Apoptotic effects of thymol, a novel monoterpene phenol, on different types of cancer.

BACKGROUND: Cancer is a major public health problem in many areas of the world. Many anticancer drugs in current clinical use have been isolated from plant species or are based on such substances. Thymol (5-methyl-2-isopropylphenol) is an oxygenated aromatic compound from monoterpene group. It is the main constituent of thyme essential oil and shows antioxidant, antiseptic and antiproliferative properties. The aim of this study is to determine the antiproliferative activity and apoptotic effect of thymol on prostate cancer (PC-3, DU145), breast cancer (MDA-MB-231), and lung cancer (KLN205) cell lines. METHODS: The cancer cells were treated with different concentrations of thymol (100, 200, 400, 600, 800 µM) at 24 h, 48 h and 72 h. The cell viability was investigated by MTT assay and analysis of apoptosis was determined with annexin V assay. RESULTS: The study showed the dose and time-dependent cytotoxic effect of thymol in PC-3, DU145, MDA-MB-231, and KLN205 cancer cell lines. Thymol significantly induced apoptosis in all groups in a dose-dependent manner. Statistical analysis showed a significant difference between thymol­treated cell lines compared to the control (p < 0.001). CONCLUSION: The data in the present study demonstrated that thymol has apoptotic and antiproliferative properties in lung, breast and prostate cancer cell lines. Thymol could serve as a potential therapeutic agent in the future (Fig. 5, Ref. 26).

Ultra-Structural Alterations in In Vitro Produced Four-Cell Bovine Embryos Following Controlled Slow Freezing or Vitrification.

Cryopreservation is the process of freezing and preserving cells and tissues at low temperatures. Controlled slow freezing and vitrification have successfully been used for cryopreservation of mammalian embryos. We investigated the effect of these two cryopreservation methods on in vitro produced four-cell stage bovine embryos which were classified according to their quality and separated into three groups. The first group was maintained as untreated controls (n = 350). Embryos of the second (n = 385) and the third (n = 385) groups were cryopreserved either by controlled slow freezing or by vitrification. Embryos in groups 2 and 3 were thawed after 1 day. Hundred embryos were randomly selected from the control group, and 100 morphologically intact embryos from the second and third group were thawed after 1 day and cultured to observe the development up to the blastocyst stage. The blastocyst development rate was 22% in the control group, 1% in the slow-freezing group and 3% in the vitrification group. Remaining embryos of all three groups were examined by light microscopy, transmission electron microscopy and immunofluorescence confocal microscopy with subsequent histological staining procedures. Cryopreservation caused degenerative changes at the ultra-structural level. Compared with vitrification, slow freezing caused an increased mitochondrial degeneration, cytoplasmic vacuolization, disruption of the nuclear and plasma membrane integrity, organelle disintegration, cytoskeletal damage, a reduced thickness of the zona pellucida and a formation of fractures in the zona pellucida. Further studies are required to understand and decrease the harmful effects of cryopreservation.

The protective effect of losartan on diabetic neuropathy in a diabetic rat model.

AIM: Involvement of the peripheral and autonomic nervous systems is possibly the most frequent complication of diabetes. Important risk factors included hyperglycemia, dyslipidemia, hypertension, and smoking. Angiotensin-converting-enzyme inhibitor (ACE) inhibitors should be beneficial in all vascular beds, including neuropathy and retinopathy. In this study we aimed to evaluate the effect of the angiotensin receptor blocker losartan on diabetic neuropathy in a diabetic rat model. MATERIAL AND METHODS: 24 male, Sprague Dawley albino mature rats were divided into 3 groups; (1) control group: No drug was administered to the remainder of rats which blood glucose levels were under 120 mg/dl, (2) diabetic control: rats were given no medication, but 4 ml per day of tap water was given by oral gavage, (3) losartan groups: rats were given 10 mg/kg/day oral of losartan for 4 weeks. Electromyography (EMG) was applied to anesthetized rats at the end of 4(th) weekend. Then, the animals were euthanized and sciatic nerve was performed for histopathological examination. RESULTS: Compound Muscle Action Potential (CMAP) amplitude of diabetic rats receiving the Saline in the EMG was significantly reduced when compared to the control group. Distal latency value and CMAP duration of diabetic rats receiving the saline were meaningfully increased when compared to the control group. CMAP amplitude and CMAP duration of diabetic rats receiving the Losartan treatment in the EMG were meaningfully reduced when compared to diabetic rats receiving the Saline.Perineural thickness in the rats receiving the Losartan treatment was found to be significantly reduced when compared to the group receiving the Saline. CONCLUSIONS: As a result, it has been shown in this study that perineural thickness of the Losartan treatment was significantly reduced when compared to saline receiving group, significantly increased the immunoexpression of NGF, and also provided a significantly recovery in EMG when compared to Saline receiving group.

