The influence of conjugation variables on the design and immunogenicity of a glycoconjugate vaccine against Salmonella Typhi.

In recent years there have been major efforts to develop glycoconjugate vaccines based on the Vi polysaccharide that will protect against Salmonella enterica Typhi infections, particularly typhoid fever, which remains a major public health concern in low-income countries. The design of glycoconjugate vaccines influences the immune responses they elicit. Here we systematically test the response in mice to Vi glycoconjugates that differ in Vi chain length (full-length and fragmented), carrier protein, conjugation chemistry, saccharide to protein ratio and size. We show that the length of Vi chains, but not the ultimate size of the conjugate, has an impact on the anti-Vi IgG immune response induced. Full-length Vi conjugates, independent of the carrier protein, induce peak IgG responses rapidly after just one immunization, and secondary immunization does not enhance the magnitude of these responses. Fragmented Vi linked to CRM197 and diphtheria toxoid, but not to tetanus toxoid, gives lower anti-Vi antibody responses after the first immunization than full-length Vi conjugates, but antibody titres are similar to those induced by full-length Vi conjugates following a second dose. The chemistry to conjugate Vi to the carrier protein, the linker used, and the saccharide to protein ratio do not significantly alter the response. We conclude that Vi length and carrier protein are the variables that influence the anti-Vi IgG response to immunization the most, while other parameters are of lesser importance.

Determination of free polysaccharide in Vi glycoconjugate vaccine against typhoid fever.

Glycoconjugate vaccines based on the Vi capsular polysaccharide directed against Salmonella enterica serovar Typhi are licensed or in development against typhoid fever, an important cause of morbidity and mortality in developing countries. Quantification of free polysaccharide in conjugate vaccines is an important quality control for release, to monitor vaccine stability and to ensure appropriate immune response. However, we found that existing separation methods based on size are not appropriate as free Vi non-specifically binds to unconjugated and conjugated protein. We developed a method based on free Vi separation by Capto Adhere resin and quantification by HPAEC-PAD. The method has been tested for conjugates of Vi derived from Citrobacter freundii with different carrier proteins such as CRM197, Tetanus Toxoid and Diphtheria Toxoid.

Preoperative PTH as a marker of risk for post-thyroidectomy hypocalcemia.

AIM: There are no common guidelines to identify the population at risk to develop hypocalcemia preoperatively or early in the postoperative course in thyroidectomized patients, therefore the authors suggest to examine the PTH value preoperatively. METHODS: We divided 391 patients in two groups according to the preoperative PTH level (normal, ≤ 72 pg/mL vs. increased >73 pg/mL). RESULTS: In 92/391 cases (23.52%) preoperative PTH was increased (mean PTH level 112.4+/-24.8 pg/mL; normal range 12-72 pg/mL). Out of these, 43 (46.7%) had hypocalcaemia postoperatively. In 18 out of the 43 patients clinical hypocalcemia also developed. The mean follow-up was of 148+/-13 days. Of the 299 patients with normal preoperative PTH, 127 (42.47%) developed postoperative hypocalcemia (mean calcium level 7.4+/-0.33 mg/dL). In 30 patients it was also clinically evident. The difference in terms of incidence of symptomatic hypocalcemia was statistically significant (increased preoperative PTH 19.5% vs. normal preoperative PTH 10.03% , P=0.036). CONCLUSION: All candidates to thyroidectomy should be investigated for preoperative PTH abnormalities.

Operative time and postoperative pain following minimally invasive video-assisted parathyroidectomy.

