Effects of denervation on the distribution of peanut agglutinin binding molecules in frog muscles

Summary The extracellular matrix has been shown to play an important role in the differentiation of neuromuscular junctions during reinnervation in frogs. Peanut agglutinin, a lectin, is known to specifically bind to some glycoconjugates in the extracellular matrix at the frog neuromuscular junction and myotendinous junction. In order to determine if innervation has any role in regulating the specific binding of peanut agglutinin at neuromuscular junctions and myotendinous junctions, the distribution of peanut agglutinin binding was examined in muscles chronically denervated for various periods. Short-term denervated muscles (2 months) showed no changes in peanut binding agglutinin binding at neuromuscular junctions and no extrajunctional binding. In contrast, long-term denervation (>2 months-7.5 months) resulted in altered peanut agglutinin distribution and a substantial reduction or a total loss in its binding at denervated neuromuscular junctions; binding at myotendinous junctions was not affected. Results of electron microscopic studies suggest that the presence of Schwann cells at denervated endplates delays the loss of peanut agglutinin binding. Reinnervation restores normal peanut agglutinin binding at neuromuscular junctions following long-term denervation. This report demonstrates that although the distribution of peanut agglutinin binding molecules is unchanged following short-term denervation, intact innervation is necessary for the long-term maintenance of these molecules at neuromuscular junctions.     

Extremity vascular trauma in civilian population: a seven-year review from North India

This is a retrospective study of the experience with extremity vascular trauma at a tertiary level referral centre in North India where the majority of the population lives in villages and the incidence of high-speed automobile accidents and civilian violence is low. The aim was to study the aetiology, pattern of injuries and the mortality and morbidity rates due to vascular trauma in our population. Data relating to 148 patients presenting with vascular trauma requiring surgical intervention other than amputation between January 1996 and December 2002 were collected retrospectively. There were 132 males and 16 females with a mean age of 39 years presenting to the casualty with a median delay of 9.3 h after injury. Blunt trauma accounted for 84% of the injuries with extremities involved in 88%. The brachial artery was the most common artery injured with the femoral next most common. Repair without graft interposition was done in 74% and autogenous vein grafts were used in 23% of cases. Comparison of our results with those that would have been obtained, had the recommendations of Mangled Extremity Severity Score (MESS) been followed, showed that this scoring system had low sensitivity and specificity for prediction of salvageability of limbs. Eight percent patients died due to associated visceral organ injuries. Complications occurred in 32% patients and amputation was required in 6% of patients after an initial surgical repair mainly due to inadequate functional recovery. Eighty-eight percent of the survivors with salvaged limbs were able to achieve full functional recovery. Judicious selection and appropriate intervention can result in satisfactory limb salvage with good functional outcomes even with delayed presentations.     

Long-term prognosis and significance of the sentinel lymph node in head and neck melanoma

Objective To report the long-term significance of sentinel lymph node (SLN) biopsy on prognosis, determine false-negative SLN occurrences, and determine risk factors for death and recurrence in a large series of patients with head and neck melanoma. Study Design Case series with tumor registry review. Setting Academic tertiary care medical center. Subjects and Methods A database review was performed of all patients who underwent SLN biopsy for head and neck melanoma from 1994 to 2009. End points assessed were SLN status, recurrence, false-negative SLN results, and survival comparing SLN-positive and SLN-negative patients and different locations. Survival curves and multivariate analyses were performed. Results SLN biopsy was performed in 365 patients. SLNs were identified in 98.6% of patients with a mean of 3.7 nodes removed from 1.6 nodal basins per patient. Median follow-up was 8 years. The SLN was positive in 40 (11%) patients. SLN-positive patients had significantly thicker melanomas, higher recurrence (P < .0001), and a significant decrease in overall survival compared with SLN-negative patients (P < .002). Scalp melanoma patients had significantly thicker melanomas and an elevated risk of SLN positivity, recurrence, and death compared with other sites. Seventeen of 365 SLN-negative patients developed regional nodal disease for a false-omission rate of 5.2% and a negative predictive value of a negative SLN to be 94.8%. Risks for false negative-SLN occurrences included thick melanomas and scalp melanomas. Conclusion SLN biopsy is accurate in head and neck melanoma and provides significant prognostic data. Scalp melanoma patients present with thicker tumors with an increase in SLN positivity and false-negative SLN occurrences.     

