Construction of nanocarriers based on nucleic acids and their applications in nanobiology delivery systems.

In recent years, nanocarriers based on nucleic acids have emerged as powerful and novel nanocarriers that are able to meet the demand for cancer-cell-specific targeting. Functional dynamics analysis revealed good biocompatibility, low toxicity and programmable structures, and their advantages include controllable size and modifiability. The development of novel hybrids has focused on the distinct roles of biosensing, drug and gene delivery, vaccine transport, photosensitization, counteracting drug resistance and functioning as carriers and logic gates. This review is divided into three parts: (i) DNA nanocarriers, (ii) RNA nanocarriers and (iii) DNA/RNA hybrid nanocarriers and their applications in nanobiology delivery systems. We also provide perspectives on possible future directions for growth in this field.

Perioperative hyperglycemia: an unmet need within a surgical site infection bundle.

BACKGROUND: The aim of this study was to determine whether perioperative stress hyperglycemia is correlated with surgical site infection (SSI) rates in non-diabetes mellitus (DM) patients undergoing elective colorectal resections within an SSI bundle. METHODS: American College of Surgeons National Surgical Quality Improvement Program data of patients treated at a single institution in 2006-2012 were supplemented by institutional review board-approved chart review. A multifactorial SSI bundle was implemented in 2009 without changing the preoperative 8-h nil per os, and in the absence of either a carbohydrate loading strategy or hyperglycemic management protocol. Hyperglycemia was defined as blood glucose level > 140 mg/dL. The primary endpoint was SSI defined by the Centers for Disease Control National Nosocomial Infections Surveillance. RESULTS: Of 690 patients included, 112 (16.2%) had pre-existing DM. Overall SSI rates were significantly higher in DM patients as compared to non-DM patients (28.7 vs. 22.3%, p = 0.042). Postoperative hyperglycemia was more frequently seen in non-DM patients (46 vs. 42.9%). The SSI bundle reduced SSI rates (17 vs. 29.3%, p < 0.001), but the rate of hyperglycemia remained unchanged for DM or non-DM patients (pre-bundle 59%; post-bundle 62%, p = 0.527). Organ/space SSI rates were higher in patients with pre- and postoperative hyperglycemia (12.6%) (p = 0.017). Overall SSI rates were higher in DM patients with hyperglycemia as compared to non-DM patients with hyperglycemia (35.6 vs. 20.8%, p = 0.002). At multivariate analysis DM, chronic steroid use, chemotherapy and SSI bundle were predictive factors for SSI. CONCLUSIONS: This study showed that non-DM patients have a postoperative hyperglycemia rate as high as 46% in spite of the SSI bundle. A positive correlation was found between stress hyperglycemia and organ/space SSI rates regardless of the DM status. These data support the need for a strategy to prevent stress hyperglycemia in non-DM patients undergoing colorectal resections.

Relationship between intrafollicular concentrations of parathyroid hormone-related peptide (PTHrP) and steroid hormones in oestrogenic and non-oestrogenic ovarian follicles in the mare.

The objective of the present study was to determine whether parathyroid hormone-related peptide (PTHrP) is present in the equine follicular fluid and if so, how it is related to the follicular development in the horse. For this purpose, ovaries were collected from 40 Thoroughbred and Thoroughbred Cross mares at slaughter during the period from February to May. Normal growing follicles were dissected from the ovaries of each mare and their diameters measured. A total of 174 follicles was used in this study. The follicular fluid was aspirated from each follicle and assayed for PTHrP, oestradiol (E), testosterone (T) and progesterone (P). The follicles were classified as either oestrogenic or non-oestrogenic if the follicular fluid content of oestradiol was >40 or <40 ng/ml, respectively. PTHrP concentrations were significantly (P<0.05) higher in oestrogenic follicles, but T and P concentrations did not differ. Furthermore, E:T ratio was significantly (P<0.05) greater in oestrogenic follicles compared to the non-oestrogenic ones. The mean diameter of oestrogenic follicles was significantly (P<0.05) greater than that of non-oestrogenic ones. The higher concentrations of PTHrP observed in the follicular fluid of healthy oestrogenic follicles suggest that it may have a role in the control of ovarian function.

