Ultrastructural and biochemical analysis of the effects of alendronate on salivary glands of young rats

IntroductionThe bisphosphonates are drugs known by their antiresorptive potency and are widely prescribed for treating and preventing osteoporosis. In the past years the employment of this class of drugs had spread to other pathologies, and it is being prescribed to patients in a wide range of ages. Some adverse effects of bisphosphonate treatment have been highlighted recently, however, little is known about its potential side effects in salivary glands.MethodsNewborn rats received daily doses of 2.5 mg/kg/day of sodium alendronate during 30 days. On the thirtieth day the animals were stimulated with pilocarpine and their parotid and submandibular glands were collected, fixed and embedded for histological and ultrastructural analysis. Some glands were collected for analysis of protein content and amylase activity.ResultsAt light microscopy, the alendronate-treated animals presented an accumulation of secretion granules in their cytoplasm, which was confirmed by the ultrastructural examination. Biochemical analysis revealed an increase in total protein content and decreased amylase levels of both glands in the alendronate-treated animals in relation to the control.ConclusionBased on the current findings, alendronate is probably interfering in secretory pathways of parotid and submandibular glands.     

Effects of hyperleptinemia in rat saliva composition,histology and ultrastructure of the major salivary glands

ObjectiveTo study the effect of the satiety hormone, leptin, in saliva proteome and salivary gland histology and ultrastructure.DesignIncreases in blood leptin levels were induced through mini-pump infusion in male Wistar rats, during a period of 7 days. Saliva was collected before and at the end of the experimental period, for proteomic analysis, and major salivary glands were collected, at the end, for biochemical, histological and ultrastructural analysis.ResultsImmunohistochemistry revealed the presence of leptin receptors in major salivary glands. Salivary amylase levels and enzymatic activity were decreased in saliva, whereas the enzymatic activity of this protein was increased in the cytosol of parotid gland cells. Transmission electron microscopy allowed the observation of high number of electron-dense granules in cytosol of parotid acinar cells, from leptin treated animals.ConclusionsIncreased levels of plasmatic leptin result in changes in saliva composition and salivary glands function. To our knowledge, this is the first study providing evidences for a potential role of leptin in salivary gland secretion and saliva composition. An understanding of how appetite/satiety factors influence saliva composition and how this composition influences food processing in mouth may be relevant in understanding ingestivebehaviour.     

血尿淀粉酶检验指标在急性胰腺炎诊断中的应用

目的：运用实验室血尿淀粉酶检验指标对急性胰腺炎的诊断提供参考。方法：观察我院2010-2011年收治的急性胰腺炎患者19N,探讨在不同的病变过程中血尿淀粉酶的变化情况。结果：19例急性胰腺炎患者血尿淀粉酶测定指标都不同程度的高出正常值,其中有2例误诊。结论：急性胰腺炎（Ap）起病急,发展迅速,演变复杂,临床表现不典型,容易误诊,导致不良后果。因此,凡遇突然发作的持续性腹痛,应详细询问病史,认真查体,结合血尿淀粉酶测定,综合分析,可减少误诊。     

What should we trust to define,predict and assess pancreatic fistula after pancreatectomy?

ObjectiveThe ISGPF postoperative pancreatic fistula (POPF) definition using amylase drain concentration is widely used. However, the interest of lipase drain concentration, daily drain output and absolute enzyme daily production (concentration x daily drain volume) have been poorly investigated.Material and methodsThese predictive on postoperative day (POD) 1, 3, 5 and 7 were analyzed in a development cohort, and subsequently tested in an independent validation cohort.ResultsOf the 227 patients of the development cohort, 17% developed a biochemical fistula and 34% a POPF (Grade B/C). Strong correlation was found between amylase/lipase drain concentration at all postoperative days (ρ = 0.90; p = 0.001). Amylase and lipase were both significantly higher in patients with a POPF (p < 0.001) presenting an equivalent under the ROC curve area (0.85 vs 0.84; p = 0.466). Combining POD1 and POD3 threefold enzyme cut-off value increased significantly POPF prediction sensibility (97.4% vs 77.8%) and NPV (97.1% vs 86.3%). These results were also confirmed in the validation cohort of 554 patients. Finally, absolute enzyme daily production and daily drain output were significantly higher in patients with a POPF (p < 0.001) but did not add clinical value when compared to drain enzyme concentration.ConclusionLipase is as effective as amylase drain concentration to define POPF. Absolute enzyme daily production or daily drain output do not help to better predict clinically significant POPF occurrence and severity. Lipase and amylase should mainly be used for their negative predictive value to predict the absence of clinically significant POPF and could allow early drain removal and hospital discharge.     

