Polystyrene nanoparticles may affect cell mitosis and compromise early embryo development in mammals.

A great interest surrounds the development of nanoparticles (NPs) for biomedical applications such as drug delivery and cancer therapy. However, the interplay between nanoscale materials and biological systems and the associated hazards have not been completely clarified yet. In this study, bovine oviductal epithelial cells (BOECs) and embryos were used as in vitro models to investigate whether cell mitosis and early mammalian embryo development could be affected by the exposure to polystyrene (PS) nanoparticles. Analysis of the karyotype performed on BOECs exposed to PS-NPs did not show chromosomal anomalies compared to the control, although more tetraploid metaphase plates were observed in the former. In vitro fertilization experiments designed to understand whether exposure to PS-NPs could affect pre-implantation development showed that incubation with PS-NPs decreased 8-cell embryo and blastocyst rate in dose-dependent fashion. The quality of the blastocysts in terms of mean cell percent blastomeres with fragmented DNA was the same in exposed blastocysts compared to controls. These results show that the exposure to PS-NPs may impair development. In turn, this may affect the rate of mitosis in embryos and yield a lower developmental competence to reach the blastocyst stage. This suggests that release in the environment and the subsequent accumulation of PS-NPs into living organisms should be carefully monitored to prevent cytotoxic effects that may compromise their reproduction rates.

Time course of carbon monoxide transfer factor after breath-hold diving.

Breath-hold divers may experience haemoptysis during diving. Central pooling of blood as well as compression of pulmonary gas content can damage the integrity of the blood-gas barrier, resulting in alveolar hemorrhage. The single-breath carbon monoxide test (DL,CO) was used to investigate the blood-gas barrier following diving. The study population consisted of 30 divers recruited from a training course. DL,CO levels were measured before diving and at 2, 10 and 25 min after the last of a series of four dives to depths of 10, 15, 20 and 30 m. When compared to pre-diving values, DL,CO values increased significantly at 2 min following diving in all subjects except one. Thereafter values progressively decreased toward baseline at 10 and 25 min in all subjects but one, while in four divers DL,CO values decreased below baseline. The early but transient increase in DL,CO levels shortly after diving supports the persistence of capillary pooling of red blood cells following emersion. Persistence at 25 min of high DL,CO values in one subject could be attributed by lung CT to extravasation of blood into the alveoli. Early or late DL,CO values >10% below baseline values suggest the presence of pulmonary edema. The relatively high prevalence of DL,CO alterations found suggests caution on the safety of breath-hold diving activities.

Giant cystic intraventricular cerebral cavernous malformation: MRI with pathologic correlation. A case report.

The purpose of this case report is to increase awareness of the spectrum of cerebral cavernous malformations (CCMs). We report the MRI findings and histological features of an uncommon case of a single giant (maximum diameter: >6 cm) cystic CCM of the left lateral ventricle occurring in a 26-year-old man who had undergone 30 Gy cranial irradiation for acute leukemia at the age of six years. Large cystic CCMs must be included in the neuroradiological differential diagnosis of intraventricular hemorrhagic cystic lesions.

Facial nerve schwannoma presenting as a mass in the middle cranial fossa: typical neuroradiological patterns for a preoperative diagnosis.

Facial nerve schwannoma is a rare primary neurogenic tumour that may originate anywhere along the VII(th) nerve course. The clinical presentation is highly dependent on the location of the lesion along the nerve course and this makes the pre-operative diagnosis difficult without radiologic examination. The most common presentation is facial palsy and even though tumours are responsible for only 5% of facial palsies, if a patient does not recover within six months a complete work-up for neoplasm is recommended. On the basis of clinical presentation and imaging characteristics radiologists should try to make a preoperative diagnosis, to help in the patient's management and possibly to plan the surgical approach. We describe the case of a successful preoperative diagnosis of facial nerve schwannoma. The aim is to describe the main CT and MRI findings which may help the radiologist to establish a correct differential diagnosis.

Physical-chemical and biological characterization of silk fibroin-coated porous membranes for medical applications.

