Virus removal from semen with a pinched flow fractionation microfluidic chip.

Nowadays pigs are bred with artificial insemination to reduce costs and transportation. To prevent the spread of diseases, it is important to test semen samples for viruses. Screening techniques applied are enzyme-linked immunosorbent assays and/or polymerase chain reaction, which are labor-intensive and expensive methods. In contrast to the current used screening techniques, it is possible to remove viruses physically from semen. However, existing methods for virus removal techniques have a low yield of spermatozoa. Therefore, we have developed a microfluidic chip that performs size-based separation of viruses and spermatozoa in boar semen samples, thereby having the potential to reduce the risk of disease spreading in the context of artificial insemination in the veterinary industry. As the head of a spermatozoon is at least twenty times larger than a virus particle, the particle size can be used to achieve separation, resulting in a semen sample with lower viral load and of higher quality. To achieve the size separation, our microfluidic device is based on pinched-flow fractionation. A model virus, cowpea chlorotic mottle virus, was used and spiked to porcine semen samples. With the proposed microfluidic chip and the optimized flow parameters, at least 84 ± 4% of the model viruses were removed from the semen. The remaining virus contamination is caused by the model virus adhering to spermatozoa instead of the separation technique. The spermatozoa recovery was 86 ± 6%, which is an enormous improvement in yield compared to existing virus removal techniques.

Polymorphic assembly of virus-capsid proteins around DNA and the cellular uptake of the resulting particles.

Virus-like particles (VLPs), i.e. molecular assemblies that resemble the geometry and organization of viruses, are promising platforms for therapeutics and imaging. Understanding the assembly and cellular uptake pathways of VLPs can contribute to the development of new antiviral drugs and new virus-based materials for the delivery of drugs or nucleic acid-based therapies. Here we report the assembly of capsid proteins of the cowpea chlorotic mottle virus (CCMV) around DNA into defined structures at neutral pH. Depending on the type of DNA used, we are able to create spherical structures of various diameters and rods of various lengths. In order to determine the shape dependency, the cellular uptake routes and intracellular positioning of these formed polymorphic VLPs in RAW264.7, HeLa and HEK 293 cells are evaluated using flow cytometry analysis with specific chemical inhibitors for different uptake routes. We observed particular uptake routes for the various CCMV-based nanostructures, but the experiments point to clathrin-mediated endocytosis as the major route for cell entry for the studied VLPs. Confocal microscopy reveals that the formed VLPs enter the cells, with clear colocalization in the endosomes. The obtained results provide insight in the cargo dependent VLP morphology and increase the understanding of shape dependent uptake into cells, which is relevant in the design of new virus-based structures with applications in drug and gene delivery.

Appendicular Mucocele: Possibilities and Limits of Laparoscopy. Brief Series and Review of the Literature.

INTRODUCTION: Appendicular mucocele, a cystic dilatation of the appendix, is a rare disease, but unfortunately about 1/10 of cases evolves into pseudomyxoma peritonei. METHODS: We performed a prospective study between 1 January 2010 to 31 December 2014 in order to track the incidence, symptoms, and circumstances of diagnosis, treatment and evolution of these rare tumors. RESULTS: A total of seven patients underwent curative surgery for a mucocele of the appendix: one woman and six men with an average age of 59.71 years. Clinical signs, present in two cases, were uncharacteristic. Ultrasound performed in all cases, could guide diagnosis in 5 cases. CT performed in 5 cases diagnosed only two cases. All cases were operated on: the open approach was used in four cases and a minimally invasive in three cases. We performed two right colectomies, an open appendectomy associated to anterior resection of the rectum, two laparoscopic appendectomies and two appendectomies and cecum resection with stapler, one by open approach and one by a minimally invasive approach. Intraoperative spillage of mucinous tumor did not occur in any case. The mean hospital stay was 5.7 days. Postoperative complications were present in 1 case (14.2%): wound infection. The average follow-up period was 40.28 months. (Range 6 to 48 months). No tumor recurrence or readmission, such as pseudomyxoma peritonei, has occurred. CONCLUSIONS: Appendicular mucocele is a rare entity; it can be found incidentally and it can mimic acute appendicitis, appendicular plastron or cecum tumor. Once diagnosed, surgical treatment is required for fear of perforation, tumor evolution and the emergence of the rule of complications. Laparoscopic approach in selected cases can be used, accompanied by safety measures to avoid iatrogenic perforation and peritoneal and parietal seeding.