奥曲肽作用大鼠急性胰腺炎胰腺病理改变及对淀粉酶分泌影响

目的：观察奥曲肽作用于大鼠急性坏死性胰腺炎(ANP)胰腺组织的病理改变及对血液、腹腔液、尿液淀粉酶的影响。方法：实验研究分A、B、C三组，每组8只，共24只，采用随机分配原则分组。A组：空白对照组；B组：ANP模型组：C组：奥曲肽治疗组。通过光镜及透射电镜观察各组胰腺组织的病理改变，同时测定各组血液、腹腔液及尿液中淀粉酶含量。结果：实验性大鼠急性坏死性胰腺炎时胰腺组织充血、水肿、坏死明显，线粒体明显肿胀，嵴消失，间质中可见酶原颗粒，使用奥曲肽治疗时上述变化明显减轻，间质中未见到酶原颗粒积聚。大鼠急性坏死性胰腺炎时血液、尿液淀粉酶水平有下降趋势，但尚无显著差异；腹腔液淀粉酶则有显著下降。结论：奥曲肽作用于大鼠急性坏死性胰腺炎时可减轻胰腺组织充血、水肿及坏死的程度，同时可减少胰腺组织腺泡细胞淀粉酶向腹腔内分泌。     

高活力胰酶的生产

将酶激活剂及稳定剂用于高活力胰酶生产.酶活力为蛋白酶4.5ku/g,淀粉酶70ku/g,脂肪酶35ku/g.胰酶总收率为7.9～1O.8%.     

Presence of cytoskeleton proteins in parotid glands and their roles during secretion

Amylase secretion is induced by the accumulation of cAMP in response to β-adrenergic stimulation and by the augmentation of intracellular Ca²⁺ in response to muscarinic-cholinergic stimulation in rat parotid glands. The roles of cytoskeleton and motor proteins in the secretory process are not yet known. We examined the effects of cytoskeleton-modulating reagents on the amylase release induced by isoproterenol (IPR) and carbamylcholine (Cch) in rat parotid acinar cells. The amylase release induced by Cch was decreased by the microtubule-disrupting reagent colchicine (Colch) and the myosin ATPase inhibitor 2,3-butanediene monoxime (BDM), but the release induced by IPR was not. The actin filament-stabilizing reagent jasplakinolide (Jasp) and actin filament-disrupting reagent cytochalasin D (CytoD) decreased the amylase release induced by both the β-adrenergic and the muscarinic-cholinergic stimulants. Pretreatment with CytoD affected the shape of the acinar cells, which showed an intermediate state between the fusion of the secretory granules with the apical membrane and the retrieval of the membranes only after stimulation with IPR. Myosin and Dynein/dynactin complex were detected in the secretory granule membrane fraction. We concluded from this study that the cytoskeleton played different roles in the β-adrenergic and the muscarinic-cholinergic secretory processes.     

Flow rate,amylase activity,and protein and sialic acid concentrations of saliva from children aged 18, 30 and 42 months attending a baby clinic

Whole-saliva samples were collected under slight suction from children enrolled in a dental-care programme at a baby clinic and analysed according to sex. The children were allocated to three age groups: 18, 30 and 42 months. No distinct sex difference was found for flow rate; there was a statistically non-significant tendency for an increased rate in females. In the males the flow rates of the 30- and 42-month groups were higher than those of the 18-month group; a similar result was obtained when findings for both sexes were combined. There was no difference in salivary protein concentration between the sexes or the age groups. Amylase activity increased significantly in males from 18 to 30 months of age. Sialic acid concentration (micromol/mg protein) showed a sex difference for the 18-month group; it was 38% higher in males than in females, and 48% higher than in the males of 42 months. This trend toward a reduction in sialic acid concentration with age suggests a reduced synthesis and secretion of glycoproteins.     

CAPD患者腹膜电荷屏障与腹膜透析液蛋白质丢失的关系

目的测定持续非卧床腹膜透析（CAPD）患者腹膜电荷屏障,并对原发病为糖尿病肾病（DN）与慢性肾小球肾炎（CGN）的CAPD患者的腹膜电荷屏障以及腹膜透析液蛋白进行比较,进而探讨CAPD患者腹膜电荷屏障与腹膜透析液蛋白丢失的相关性。方法选择32例CAPD患者,收集血清以及腹膜透析液,采用清除法测定胰淀粉酶清除率（Cpam）以及唾液淀粉酶的清除率（Csam）的比值（Cpam/Csam）用以评价腹膜电荷屏障;同时测定腹膜透析液蛋白的丢失量。结果①32例CAPD患者腹膜Cpam/Csam的比值为（6.296±21.514）;腹膜透析液蛋白为（4.14±1.91）g;②CGN组腹膜透析液蛋白显著低于DN组[（4.35±1.88）g比（5.61±0.86）g,P=0.011〈0.05];而CGN组Cpam/Csam的比值显著高于DN组[（9.94±28.35）比（0.68±0.86）,P=0.017〈0.05]。③所有患者Cpam/Csam与腹膜透析液蛋白丢失量之间具有显著负相关,相关系数为-0.584,P〈0.01。而且其自然对数与腹膜透析液蛋白丢失量呈直线负相关。结论CAPD患者腹膜电荷屏障的丢失可增加腹膜透析液蛋白的漏出。