Molecular, biochemical, and proteomic analyses of transplastomic tobacco plants expressing an endoglucanase support chloroplast-based molecular farming for industrial scale production of enzymes.

The successful production of recombinant enzymes by tobacco transplastomic plants must maintain compatibility of the heterologous enzyme with chloroplast metabolism and its long-time enzyme stability. Based on previous reports, it has been taken for granted that following biolistic-transformation, homoplasticity could be obtained from the initially heteroplastic state following successive rounds of selection in the presence of the selection agent. However, several studies indicated that this procedure does not always ensure the complete elimination of unmodified wild-type plastomes. The present study demonstrates that CelK1 transplastomic plants, which were photosyntetically as active as untransformed ones, remain heteroplastomic even after repeated selection steps and that this state does not impair the relatively high-level production of the recombinant enzyme. In fact, even in the heteroplastomic state, the recombinant protein represented about 6% of the total soluble proteins (TSP). Moreover, our data also show that, while the recombinant endoglucanase undergoes phosphorylation, this post-translation modification does not have any significant impact on the enzymatic activity. Biomass storage might be required whenever the enzyme extraction process could not be performed immediately following the harvest of tobacco mature plants. In this respect, we have observed that enzyme activity in the detached leaves stored at 4 °C is maintained up to 20 weeks without significant loss of activity. These findings may have major implications in the future of chloroplast genetic engineering-based molecular farming to produce industrial enzymes in transplastomic plants.

The production of alpha/beta and gamma/delta double negative (DN) T-cells and their role in the maintenance of pregnancy.

The ability of the thymus gland to convert bone marrow-derived progenitor cells into single positive (SP) T-cells is well known. In this review we present evidence that the thymus, in addition to producing SP T-cells, also has a pathway for the production of double negative (DN) T-cells. The existence of this pathway was noted during our examination of relevant literature to determine the cause of sex steroid-induced thymocyte loss. In conducting this search our objective was to answer the question of whether thymocyte loss is the end product of a typical interaction between the reproductive and immune systems, or evidence that the two systems are incompatible. We can now report that "thymocyte loss" is a normal process that occurs during the production of DN T-cells. The DN T-cell pathway is unique in that it is mediated by thymic mast cells, and becomes functional following puberty. Sex steroids initiate the development of the pathway by binding to an estrogen receptor alpha located in the outer membrane of the mast cells, causing their activation. This results in their uptake of extracellular calcium, and the production and subsequent release of histamine and serotonin. Lymphatic vessels, located in the subcapsular region of the thymus, respond to the two vasodilators by undergoing a substantial and preferential uptake of gamma/delta and alpha/beta DN T- cells. These T- cells exit the thymus via efferent lymphatic vessels and enter the lymphatic system.The DN pathway is responsible for the production of three subsets of gamma/delta DN T-cells and one subset of alpha/beta DN T-cells. In postpubertal animals approximately 35 % of total thymocytes exit the thymus as DN T-cells, regardless of sex. In pregnant females, their levels undergo a dramatic increase. Gamma/delta DN T-cells produce cytokines that are essential for the maintenance of pregnancy.

An eight-year experience of implementing a pro-active approach to early detection of breast cancer in a community

OBJECTIVES: The objectives of this study were to determine the demographic as well as clinical profile of patients participating in the breast cancer community outreach program and determine if the Breast Cancer Community Outreach Program can successfully find breast cancer cases in the community.METHODS: In 2001, a breast cancer community outreach program was organized by the Vicente Sotto Memorial Center - Breast Center. The program components included breast cancer lay forum, breast self-examination (BSE) classes and breast clinic. Twenty seven breast cancer community outreach program were conducted in a span of 8 years.RESULTS: There were 1719 participants with a mean age of 32.5 years. The most common reason for consultation was breast check-up (79.6%), majority had normal breast examination findings (75.9%) and breast cancer was also identified in only 19 out of 1719 (1.1%). Most of these were early breast cancer.CONCLUSION: The Breast Cancer Community Outreach Program was not only vital in providing an-site breast cancer cases among women consulting at the community breast clinic.

Secondary prevention of latex allergy in children: analysis of results.

OBJECTIVE: Latex allergy has become an increasing and clinically important problem. Several recommendation for secondary preventive measures have been advised. The aims of the study were to illustrate the results of the latex-safe protocol and to evaluate in allergic patients the role of risk factors for the development of latex allergy. METHODS: Latex-safe treatment was divided into the following phases: anamnestic identification, allergologic assessment, patient selection, intervention programme, preventive medication, operating room equipment, postoperative management, patient and family training, follow-up. RESULTS: Between 1998 and 2004, 6.832 patients underwent 7.333 operations. Anamnestic and diagnostic tests showed that 26 patients had latex allergy. 44 secondary perioperative latex-safe management have been accomplished in 26 children. No allergic event or complications linked to the procedure occurred. Atopy, congenital malformations frequently associated with latex allergy and the presence of 5 or more surgical procedures were the major risk factors recognized. Six out of the 26 patients (23%) had only one risk factor (atopy). Twenty out of 26 children (77%) had several associated risk factors: 8 of them had simultaneously 9 of the 10 analysed risk factors. Our data shows that, the higher their number, the higher the gravity of the allergy. CONCLUSIONS: Although latex allergy is a limited phenomenon, it is nevertheless quite frequent within risk groups. Most patients have simultaneously many risk factors for the development of such an allergy, and the occurrence of several risk factors increases severity of the allergy. Latex-safe perioperative management offers guarantees of safety against latex allergy phenomena.

