Surgical approaches to cavernous haemangiomas of the orbit

Cavernous haemangiomas are the most common orbital masses and the second most common cause of unilateral proptosis after thyroid ophthalmopathy. We retrospectively analysed 19 patients with retrobulbar cavernous haemangiomas, 9 of whom had lateral orbitotomy to remove retrobulbar cavernous haemangiomas located superior (n=4), inferior (n=2) or lateral (n=3) to the optic nerve. Seven patients had lateral orbitotomy together with an anterior medial approach to gain access to retrobulbar cavernous haemangiomas located medially to the optic nerve in the posterior half of the orbit. An anterior approach was used in 3 patient with an anteriorly located cavernous haemangioma. We describe here the planning of surgical treatment based on the site of the lesion.     

Pathways for proton release during ubihydroquinone oxidation by the bc1 complex

Quinol oxidation by the bc(1) complex of Rhodobacter sphaeroides occurs from an enzyme-substrate complex formed between quinol bound at the Q(o) site and the iron-sulfur protein (ISP) docked at an interface on cytochrome b. From the structure of the stigmatellin-containing mitochondrial complex, we suggest that hydrogen bonds to the two quinol hydroxyl groups, from Glu-272 of cytochrome b and His-161 of the ISP, help to stabilize the enzyme-substrate complex and aid proton release. Reduction of the oxidized ISP involves H transfer from quinol. Release of the proton occurs when the acceptor chain reoxidizes the reduced ISP, after domain movement to an interface on cytochrome c(1). Effects of mutations to the ISP that change the redox potential and/or the pK on the oxidized form support this mechanism. Structures for the complex in the presence of inhibitors show two different orientations of Glu-272. In stigmatellin-containing crystals, the side chain points into the site, to hydrogen bond with a ring hydroxyl, while His-161 hydrogen bonds to the carbonyl group. In the native structure, or crystals containing myxothiazol or beta-methoxyacrylate-type inhibitors, the Glu-272 side chain is rotated to point out of the site, to the surface of an external aqueous channel. Effects of mutation at this residue suggest that this group is involved in ligation of stigmatellin and quinol, but not quinone, and that the carboxylate function is essential for rapid turnover. H(+) transfer from semiquinone to the carboxylate side chain and rotation to the position found in the myxothiazol structure provide a pathway for release of the second proton.     

Oral contraceptives highlight the genotype-specific association between serum phospholipids and activated factor VII.

The present analysis was undertaken to study the effect of oral contraceptive (OC) use on activated factor VII (FVIIa) in subjects characterized by FVII genotypes, with the further aim of evaluating the role of lipids in this pharmacological interaction. In OC users (n=42) and nonusers (n=130) of comparable age, we examined the FVII phenotypic variables (FVII coagulant activity [FVIIc], FVII antigen, and FVIIa), FVII genotypes (the 353R/Q and 5'F7 polymorphisms analyzed in combination; alleles M1/M2 and A1/A2, respectively), and a number of lipid and lipoprotein parameters: serum concentrations of total cholesterol (chol), low density lipoprotein and high density lipoprotein-chol, triglycerides, phospholipids (PhLs), apolipoprotein A1, and lipoprotein(a). PhLs, triglycerides, apolipoprotein A1, chol, FVII antigen, FVIIc, and high density lipoprotein-chol levels were shown to be statistically higher in users than nonusers. FVII levels, particularly those of FVIIa and FVIIc, were much higher in homozygotes for the A1 and M1 alleles (A11 M11), especially in OC users. A strong association was found between PhL and FVIIa: in the multiple regression analysis, women taking OCs who had elevated PhL concentrations also had very high levels of FVIIa, but only if their genotype was A11 M11. These results indicate that the increased FVII levels in OC users depend on the FVII genotype and that high PhL concentrations predict very high levels of FVIIa and FVIIc.     