I. Juvara (1913-1996), a surgeon for all seasons.

Ion Juvara is a creator of a genuine Romanian school of surgery. If I. Juvara had lived and worked in a country that had been the proud owner of a Pantheon, undoubtedly he would have rested there to the end of time, among the eternal ones i.e. those who triumphed against time and who had never been defeated. If the first generation of Romanian surgeons represented mainly by Thoma Ionesco (1860-1926) was educated mainly abroad, the next generation was trained professionally at the Romanian school of surgery. Despite of the hostile period he lived in, I. Juvara kept intact his freedom of thinking and opposed with all his powers abuses of any kind, always preserving his integrity, avoiding with elegance the traps of the totalitarian regime. But even so nobody can state that the professor, who came from -Moldavia, as his master, N. Hortolomei, could not say at the end of his life that he hadn't had a fulfilled professional life, as he is considered even today the greatest surgeon of Romania in the latter half of the 20th century. He was, after all, a surgeon for all seasons.

Antoine Depage's relationship with Queen Elisabeth of Belgium.

The article describes the intimate relationship between H.M. Queen Elisabeth of Belgium and the great Belgian surgeon Dr. Antoine Depage. The brilliant academic career of Depage was followed during World War I by his prominent role in the 'Océan'-hospital in De Panne at the Flemish coast. His close connection with Queen Elisabeth, working as a nurse in the hospital, resulted in an intimate friendship, which was particularly hearty when Depage lost his wife in 1915, and during his illness in 1923-1925. The letters of Depage, present in the Archives of the Royal Palace, give an insight in this intimate relationship.

Assault induced stab injuries: epidemiology and actual treatment strategy.

OBJECTIVES: To investigate and analyse epidemiology, demographics and patterns of presentation of assault induced stab injuries in a main Belgian trauma centre. To evaluate surgical management, complications and postoperative follow-up of the stab wound victims. METHODS: One hundred and seventy assaulted patients, hospitalised because of stab injuries from January 2000 to June 2007 are studied retrospectively. RESULTS: Ninety-five percent of the assaults occurred on men and the mean age of the patients was 31.1 +/- 9.7 years. Ethnic minorities represent 77% of the patients hospitalised for assaults and 26.5% of all patients proved to be under toxic influence, predominantly from alcohol (21.8%). A decline of admissions of patients with stab injuries during the period 2002-2004 is recorded. However, the incidence doubled in the next two-year period. A weekend peak and circadian rhythm is apparent with more than 20% of the patients admitted between 4 and 6 am. The trunk is most frequently stabbed (54.5%) resulting in a laparotomy rate of 51%. One third of the patients who underwent thoraco-abdominal surgery revealed diaphragmatic injuries. Seventy-five percent of the patients left the hospital in a good condition while 2.4% had neuromuscular lesions. Two patients had serious vascular complications during follow-up. During the study period, no mortality was recorded. CONCLUSIONS: Stab wounds were recorded mainly in young and middle-aged men from ethnic minorities, whereas almost 27% were under the influence of drugs. A conservative approach was generally used resulting in a low laparotomy and thoracotomy rate without affecting mortality. Neuromuscular lesions are important long-term complications of stab injuries.

Effects of calcium-fortified ice cream on markers of bone health.

UNLABELLED: Premenopausal women with low calcium intakes consumed calcium-fortified ice cream daily for 28 days. Bone markers, NTX, CTX and PTH decreased significantly by 7 days, with some evidence of a calcium dose-dependent effect. Bone marker responses were observed within 1 h of consuming ice cream. Body weight remained constant over 28 days. INTRODUCTION: Dietary calcium is important for lifelong bone health. Milk is a good source of bioavailable calcium, but consumption has declined among young adults. The aims were to determine whether calcium-fortified ice cream, a palatable source of calcium, produces significant, sustainable changes in bone turnover markers and parathyroid hormone (PTH) in premenopausal women with calcium intake below recommended UK levels. METHODS: Eighty women, ages 20-39 years (calcium intake <750 mg/day) were randomised to consume lower saturated fat/sugar ice cream containing 96, 244, 459 or 676 mg calcium daily for 28 days. Urinary NTX/Cr, serum CTX, PINP, 1,25D and PTH were measured (baseline, days 1, 7 and 28). Acute changes in CTX and PTH were measured over 5 h (n = 29 women). RESULTS: There were significant mean decreases by 7 days in NTX/Cr, CTX, PTH and 1,25D and increases in PINP (one sample t tests), with a significant dose-dependent effect on CTX analysis of covariance. Only CTX remained suppressed at 28 days. Serum CTX and PTH decreased within 1 h. Body weight did not change significantly between baseline and 28 days. CONCLUSIONS: Daily consumption of calcium-fortified ice cream by premenopausal women may significantly reduce levels of the bone resorption marker serum CTX, without stimulating weight gain. The ice cream could be incorporated into the diet to replace low-calcium snacks and thus help individuals with habitually low calcium intakes to meet recommended intakes. The 244 mg calcium preparation would provide more than a quarter of the UK daily recommended nutrient intake for premenopausal women.