BACKGROUND: Mininvasive surgical techniques have been proposed to treat the patients affected by parathyroid adenoma starting by endoscopically-assisted parathyroidectomy up to video-assisted and radio-guided approaches. PATIENTS AND METHODS: Our technique, minimally invasive video-assisted parathyroidectomy (MIVAP), in accord with Miccoli's technique, has been introduced in our center since 2006 after extensive experience with MIVAT (minimally invasive videoassisted thyroidectomy). From September 2006 to October 2008 we performed 32 MIVAP on 32 patients, 21 female and 11 males with a mean age of 53.4 years (range 25-77) affected by parathyroid adenoma. Patients have been divided in two groups in chronological order: Group A included the first 15 patients, Group B the second ones. RESULTS: Mean operative time from incision to skin closure has been 47.4+/-14.2 minutes for group A and 34+/-10.3 minutes for group B (p<0.01). Postoperative pain, scored from 0 to 10 evaluated at time 0 and after 24 hours from the procedure, has been of 2.6+/-0.5 and 1.4+/-0.5 in group A (p<0.001) while in group B of 2.58+/-0.51 and 1.16+/-0.38 (p<0.001) respectively. The difference in postoperative pain was not significant between the two groups and the pain was controlled by the administration of paracetamol 1 g. On the other side, the comparison between postoperative pain in patients operated via traditional bilateral cervical exploration and MIVAP (2.61+/-0.5 vs 3.55+/-0.51 and 1.38+/-0.5 vs 2.16+/-0.61 at 0 and 24 hours respectively), was statistically significant (p<0.001) and in favour of MIVAP. CONCLUSIONS: We showed a shorter operative time between the A group and B group. 15 cases are sufficient as good learning curve if the surgeon is experienced in videoassisted neck procedure. The postoperative pain is lower in videoassisted procedure than cervical bilateral approach.

Laparoscopic treatment of sigmoid colon intussusception by large malignant tumor. Case report.

Intestinal intussusception is rare in adults, but common in children. The ileocolic or appendiceal types are more frequent compared to the colo-colic one. We report successful laparoscopic left hemicolectomy in a patient with intussusception caused by a sigmoid tumor. Abdominal CT demonstrated a colo-colic intussusception at the level of the tumoral lesion with dilation of the proximal colon. The patient underwent urgent laparoscopic oncologically radical left hemicolectomy. A 10 cm Pfannenstiel incision allowed the removal of the resected segment. The laparoscopic approach was feasible because the dilation was moderate; however, if intussusception is due to cancer, laparoscopy can be safely performed if a correct and prompt diagnosis is achieved following oncologic criteria.

Can we use ionized calcium in the evaluation of post-thyroidectomy hypocalcemia?

AIM: The postoperative hypocalcaemia has the higher incidence as complications related to thyroidectomy. METHODS: From 1 June 2006 to 30 June 2008, we examined 492 patients operated on consecutively in our unit with a total thyroidectomy for thyroid disease. We evaluated the values of ionized calcium in all the cases, matching these with the preoperative and postoperative values of serum calcium. The pre- and postoperative (24 hours after treatment) data for ionized calcium and serum calcium were examined statistically with the Student's t-test; results with a P-value <0.05 were considered to be statistically significant. RESULTS: Two-hundred-and-twenty-three of the 492 patients (45.2%) treated with total thyroidectomy had preoperative values of ionized calcium lower than 1.13 mmol/L (normal values 1.13-1.32 mmol/L), while the ionized calcium values were lower than 1.10 mmol/L in 154 of the 223 patients. The mean value of ionized calcium in all 223 cases was 1.04+/-0.07 mmol/L. The mean serum calcium value in these patients was 9.13+/-0.291 mg/dL (normal values 8.3-10.5 mg/dL). In the other 259 cases, the values of ionized calcium and serum calcium were 1.21+/-0.03 mmol/L and 9.1+/-0.29 mg/dL, respectively. In 75 cases on 223 with symptomatic hypocalcemia, the mean value of ionized calcium was 0.88+/-0.05 mmol/L, while, in the remaining 148 cases, the mean value was equal to 0.97+/-0.08 mmol/L (P<0.001). We compared this, in both groups, with the values of postoperative serum calcium; in the 75 cases with clinical hypocalcemia, the value of serum calcium was 7.32+/-0.35 mg/dL, while the value was equal to 8.4+/-0.34 mg/dL in the other cases (P<0.001). CONCLUSIONS: The values of ionized calcium must not be used as marker of hypocalcemia but must be seen as a diagnostic aid linked to others laboratory values, such as serum calcium.

Hemiclamshell incision in the treatment of mediastinal goiter.

BACKGROUND: The thyroid disease can appear in 0.16-3.3% of cases as mediastinal goiter. The treatment is difficult and requires a mediastinal approach. MATERIALS AND METHODS: We have analyzed our experience from September 1995 to September 2007 among 2,439 thyroidectomies conducted for thyroid disease; 16 cases required a sternotomy, in seven patients conducted as hemiclamshell approach (median sternotomy associated to a fourth intercostals space incision). Preoperative evaluation included otorhinolaryngology evaluation and computed tomography. All the cases were followed up to 12 months. RESULTS: For seven cases treated trough hemiclamshell, the mean age was 57.8 years; hospital stay is 5.2 days. All the patients were discharged after respiratory evaluation; two patients required a pneumological admittance to physio-kinesi-respiratory. There were no cases of hemorrhage, nerve injury, permanent hypocalcemia, and chylothorax. There was no mortality at 30 days. CONCLUSIONS: The hemiclamshell is a safe procedure to treat mediastinal goiter and permit a good exposure of subclavian vessels and mediastinal nodes.

