Transporting bovine oocytes in a medium supplemented with different macromolecules and antioxidants: Effects on nuclear and cytoplasmic maturation and embryonic development in vitro.

We investigated whether supplementing the medium used to transport bovine oocytes with different macromolecules [foetal calf serum (FCS) or bovine serum albumin (BSA)] or a mixture of antioxidants (cysteine, cysteamine and catalase) affects their nuclear and cytoplasmic maturation and thereby affects their subsequent embryonic development and cryotolerance. Oocytes were transported for 6 hr in a portable incubator and then subjected to standard in vitro maturation (IVM) for 18 hr. The oocytes in the control groups were cultured (standard IVM) for 24 hr in medium containing 10% FCS (Control FCS) or 10% FCS and the antioxidant mixture (Control FCS+Antiox). The intracellular concentrations of reactive oxygen species (ROS) at the end of IVM period were lower in the oocytes subjected to simulated transport in the presence of a macromolecular supplement or the antioxidant mixture than that of the control group (FCS: 0.62 and BSA: 0.66 vs. Control FCS: 1.00, p < .05; and Transp: 0.58 and Transp Antiox: 0.70 vs. Control FCS: 1.00, p < .05). After IVM, the mitochondrial membrane potentials of the transported oocytes were lower than those of the non-transported oocytes (FCS: 0.41 and BSA: 0.57 vs. Control FCS: 1.00, p < .05; and Transp: 0.48 and Transp Antiox: 0.51 vs. Control FCS: 1.00 and Control Antiox: 0.84, p < .05). The blastocyst formation rates (36.9% average) and the re-expansion rates of vitrified-warmed blastocysts (53%, average) were unaffected (p > .05) by the treatments. In conclusion, supplementing the medium in which bovine oocytes are transported with antioxidants or different macromolecules did not affect their in vitro production of embryos or their cryotolerance.

Health and security risks of humanitarian aid workers during field missions: Experience of the International Red Cross.

BACKGROUND: Humanitarian disaster environments put relief workers at risk, both physically and mentally. Research looking at the growing humanitarian workforce's health and working conditions is lacking. METHODS: A questionnaire was presented to ICRC (International Committee of the Red Cross) workers during debriefing between 2016 and 2018. Participants were queried about their experience abroad including security incidents (the primary objective), health, and malaria prevention. Descriptive statistics and multivariate logistic regressions were used for data analysis. RESULTS: Security incidents were reported by 12% (95/796) of expatriates, mainly armed threats and attacks (83%, 79/95). Accidents or injuries occurred for 7.5% (60/796), of which 42% (25/60) were due to sporting activities and 22% (13/60) to road accidents. Many engaged in risky behaviours (15%, 119/795), such as driving too fast or inebriated (37%, 44/119) and/or engaging in unsafe sex, (unfamiliar partner, no condom) (71%; 84/119). Compliance to malaria prophylaxis was low 43% (113/263). Over 40% (324/795) found the mission more stressful than expected. Almost a third (28%, 219/796) reported worsened health on their return. CONCLUSIONS: Humanitarian aid workers display a worsening of their health when on mission due to violence, accidents and risky behaviours. To preserve their staff's wellbeing, humanitarian organizations should pay attention to medical history, pre-travel training specific to assignment, and continuous medical and psychological support.

Effects of epigallocatechin-3-gallate (EGCG) on in vitro maturation and fertilization of porcine oocytes.