Tracheal agenesis: management of the first 10 months of life.

Tracheal agenesis is a potentially lethal congenital anomaly, appearing only at birth. We describe a newborn preterm infant who presented with immediate respiratory distress and no audible cry. There was almost complete tracheal agenesis with a very short segment of distal trachea (only two tracheal rings) arising from the anterior wall of the esophagus, before dividing into the mainstem bronchi. The anomaly was unsuspected prenatally, as the scan showed pyloric atresia and complex congenital cardiac disease. Despite the patient's difficult course, with correction of the rare-associated malformations (cardiac and gastrointestinal tract anomalies), the fact that the child is lively and neurologically normal for her age, requires that we now consider the patency of the airway and the possibility of surgical correction, in accordance with a good quality of life.

Peripherally inserted central venous catheters in preterm newborns: two unusual complications.

This report describes the case of two newborns who suffered unusual complications after peripheral insertion of a central venous catheter. In one baby a fragment of the catheter tip became embolized in a peripheral branch of the left pulmonary artery. In the other baby, the catheter perforated a peripheral branch of the pulmonary artery, giving rise to chemical pneumonitis with extensive pleural effusion. The outcome was positive for both babies. A large clinical series is necessary to establish the complications of this procedure, their prevention and management.

[Thoracoscopic management of suspected thoraco-pulmonary malignant diseases in pediatric age]. / Management toracoscopico delle patologie toraco-polmonari di sospetta natura tumorale in età pediatrica.

Recent improvements and miniaturization of instruments have encouraged a wider use of thoracoscopy and laparoscopy as a modality for diagnostic and operative procedures in pediatric age. The utility of thoracoscopy in pediatric patients with suspected thoracopulmonary oncological diseases is shown by diagnostic accuracy and, if necessary, the possibility to perform at the meantime a mininvasive surgery. We report the experience of our Institution in 16 patients with suspected thoraco-pulmonary oncological diseases and treated for this reason with thoracoscopy. Thoracoscopic is indicated in cases of suspected oncological diseases in children both for diagnosis and treatment.

[In-hospital pediatric cardiopulmonary resuscitation]. / Emergenze intraospedaliere pediatriche.

Between 0,7-3% of pediatric patients may require resuscitation during hospital stay. The physicians of the Pediatric Intensive Care Unit of the C.O.U. Anesthesia and Intensive Care-Baroncini developed a plan for the management of pediatric emergencies inside the Department of Pediatric Medical and Surgical Sciences. The plan consisted of: the drawing up of a PI 34-Procedure; the preparation and implementation of a training course for doctors and nurses; the purchase of 12 emergency-trolleys according to the Broselow Pediatric Resuscitation Measuring Tape and the implementation of a specific system for the emergency-call. Precise duty for anesthesiologists and intensivists is the emergency- planning and management, in order to diffuse the medical knowledge needed to assist patients requiring vital functions support. The management of the intra-hospital pediatric emergencies is strictly dependent on the training of the staff, based on a specific support algorithm, and specific equipment for the different ages.

Anaesthesia for laparoscopic surgery in paediatrics.

Laparoscopic surgery is an emerging procedure in the treatment of many surgical pathologies. Laparoscopy in the paediatric patient reduces surgical trauma and improves cosmetic RESULTS. Physiological changes during laparoscopic surgery are mainly related to the increased intra-abdominal pressure (IAP) associated with CO2 insufflation of the abdomen, the patient's postural modifications (head-up or head-down) and CO2 absorption and its general effects. Increases in IAP affect both ventilation and circulation. Increased IAP induces a mechanical compression of the diaphragm that reduces pulmonary compliance, vital capacity, functional residual capacity (FRC) and total lung volume. Pneumoperitoneum in children has a major impact on cardiac volumes and function, mainly through the effect on ventricular load conditions. The acute increase in IAP affects both preload and afterload, while the systolic cardiac performance remains unchanged. During anaesthesia for videolaparoscopy it is important not to exceed an intrabdominal pressure of 6 mmHg in newborns and infants and 12 mmHg in older children. In our clinical experience the respiratory, cardiocirculatory and temperature parameters have been slightly influenced during laparoscopy, but have always been maintained within the normal ranges. Laparoscopic videosurgery in newborns, infants and paediatric age group patients can be performed safely and with satisfactory clinical results.