The relaxant effects of parathyroid hormone(1-34) and parathyroid hormone-related protein(1-34) on ovine reticulo-ruminal smooth muscle in vivo.

The motility of the reticulo-rumen has been measured in trained, conscious sheep using inflated balloons temporarily introduced to selected regions of that forestomach. The frequency and amplitude of the contractions of the reticulum and both the A and B waves of contraction of the rumen were measured under the same conditions before, during and after the administration of an i.v. bolus of either parathyroid hormone (PTH(1-34)) or PTH-related protein (PTHrP(1-34)) followed by its i.v. infusion. These two peptides are known to share a common receptor in other organs, e.g. the kidney. In this study they both showed an inhibitory effect on reticulo-ruminal motility. The effect of PTHrP(1-34) on the rate of ruminal blood flow was also examined and a significant reduction observed, after a transient increase. The secretion of endogenous PTH(1-34) was stimulated by a 32% reduction in the plasma calcium ion concentration induced by an i.v. infusion of sodium citrate. Associated with this were significant reductions in reticulo-ruminal motility, e.g. the reduction in the mean amplitude of the reticular contractions reflected the reduction in plasma calcium ion concentration. When the PTH(1-34)/PTHrP(1-34) receptor was blocked with [Asn10,Leu11,D-Trp12]PTHrP(7-34) before and during the induction of hypocalcaemia, all but one of the parameters of reticulo-ruminal motility were normalized. Indeed, by the day following the administration of this blocking agent, all these parameters had returned to their normal range. It is concluded that stimulation of the PTH(1-34)/PTHrP(1-34) receptor in reticulo-ruminal smooth muscle reduces the motility of this tissue and may play a role in the depression of motility of the digestive tract which is characteristic of clinical milk fever in the dairy cow.

Stimulatory effect of insulin on calcitonin secretion.

Infusion of insulin directly into thyroid arterial blood perfusing the surgically isolated in situ pig thyroid gland produced an increase in the secretion rate of calcitonin (CT) measured by immunoassay in thyroid venous effluent blood. Insulin in concentrations ranging from approximately 1 to 400 ng/ml produced a maximal stimulation of 4-5 fold. The stimulatory effect of insulin on CT could not be duplicated by infusion of either IGF-I or amylin. Specific binding of radiolabeled insulin was demonstrated using isolated pig thyroid plasma membranes and both rat (6-23) and human (TT) medullary thyroid carcinoma C-cells. Increased CT release was observed from C-cells exposed to a high concentration of insulin. The administration of glucose iv to pigs in order to stimulate secretion of endogenous insulin produced an increase in circulating insulin, which was accompanied by an increase in the secretion of CT. The results show that insulin, delivered directly to the pig thyroid gland, can stimulate CT release. The in vitro binding and secretion studies indicate that C-cells can bind insulin and respond with an increase in CT secretion, and the iv glucose experiments suggest that endogenous insulin is capable of stimulating CT secretion. The findings imply that insulin is capable of acting as a CT secretagogue and suggest that changes in CT secretion may accompany altered states of insulin production such as diabetes or insulin-secreting tumors.

The relationship between the concentration of ionised calcium and parathyroid hormone-related protein (PTHrP[1-34]) in the milk of mares.

Once lactation is established in mares, there is little change in the ionised calcium concentration in their milk. In contrast, the concentration of PTHrP(1-34) in the milk increases to a maximum level by the end of the second week of lactation, near which it remains for the rest of the lactation. As found in other species, the concentration of PTHrP(1-34) in mare's milk is considerably higher than that in plasma, sampled at the same time. No significant correlation could be demonstrated between the concentrations of PTHrP(1-34) and ionised calcium in the milk except during the last 10 weeks of lactation.

Structural and physiologic characterization of the mid-region secretory species of parathyroid hormone-related protein.