The effect of pioglitazone treatment on 15-epi-lipoxin A4 levels in patients with type 2 diabetes

ObjectivesArachidonic acid-derived eicosanoids (lipoxins and 15-epilipoxins) have a major role in resolution of inflammation. 15-epi-lipoxin A₄ (15-epi-LXA₄) is a lipid mediator with strong anti-inflammatory and inflammation-resolving effects. We examined the effect of pioglitazone therapy on plasma 15-epi-LXA₄ in patients with type 2 diabetes (T2DM).MethodsT2DM patients (Age = 56 ± 2 y, BMI = 33 ± 1.8, HbA1c = 7.8 ± 0.3%) not on thiazolidinedione therapy for at least 12 months were randomized to receive either pioglitazone 15 mg/daily for two months (PIO 15) or pioglitazone 15 mg/day for one month followed by a dose escalation to 30 mg/day for an additional one month (PIO 30).ResultsPIO 15 increased plasma 15-epi-LXA₄ levels (0.63 ± 0.06–1.05 ± 0.08 ng/mL, p < 0.01) and adiponectin levels (6.4 ± 0.3–10.1 ± 0.7 μg/mL, p < 0.001) and decreased fasting plasma glucose (125 ± 8–106 ± 9 mg/dL, p < 0.05), free fatty acids (FFA) (414 ± 46–320 ± 38 μmol/l, p < 0.05) and HOMA-IR (5.3 ± 0.4 to 4.0 ± 0.4, p < 0.05). Body weight (Δ = 0.2 kg) and HbA1c (7.4 ± 0.2–7.1 ± 0.2%) did not change significantly. PIO 30 treated patients had similar increase in plasma 15-epi-LXA₄ (0.64 ± 0.10–1.08 ± 0.09 ng/mL, p < 0.01), and decrease in plasma FFA (423 ± 42–317 ± 40 μmol/l, p < 0.05) despite a greater increase in plasma adiponectin (6.5 ± 0.4–15.5 ± 0.7 ug/mL, p < 0.001) and a greater reduction in HbA1c (8.7 ± 0.5–7.4 ± 0.3%, p < 0.01), FPG (159 ± 16–120 ± 10 mg/dL, p < 0.01), and HOMA-IR (6.6 ± 0.8–4.4 ± 0.4, p < 0.005). Furthermore, PIO 30 treated patients had a significant increase in body weight (Δ = 1.7 kg, p < 0.02).ConclusionIn T2DM, low dose pioglitazone (15 mg/day) increases 15-epi-LXA₄ and adiponectin levels in the absence of significant changes in body weight. Dose escalation of pioglitazone to 30 mg/day is associated with a similar increase in 15-epi-LXA₄ despite a greater increase in plasma adiponectin concentrations.     

Dose-dependent effect of oregano (Origanum vulgare L.) on lipid peroxidation and antioxidant status in 1,2-dimethylhydrazine-induced rat colon carcinogenesis

Colon cancer is a major cause of morbidity and mortality in developed and developing countries. Diet and dietary constituents play a major role in the aetiology of colon cancer. We have investigated the effect of an aqueous extract of oregano (Origanum vulgare. L.) on lipid peroxidation and anti-oxidant status in 1,2-dimethylhydrazine (DMH)-induced rat colon carcinogenesis. We aimed to identify the important antioxidants present in Indian oregano using RP-HPLC. DMH (20 mgkg(-1)) was administered subcutaneously once a week for the first four weeks and then discontinued. Oregano was supplemented every day orally at a dose of 20, 40 or 60 mgkg(-1) to different groups of rats for 15 weeks. After this time the rats were killed and the colons were examined visually and evaluated biochemically. The levels of lipid peroxidation products, such as thiobarbituric acid reactive substances and conjugated dienes were significantly higher in the liver whereas in caecum and colon the levels were lower in DMH-treated animals as compared with control rats. The levels of the anti-oxidants superoxide dismutase, catalase, reduced glutathione, glutathione reductase, glutathione peroxidase and glutathione-S-transferase were decreased in DMH-treated rats, but were significantly reversed on oregano supplementation. Oregano supplementation (40 mgkg(-1)) had a modulatory role on tissue lipid peroxidation and antioxidant profile in colon cancer-bearing rats, which suggested a possible anti-cancer property of oregano.     

Selective Sentinel Lymph Node Dissection for Melanoma: Importance of Harvesting Nodes with Lower Radioactive Counts without the Need for Blue Dye

Background

Determining how many sentinel lymph nodes (SLNs) should be removed for melanoma is important. The purpose of this study is to determine the frequency at which nodes that are less radioactive than the “hottest” node (which is negative) are positive for melanoma, how low of a radioactivity should warrant harvest, and if isosulfan blue is necessary.

Methods

We reviewed 1,152 melanoma patients who underwent lymphoscintigraphy with technetium, with or without blue dye, and SLN dissection from 1996 to 2008. SLNs with radioactivity ≥10% of the “hottest” SLN, all blue nodes, and all suspicious nodes were removed and analyzed. The miss rate was calculated as the proportion of node positive cases in which the “hottest” SLN was negative.