Membranes in artificial organs and scaffolds for tissue engineering are often coated with biomimetic molecules (e.g., collagen) to improve their biocompatibility and promote primary cell adhesion and differentiation. However, animal proteins are expensive and may be contaminated with prions. Silk fibroin (SF) made by Bombyx Mori silk worms, used as a scaffold or grafted to other polymers, reportedly promotes the adhesion and growth of many human cell types. This paper describes how commercial porous membranes were physically coated with SF, and their physical-chemical properties were characterized by SEM, AFM, tensile stress analysis and dynamic contact angle measurements. The effect of the SF coating on membrane biocompatibility and resistance to bacterial colonization is also examined. The proposed technique yields SF coats of different thickness that strengthen the membranes and make their surface remarkably more wettable. The SF coat is not cytotoxic, and promotes the adhesion and proliferation of an immortalized fibroblast cell line. Similarly to collagen, SF-coated membranes also exhibit a much better resistance to the adhesion of S. epidermidis bacteria than uncoated membranes. These preliminary results suggest that SF is a feasible alternative to collagen as a biomimetic coating for 3D scaffolds for tissue engineering or bioartificial (as well as artificial) prosthesis.

New generation ceramic membranes have the potential of removing endotoxins from dialysis water and dialysate.

Poor water properties, use of concentrated bicarbonate, and biofilm growth in pipes and storage tanks often cause dialysis water and dialysate contamination with bacteria and endotoxins. High-flux dialysis with bicarbonate may favor endotoxin transfer from the dialysate into the blood exposing patients to serious short-and long-term side effects. Ultrafiltration across hydrophobic synthetic membranes effectively removes endotoxins from dialysis water by combined filtration and adsorption. However, repeated sterilization worsens the membrane separation properties,and limits their use. Ceramic membranes are generally more resistant to harsh operating conditions than polymeric membranes, and may represent an alternative for endotoxin removal. Previously, we proved that the ceramic membranes commercially available at that time were not retentive enough to ensure production of endotoxin-free dialysis water. In this paper, we investigated the endotoxin removal capacity of new generation commercial ceramic membranes with nominal molecular weight cut-off down to 1,000. In dead-end filtration, all investigated membranes produced water meeting, the European standards, or close to,when challenged with low endotoxin concentrations, but only one membrane type succeeded at high endotoxin concentrations. In cross-flow filtration, none produced water meeting the European standard. Moreover, sterilization and rinsing procedures altered the separation properties of two out of three membrane types.

Feasibility of continuous CO2 removal with hydrophilic membranes at low blood flow rates.

In the conventional treatment of acute respiratory distress syndrome (ARDS), high O2 concentrations and mechanical ventilation may damage the lung tissue. Extracorporeal membrane oxygenation limits damage, provides the needed O2 supply and improves survival of ARDS neonates, but not of adults. Hydrophilic membranes used in hemodialysis are more non-thrombogenic and biocompatible than those used in blood oxygenation, but their O2 transport capacity is not as high. In recent years, CO2 removal at low blood flow rates combined with apneic oxygenation and low frequency ventilation has proved promising in the treatment of ARDS. This approach makes O2 supply across ECMO membranes unnecessary; it also makes hydrophilic membranes candidates for extracorporeal CO2 removal to minimize anticoagulation and immune system activation. This paper reports on the in vitro capacity of hydrophilic polysulphone membranes to remove CO2 from carbonated pig blood into an oxygen-rich gas stream. Experiments were performed on clinical-size dialysis modules and their capacity to remove CO2 as a function of blood flow rate and membrane surface area was investigated. Membranes effectively removed CO2 , more so at increasing blood flow rates and membrane surface areas, at rates of up to 15% of the CO2 metabolic production rate. The specific CO2 removal rate was comparable to that of blood oxygenators equipped with microporous hydrophobic membranes. It is concluded that CO2 removal from slowly flowing blood with hydrophilic membranes is feasible.

Adhesion and function of rat liver cells adherent to silk fibroin/collagen blend films.