Phialemonium Fungemia: Two Documented Nosocomial Cases

Two fungal isolates recovered from the blood of two immunosuppressed patients are described as Phialemonium curvatum. One patient died, while the other, who was infected with Exophiala jeanselmei at the same time, survived after successful treatment with itraconazole. Analysis of internal transcribed spacer sequences demonstrated that the isolates belonged to the same strain and that the source of infection was probably a catheter. The taxonomic position of P. curvatum is discussed, and Phialemonium dimorphosporum is considered a synonym. The in vitro inhibitory activities of six antifungal agents (amphotericin B, itraconazole, ketaconazole, miconazole, flucytosine, and fluconazole) were determined against seven isolates of Phialemonium. Except for flucytosine, all of them were remarkably effective. Phialemonium should be added to the list of potential causes of nosocomial fungemia in cancer patients.     

Interaction of Leishmania mexicana promastigotes with the plasminogen–plasmin system

The binding of human plasminogen and plasmin to the promastigote form of Leishmania mexicana was investigated. L. mexicana was capable to bind both molecules, the binding being inhibited by -aminocaproic acid. Scatchard plot analysis revealed a dissociation constant (K_d) value of 2.4±0.8 μM and 0.9±0.1×10⁴ binding sites per cell for plasminogen and a K_d value of 1.2±0.4 μM and 1.6±0.2×10⁵ binding sites per cell for plasmin. C-terminal lysine residues are involved in plasminogen binding to cells, since carboxypeptidase B treatment reduced this binding by 34%. Ligand blotting analysis showed a group of proteins, with molecular masses between 105 and 115 kDa, capable to interact with plasminogen. Zymogram analysis showed that the protease activity acquired by L. mexicana, due to the interaction with either plasminogen or plasmin, comprises an important fraction of the total protease activity at pH 7.7. Plasminogen activation by tissue-type plasminogen activator (t-PA) was enhanced by the presence of L. mexicana promastigotes. These results raise the question whether the interaction of L. mexicana with components of the fibrinolytic system is involved in the virulence of the parasite.     

Leucocyte recruitment induced by type II phospholipases A2 into the rat pleural cavity

Bothropstoxin-I (BthTX-I) and bothropstoxin-II (BthTX-II) are Lys-49 and Asp-49 phospholipases A₂ (PLA₂s), respectively, isolated from Bothrops jararacussu venom. Piratoxin-I (PrTX-I) is a Lys-49 PLA₂ isolated from Bothrops pirajai venom. In this study, the ability of BthTX-I, BthTX-II and PrTX-I to recruit leucocytes into the rat pleural cavity and potential mechanisms underlying this effect were investigated. Intrapleural injection of either BthTX-I or PrTX-I (10–100 μg/cavity each) caused a significant leucocyte infiltration at 12 h after injection. The maximal cell migration was observed with the dose of 30 μg/cavity (14.9±15.5 and 17.6±1.6×10⁶ cells/cavity, respectively). Leucocyte counts consisted mainly of mononuclear cells, but significant amounts of neutrophils and eosinophils were also observed. Intrapleural injection of BthTX-II (10–100 μg/cavity) caused a marked leucocyte infiltration at 6 and 12 h after injection. The maximal response was observed with the dose of 100 μg/cavity (57.3±3.4×10⁶ cells/cavity, 6 h). The leucocyte counts were mainly composed of neutrophils and mononuclear cells. The treatment of either BthTX-I (30 μg/cavity, 12 h) or BthTX-II (30 μg/cavity, 6 h) with the PLA₂ inhibitor p-bromophenacyl bromide (p-BPB) had no effect on the total and differential leucocyte counts induced by these proteins. The same treatment partially reduced the PrTX-I-induced pleural leucocyte infiltration. In rats depleted of the histamine and 5-hydroxytryptamine (5-HT) stores by chronic treatment with compound 48/80, the total leucocyte counts in response to BthTX-I, BthTX-II and PrTX-I was not significantly affected compared to control animals. In addition, BthTX-I, BthTX-II and PrTX-I (100 μg/ml each) significantly degranulated pleural mast cells in vitro leading to the release of [¹⁴C]5-hydroxytryptamine ([¹⁴C]5-HT). p-BPB and heparin (50 IU/ml) significantly reduced the [¹⁴C]5-HT release induced by these PLA₂s. Our results demonstrate that BthTX-I, BthTX-II and PrTX-I recruit leucocyte into the pleural cavity of the rat by mechanisms unrelated to enzymatic activity and pleural mast cell degranulation.     