Results

SLNs were identified in 1,520 nodal basins in 1,152 patients. SLN micrometastases were detected in 218 basins (14%) in 204 patients (18%). In 16% of SLN-positive patients (33/204 patients), the positive SLN was found to have a lower radioactive count than the “hottest” SLN, which was negative. In 21 of these cases, the positive SLNs had radioactivity ≤50% of the “hottest” SLN. The 10% rule significantly reduced the miss rate to 2.5% compared with removal of only the “hottest” SLN (miss rate = 16%). Also, blue dye did not significantly decrease the miss rate compared with radiocolloid alone using the 10% rule.

Conclusions

To decrease the miss rate, all SLNs with ≥10% of the ex vivo radioactivity of the “hottest” SLN should be removed and blue dye is not essential.     

R723, a selective JAK2 inhibitor, effectively treats JAK2V617F-induced murine myeloproliferative neoplasm

The activating mutations in JAK2 (including JAK2V617F) that have been described in patients with myeloproliferative neoplasms (MPNs) are linked directly to MPN pathogenesis. We developed R723, an orally bioavailable small molecule that inhibits JAK2 activity in vitro by 50% at a concentration of 2nM, while having minimal effects on JAK3, TYK2, and JAK1 activity. R723 inhibited cytokine-independent CFU-E growth and constitutive activation of STAT5 in primary hematopoietic cells expressing JAK2V617F. In an anemia mouse model induced by phenylhydrazine, R723 inhibited erythropoiesis. In a leukemia mouse model using Ba/F3 cells expressing JAK2V617F, R723 treatment prolonged survival and decreased tumor burden. In V617F-transgenic mice that closely mimic human primary myelofibrosis, R723 treatment improved survival, hepatosplenomegaly, leukocytosis, and thrombocytosis. R723 preferentially targeted the JAK2-dependent pathway rather than the JAK1- and JAK3-dependent pathways in vivo, and its effects on T and B lymphocytes were mild compared with its effects on myeloid cells. Our preclinical data indicate that R723 has a favorable safety profile and the potential to become an efficacious treatment for patients with JAK2V617F-positive MPNs.     

Strangulated Interstitial Hernia with Strangulated Cryptorchid Testis—First Case Report in the Medical Literature

Interstitial hernias are a rare subtype of abdominal hernias, where sac is present in between the abdominal wall layers. Although, difficult to diagnose clinically, they are usually detected on imaging studies. Interstitial hernias presenting with undescended testis are common. Even after a meticulous online search of the medical literature, we could not find a case report of 'Strangulation of Interstitial hernia, with a strangulated undescended testis in an elderly man. We hereby present this extremely rare case, and possibly, this could be the first of its kind, case report in the medical literature. Our aim is to present a combination of rare presentations in an elderly cryptorchid person with strangulated interstitial hernia. A 65-year male patient presented with 4 days of pain abdomen and vomiting. He was having features of toxaemia. Ultrasonography revealed interstitial hernia with bowel loops. During surgery, gangrenous bowel loops with the sac between the interstitial planes, with strangulated cryptorchid testis, were found. Strangulated interstitial hernia in elderly cryptorchid person, leading to the strangulation of the undescended testis, is extremely rare, and this could be the first case reported in the medical literature.     

Development and Validation of a Stability-Indicating RP-UPLC Method for the Estimation of Impurities in Cinacalcet Hydrochloride API and its Formulation

A sensitive, stability-indicating, gradient reversed-phase ultra-performance liquid chromatography method has been developed for the quantitative estimation of cinacalcet hydrochloride impurities in active pharmaceutical ingredients and pharmaceutical formulations. Efficient chromatographic separation was achieved on an Acquity BEH Shield RP18, 100 × 2.1 mm, 1.7 µm column with the mobile phase containing pH 6.6 phosphate buffer and acetonitrile. The flow rate of the mobile phase was 0.3 mL min⁻¹ with a column temperature of 35°C and detection wavelength at 223 nm. The relative response factor values of (+)-R-1-(1-Naphthyl)ethylamine, regioisomer, diastereomer isomer-1, and diastereomer isomer-2 were 1.79, 0.99, 0.89, and 0.88, respectively. The cinacalcet hydrochloride formulation sample was subjected to the stress conditions of acid, base, oxidative, hydrolytic, thermal, humidity, and photolytic degradation. Cinacalcet hydrochloride was found to degrade significantly under the peroxide stress conditions. The degradation products were well-resolved from cinacalcet hydrochloride and its impurities. The peak purity test results confirmed that the cinacalcet hydrochloride peak was homogenous in all stress samples and the mass balance was found to be more than 96%, thus proving the stability-indicating power of the method. The developed method was validated according to ICH guidelines.