Transformation of Rhodococcus Pigment Production Hydroxylase (PPH) gene into Camelina sativa: an alternative marker for the detection of transgenic plants.

Production of transgenic plants with desired agronomic and horticultural traits has gained great importance to fulfill demands of the growing population. Genetic transformation is also a fundamental step to study basics of plant sciences. Different transformation protocols have been developed and used which are reliable and efficient. These protocols used antibiotic or herbicide resistance genes incorporated along with gene of interest to identify transformed plants from non-transformed ones. These marker genes may pose a threat to human and environment. Use of visual markers enables direct and easier observation of transformed plants with more precision. In current study a gene cassette with 'pigment production hydroxylase (PPH) gene under fiber specific promoter (GhSCFP) and downstream Nos-terminator was designed. After checking the structural and functional efficiency of codon optimized gene using bioinformatics tools, the cassette was sent for chemical synthesis from commercial source. The pigment gene cassette (PPH_CEMB), cloned in pCAMBIA-1301, was transformed into Agrobacterium through electroporation. Agrobacterium-mediated floral dip method was used to transform Camelina sativa inflorescence. After seed setting a total of 600 seed were observed for change in color and out of these, 19 seeds developed a reddish-brown coloration, while the remaining 581 seeds remained yellow. The transformation efficiency calculated on basis of color change was 1.0%. PCR analysis of leaves obtained after sowing reddish seeds confirmed the transformation of pigment production gene, while no PCR amplification was observed in leaves of plants from wild type seeds. From the results it is evident that Agrobacterium-mediated transformation of C. sativa inflorescence is very efficient and environment friendly technique not only for detection of transformed plants but also to study basic cellular processes.

E. coli expression and immunological assessment of expressed recombinant Newcastle disease virus hemagglutinin-neuraminidase protein in chickens.

Every year, the poultry industry experiences significant economic losses due to epidemics of Newcastle disease virus (NDV). Developing new vaccines by identifying and using the immunogenic hemagglutinin-neuraminidase (HN) protein can protect the poultry industry. In the present study, the full-length HN protein was expressed in Escherichia coli (E. coli) BL21 (DE3) cells, purified via affinity chromatography and detected via western blot analysis using His-specific antibodies. The purified HN protein was further evaluated in chickens to study the immune response against NDV. The successful production of HN-specific IgY proved the activity of the purified HN protein. IgY was present in the serum of immunized chickens. However, the immune response was higher in chickens immunized with purified HN protein along with complete and incomplete adjuvants than in chickens immunized with only the HN protein. Keywords: protein; Newcastle disease virus; poultry; infectious diseases; vaccines.

Role of oral vaccines as an edible tool to prevent infectious diseases.

Plants have been as medicinal mediators for centuries. Recent trends in agro-biotechnology however, improved the therapeutic roles of plants to a significant level and introduced plant-based oral vaccine which can arouse an immune response in consumer. Although conventional vaccines against infectious diseases have been administrated for years the discovery of plant-based oral vaccines can potentially replace them completely in the future. The probable limitations in conventional vaccines are found to be overcome by plant-based oral vaccines. Humans and animals will no longer be dependent upon local or systemic administration of vaccines but they will just receive the vaccines as a routine food. For the purpose, gene of interest is introduced into plant through transformation, and expression of specific antigen is obtained in plant products which are then consumed by humans or animals. Therefore, plants can serve as bioreactors or bio-factories for production of edible vaccines. A detailed overview about edible vaccines, methods for edible vaccine production, candidate bioreactors and future perspectives of edible vaccines has been summarized in current article. The future of vaccination seems to be present within plant-based vaccination system. Keywords: vaccine; edible vaccine; infectious diseases; antigen; edible crops; oral immunization.

Spontaneous bladder rupture of a urinary bladder with non-muscle invasive bladder cancer.

We describe the case of a 65-year-old man who developed spontaneous bladder rupture after picking up his suitcase from a squatting position. He was known to have non-muscle invasive bladder cancer (NMIBC), managed previously with transurethral resections and intravesical chemotherapy. CT scan showed a large amount of free intraperitoneal fluid anterior to the bladder, suggestive of a urinoma. Management was initially conservative, with insertion of urethral catheter, intravenous antibiotics and fluid resuscitation. Follow-up CT scan showed resolution of the urinoma with the patient making a full recovery after 3 weeks.

Live surgical education: a perspective from the surgeons who perform it.