The beneficial properties of green tea and especially of its principal active polyphenol, epigallocatechin-3-gallate (EGCG), have led to an increased demand for dietary supplements with highly enriched EGCG concentrations. In order to investigate the possible reproductive-related consequence of EGCG supplementation, the effects of this catechin on in vitro maturation (IVM) and fertilization (IVF) of oocyte, using the pig as experimental model, were examined. In the first series of experiments EGCG, at concentrations ranging from 0 to 25 microg/ml, was added during in vitro maturation of pig oocytes. EGCG had no effect on nuclear maturation of pig oocytes and on fertilization traits considered after IVF at any of the doses tested. By contrast, a significant (p<0.05) decrease in the number of embryos that developed to blastocysts following parthenogenetic activation was recorded when 25 microg/ml EGCG was added to IVM medium; in addition this catechin concentration significantly (p<0.05) inhibited progesterone production by cumulus cells after 48 h of culture. When induction of sperm capacitation was performed in presence of EGCG, a significantly lower percentage of spermatozoa showing a Hsp70-capacitated pattern and a significant reduction of sperm H(2)O(2) production were evident at a concentration of 25 microg/ml EGCG (p<0.05). During gamete coincubation EGCG reduced, in a dose response manner, the number of reacted spermatozoa suspended in fertilization medium and increased the number of sperm bound to ZP. Supplementation of 10 microg/ml EGCG during IVF significantly increased the fertilization rate while higher EGCG concentrations (25 microg/ml) decreased the percentage of fertilized oocytes (p<0.05). In conclusion, our data suggest that high EGCG concentrations could affect in vitro maturation and fertilization in pig; it cannot be totally excluded that excessive EGCG concentrations could induce reproductive-related consequences also in vivo.

IIIB-T4 non-small cell lung cancer: indications and results of surgical treatment.

AIM: T4 non-small cell lung cancer (NSCLC) is commonly considered a contraindication to surgery, indeed chemo-radiotherapy achieves a poor survival rate. We have reviewed our experience with T4 NSCLC patients who underwent surgery with the aim of debating the indications and results of surgical treatment in this highly selected group of patients. METHODS: We investigated a cohort of 60 patients, 49 men and 11 women, who underwent surgery for NSCLC from January 1998 to December 2002 and whose pathological staging was T4N0-2M0. The median age was 65 years (range 46-82). The tumors were classified T4 for the following reasons: intralobar satellite metastasis in 24 cases, direct mediastinum invasion in 18 cases, malignant pleural effusion in 7 cases, involvement of the superior vena cava in 4 cases, marginal invasion of the vertebral body in 3 cases, involvement of the carena in 3 cases and invasion of the left atrium in 1 case. Thirteen patients had undergone neo-adjuvant chemotherapy while 39 underwent adjuvant therapies. RESULTS: Thirty-two patients resulted with N0 disease, 5 with N1 and 23 with N2 disease. Forty patients relapsed (27 systemic and 13 local relapses). The mean survival was 20 months. Of the examined parameters only metastatic nodal involvement was significantly associated with a worse prognosis (P=0.007). CONCLUSION: Surgery for T4 NSCLC may be effective in those patients without mediastinal (N2) lymph node involvement. The prognosis was neither affected by the subtype of T4 tumor nor by the use of adjuvant therapies and/or neoadjuvant chemotherapy but only by the N status. In the preoperative work-up, every possible effort should be made to achieve a careful evaluation of lymph-nodal status (primarily by mediastinoscopy and video operative staging).

Role of leptin on growth hormone and prolactin secretion by bovine pituitary explants.

Leptin is an important hormone regulating nutritional status in humans and animals. Its most relevant activity is at the hypothalamic level, where it modulates food behavior, thermogenesis, and secretion of several pituitary hormones. The exact mechanisms underlying these processes are unclear. The purpose of this study was to verify whether leptin could modulate growth hormone (GH) and prolactin (PRL) secretion acting directly on bovine pituitary cells. Adenohypophyseal explants were cultured with different concentrations of leptin (50, 250, and 500 ng/mL); GH and PRL concentrations in culture media were determined by RIA. On tissues treated with 250 ng/mL of leptin, GH and PRL mRNA, as well as protein content, were estimated by reverse transcription-PCR and Western immunoblotting, respectively. Concentrations of GH in culture media containing 250 and 500 ng/mL of leptin were significantly higher than in controls: 1,063.5 +/- 141.2 (mean +/- SEM) and 1,018.8 +/- 88.4 vs. 748.9 +/- 74.0 ng/mg of tissue, respectively, after 1 h of treatment. Prolactin concentrations were significantly higher in culture media containing 50, 250, and 500 ng/mL of leptin than in controls after 2 h of treatment (547.1 +/- 50.3, 547.5 +/- 58.8, and 577.0 +/- 63.7 vs. 406.8 +/- 43.9 ng/mg of tissue, respectively). Tissues cultured with 250 ng/mL of leptin had significantly higher GH mRNA and lower GH protein content than controls (389.7 +/- 17.9 vs. 289.7 +/- 16.7; 1,601.5 +/- 90.1 vs. 2,212.7 +/- 55.6 arbitrary units, respectively) after 5 h of treatment. In contrast, no significant differences were found for PRL mRNA and protein content, possibly because of a delay in the leptin stimulation of PRL secretion. The results suggest that GH and PRL secretion in bovine pituitary explants can be directly regulated by leptin.