Collagen is often used in bioartificial livers as a biomimetic coating to promote liver cell adhesion and differentiation. Animal proteins are expensive and expose the host to risks of cross-species infection due to contamination with prions. Silk fibroin (SF) is a biocompatible protein produced by Bombyx mori silk worms and possibly an alternative to collagen. We prepared SF-collagen blend films with different SF content adherent to the bottom of standard tissue culture dishes, and characterized their surface morphology by SEM, their wettability and examined them for their capacity to support rat liver cell adhesion and metabolism. Cell metabolism was characterized by estimating the rate at which cells eliminated ammonia and synthesized urea for up to 48h of culture. SF-containing films were smooth, clear and more wettable than collagen. Cells readily adhered, formed junctions and small size aggregates on all films. As many cells adhered on SF as on collagen films. Cell adhesion to high collagen content blend films could not be reliably estimated because cells dwelt in the large cavities in the film. The effect of SF on cell metabolism differed with the investigated metabolic pathway. However, cells on SF-containing films eliminated ammonia and synthesized urea at rates generally comparable to, for urea synthesis at times higher than, that of cells on collagen. These results suggest that silk fibroin is a suitable substratum for liver cell attachment and culture, and a potential alternative to collagen as a biomimetic coating.

Combined effect of oxygen and ammonia on the kinetics of ammonia elimination and oxygen consumption of adherent rat liver cells.

Oxygen is essential for the survival of isolated liver cells and its concentration is known to affect their viability and function. Recent reports have also shown that ammonia is eliminated at a rate depending on its concentration and that high ammonia concentrations may be cytotoxic to rat liver cells. Nonetheless, little quantitative information on the effect of either metabolite on liver cell reaction kinetics is available although important to the design of bioreactors for bioartificial livers (BALs). In this investigation, we characterized the dependence of the rate of oxygen consumption (OCR), ammonia elimination (AER) and urea synthesis (USR) on ammonia concentration at physiological (i.e., 43 and 72 mmHg) and supra-physiological (i.e., 134 mmHg) dissolved oxygen tensions. To this purpose, isolated rat liver cells were cultured in adhesion on collagen in a continuous-flow bioreactor optimised for the kinetic characterisation of liver cell metabolic reactions. Rates of the investigated reactions generally increased with increasing ammonia concentrations. OCR and USR significantly increased with increasing dissolved oxygen tensions, particularly at high ammonia concentrations. The actual dissolved oxygen tension significantly influenced also OCR and USR dependence on ammonia concentration. The best-fit rate equations were used to show that, at the beginning of the treatment with a bioreactor packed with primary liver cells, high ammonia concentration in the blood may cause large hypoxic zones in the bioreactor as a result of its effect on OCR. This suggests that plasma (or blood) detoxification prior to entering the bioreactor might enhance BAL efficacy by preserving a large fraction of the available cell activity for longer times.

Propofol EEG burst suppression in carotid endarterectomy.

BACKGROUND: The results of randomized trials indicate that carotid endarterectomy, performed with a low morbidity-mortality perioperative risk, is the best therapeutic option both for patients with high-grade symptomatic and asymptomatic stenosis. Since the main operative risk is represented by embolic or hemodynamic cerebral ischemia, it appears necessary to maintain an adequate intraoperative cerebral blood flow and to carry out a meticulous endarterectomy. METHODS: On the basis of these considerations we prospectively studied a series of 100 consecutive patients operated on for high-grade carotid stenosis, by using a protocol based on: 1) an accurate selection of patients for surgery; 2) meticulous surgical technique without any shunt; 3) perioperative cerebral protection by barbiturate or propofol; 4) pre- and postoperative medical treatment of risk factors. All patients of our series performed preoperatively brain CT scan, transcranial Doppler, carotid duplex scanning, four vessel angiography, brain 99mTc-HMPAO SPECT. Eighty-two patients had symptomatic carotid stenosis ranged between 70 and 90%, 18 had carotid stenosis higher than 90%. RESULTS: In this series there have been one postoperative death due to myocardial infarction and one major stroke. CONCLUSIONS: We think that this protocol can significantly minimize risks of endarterectomy and probably maximize the benefits of surgery, also in patients with asymptomatic high-grade carotid stenosis.