Parathyroid hormone-related protein (PTHrP) is initially translated as a preprohormone which is posttranslationally processed to yield a family of mature secretory forms. Most attention has focused on the amino-terminal portion of the molecule which is homologous to parathyroid hormone. It is clear, however, that a mid-region species of PTHrP is posttranslationally cleaved from the highly conserved mid-region of PTHrP, and that the amino terminus of this peptide is Ala38. The purposes of the current study were three: 1) to confirm that Arg37 immediately preceding Ala38 serves as a posttranslational processing site in the PTHrP precursor, 2) to determine the carboxyl terminus of the mid-region secretory species of PTHrP, and 3) to synthesize this authentic mid-region secretory form of PTHrP and determine whether it is biologically active. The results indicate that: 1) Arg37 is indeed a processing site in the PTHrP precursor; 2) three distinct mid-region PTHrP species are generated by posttranslational processing, PTHrP(38-94)amide, PTHrP(38-95), and most likely, PTHrP(38-101); and 3) synthetic mid-region PTHrP(38-94)amide is active in four different biological systems. These studies confirm the finding that PTHrP is a prohormone. More importantly, they define a novel, biologically active highly conserved mid-region secretory form of PTHrP.

The secretion of parathyroid hormone-related protein in the saliva of sheep and its effects on the salivary clearance of phosphate, calcium, magnesium, potassium and sodium ions.

Parathyroid hormone-related protein (PTHrP(1-34)) was infused into five sheep, each fitted with a large rumen cannula. After infusion, significant increases were observed in the total and ionized calcium concentrations in plasma but not in saliva. In contrast, significant decreases in the plasma concentrations of phosphate and potassium and corresponding increases in their salivary concentrations and clearance rates were observed. The salivary concentration of endogenous PTH1P(1-34) was significantly greater than that in plasma sampled simultaneously, but during the infusion of PTHrP(1-34) both plasma and salivary concentrations of PTHrP(1-34) increased.

Local control of parathyroid hormone-related protein secretion by the mammary gland of the goat.

In late-pregnant goats, daily removal of secretion from one mammary gland stimulated an early secretion of fluid by that gland in five of the six animals studied. This early secretion of fluid was accompanied by an early increase of parathyroid hormone-related protein (PTHrP) 1-86 concentration in the secretion of that gland alone. When lactation was established, glands emptied thrice daily secreted PTHrP (1-86) in greater quantities and concentrations than glands emptied once daily concurrently in the same animals. Toward the end of lactation, abrupt cessation of emptying one gland caused levels of PTHrP (1-86) to decline in the fluid in that gland but not the contralateral, emptied gland. We conclude that, in the goat, PTHrP (1-86) secretion into milk is linked to autocrine factor(s) that stimulate milk secretion when fluid is removed from the gland.

The role of parathyroid hormone-related protein in calcium homeostasis in the fetal pig.

The concentrations of plasma parathyroid hormone-like bioactivity and parathyroid hormone-related protein (1-86) (PTHrP) immunoreactivity were both higher in fetal pigs than in their mothers during the last 3 weeks of gestation. Both activities changed inversely with alterations in the plasma ionized calcium concentration. The data suggest that PTHrP may have a role in calcium homeostasis in the fetal pig, similar to its postulated role in sheep in the stimulation of calcium transport across the placenta.

Effects of parathyroid hormone and parathyroid hormone-related protein on the rates of absorption of magnesium, calcium, sodium, potassium and phosphate ions from the reticulo-rumen of sheep.

