Structural characteristics of septal cartilage and mucoperichondrium.

AIM: During nasal septum surgery, elevation of mucoperichondrium from the anterior nasal septum may be more difficult than from the medial and posterior septum. This study aimed to evaluate any histological structural differences between the anterior and posterior nasal septum cartilage, mucoperichondrium and intervening tissue. MATERIAL AND METHOD: Unilateral mucoperichondrial flap elevation without infiltration was performed, after nasal tip and dorsum decortication, in four patients undergoing open septorhinoplasty. Full-thickness samples, including cartilage and mucoperichondrium, were removed from the anterior and posterior nasal septum and examined under light and electron microscopy. RESULTS: Light microscopy showed no difference between anterior and posterior septum specimens regarding perichondrial thickness and subperichondrial cell density. Demarcation between cartilage and perichondrium and between perichondrium and lamina propria was more regular in the posterior versus the anterior septum. Electron microscopy showed no difference in chondroblast activity at the two sites. CONCLUSION: The observed tissue demarcation irregularities may explain the greater reported difficulty in elevating anterior versus posterior nasal septum mucoperichondrium. Immunohistochemical examination would further elucidate these interstructural connections.

Êxpression of inducible nitric oxide synthase (iNOS) in the azoospermic human testis.

Azoospermia, which is the absence of spermatozoa in the ejaculate, is not a rare cause of male infertility. Inducible nitric oxide synthase (iNOS) is a calcium-independent NOS, which is present in the testis and involved in spermatogenesis, and apoptosis of Sertoli and germ cells. Twenty idiopathic infertile men presenting nonobstructive azoospermia were enrolled in this study, and testicular sperm extraction procedures were performed. Tissue extracts were dissected, and the fluid samples were investigated to determine the presence of spermatozoa. Histologic evaluation of the spermatozoa-present samples revealed that seminiferous tubules were normal and were lined by Sertoli cells and spermatogenic cells. However, in the spermatozoa-absent samples, the diameter of the seminiferous tubules was small, and Sertoli-cell-only syndrome was determined in most of the tubules. iNOS expression was very weak in Sertoli cells, germ cells, and in Leydig cells in the spermatozoa-present group. In the spermatozoa-absent group, the immunostaining was very intense in Sertoli and Leydig cells. Electron microscopy findings were supported the histologic results. In conclusion, complete germ cell loss and intense expression of iNOS in the Sertoli and Leydig cells in the spermatozoa-absent groups of azoospermic human testis suggest an essential role of iNOS in spermatogenesis.

Oculopharyngodistal myopathy is a distinct entity: clinical and genetic features of 47 patients.

BACKGROUND: Oculopharyngodistal myopathy (OPDM) has been reported as a rare, adult-onset hereditary muscle disease with putative autosomal dominant and autosomal recessive inheritance. Patients with OPDM present with progressive ocular, pharyngeal, and distal limb muscle involvement. The genetic defect causing OPDM has not been elucidated. METHODS: Clinical and genetic findings of 47 patients from 9 unrelated Turkish families diagnosed with OPDM at the Department of Neurology, Istanbul Faculty of Medicine, between 1982 and 2009 were evaluated. RESULTS: The mean age at onset was around 22 years. Both autosomal dominant and autosomal recessive traits were observed, without any clear difference in clinical phenotype or severity. The most common initial symptom was ptosis, followed by oropharyngeal symptoms and distal weakness, which started after the fifth disease year. Intrafamilial variability of disease phenotype and severity was notable in the largest autosomal dominant family. Atypical presentations, such as absence of limb weakness in long-term follow-up in 9, proximal predominant weakness in 4, and asymmetric ptosis in 3 patients, were observed. Swallowing difficulty was due to oropharyngeal dysphagia with myopathic origin. Serum creatine kinase levels were slightly increased and EMG revealed myopathic pattern with occasional myotonic discharges. Myopathologic findings included rimmed and autophagic vacuoles and chronic myopathic changes. Importantly, a considerable proportion of patients developed respiratory muscle weakness while still ambulant. Linkage to the genetic loci for all known muscular dystrophies, and for distal and myofibrillar myopathies, was excluded in the largest autosomal dominant and autosomal recessive OPDM families. CONCLUSIONS: We suggest that OPDM is a clinically and genetically distinct myopathy.

TGF-beta1 neutralizing antibodies decrease the fibrotic effects of ischemic priapism.