Laparoscopic retrieval of gossypibomas--short series and review of literature.

UNLABELLED: Laparoscopic removal of retained abdominal foreign bodies represents an obscure domain of minimally invasive surgery. Although not an infrequent situation in general surgical practice, there are very few papers presenting laparoscopic approache in these circumstances. An iatrogenic foreign body following surgery, is a serious complication that may lead to medico-legal problem. We present our experience and a literature review. METHODS: In the last 20 years 48 patients with abdominal foreign bodies were referred to us. Out of these four gossypibomas were managed laparoscopically. RESULTS: Retained swabs represent the most common iatrogenic abdominal foreign bodies. Removal of gossypiboma present more problems in laparoscopic environment due to encapsulation and difficulties in localisation, as retained swabs unusually display radio-opaque markings. There was one conversion due to dense adhesion to the gastric wall. Postoperative recovery was uneventful in all cases. CONCLUSIONS: Gossypibomas, among abdominal foreign bodies, represent a certain reality with significant legal implications. Prevention should prevail and all efforts should be made in such respect. Laparoscopic approach is possible in selected cases (small swabs, encapsulated, no complications).

Some notes on medical liability in ancient times.

Already in ancient times did medical liability occupy mankind. Various civilizations did give their own interpretation on the subject and proposed solutions. Original writings are rare and articles concerning ancient medical liability equally are hard to find. The only relatively trustworthy sources are of legal nature and find their origin in Greek philosophy and Roman Law. At a later stage, Arabic philosophers gave a renewed view on the statements of these previous civilizations and added their own way of thinking. All these influences still reflect in our modern western way of medical acting. Some of these ancient customs concerning medical liability will be discussed in this article.

Dissection of the common carotid artery: a case report.

Carotid artery dissection is an important cause of stroke in patients under forty years of age. A dissection of the common carotid artery is relatively rare. This condition can easily be overlooked because symptoms may be lacking or may seem trivial. Carotid artery dissection occurs mostly in previously healthy patients. Urgent treatment however, by means of coumarines or antiplatelet agents, is of life-saving importance, because this disease can lead to permanent neurological deficits or even death. We report a case of a 36-year-old patient who was admitted to the intensive care unit with hemi-paralysis and acute abdomen after a period of severe vomiting. A perforated bulbar ulcer was visible on abdominal CT scan. The patient underwent laparoscopic repair of the perforation. CT scan of the brain showed an extensive area of infarction at the region of the left medial cerebral artery. The cause of the infarction was revealed by angio CT of the supra-aortic arteries, showing a floating thrombus and a possible dissection of the left common carotid artery. Under Clopidogrel therapy, the right-sided paralysis partially recovered. Ten days post-admission, an endarterectomy of the left carotid artery was successfully performed. The patient recovered well and was discharged from the ICU 28 days after admission.

Acute colonic pseudo-obstruction in vascular patients (Ogilvie syndrome).

Two patients with acute colonic pseudo-obstruction are presented. Acute colonic pseudo-obstruction, first described by Ogilvie in 1948, is characterised by signs of colonic obstruction, but without mechanical obstruction to the intestinal flow. The current hypothesis states that it is caused by an imbalance between sacral parasympathetic excitatory nerves and sympathetic inhibitory nerves, thus inducing functional obstruction. It mostly develops in hospitalised patients with a variety of medical and surgical conditions. Two such patients with extensive co-morbidities, developed progressive colon dilatation with signs of acute peritonitis, and were operated on urgently. In both patients, right hemicolectomy and postoperative medical treatment resulted in complete recovery.