Not only parathyroid hormone (PTH) but also parathyroid hormone-related protein (PTHrP) may play a role in calcium and phosphate homeostasis in ruminants. In five trained sheep, each with a large rumen cannula, the isolated rumen wash technique was used to measure electrolyte absorption rates from the rumen. After two control periods of measurement, 20 micrograms PTH(1-34) (n = 4), or PTHrP(1-34) (n = 5), was injected intravenously as a loading dose, followed by an infusion of 0.67 micrograms min-1 over 2 h. Both PTH(1-34) and PTHrP(1-34) significantly increased the absorption rates of calcium and phosphate from the reticulo-rumen. The increases in the absorption rates of magnesium, sodium and potassium observed were not significant. The same technique was also used to demonstrate that increasing the intraruminal calcium concentration from 1 to 4 mmol l(-1) caused corresponding increases in the net rate of absorption of both calcium and inorganic phosphate from the reticulo-rumen. Rumen epithelium was taken from four sheep and mounted in Ussing chambers so that fluxes of calcium could be measured in both directions using 45Ca. It was found that the addition of 100 ng ml-1 PTH(1-34) or PTHrP(1-34) to the serosal side increased the net calcium flux rates across the ruminal epithelium. It is concluded that both PTH and PTHrP can influence calcium and phosphate homeostasis in sheep not only by their recognized actions on bone and kidney but also on the absorption of these ions from the forestomachs.

Parathyroid hormone-related peptide and milking-induced phosphaturia in dairy cows.

Five intact and four thyroparathyroidectomized (TPTX) lactating dairy cows were fitted with polyvinyl catheters implanted into the left carotid artery and the left mammary vein (for blood sampling) and into the right external jugular vein (for injections), and with a Foley catheter inserted into the urinary bladder. In these nine cows, daily morning and evening milking induced a prompt and transient rise in mammary venous blood plasma parathyroid hormone-related peptide (PTHrP)(1-34) concentrations, followed by an increase in both urinary phosphate concentration and phosphate renal clearance. Such a milking-induced phosphaturia was not observed when (tyr)34-bPTH(7-34)-NH2 (5 pmol/kg body wt) was infused during milking into the TPTX cows. These results indicate that PTHrP released systematically by the mammary gland during milking might be responsible for the observed phosphaturia.

Clearance of exogenous parathyroid hormone-related protein in pregnant, non-pregnant and fetal sheep, goats and pigs.

In the sheep, goat and pig, radiolabelled parathyroid hormone related protein (PTHRP) and immunoreactive PTHRP(1-34) and (1-86) were rapidly cleared from the circulation. Metabolic clearance rates (MCR) were in the range of 1.25-7.5 ml/min per kg and were slightly slower than that of intact PTH in man (10 ml/min per kg); while the mean MCR of labelled PTHRP(1-86) in fetal sheep and goats was significantly faster than that in their respective mothers (14.4 vs 4.0 ml/min per kg respectively). This may reflect increased metabolism of PTHRP by fetal tissues, e.g. the placenta. Similar rates of clearance of radiolabelled PTHRP(1-141), (1-86) and (1-34) suggest that clearance involves the amino terminus of the molecule.

The effects in the rat of two fragments of parathyroid hormone-related protein on uterine contractions in situ.

Synthetic parathyroid hormone fragment PTH(1-34) has been reported recently to inhibit uterine contractions stimulated by a variety of agonists. We have studied the effect in this system of the parathyroid hormone-related protein fragment PTHrP(1-34) which shows 60% homology with PTH over the first thirteen amino acid residues. The effects of two different PTHrP fragments on acetylcholine-stimulated uterine contractions in vitro were studied. Whereas synthetic hPTHrP(75-86 amide) (10(-9)-10(-7) M) was without effect, synthetic hPTHrP(1-34) (10(-9)-10(-7) M) was capable of inhibiting, in a dose-related fashion, uterine muscle contractions precontracted with 10(-6) M-acetylcholine. In a second series of experiments the bovine PTH(3-34) fragment itself was shown to have no stimulatory effect on acetylcholine-stimulated contractions. Also this fragment in an equimolar concentration (10(-7) M) failed to antagonize the effects of PTHrP(1-34) on acetylcholine-stimulated uterine contractions. However, a 100-fold excess molar concentration of bPTH(3-34) (10(-6) M) completely abolished the inhibitory action of hPTHrP(1-34) (10(-8) M) on acetylcholine-stimulated uterine contractions. These results clearly show that the inhibitory action of PTH(1-34) and PTHrP(1-34) on uterine contractions depends on the integrity of the amino-terminal region of the molecule.