Asymmetrically increased femoral version with high prevalence of moderate and severe femoral anteversion in unilateral Legg-Calvé-Perthes disease

PurposeTo determine and stratify femoral version in Legg-Calvé-Perthes disease (LCPD), and to compare the femoral version between the LCPD hip and the contralateral unaffected hip.MethodsWe performed a retrospective review of 45 patients with unilateral LCPD who had available CT scan through the hips and knees between January 2000 and June 2017. There were 34 (76%) male cases with a mean age of 14 years (sd 4.69). Two independent readers measured femoral version on the affected and the unaffected contralateral femur. Femoral version was classified as follows: severely decreased version (< 10°); moderately decreased (10° to 14°); normal femoral version range (15° to 20°); moderately increased (21° to 25°); and severely increased version (> 25°).ResultsLCPD hips had predominantly increased femoral version (38% severely increased anteversion, 24% moderately increased anteversion), while 51% of the contralateral unaffected hips had normal femoral version (p < 0.001). LCPD hips had higher mean femoral version than the contralateral, unaffected side (mean difference = 13^o; 95% confidence iterval 10^o to 16^o; p < 0.001). As the version of the affected hip increased, so did the discrepancy between sides. No effect of sex on the LCPD femoral version was detected (p = 0.34).ConclusionThis study included a selected group of patients with unilateral LCPD and available CT scans obtained for surgical planning. The femoral version was asymmetric, with a high proportion of excessive anteversion observed at later stages of disease in the affected hips. Future studies will be necessary to determine the pathogenesis of increased femoral version associated with LCPD.Level of EvidenceLevel IV, retrospective study.     

Effects of the Manufacturing Methods on the Mechanical Properties of a Medical-Grade Copolymer Poly(L-lactide-co-D,L-lactide) and Poly(L-lactide-co-ε-caprolactone) Blend

Biocompatible and biodegradable polymers represent the future in the manufacturing of medical implantable solutions. As of today, these are generally manufactured with metallic components which cannot be naturally absorbed within the human body. This requires performing an additional surgical procedure to remove the remnants after complete rehabilitation or to leave the devices in situ indefinitely. Nevertheless, the biomaterials used for this purpose must satisfy well-defined mechanical requirements. These are difficult to ascertain at the design phase since they depend not only on their physicochemical properties but also on the specific manufacturing methods used for the target application. Therefore, this research was focused on establishing the effects of the manufacturing methods on both the mechanical properties and the thermal behavior of a medical-grade copolymer blend. Specifically, Injection and Compression Molding were considered. A Poly(L-lactide-co-D,L-lactide)/Poly(L-lactide-co-ε-caprolactone) blend was considered for this investigation, with a ratio of 50/50 (w/w), aimed at the manufacturing of implantable devices for tendon repair. Interesting results were obtained.     

Glucoamylase activity from the thermophilic fungus Scytalidium thermophilum. Biochemical and regulatory properties

A glucoamylase activity produced by the thermophilic fungus Scytalidium thermophilum was purified 6.8‐fold by ion exchange chromatography. The protein exhibited a molecular mass of about 86 kDa (7% PAGE and SDS‐PAGE), or 68.5 kDa (Bio Sil SEC‐400 FPLC). The pI of the enzyme was 8.4. Optima of pH and temperature, with starch or maltose as substrates were, 6.5/60 °C and 5.0/55 °C, respectively. The enzyme had a half‐life of 22 min at 55 °C with starch as substrate, and it was fully stable with maltose. Maltase activity was activated by 10 mM Ba⁺⁺ (7.8%); Mn⁺⁺ (12.4%) or Mg⁺⁺ (28%). The enzyme contained approximately 25.5% carbohydrate. K_m and V_max values for starch and maltose were 0.28 mg/ml, 67.2 U/mg protein and 1.40 mg/ml, 5.61 U/mg protein, respectively. The products of hydrolysis of starch, detected by thin layer chromatography, showed only glucose after 30 min, indicating a glucoamylase activity. The amino terminal sequence of the purified protein showed 93% homology with a glucoamylase activity purified from Humicola grisea var. thermoidea.