The objective of this study was to evaluate the possible role of transforming growth factor beta 1 (TGF-beta1) antibodies (ab) for the prevention of fibrotic effects of priapism in a rat model. In total, 30 adult Sprague-Dawley rats were divided into five groups. Priapism with 6 h (group 1), priapism with 6 h+ab (group 2), priapism with 24 h (group 3), priapism with 24 h+ab (group 4) and control (group 5). Priapism was induced with a vacuum erection device and a rubber band was placed at the base of the erect penis. At 1 h after the initiation of priapism, TGF-beta1 antibodies were given intracavernosaly. All rats underwent electrical stimulation of the cavernous nerve after 8 weeks. Intracavernous and systemic blood pressures were measured during the procedure. Rats in group 1 showed significantly higher (intracavernosal pressure (ICP) pressures to cavernous nerve stimulation and had higher ICP/BP ratios when compared to other groups. Similarly, histopathologic examination revealed less fibrosis in group 2, compared with the other groups. Consequently, TGF-beta1 antibodies antagonise the fibrotic effects of TGF-beta1, especially in cases with duration of priapism less than 6 h.

The relationship between neutrophils and incisional wound healing.

The systemic administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) is used clinically to increase circulating neutrophils, but its wound healing effects after intraperitoneal treatment have not been studied yet. We planned to investigate the effect of neutrophils on wound healing under cyclophosphamide and GM-CSF treatment. Forty rats were divided into three groups: control group (group I, n = 12) receiving saline, group II (n = 14) receiving cyclophosphamide and group III (n = 14) receiving GM- CSF. The rats in all groups underwent incisional wounding and were euthanized after 7 days. Blood neutrophil counts and functions, tensile strengths and the hydroxyproline level of skin were determined, and a histopathological evaluation of healing was made. Neutrophil counts and phagocytosis significantly increased in group III and decreased in group II. Although the skin hydroxyproline level did not differ, there was a difference in tensile strength of the wounded skin between group II and group III. The wound score in group II was lower than that in groups III and I. As a result we suggest that systemically given GM-CSF - by increasing the neutrophil count and neutrophil phagocytosis index - can enhance the tensile strength of surgical incisions.

The role of L-arginine and neutrophils on incisional wound healing.

Although arginine plays an important role in many aspects of inflammation and wound healing, the mechanism is not clear. We aimed to evaluate the effect of L-arginine administration on wound healing and neutrophil activity and on the interaction of these effects. Sixteen rats were divided into two groups: control group and L-arginine group. L-arginine was given intraperitoneally. The rats underwent incisional wounding and were killed on the 7th day of wounding. Blood neutrophil counts, neutrophil adhesivity index, tensile strengths and hydroxyproline level of skin were determined, histopathological and electron microscopical evaluation of healing was performed. Wound scores in the control group were significantly lower (p < 0.05). Hydroxyproline and collagen levels of skin were significantly increased in the L-arginine group (p < 0.05). Blood neutrophil counts and neutrophil adhesivity index in the L-arginine group were significantly increased (p < 0.05), as were the inflammatory cells in the skin. L-arginine may be used during the first phase of healing to induce inflammation in high risk patients.

Effects of early pre- and postoperative irradiation on the healing of microvascular anastomoses.

A comparative study was undertaken to evaluate the possible effects of time selection of irradiation on the healing of microvascular anastomoses. In this experimental study, 40 adult male Sprague-Dawley rats were divided into four groups, 10 animals in each. In Group 1, the rats were exposed to radiotherapy with a dose of 20 Gy in a single fraction at a 170,916 cby/min dose rate, at an 80-cm source to a skin distance from 3 to 4 cm in the right groin fields involving the femoral artery, using a Co60 megavoltage radiotherapy machine. The animals were operated on under general anesthesia 2 weeks later. During the operation, femoral microarterial anastomoses were performed. In Group 2, the rats were irradiated and operated on as in Group 1, on the same day. In Group 3, the rats were operated on and, 2 weeks later, irradiated, as in Group 1. In Group 4 (control group), the rats were operated on as in Group 1. No radiotherapy was performed. All rats were explored under general anesthesia after 1 month. Patency was evaluated, and the anastomotic sites were taken for histopathologic and electron microscopy studies. The authors found that there were no major differences in the healing of microvascular anastomoses in the early preoperative, immediate preoperative, and early postoperative irradiated groups. The patency rates of all radiotherapy groups were not statistically different from control arteries (p > 0.05).