OBJECTIVE: To evaluate the experience and views regarding live surgical broadcasts (LSB) among European urologists attending the European Association of Urology Robotic Urology Society (ERUS) congress in September 2012. MATERIALS AND METHODS: An anonymous survey was distributed via email inviting the participants of the ERUS congress with experience of LSB to share their opinions about LSB. The outcomes measured included; personal experience of LSB, levels of anxiety faced and the perceived surgical quality. The impact of factors, such as communication/team-working, travel fatigue and lack of specific equipment were also evaluated. RESULTS: In all, 106 surgeons responded with 98 (92.5%) reporting personal experience of LSB; 6.5% respondents noted 'significant anxiety' increasing to 19.4% when performing surgery away from home (P < 0.001). Surgical quality was perceived as 'slightly worse' and 'significantly worse' by 16.1% and 2.2%, which deteriorated further to 23.9% and 3.3% respectively in a 'foreign' environment (P = 0.005). In all, 10.9% of surgeons 'always' brought their own surgical team compared with 37% relying on their host institution; 2.4% raised significant concerns with their team and 18.8% encountered significantly more technical difficulties. Lack of specific equipment (10.3%), language difficulties (6.2%) and jet lag (7.3%) were other significant factors reported. In all, 75% of surgeons perceived the audience wanted a slick demonstration; however, 52.2% and 42.4% respectively also reported the audience wished the surgeon to struggle or manage a complication during a LSB. CONCLUSIONS: A small proportion of surgeons had significantly heightened anxiety levels and lower perceived performance during LSB, which in a 'foreign' environment seemed to affect a greater proportion of surgeons. Various factors appear to impact surgical performance raising concerns about the appropriateness of unregulated LSB as a teaching method. To mitigate these concerns, surgeons' performing live surgery feel that the operation needs to be well planned using appropriate equipment; with many considering bringing their own team or operating from home on a video link.

Live surgery at conferences - Clinical benefits and ethical dilemmas.

Live surgical broadcasts (LSBs) are becoming increasingly popular in urological conferences. These activities can provide excellent training opportunities, as they allow the audience to view an operation conducted by world-renowned surgeons, and have the ability to interact with them in real time. However, several ethical considerations have been raised with this practice, which the participating surgeons and conference organisers must appreciate and address carefully. In this article we highlight the ethical considerations related to LSBs and advise on how these should be addressed. We also present the latest recommendations made by the European Association of Urology Live Surgery Committee and discuss alternatives to LSB.

Primary carcinoid tumour of the testis.

A left radical orchidectomy was performed on a 32-year-old man presenting with a testicular mass. Histology revealed a tumour comprising of cells with monotonous nuclei and granular cytoplasm arranged in an insular pattern. Immunohistochemical expression for NSE, synaptophysin and chromogranin A were positive. No teratomatous elements were seen. Postoperatively, a staging CT and an octreotide scan were normal. Urinary 5-HIAA levels were not elevated. Based on these investigations the tumour was diagnosed as a pure primary carcinoid tumour of the testis. The patient has been followed up for 3 years with no evidence of recurrence.

Violence against nurses in hospitals: prevalence and effects.

There is abundant evidence to suggest that violence in healthcare settings, especially in emergency departments (EDs) and psychiatry, is escalating and that nurses are particularly vulnerable. The authors, therefore, investigated the prevalence and effects of violence against nurses in an ED in a general hospital setting in Kuwait. A 12-item frequency-weighted questionnaire was used to measure rates, frequency and severity of violence. The questions related to the experience of violent incidents during the past year. Seventy out of 81 nurses experienced verbal insults or threats of imminent violence and 13 were also physically attacked during the 1-year period. Sixty-seven out of 70 nurses suffered from one or more after-effects, including flashbacks, sleeplessness, fearfulness, depression or taking time off work. Violence in healthcare services often reflects the community in which service is provided. Our findings suggest that doctors experience more violence but nurses suffer from more after-effects of violence at work.

Violence towards doctors: prevalence and effects.

Violence against doctors has already been identified as a matter of particular concern. Both the severity and the psychological sequelae of violence are difficult to quantify. This study was aimed at measuring the rates and severity, and determining the effects, of violence in doctors working in a general hospital.

Violence against doctors: 1. A study of violence against doctors in accident and emergency departments.

The risk of violence directed at health care professionals in their working environment has aroused widespread concern in recent years. Clinical areas most associated with violence are accident and emergency departments, psychiatry, and general practice. Surveyed physicians reported rates of violence against them to vary from 54% to 79%. Violence, however, is difficult to quantify and there is wide variation between the severity of incidents recorded by different workers. This study was an attempt to measure the incidence and the severity of violence against doctors in accident and emergency departments in Kuwait. Eighty-seven (86%) out of 101 of our doctors reported having experienced verbal insults or imminent threat of violence; in addition, 28% had also experienced physical attacks, and 7% had experienced physical assaults likely to have caused serious or fatal injury. Similarly, out of a total of 781 violent incidents reported by our doctors, 73 involved physical attacks, and eight involved physical assaults likely to have caused serious or fatal injury.

Violence against doctors: 2. Effects of violence on doctors working in accident and emergency departments.

There is abundant evidence to suggest that doctors are increasingly being exposed to violent incidents at their workplace. The possible effects of aggression on an individual are varied and likely to depend on the severity and frequency of episodes and the perceived vulnerability to further episodes. The reported sequaelae of violent incidents towards doctors include varied psychological disturbances, and changes in behaviour, such as increasing prescribing, ongoing fear of violence at work, and poor staff morale. We investigated the effects of violence against doctors in the accident and emergency departments in Kuwait. Seventy-five (86%) out of 87 doctors exposed to violent incidents reported one or more of the symptoms consisting of: depression, reliving experience (flashbacks), insomnia, and taking 'time off'. The effects lasted for more than 4 weeks in 25, for 3-4 weeks in 17, and for 2-3 weeks in 21. The duration of symptoms was longer in doctors exposed to verbal insults or threats of imminent violence coupled with incidents involving single acts of violence. Out of a total of 101 doctors; 90 (89%) remained worried about violence at work and 72 (71%) thought training to deal with potentially violent situations would be useful.