Effects of the common pesticide methoxychlor on ovarian steroidogenesis

Methoxychlor, an organochlorine pesticide, is released into the environment as a result of its agricultural, home and veterinary use. Following in vivo administration, it is quickly metabolized by liver to the proposed active agent, HPTE. Both methoxychlor and HPTE have been shown to exhibit weak estrogenic and antiandrogenic activities, and they are thought to exert their effects through estrogen and androgen receptors. In vivo exposure to methoxychlor in laboratory animals causes ovarian atrophy and impaired ovarian steroid production. The effect of methoxychlor and its mechanism(s) of action on ovarian steroidogenesis are not clearly defined. The central hypothesis of these studies is that exposure to HPTE directly inhibits ovarian steroid biosynthesis, which is able to cause an imbalance in circulating progesterone and estradiol levels. The specific aims of the project are: (1) To pinpoint the potential effect of HPTE on ovarian steroidogenesis, (2) To examine the possible mechanism(s) of action of HPTE on ovarian steroidogenesis, (3) To evaluate the potential effect of HPTE on ovarian luteal steroidogenesis, (4) To determine whether in vivo exposure to methoxychlor lowers serum progesterone or estradiol levels and/or inhibits ex vivo ovarian steroidogenesis. Results obtained from the in vitro studies demonstrated that HPTE directly inhibits progesterone formation by PMSG-primed rat ovarian theca, granulosa and luteal cells, and this decline in progesterone corresponded to a decline in P450 cholesterol side-chain cleavage activity, which converts cholesterol to pregnenolone in the pathway of progesterone and estrogen biosynthesis. In addition, the percentage decrease in progesterone production was greater than the declines in estradiol and androgen formation suggesting that HPTE may increase the estrogen/progesterone ratio. Further, this effect of HPTE action could be observed at the environmentally relevant concentrations of 10-50 nM and does not appear to be mediated through ER or AR signaling pathways. Consistent with the HPTE action in the in vitro studies, in vivo exposure to methoxychlor lowers circulating progesterone levels in the immature rats not treated with gonadotropins; however, the exposure of animals to gonadotropins leads to an ex vivo increase in progesterone production by isolated ovarian theca cells and no change in circulating progesterone levels. This suggests that the pattern of active steroids secreted ex vivo by ovarian cells in response to methoxychlor exposure differs from that observed following in vitro exposure to methoxychlor/HPTE

The effect of continuous diffusion of oxygen treatment on cytokines, perfusion, bacterial load, and healing in patients with diabetic foot ulcers

To evaluate continuous diffusion of oxygen therapy (CDO) on cytokines, perfusion, and bacterial load in diabetic foot ulcers we evaluated 23 patients for 3 weeks. Tissues biopsies were obtained at each visit to evaluate cytokines and quantitative bacterial cultures. Perfusion was measured with hyperspectral imaging and transcutaneous oxygen. We used paired T tests to compare continuous variables and independent T tests to compare healers and nonhealers. There was an increase from baseline to week 1 in TGF-β (P =.008), TNF-α (P =.014), VEGF (P =.008), PDGF (P =.087), and IGF-1 (P =.058); baseline to week 2 in TGF-β (P =.010), VEGF (P =.051), and IL-6 (P =.031); and baseline to week 3 with TGF-β (P =.055) and IL-6 (P =.054). There was a significant increase in transcutaneous oxygen after 1 week of treatment on both medial and lateral foot (P =.086 and.025). Fifty-three percent of the patients had at least a 50% wound area reduction (healers). At baseline, there were no differences in cytokines between healers and nonhealers. However, there was an increase in CXCL8 after 1 week of treatment (P =.080) and IL-6 after 3 weeks of treatment in nonhealers (P =.099). There were no differences in quantitative cultures in healers and nonhealers

Adipose tissue hyaluronan production improves systemic glucose homeostasis and primes adipocytes for CL 316,243-stimulated lipolysis

Plasma hyaluronan (HA) increases systemically in type 2 diabetes (T2D) and the HA synthesis inhibitor, 4-Methylumbelliferone, has been proposed to treat the disease. However, HA is also implicated in normal physiology. Therefore, we generated a Hyaluronan Synthase 2 transgenic mouse line, driven by a tet-response element promoter to understand the role of HA in systemic metabolism. To our surprise, adipocyte-specific overproduction of HA leads to smaller adipocytes and protects mice from high-fat-high-sucrose-diet-induced obesity and glucose intolerance. Adipocytes also have more free glycerol that can be released upon beta3 adrenergic stimulation. Improvements in glucose tolerance were not linked to increased plasma HA. Instead, an HA-driven systemic substrate redistribution and adipose tissue-liver crosstalk contributes to the systemic glucose improvements. In summary, we demonstrate an unexpected improvement in glucose metabolism as a consequence of HA overproduction in adipose tissue, which argues against the use of systemic HA synthesis inhibitors to treat obesity and T2D

Antioxidant Protects against Increases in Low Molecular Weight Hyaluronan and Inflammation in Asphyxiated Newborn Pigs Resuscitated with 100% Oxygen

