Store-operated calcium entry mediates intracellular alkalinization, ERK1/2, and Akt/PKB phosphorylation in bovine neutrophils

Neutrophil's responses to G protein-coupled chemoattractants are highly dependent on store-operated calcium (Ca(2+)) entry (SOCE). Platelet-activating factor (PAF), a primary chemoattractant, simultaneously increases cytosolic-free Ca(2+), intracellular pH (pH(i)), ERK1/2, and Akt/protein kinase B (PKB) phosphorylation. In this study, we looked at the efficacy of several putative SOCE inhibitors and whether SOCE mediates intracellular alkalinization, ERK1/2, and Akt/PKB phosphorylation in bovine neutrophils. We demonstrated that the absence of external Ca(2+) and the presence of EGTA reduced the intracellular alkalinization and ERK1/2 phosphorylation induced by PAF, apparently via SOCE influx inhibition. Next, we tested the efficacy of several putative SOCE inhibitors such as 2-aminoethoxydiphenyl borate (2-APB), capsaicin, flufenamic acid, 1-{beta-[3-(4-methoxy-phenyl)propoxy]-4-methoxyphenethyl}-1H-imidazole hydrochloride (SK&F 96365), and N-(4-[3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide (BTP2) on Ca(2+) entry induced by PAF or thapsigargin. 2-APB was the most potent SOCE inhibitor, followed by capsaicin and flufenamic acid. Conversely, SK&F 96365 reduced an intracellular calcium ([Ca(2+)](i)) peak but SOCE partially. BTP2 did not show an inhibitory effect on [Ca(2+)](i) following PAF stimuli. 2-APB strongly reduced the pH(i) recovery, whereas the effect of flufenamic acid and SK&F 96365 was partial. Capsaicin and BTP2 did not affect the pH(i) changes induced by PAF. Finally, we observed that 2-APB reduced the ERK1/2 and Akt phosphorylation completely, whereas the inhibition with flufenamic acid was partial. The results suggest that 2-APB is the most potent SOCE inhibitor and support a key role of SOCE in pH alkalinization and PI-3K-ERK1/2 pathway control. Finally, 2-APB could be an important tool to characterize Ca(2+) signaling in neutrophils.     

Regulation of carbohydrate metabolism by 2,5-anhydro-D-mannitol.

In hepatocytes isolated from fasted rats, 2,5-anhydromannitol inhibits gluconeogenesis from lactate plus pyruvate and from substrates that enter the gluconeogenic pathway as triose phosphate. This fructose analog has no effect, however, on gluconeogenesis from xylitol, a substrate that enters the pathway primarily as fructose 6-phosphate. The sensitivity of gluconeogenesis to 2,5-anhydromannitol depends on the substrate metabolized; concentrations of 2,5-anhydromannitol required for 50% inhibition increase in the order lactate plus pyruvate less than dihydroxyacetone less than glycerol less than sorbitol less than fructose. The inhibition by 2,5-anhydromannitol of gluconeogenesis from dihydroxyacetone is accompanied by an increase in lactate formation and by two distinct crossovers in gluconeogenic-glycolytic metabolite patterns-i.e., increases in pyruvate concentrations with decreases in phosphoenolpyruvate and increases in fructose-1,6-bisphosphate concentrations with little change in fructose 6-phosphate. In addition, 2,5-anhydromannitol blocks the ability of glucagon to stimulate gluconeogenesis and inhibit lactate production from dihydroxyacetone. 2,5-Anhydromannitol decreases cellular fructose 2,6-bisphosphate content in hepatocytes; therefore the effects of the fructose analog are not mediated by fructose 2,6-bisphosphate, a naturally occurring allosteric regulator. 2,5-Anhydromannitol also inhibits gluconeogenesis in hepatocytes isolated from fasted diabetic rats, but higher concentrations of the analog are required.     

Clinical management of patients receiving cell‐based immunoregulatory therapy

Administering immunoregulatory cells as medicinal agents is a revolutionary approach to the treatment of immunologically mediated diseases. Isolating, propagating, and modifying cells before applying them to patients allows complementation of specific cellular functions, which opens astonishing new possibilities for gain‐of‐function antigen‐specific treatments in autoimmunity, chronic inflammatory disorders, and transplantation. This critical review presents a systematic assessment of the potential clinical risks posed by cell‐based immunotherapy, focusing on treatment of renal transplant recipients with regulatory macrophages as a concrete example.     