Comparison of nephron-protective effects of enalapril and GLP analogues (exenatide) in diabetic nephropathy.

BACKGROUND: One of the major concerns is a nephropathy in diabetes, which applies many different kinds of medicines. However, required level of the treatment of renal disease has not been achieved. AIM: To investigate and compare the effect of the enalapril and the exenatide on diabetic nephropathy in rats developed diabetes by streptozosin. MATERIAL AND METHODS: 32 male Sprague Dawley rats were divided into 4 groups: (1) Control, (2) Diabetic (DM), (3) DM+ Enalapril, and (4) DM+ exenatide groups. Then, the animals were euthanized and their blood samples were collected by cardiac puncture for blood glucose; blood urea nitrogen (BUN), creatinin, and nephrectomy were performed for histopathologic examination, and urine samples were taken on stick for proteinuria. RESULTS: Administration of the enalapril or the exenatide in diabetic rats resulted in a significant reduction both fibronectin, induced nitric oxide synthase (i-NOS) expression in glomerular area and urine protein levels. It was shown that both of enalapril and exenatide protected the renal glomerulus more than diabetic group in the nephropathy histopathologically. CONCLUSION: The beneficial effects of enalapril and exenatide which reduces fibronectin, i-NOS expression and urine protein levels or increases recovery of glomerules, might be used for preventing the harmful effects of diabetic nephropathy.

Incision wound healing activity of pine bark extract containing topical formulations: a study with histopathological and biochemical analyses in albino rats.

The present study was designed to identify and compare the in vivo wound healing capacity of a bark extract from Pinus brutia and Pycnogenol in an incision wound model in rats. O/W cream formulations were prepared incorporating 2% Pycnogenol and P. brutia bark extract. The rats were divided into three groups (n = 8). Subsequently placebo and test formulations were applied to animals once a day from day "0" until the 9th day. Malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were studied in addition to histopathological examinations. Treatment with F. brutia extract containing cream inhibited lipid peroxidation by a 35% decrease in MDA and 46.8% increase in SOD activity, whereas 19.3% decrease in MDA and 34.7% increase in SOD activity were attained with Pynogenol compared to control. The histological data revealed a better performance of P. brutia extract enriched formulation in terms of degeneration of hair roots, increased vascularization and a decrease in necrotic area. Consequently, a high wound healing activity was observed in animals treated with P. brutia extract significantly accelerating the wound healing process.

Auricular cartilage repair using cryogel scaffolds loaded with BMP-7-expressing primary chondrocytes.

The loss of cartilage tissue due to trauma, tumour surgery or congenital defects, such as microtia and anotia, is one of the major concerns in head and neck surgery. Recently tissue-engineering approaches, including gene delivery, have been proposed for the regeneration of cartilage tissue. In this study, primary chondrocytes were genetically modified with plasmid-encoding bone morphogenetic protein-7 (BMP-7) via the commercially available non-viral Turbofect vector, with the aim of bringing ex vivo transfected chondrocytes to resynthesize BMP-7 in vitro as they would in vivo. Genetically modified cells were implanted into gelatin-oxidized dextran scaffolds and cartilage tissue formation was investigated in 15 × 15 mm auricular cartilage defects in vivo in 48 New Zealand (NZ) white rabbits for 4 months. The results were evaluated via histology and early gene expression. Early gene expression results indicated a strong effect of exogenous BMP-7 on matrix synthesis and chondrocyte growth. In addition, histological analysis results exhibited significantly better cartilage healing with BMP-7-modified (transfected) cells than in the non-modified (non-transfected) group and as well as the control.