Relationship between extent of pulmonary emphysema by high-resolution computed tomography and lung elastic recoil in patients with chronic obstructive pulmonary disease.

We investigated the relationship between the extent of pulmonary emphysema, assessed by quantitative high-resolution computed tomography (HRCT), and lung mechanics in 24 patients with chronic obstructive pulmonary disease (COPD). The extent of emphysema was quantified as the relative lung area with CT numbers < -950 Hounsfield Units (HU). Patients with COPD had severe airflow obstruction (FEV(1) 35 +/- 15% pred) and severe reduction of CO diffusion constant (DCO/VA 37 +/- 19% pred). Maximal static elastic recoil pressure (Pst(max)) averaged 54 +/- 24% predicted, and the exponential constant K of pressure-volume curves was 258 +/- 116% predicted. Relative lung area with CT numbers < -950 HU averaged 21 +/- 11% (range 1 to 38%). It showed a highly significant negative correlation with DCO/VA (r = -0.84, p < 0.0001), a weak correlation with FEV(1)% predicted, and no correlation with either Pst(max) or constant K. A significant relationship was found between the natural logarithm of K and the full width at half maximum of the frequency distribution of CT numbers, taken as an index of the heterogeneity of lung density (r = 0.68, p < 0.0005). We conclude that currently used methods of assessing the extent of emphysema by HRCT closely reflect the reduction of CO diffusion constant, but cannot predict the elastic properties of the lung tissue.

Morphology and metabolism of hepatocytes cultured in Petri dishes on films and in non-woven fabrics of hyaluronic acid esters.

Polymers of hyaluronic acid (Hyal) esters exhibit good tissue compatibility and are available in various geometrical configurations. These properties can be exploited for the design of innovative bioartificial liver support devices (BALSDs) using primary hepatocytes. In this paper, we report a preliminary investigation of the polymer feasibility of the ethyl and the benzyl Hyal ester in the form of films and non-woven fabrics for the in vitro culture of primary rat hepatocytes. Cell function was evaluated daily in Petri dishes with respect to the rate of ammonia elimination (AER) and urea synthesis (USR). Cells cultured in non-woven fabrics of the ethyl ester of Hyal (HYAFF7nw) exhibited an initial AER about 32% lower and synthesised urea 33% faster than that of cells on collagen films. After a week in culture, cells on collagen films retained only a minor fraction of their initial rates. Cells cultured in non-woven fabrics of HYAFF7nw retained about 62 and 44% of their initial AER and USR, respectively, and exhibited an AER approximately equal to and a USR 3.6 times greater than those of cells adherent to collagen. These results suggest that non-woven fabrics of HYAFF7nw are promising substrata for hepatocyte culture in BALSDs.

Characterization of the distribution of matter in hybrid liver support devices where cells are cultured in a 3-D membrane network or on flat substrata.

Bioreactors for liver assist tested on small animal models are generally scaled-up to treat humans by increasing their size to host a given liver cell mass. In this process, liver cell function in different culture devices is often established based on the metabolite concentration difference between the bioreactor inlet and outlet irrespective of how matter distributes in the bioreactor. In this paper, we report our investigation aimed at establishing whether bioreactor design and operating conditions influence the distribution of matter in two bioreactors proposed for liver assist. We investigated a clinical-scale bioreactor where liver cells are cultured around a three-dimensional network of hollow fiber membranes and a laboratory-scale bioreactor with cells adherent on collagen-coated flat substrata. The distribution of matter was characterized under different operating modes and conditions in terms of the bioreactor residence time distribution evaluated by means of tracer experiments and modeled as a cascade of N stirred tanks with the same volume. Under conditions recommended by the manufacturers, matter distributed uniformly in the clinical-scale bioreactor as a result of the intense backmixing (N=1) whereas axial mixing was negligible in the laboratory-scale bioreactor (N=8). Switching from recycle to single-pass operation definitely reduced axial mixing in the clinical-scale bioreactor (N=2). Increasing feed flow rate significantly enhanced axial mixing in the laboratory-scale bioreactor (N=4). The effects of design, operating mode and conditions on matter distribution in bioreactors for liver cell culture suggest that characterization of the distribution of matter is a necessary step in the scale-up of bioreactors for liver assist and when function of liver cells cultured in different bioreactors is evaluated and compared.