BACKGROUND: Newborn resuscitation with 100% oxygen is associated with oxidative-nitrative stresses and inflammation. The mechanisms are unclear. Hyaluronan (HA) is fragmented to low molecular weight (LMW) by oxidative-nitrative stresses and can promote inflammation. We examined the effects of 100% oxygen resuscitation and treatment with the antioxidant, N-acetylcysteine (NAC), on lung 3-nitrotyrosine (3-NT), LMW HA, inflammation, TNFα and IL1ß in a newborn pig model of resuscitation. METHODS & PRINCIPAL FINDINGS: Newborn pigs (n = 40) were subjected to severe asphyxia, followed by 30 min ventilation with either 21% or 100% oxygen, and were observed for the subsequent 150 minutes in 21% oxygen. One 100% oxygen group was treated with NAC. Serum, bronchoalveolar lavage (BAL), lung sections, and lung tissue were obtained. Asphyxia resulted in profound hypoxia, hypercarbia and metabolic acidosis. In controls, HA staining was in airway subepithelial matrix and no 3-NT staining was seen. At the end of asphyxia, lavage HA decreased, whereas serum HA increased. At 150 minutes after resuscitation, exposure to 100% oxygen was associated with significantly higher BAL HA, increased 3NT staining, and increased fragmentation of lung HA. Lung neutrophil and macrophage contents, and serum TNFα and IL1ß were higher in animals with LMW than those with HMW HA in the lung. Treatment of 100% oxygen animals with NAC blocked nitrative stress, preserved HMW HA, and decreased inflammation. In vitro, peroxynitrite was able to fragment HA, and macrophages stimulated with LMW HA increased TNFα and IL1ß expression. CONCLUSIONS & SIGNIFICANCE: Compared to 21%, resuscitation with 100% oxygen resulted in increased peroxynitrite, fragmentation of HA, inflammation, as well as TNFα and IL1ß expression. Antioxidant treatment prevented the expression of peroxynitrite, the degradation of HA, and also blocked increases in inflammation and inflammatory cytokines. These findings provide insight into potential mechanisms by which exposure to hyperoxia results in systemic inflammation

Changes of Large Molecular Weight Hyaluronan and Versican in the Mouse Pubic Symphysis Through Pregnancy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)During pregnancy, the mouse pubic symphysis undergoes expansion and remodeling resulting in formation of a flexible and elastic interpubic ligament allowing passage of a term fetus. In the current study, we sought to identify and characterize components of the extracellular matrix that likely play an important role in elongation and flexibility of the interpubic ligament during parturition. Mouse pubic symphyses and interpubic ligaments collected at time points during pregnancy and postpartum were utilized to evaluate collagen type, collagen content, processing and solubility, matricellular protein, and proteoglycan expression and quantitative assessment of all glycosaminoglycans. These studies revealed increased gene expression for hyaluronan synthase 1, hyaluronan synthase 2, and versican on Gestation Day 18 as well as a decline in protein expression for the versican-degrading protease a disintegrin-like and metalloprotease with thrombospondin type 1 (ADAMTS1) motif. These findings suggest that the primary mediators of increased elongation and flexibility of the interpubic ligament at term result from increased synthesis and reduced metabolism of viscoelasticity-promoting molecules such as high molecular weight hyaluronan and versican.862Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)U.S. National Institutes of Health [P01 HD011149]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

<i>&#x2032;Stepped procedure</i> &#x2032; in laparoscopic cyst decortication during the learning period of laparoscopic surgery: Detailed evaluation of initial experiences

<b>Background:</b> We evaluated the importance and efficacy of &#x2032;<i>stepped procedure</i>&#x2032; in laparoscopic cyst decortication as an initial experience in it. <b> Materials and Methods: </b>A 36 renal cyst cases were included. The stepped retroperitonoscopic cyst excision divided into three groups. First step, doing the incisions to place the ports and expanding the retroperitoneal space with balloon distension, second step, placement of trocars and reach to the cyst, third step, aspiration and decortication of the cyst. The difficulty of the sessions was measured with the Visual Analog Scale (VAS) scoring system. Score was determined according to the difficulty of the surgical step ranging from &#x2032;0&#x2032; to &#x2032;10&#x2032;, &#x2032;0&#x2032;, too easy, &#x2032;10&#x2032; too difficult&#x2032;. The durations were measured. One-way ANOVA test was used for statistical analysis.<b> Results: </b> The<b> </b>mean age was 52.0 (20-75) years. The mean operation time was 52.0 min. The mean duration of the first step was 12.5, second, 26.0 and third, 22.5 min. The mean VAS of first step, 3.2, second, 6.0 and third, 3.6 There were only significant differences in duration time and VAS score for second step among the surgeons (<i>P</i>&#60; 0.05). <b> Conclusions:</b> Laparoscopic cyst decortication may provide gaining experience to approach the kidney laparoscopically. The side, size and localization of cysts were not found associated with the difficulty of the method

The results of low profile locking anatomical plate application for the treatment of Edinburg type 2 clavicle diaphysis fractures

BACKGROUND: Although conservative measures are the general choice of treatment for clavicle fractures; surgery is advised for displaced and multifragmentary fractures. Open reduction and osteosynthesis with a plate-screw combination are used widely as surgical treatment options