El osteoma en la mastoides, ¿un hallazgo incidental?

Osteoma in the mastoid is a rare benign osteogenic tumour that has been described in literature in only 137 cases. It usually appears in asymptomatic patients, although a few cases are described associated with clinical manifestations. We report three cases of mastoid osteoma: a pedunculated osteoma in the aditus ad antrum (associated with a cholesteatoma), a superficial osteoma of the mastoid surface and a sessile osteoma that progressed to the temporal lobe (associated with vertigo). A brief review of this rare entity is presented and a possible association between mastoid osteoma, cholesteatoma otitis and vertigo is posed.     

Laparoscopic extravesical transperitoneal approach for vesicoureteral reflux

INTRODUCTION: Laparoscopy may have a place in the treatment of vesicoureteral reflux. In this study, we present the results of laparoscopic extravesical transperitoneal treatment in 15 children (19 ureters) of vesicoureteral reflux. MATERIALS AND METHODS: Between January 2001 and February 2004, 15 children (11 with unilateral and 4 with bilateral vesicoureteral reflux) were treated with extravesical reimplantation (Lich- Gregoir technique) via a transperitoneal laparoscopic approach. The mean age was 48.2 months (range 12-62), and there were 14 females and 1 male. Two patients had a double total collector system associated with reflux without ureterocele. RESULTS: The mean surgical time was 110 minutes in unilateral and 180 in bilateral vesicoureteral reflux. All procedures were successfully completed laparoscopically and the reflux was corrected in all but one patient, whose grade III vesicoureteral reflux changed to grade I. We had 3 mucosal perforations without leakage. The longest hospital stay was 72 hours. After follow-up ranging 15-49 months, only one patient had urinary tract infection. CONCLUSION: Laparoscopic extravesical transperitoneal reimplantation for vesicoureteral reflux is a safe and effective approach even in bilateral simultaneous and duplex ureters, with success rates similar to the open technique, and a dramatic reduction in postoperative stay. Mucosal perforation was treated by maintaining a Foley catheter for 3-4 days postoperatively.     

Psychological care: social and family support for patients awaiting a liver transplant

Introduction

In patients awaiting a transplant, the help received from friends and/or family members is considered to be an important factor in the transplantation process. Our objective was to determine the level of social/family support for patients on the liver transplant waiting list and to determine the relationship between clinical psychopathology and the level of social/family support.

Materials and methods

The study population consisted of 70 patients on the liver transplant waiting list. We used the following instruments: (1) Medical Outcomes Study-Social Support Survey (MOS) Questionnaire. For size of the social network, four support dimensions and a global support index for emotional support, material/instrumental support, leisure/free time activities, and love/care; (2) Family Apgar Questionnaire for personal perception of family function; (3) SA-45 questionnaire of psychopathologic symptoms for somatizations, obsessive-compulsivity, interpersonal sensitivity, depression, anxiety, hostility, phobic anxiety, paranoid ideation, and psychoticism.

Results

The MOS showed that the mean size of the social network of these patients was 12 people. In these patients, social/family support was nonfunctional in 20% of the cases. By dimensions, the patients had the following percentage of nonfunctional support: 24% in emotional support; 10% material/instrumental support; 23% social relations of leisure/free-time activities; and 11% compassionate love/care support. The patients with nonfunctional support had the following associated psychopathologic symptoms (SA-45): depression (79% vs 39%; P = .008), anxiety (86% vs 46%; P = .008), hostility (43% vs 12%; P = .009), and psychoticism (14% vs 2%; P = .039) compared to functional patients. The Family Apgar showed that 27% of patients perceived a family dysfunction. These patients had greater interpersonal sensitivity-type emotional symptoms (32% vs 8%; P = .012), depression (79% vs 35%; P = .001), anxiety (79% vs 45%; P = .011), and hostility (42% vs 10%; P = .002) compared to normally functioning patients.

Conclusions

Nearly a quarter of patients on the liver transplant waiting list have social/family support that is nonfunctional, which leads to greater emotional psychopathologic symptoms that would need to be treated.