Membranes for endotoxin removal from dialysate: considerations on feasibility of commercial ceramic membranes.

As the quality of water in dialysis fluid varies considerably, dialysate is often contaminated by large amounts of bacteria and endotoxins. Membrane properties and operating pressures are acknowledged to give high-flux dialysis with bicarbonate the bacteriological potential to favor passage of endotoxin fragments from the dialysate into the blood stream. Therefore, a sterile dialysate will have to become a standard. Ultrafiltration across hydrophobic synthetic membranes was shown to remove endotoxins (and their fragments) from dialysis water by the combined effect of filtration and adsorption. However, each module can be used for a limited time only. Ceramic membranes may represent an alternative to polymeric membranes for endotoxin removal. In this article, we tested the capacity of different commercial ceramic membranes with nominal molecular weight cut-off down to 1,000 to retain endotoxins from Ps. aeruginosa. The tested membranes did not generally produce dialysate meeting the Association for the Advancement of Medical Instrumentation standard. When using aluminum-containing membranes, we detected aluminum leaking into the dialysate that could possibly be transported into the blood stream.

Comparative analysis of metabolism of medium- and plasma perfused primary pig hepatocytes cultured around a 3-D membrane network.

Culture media are frequently used in the evaluation of metabolical functions of hepatocytes in hybrid liver support systems (hLSS). However, media compositions differ substantially from those of plasma. Therefore, our study was designed to investigate whether current in vitro studies with medium are suitable to assess the metabolical competence of hLSS-cultures during clinical application as well as to explore whether the cell nutrition with medium provides a suitable modus operandi for stand by cultivation. Paired bioreactor cultures were perfused with either Williams' Medium E (MPB) or human plasma (PPB). About 6x108 primary pig hepatocytes (>97% viability) were cultured in three laboratory scale bioreactors designed according to Gerlach's bioreactor-concept. Different perfusion protocols were initiated after a standardised period allowing for cell attachment and reorganisation in aggregates. Whereas patterns of enzyme release were similar in both protocols the metabolical behaviour was different between MPB (anabolic state) and PPB (catabolic state). Furthermore, compared to MPB the lidocaine-MEGX-tests for PPB demonstrated lower MEGX-concentrations and a different reaction pattern. We conclude that the nutrition of hepatocytes with medium during the stand by period itself might influence the cell function and subsequently the efficacy of the hLSS-treatment during clinical application.

The effect of surface roughness of microporous membranes on the kinetics of oxygen consumption and ammonia elimination by adherent hepatocytes.

In membrane hybrid liver support devices (HLSDs) using isolated hepatocytes where oxygen is transported only by diffusion to the cells, about 15-40% of the cell mass is likely to be in direct contact with the semipermeable membranes used as immunoselective barriers: quantitative effects of membrane surface properties on the kinetics of hepatocyte metabolic reactions may also affect HLSD performance. In this paper, we report our investigation of the effects of surface morphology of two microporous commercial membranes on the kinetics of oxygen consumption and ammonia elimination by primary hepatocytes in adhesion culture. Isolated rat hepatocytes were cultured on polypropylene microporous membranes with different surface roughness and pore size in a continuous-flow bioreactor whose fluid dynamics was optimized for the kinetic characterization of liver cell metabolic reactions. Collagen-coated membranes were used as the reference substratum. Hepatocyte adhesion was not significantly affected by membrane surface morphology. The rates of the investigated reactions increased with ammonia concentration according to saturation kinetics: the values of kinetic parameters Vmax and K(M) increased as cells were cultured on the membrane with the greatest membrane surface roughness and pore size. For the reaction of oxygen consumption, Vmax increased from 0.066 to 0.1 pmol h(-1) per cell as surface roughness increased from 70 to 370 nm. For the kinetics of ammonia elimination. K(M) increased from 0.23 to 0.32 mM and Vmax increased from 1.49 to 1.79 pmol h(-1) per cell with membrane surface roughness increasing from 70 to 370 nm. Cells cultured on collagen-coated membranes consistently yielded the highest reaction rates. The Vmax values of 0.18 and 2.84 pmol h(-1) per cell for oxygen consumption and ammonia elimination, respectively, suggest that cell functions are also affected by the chemical nature of the substratum.