[Severe obstruction of upper airway by cervicomediastinal goitre in patients with cardiopulmonary insufficiency]. / Grave ostruzione delle alte vie respiratorie da gozzo cervico-mediastinico inveterato in paziente cardiopneumopatica.

The symptoms of the retrosternal goitre are due to the compression especially on airway, by thyroid growth in mediastinum. We present a case of an old woman affected by chronic obstructive broncopneumopathy, atrial fibrillation and mediastinal goitre. The previous growth in mediastinum was the cause of compression on airway with unexpected worsening of respiratory function. The risk of total thyroidectomy was related to the possible tracheomalacia. The surgery improved respiratory performance.

Localization of pathological gland's site in primary hyperparathyroidism: ten years experience with MIBI scintigraphy.

BACKGROUND: Primary hyperparathyroidism (PHPT) is characterised by pathological hyperfunctioning of one or more of the parathyroid glands leading to excessive parathyroid hormone (PTH) secretion. The aim of this study was to assess the diagnostic capacity of scintigraphy with MIBI, considering the surgical findings and the level of agreement with the result of cervical ultrasonography. PATIENTS AND METHODS: In the period January 1996 to January 2006, 84 cases with PHPT were included in the study, in which scintigraphy with MIBI was used in addition to cervical ultrasonography. All were hospitalised as "short stay surgery" patients and then seen as outpatients at follow-up visits (at 7, 14 and 30 days), during which calcium, phosphorus and PTH values were measured. RESULTS: In 55 cases (65.5%), the site of the pathology was suspected on the basis of the cervical ultrasonography. Scintigraphy with MIBI was positive in 76 cases (90.4%) and negative in the other eight (9.6%). In six of these eight cases the site had been detected by the cervical ultrasonography. In the remaining two cases (2.4%), neither of these examinations gave positive diagnostic findings. In 58 patients we proceeded with the removal of a single adenoma, in 19 cases with the removal of two out of the four glands, and in seven cases with the removal of three out of the four glands, the single formations removed not showing clearly-defined macroscopic characteristics; in two of the seven patients in whom three of the four glands were removed, the preoperative diagnostic examinations had not shown any evidence of gland pathology. CONCLUSIONS: We regard scintigraphy with MIBI as a preoperative diagnostic examination that has modified the surgical approach to PHPT, inclining surgeons towards a mini-invasive surgical procedures. We consider CT, MRI and SPECT techniques to be indicated only in cases of relapse of PHPT, possibly associated with ectopic localisation of the parathyroid gland not identified in the course of previous surgical procedures.

Minimally invasive video-assisted thyroidectomy: the learning curve.

BACKGROUND: MIVAT (minimally invasive video-assisted thyroidectomy) is a recent technique that requires a learning curve. MATERIALS AND METHODS: From July 2005 to December 2006, we treated 100 from a total of 467 thyroidectomy patients with MIVAT. We divided the patients into 3 groups. The first 2 groups consisted of 25 patients each: group A (cases 1-25) and group B (26-50). We also divided patients into 2 groups based on our surgical experience: group A + B (cases 1-50) and group C (cases 51-100). RESULTS: The operative times for groups A and B were 101.7 and 84.6 min, respectively (p < 0.03); those for groups A + B and C were 91.07 and 63.06 min, respectively (p < 0.004). Complications of hypocalcemia were observed in 6 cases (4 in the first 50 cases and 2 in the second 50), and complications of nerve palsy were observed in 2 cases from group A. CONCLUSIONS: After 25 cases, we observed that the MIVAT procedure allows for a lower mean operative time and a reduction of complications.

The association between papillary carcinoma and chronic lymphocytic thyroiditis: does it modify the prognosis of cancer?