Antiproliferative effects and insulin-like growth factor-I expression in Balb-C 3T3 fibroblasts after treatment with somatostatin and gonadotropin-releasing hormone analogs.

The present study was aimed to compare antiproliferative effects of somatostatin (SS) and gonadotropin-releasing hormone analogs (GnRHa) on a fibroblast cell line. Proliferation index, cell count, viability of the cells and insulin-like growth factor-I (IGF-I) immunoreactivity were determined after treatment with either SS (100 microM/ml), GnRHa (35 nM/ml) or SS and GnRHa of Balb-C 3T3 mouse fibroblasts. It was found that the proliferation index, cell count, viability and IGF-I immunoreactivity were not affected by GnRHa treatment as compared with no treatment (p > 0.05). Application of SS to the fibroblasts resulted in a significant reduction in proliferation index, cell count, and IGF-I immunoreactivity as compared with GnRHa treatment and no treatment, but it had no effect on cell viability. The labelling index in SS-treated cells was significantly reduced as compared with combined treatment with SS and GnRHa. In conclusion, a direct effect of GnRHa on fibroblast cells in culture could not be demonstrated. SS had direct inhibitory effects on cell proliferation possibly via inhibition of IGF-I effects without affecting cell viability.

The role of neutrophils in the formation of peritoneal adhesions.

The most common cause of intraperitoneal adhesions which may result in infertility and intestinal obstruction is previous abdominal surgery. Surgical trauma of the peritoneum in the absence of infection elicits a rapid and transient influx of polymorphonuclear leukocytes (PMN) into the peritoneal cavity. The role of neutrophils in intraperitoneal adhesion formation has not been studied. We aimed to study the effects of PMN counts and PMN functions on peritoneal adhesion formation. Forty peritoneal adhesion-induced rats were randomly divided into three groups; group I, receiving saline; group II, receiving cyclophosphamide; and group III, receiving granulocyte-macrophage colony-stimulating factor (GM-CSF). In all groups, peritoneal lavage was performed to determine PMN counts the day after adhesion induction. Blood neutrophil counts and neutrophil functions were also determined. Adhesions were evaluated blindly 14 days after the operation. Adhesion tissue samples were microscopically evaluated. Tissue hydroxyproline and collagen concentrations were measured. The neutrophil counts and phagocytosis significantly increased in group III and neutrophil counts decreased in group II (P < 0.05). The score of adhesion formation in group II was significantly less than that in groups I and III (P < 0.05). Hydroxyproline concentrations of adhesion tissue were significantly decreased in group II when compared with group III (P < 0.05). The present study shows that neutropenia lowers the degree of postoperative adhesion formation. It is concluded that PMN may have a role to play in modulating post-operative adhesion formation.

Effects of granulocyte-macrophage colony-stimulating factor on incisional wound healing in an experimental diabetic rat model.

The exact nature of poor wound healing in diabetes is uncertain. Neutrophils play a critical role in the host defense mechanism, and it is suggested that impaired neutrophil functions cause healing difficulties with or without infections in diabetic patients. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is used clinically when given systematically to increase the circulating neutrophils, but its wound-healing effects have not been systematically studied. This study was undertaken to examine the effects of GM-CSF on incisional wound healing in an experimental diabetic rat model. Forty rats were randomly divided into three groups, group I receiving saline as control, diabetes-induced group II receiving saline and diabetes-induced group III receiving GM-CSF. The anesthetized rats in all groups were wounded 21 days after diabetes induction by streptozotocin. Blood neutrophil counts and neutrophil fractions were also determined three days after wounding. Tensile strengths of wounded skin and the hydroxyproline (hyp) level of the wound were determined and wound healing processes were evaluated by light and electron microscopy, fourteen days after wounding. Neutrophil counts and phagocytosis were significantly increased in group III and neutrophil counts decreased in group II (p < 0.05). Although the hydroxyproline level of wound tissue significantly decreased in group II as compared with group III (p < 0.05), there was no differences of tensile strength between group II and III (p < 0.05). Wound score in group II was less than that in groups I and III (p < 0.05). It is concluded that PMN may have a role in modulating wound healing. GM-CSF may be useful for creating better wound healing healing. GM-CSF may be useful for creating better wound healing in risky patients such as diabetics.