Cerebrovascular reactivity before and after carotid endarterectomy.

BACKGROUND: The hemodynamic relevance of internal carotid artery (ICA) stenosis often does not correlate with anatomic features, as angiographically defined. The cerebrovascular reactivity (CVR) has been advocated as a means of defining the cerebral hemodynamic impairment. METHODS: We assessed the results of pre- and postoperative CVR evaluation, using the CO2 transcranial doppler method, in 25 patients with high-grade ICA stenosis. The patients with history of stroke, evidence of cerebral CT infarction or symptoms from the contralateral circulation or the brain stem were excluded to avoid the effects of cerebral infarction on the hemodynamic studies. Statistical analysis was used to evaluate the CVR changes after carotid endarterectomy. RESULTS: Preoperative evaluation showed that CVR was generally well correlated with the degree of ICA stenosis and concomitant contralateral carotid steno-occlusion. Before endarterectomy the mean CVR value was 66.5% (moderately reduced). After surgery the overall mean value of CVR was 84.1% (normal), with a statistically significant improvement. CONCLUSION: The results of this study suggest that the CVR evaluation allows one to obtain hemodynamic information of clinical interest in the patients with ICA stenosis and that carotid endarterectomy is effective to restore the CVR in patients with cerebral hemodynamic impairment.

Congenital absence of the internal carotid artery.

We report three cases of congenital absence of an internal carotid artery (ICA), diagnosed incidentally by digital subtraction angiography. The analysis of the cases is based on the classification of segmental ICA agenesis proposed by Lasjaunias and Berenstein. Usually the patients with this rare vascular anomaly are asymptomatic; some may have symptoms related to cerebrovascular insufficiency, compression by enlarged intracranial collateral vessels, or complications associated with cerebral aneurysms. Diagnosis of congenital absence of ICA is made by skull base computed tomography (CT) scan, CT and magnetic resonance angiography, and conventional or digital subtraction angiography.

Treatment of symptomatic intracranial arachnoid cysts by stereotactic cyst-ventricular shunting.

BACKGROUND: The optimal treatment of symptomatic intracranial arachnoid cysts is controversial and different surgical approaches have been described. The aim of the treatment is to reduce the intracystic pressure. To accomplish this goal, many techniques have been proposed, each with specific limitations or failures. METHODS: Nine patients with symptomatic arachnoid cysts were treated by stereotactic cyst-ventricular shunting. One patient with suprasellar arachnoid cyst showed signs related to intracranial pressure, the remaining 8 patients complained of headache and/or drug-resistant epilepsy. All the intracranial cysts were supratentorial. The surgical procedure was performed under general anesthesia in 6 cases and local anesthesia in 3 cases. The CRW Stereotactic System (Radionics) was used. The acquisition of both target points (cyst and ventricle) was always realized by means of CT scan slices. A right precoronal burr hole was made and a silicon catheter was stereotactically inserted into the lateral ventricle. Another burr hole was subsequently performed close to the cyst; a silicon catheter was placed in the middle of the cavity in the shortest intracerebral crossing. Then both catheters were connected to a subcutaneous burr hole Ommaya reservoir. RESULTS: All patients tolerated the procedure well and the preoperative clinical signs progressively disappeared or improved. The CT scan at 1, 3, 6 and over 12 months showed progressive reduction in size of the cysts. No clinical recurrence was found at the follow-up (14-73 months). CONCLUSIONS: The successful outcome of the above cases suggests that, in carefully selected symptomatic intracranial arachnoid cysts, stereotactic cyst-ventricular shunting is likely to prove an effective operative method.