AIM: The association between papillary thyroid carcinoma (PTC) and chronic lymphocytic thyroiditis (CLT) has been reported in literature. The aim of this study was to examine this association evaluating the lesser or greater neoplastic aggressiveness. METHODS: One hundred and eighty-nine patients were examined from June 2004 to June 2007; they were divided into two groups: Group A included all the patients affected by PTC without CLT and Group B all the patients affected by PTC with CLT association. The surgical treatment was in all the patients the total thyroidectomy (TT). For the study age, sex, tumour features (dimensions, angioinvasion, capsular infiltration, multifocality and lymphnode metastases) were taken into consideration. The analysis was carried on with Student t test and chi squared analysis (statistically significant P<0.05). RESULTS: Group A included 117 patients; Group B 72 patients. No statistical difference in sex (P=0.989), age (P=0.480); tumour dimension (P=0.832). The capsular infiltration was present in 23 cases in Group A and 19 in Group B (P=0.368). The difference in average diameter was found to be 1.161+/-0.5812 and 1.485+/-1.082 cm in Group A and in Group B (P=0.290), respectively. The angioinvasion was found in 9 cases of Group A and in 3 cases in Group B (P=0.510). Multifocality was found in 35 patients in Group A and in 26 in Group B (P=0.469). CONCLUSION: The CLT may have only a minimum impact in the development of the tumour. In this study the association does not modify the aggressiveness.

Pain after minimally invasive videoassisted and after minimally invasive open thyroidectomy--results of a prospective outcome study.

BACKGROUND: Substantial modifications in surgical treatment of thyroid disease have changed the postoperative management of thyroidectomized patients. The reduction of postoperative pain permit a short-stay surgery. MATERIALS AND METHODS: We have analyzed the patients treated in our Unit from July 2006 to December 2006, with minimally invasive cervicotomy and mini-invasive video-assisted thyroidectomy. We have registered the postoperative pain applying an evaluation protocol numeric scale. The results were analyzed by t test. RESULTS: One hundred thirteen patients were divided in two groups: group A, minimally invasive cervicotomy (15 male and 46 female patients); group B, mini-invasive video-assisted thyroidectomy (9 male and 43 female patients). Upon returning to the ward, the pain scale group A vs B was 2.77 +/- 1.16 vs 2.5 +/- 0.762 (p = 0.22) .At 24 h after surgery, the pain scale in group A was 1.82 +/- 1.258 vs 1.031 +/- 0.8608 (p < 0.005). CONCLUSIONS: Both methods are safe, but mini-invasive video-assisted thyroidectomy gives not only a better cosmetic result but a reduction of postoperative pain especially at 24 h.

Giant malignant phylloides tumor: case report.

The incidence of phylloides breast tumors is less than 1% in the population affected by breast cancers. The age at higher risk is between 35 and 45 years. These neoplasms are characterized by a proliferation of mesenchimal and epithelial cells. We present a rare case of giant malignant phylloides tumor (28 x 21 x 15 cm) with a complet substitution of the gland. The clinical presentation of phylloides tumors is heterogenous; the surgical treatment is a conservative one of the gland if the neoplastic lesion size is less than 5 cm with a free margin of 1 cm and a mastectomy if the diameter of lesion is more than 5 cm. Complementary therapies still remain controversial.

Complications of percutaneous endoscopic gastrostomy: a surgical experience.

AIM: Percutaneous endoscopic gastrostomy (PEG) is a practical and safe option to place an alimentary gastrostomy. We observed that a relevant rate of complications are related to management of PEG. PATIENTS AND METHODS: We registered the patients treated in our Unit from September 1994 to December 2005. We placed 293 PEG (243 pts). Preferably using a tube 16 Fr, in 7 cases 18 Fr, in 21 cases 20 Fr and only in 3 cases 9 Fr. The median age was 69.8 years; ratio female:male 3:1. In 67 cases the treatment was carried out in not hospitalized patients. RESULTS: The incidence of late and early complications is statistically higher in hospitalized patients than at home. CONCLUSION: We think that a correct management of PEG (nurses correct information) and the experience of endoscopist and a dietician can significantly reduce the rate of complications.

Standardized laparoscopic right hemicolectomy technique for colon cancer.