Venous vascular malformations of the craniofacial region: pre-operative embolisation with direct percutaneous puncture and N-butyl cyanoacrylate.

Craniofacial venous vascular malformations cause severe cosmetic problems and yet these lesions are not candidates for transcatheter embolisation owing to the lack of arterial feeders. The purpose of this study was to evaluate the effectiveness of pre-operative embolisation of these lesions with N-butyl 2-cyanoacrylate (NBCA) via direct puncture. Between September 2003 and April 2006, 13 patients (7 female; age range, 6-64 years; mean, 16.7 years) were embolised with direct puncture and injection of NBCA. All of the patients were referred from plastic surgery with an operational plan. Angiography performed in all patients showed no or little arterial staining. NBCA diluted with iodized oil at a ratio of 1:6 (18%) was injected via a percutaneously placed 21 gauge needle. Complete embolisation was achieved in 8 patients and partial embolisation in the remaining 5. A total of 18 sessions of embolisation were performed on 13 patients. Nine patients underwent only one embolisation session, three patients underwent two sessions and only one patient underwent three sessions. The mean volume of NBCA used per session was 5.8 ml, ranging from 1-12 ml. All patients underwent a successful surgical resection to improve cosmetic disfigurement within 10-15 days after the embolisation procedure. Mean follow-up time was 22 months. One patient experienced skin necrosis on her nose after embolisation. No other complications related to the procedure were observed. In conclusion, pre-operative NBCA embolisation with direct puncture is a safe and easy procedure. It can increase the success of the surgical treatment of these lesions.

W-plasty technique in tracheal reconstruction: a new technique? An experimental study.

BACKGROUND: Tracheal stenosis and dehiscence of anastomosis due to excessive tension are well-known problems after long-segment tracheal resections. The aim of this study was to evaluate the efficacy of the W-plasty technique to prevent these two complications. METHODS: Animals were divided into a study and a control group. Each group consisted of 6 animals. In the control group, we performed a 5-cm tracheal segment resection, and then reconstruction was performed with an interrupted technique with 6/0 Prolene sutures. In the study group, we used the W-plasty technique with 6/0 Prolene interrupted sutures. The animals were sacrificed on the 30th day postoperatively and tracheal resection including the entire anastomosis site was performed. The traction and pullout test was applied to each specimen and all the specimens were analysed histopathologically. The intraluminal diameter and the thickness of the tracheal wall at the level of anastomoses were measured by using a micrometer. The pattern of the reaction and localization were recorded. RESULTS: The traction and pullout test results were 131.6 +/- 4.3 g and 187.5 +/- 6.4 g in the control and the study group, respectively, which was a significant difference (p = 0.004). The intraluminal diameters were 3.3 +/- 1.2 mm and 4.3 +/- 0.9 mm in the control and study group, respectively (p = 0.134). In contrast to the control group, early inflammatory and late fibroblastic reactions were negative in the study group. CONCLUSION: Considering the outcomes of this study, we think that the W-plasty technique has much more advantages than the standard techniques in terms of anastomosis durability and development of stenosis.

Dermatofibrosarcoma protuberans of the breast.

Dermatofibrosarcoma protuberans is a rare soft tissue cancer of the skin with the potential for intermediate malignity, characterized by local invasion and recurrence. Presenting with local atrophic and sometimes erythematous plaques, these lesions are best treated with surgery in which large excisions are necessary to reduce the risk of recurrence and to prevent rarely seen metastases. We report on a 26-year-old female patient with dermatofibrosarcoma protuberans on her breast, an very unusual site. As a novel surgical approach to treat the patient's tumor, a keyhole pattern was used, allowing wide safety margins during resection, while preserving the cosmetic appearance of the breast.

