one of these 25 patients needed an additional general anaesthetic, as he did not tolerate the Esmarch cuff. The postoperative analgesia in 24 patients lasted 268 +/- 55 min; the patient who had a general anaesthetic in addition had a postoperative analgesia of 315 min. Group I showed levels of mean 2.04 +/- 0.64 microns/ml, group II 1.84 +/- 0.38 microns/ml and group III 1.69 +/- 0.63 microns/ml. There were no statistical differences between the three groups. CONCLUSION. Increasing the dosage of lidocaine from 500 to 650 mg makes the block very successful without any toxic side-effects.

[Endotracheal intubation in thoracic catheter peridural anesthesia]. / Zur Frage der endotrachealen Intubation bei thorakaler Katheter-Periduralanaesthesie.

After standard preparation, the catheter is preferably inserted in a sitting position. A test dose of 4 ml etidocaine 1% is injected, initially followed by 1 ml per segment of the total calculated analgesic dose. Analgesic spread, blood pressure, and pulse are checked every 2 min. If a sufficient effect is not reached within 20 min, a further 2 ml per segment is injected. The patient is sedated in the operating room with diazepam, droperidol, or chlorpromazine and receives 6-8 l O2 during spontaneous respiration via a Hudson mask. The epidural catheter is left in place for postoperative analgesia. Intubation is used only as required. Of a total of 52 patients, 31 (60%) did not require intubation, 24 (46%) were age 70 or older, and 31 (60%) were classified as ASA risk group III-IV. Thirty-three of all the procedures (63%) involved upper abdominal surgery (liver, gallbladder) and the majority of these received thoracic (Th 9-10) punctures, with the catheter inserted 5 cm in the epidural space; 88% were punctured medially and 12% laterally. The most frequent complications were bradycardia and hypotension. Respiratory arrest and aspiration were not seen. Thoracic epidural catheter anesthesia can be carried out with or without intubation and with a minimum of complications in selected procedures. Postoperative analgesia is an important advantage.