AIM: We analysed our experience of laparoscopic surgical treatment vs traditional surgery of right colon cancer. METHODS: A series of 27 patients was treated from January 2001 to December 2005 out of a total of 927 surgical colorectal operations in the same period (147 with the laparoscopic approach). Inclusion and exclusion criteria are reported. We compared this group with 25 patients treated by the same surgical group with open surgery. The mean operative time, the distance from the distal margin of resection, the number of lymphnodes, the mean period of canalization and the mean hospital stay are reported. RESULTS: In the laparoscopic group, the mean operative time was 124.8+/-36.3 min vs open surgery group of 94+/-23.6 min; the distance from the distal margin was 6.7+/-3.1 cm vs 6.4+/-2.1 cm; number of lymphnodes was 15.2+/-4.3 vs 18.7+/-2.9 nodes; and canalization 1.7+/-0.9 vs 2.7+/-0.7 days. The hospital stay was 6.8+/-1.7 vs 7.2+/-0.8 days. CONCLUSIONS: We consider laparoscopic right colon resection a safe procedure but it needs good laparoscopic practice and the observance of inclusion criteria.

Efficient immunization of rhesus macaques with an HCV candidate vaccine by heterologous priming-boosting with novel adenoviral vectors based on different serotypes.

Efficient vaccination against viral agents requires a strong T-cell-mediated immune response to clear viral-infected cells. Optimal vaccination can be achieved by administration of recombinant viral vectors encoding phatogen antigens. Adenoviral vectors have attracted considerable attention as potential viral vectors for genetic vaccination owing to their favorable safety profile and potent transduction efficiency following intramuscular injection. However, the neutralizing antibody response against adenoviral capsid proteins following adenoviral vectors injection limits the success of vaccination protocols based on multiple administrations of the same adenoviral serotype. In this work, we describe efficient immunization of rhesus macaques, the preferred model for preclinical assessment, with an HCV candidate vaccine by heterologous priming-boosting with adenoviral vectors based on different serotypes. The induced responses are broad and show significant cross-strain reactivity. Boosting can be delayed for over 2 years after priming, indicating that there is long-term maintenance of resting memory cells.

Stability of immobilized D-hydantoinase from Vigna angularis and repeated cycles of highly enantioenriched production of N-carbamoyl-D-phenylglycines.

D-hydantoinase from Vigna angularis was immobilized by covalent linkage to aminopropyl glass beads. Thermal stability, resistance to storage at different pH values and temperatures of this biocatalyst were studied. This enzyme preparation was used as a catalyst to prepare enantioenriched N-carbamoyl-D-phenylglycine, N-carbamyl-D-p-fluorophenylglycine and N-carbamoyl-D-p-trifluoromethylphenylglycine, using a stirred batch reactor. Reactions were conducted during eight repeated reaction cycles, without loss of enzymatic activity or variation of the enantiomeric excess of the respective product (>98%).

[Laparoscopic colectomy in the treatment of colon cancer: prospective study]. / La colectomia laparoscopica nel trattamento del carcinoma del colon: studio prospettico.

The Authors report their experience on laparoscopic colectomy in 38 patients treated between June 2001-September 2003 in General Surgery and Organ Transplantation Department of University of Parma, Italy. The patients were 23 male and 15 female, with average age 58.4 years. All patients were studied with TC and colonoscopy performed by the surgeon. The conversion rate was 15.8% and the average hospital stay 6.9 days (range 6-15 days). The patient's general clinical conditions and the results showed that the laparoscopic colectomy is a safe surgical option.

Laparoscopic technique in enlarged cystic duct.

The risk of dislodgment of endoclips placed during laparoscopic cholecystectomy in enlarged cystic duct is higher with minor bile leak. From January 2000 to April 2004, we performed 1013 procedures; in 12 patients we have showed a enlarged duct ligated with 4 laparoscopic cholecystectomy endoclips after a 180 degrees rotation of the gallbladder during a retrograde cholecystectomy. We haven't registered complications and all the cases were discharged the first postoperative day. The method is safe and economically sound.

[Parathyroid carcinoma: clinical aspects and therapy]. / Il carcinoma delle paratiroidi: clinica e terapia.

Parathyroid carcinoma is a rare entity with an 0,5-1% of incidence on primary hyperparathyroidism (HPP) in literature. The very high values of calcium and parathormone (PTH) and the clinical aspects of hypercalcemia are the characteristics findings in these patients. We present our experience on 6 pts on 153 cases with HPP treated (3,9%). The clinical and diagnostic suspects are frequently intraoperative findings and the decision making for the surgeon is not always easy (parathyroidectomy with or without hemithyroidectomy, lymphadenectomy, surgical resection of other tissues). The mortality rate is high and we have registered three deaths at 8,14 and 64 months.