Treatment of intertrochanteric fractures by external fixation.

This study evaluates the results of intertrochanteric fractures of the femur treated by external fixation. One hundred and four intertrochanteric fractures of the femur were stabilized by external fixation over an 8 year period. Complete fracture healing was detected in all patients within 11.2 weeks. Varus malunions and more than 2 cm limb shortening were evaluated in 8 patients. Eight patients had permanent knee stiffness. A pin tract infection occurred in 13 patients. This technique is simple and safe. It allows the patients to mobilize earlier. External fixation in intertrochanteric fractures of the femur can be performed especially in elderly patients with stable and unstable fractures that can be reduced to anatomical or nearly anatomical position by closed methods.

Surgical treatment of urethral fistulas following hypospadias repair.

Development of urethral fistulas is one of the most common late complications of hypospadias surgery. A total of 161 male patients who had 186 urethrocutaneous fistulas were first classified according to the fistula classification of Horton and colleagues and then treated with three types of procedures: simple closure, local rotation flaps, or tube graft reconstruction. With initial surgical intervention, 156 of 186 fistulas were treated successfully. The remaining 30 fistulas (16.1%) recurred during the follow-up period. In the recurrent cases, immediate closure was not preferred, and an average of 6 months was waited before considering any additional surgical attempt. Distal cases had a higher failure rate, and the simple closure technique failed to show a success rate as high as local flap or tube graft repair. The high recurrence of distal cases was attributed mainly to the lack of adequate soft tissue adjacent to the fistula, which is vital for safe closure. In addition, the traction effect of erection on the skin and urethra, which is more prominent distally than proximally, is also believed to play an additive role. To increase success, the selection of appropriate treatment modality and customization of techniques for each patient cannot be overemphasized. However, the authors conclude that careful presurgical assessment of the patient, a 6-month delay before any secondary surgical attempt, inversion of the urethral mucosa, avoidance of any overlapping suture lines, urinary diversion proximal to the repair site for 5 to 11 days, and usage of thin, absorbable suture materials are the main criteria that should be met for a satisfactory hypospadias fistula repair.

Post-thyroidectomy hypocalcemia: the role of calcitonin, parathormone and serum albumin.

To investigate the pathogenesis of post-thyroidectomy hypocalcemia calcium, phosphate, calcitonin, parathormone, albumin, triiodothyronine and thyroxine levels were monitored during operation and postoperatively in 25 female patients undergoing bilateral subtotal thyroidectomy for non-toxic nodular goiter. A highly significant fall in uncorrected serum calcium and albumin levels started with the completion of lobectomies and a significant correlation between the course of serum calcium and albumin levels were seen, an early peak in calcitonin corresponded well with the a drop in corrected calcium and inorganic phosphate levels. An overt parathyroid response to hypocalcemia was not observed. We conclude, on the basis of the postoperative hypoalbuminemia, that a calcitonin leak triggered the early onset of hypocalcemia while an insufficient parathyroid response contributed to the post-thyroidectomy hypocalcemia.

Primary colonic non-Hodgkin lymphomas: a retrospective clinicopathologic study of 14 cases.

Fourteen cases of primary colonic non-Hodgkin lymphomas (NHL) with a mean age of 51.5 yrs and 64.3% of them female, are reported. While diagnoses were only obtained by cytologic or histopathologic means, 35.5% of the cases were in Stage 1e (S1e) and a further 42.6% in Stage 2e (S2e) and 7.1% in Stage 3e (S3e) according to the modified Manchester classification. 63.9% were of immunoblastic and 21.3% lymphoblastic type according to the Kiel classification. 85.2% of the tumours were located at the caecum. While acute abdomen required surgery in two patients, 85.2% of the series underwent radical interventions. 14.2% were able to receive chemotherapy with a subsequent total morbidity and mortality figures of 21.3% each. It is the authors' argument that prognosis is not solely dependent on the age, sex or the malignancy state of the tumour but more on its infiltrative stage and on the advent of treatment, whether by radical surgery and/or medical means.