[Urinary 1-naphthol in the general population of Umbria]. / 1-Naftolo urinario nella popolazione generale umbra. Risultati preliminari.

INTRODUCTION: Naphthalene, the most volatile polycyclic aromatic hydrocarbon (PAH), was recently classified as possible human carcinogen by International Agencies for Research on Cancer Humans may be exposed to naphthalene from a wide variety of sources, including occupation, environment, personal habits. We assessed urinary excretion of 1-naphthol (1-NAF), biomarker of naphthalene exposure, in non-occupationally exposed subjects. MATERIALS AND METHODS: Urinary 1-NAF, 1-hydroxypyrene (1-OHP), biomarker of exposure to pyrene and cotinine, biomarker of smoking habits, were measured in 104 adults (53 men, 51 women). RESULTS: 1-NAF concentrations overlapped in males and females (median: men 0.35 Microg/g creat; women: 0.46 microg/g creat). Median concentration of 1-NAF was 6-fold higher in smokers compared to nonsmokers (respectively, 7.7 microg/g creatinine vs 1.3 microg/g creatinine). Between smokers, urinary cotinine was positively correlated to 1-naphthol (rho: 0.69; p < 0.01) and 1-OHP (rho: 0.53; p < 0.01). Higher 1-OHP concentrations were found in smokers (median: smokers 0.16 microg/g creatinine, not-smokers 0.05 microg/g creatinine;). CONCLUSIONS: In our study population, we found that 1-NAF excretion is much higher as compared to 1-OHP excretion. This is due to the ubiquitous presence of naphthalene in the environment. Smoking considerably increase the exposure to naftalene.

Is less also better? A single-institution experience on treatment of early stage Malignant Pleural Mesothelioma.

OBJECTIVES: No clear evidence of which surgical procedure should be performed for early stage mesothelioma is available to date. We analyzed our 10-year experience in the treatment of early stage mesothelioma with surgery and Hyperthermic IntraTHOracic Chemotherapy. METHODS: We retrospectively analyzed all cases of histologically proven epithelioid or biphasic IMIG stage I and II mesothelioma that we operated between 2005 and 2014. We performed an open pleurectomy and partial decortication of any visible lesion on the visceral pleura in all cases and both diaphragm and pericardium were always spared; Hyperthermic IntraTHOracic Chemotherapy was ran using Cisplatin 80 mg/m2 and Doxorubicin 25 mg/m2 at a target temperature of 42.5 °C for 60 min. RESULTS: We operated on 26 patients (23 male and 3 female); epithelioid tumor was diagnosed in 23 cases. Twelve patients were in IMIG stage I and 14 in IMIG stage II; median overall survival for all patients, stage I and II were 35.6, 46 and 23 months respectively and disease free survival was 18, 18 and 16 months respectively. Our results for stage I were better than those reported in literature and were similar for stage II. We observe no 30- and 90- mortality and the rate of severe complication (all CTCAE stage 3) were 30%; the median postoperative stay was 7.5 days. CONCLUSIONS: Our lung sparing approach for the treatment of pleural mesothelioma in early stages allows promising long term outcomes with a complete sparing of pulmonary and diaphragmatic function. Larger studies are needed to confirm our good results.

Thymomas: clinical-pathological correlations.

AIM: Since World Health Organization (WHO) histologic typing of tumors of the thymus publication in 1999 only a few studies correlated this classification with the clinical features of the patients. We present the results of a retrospective analysis on patients, operated on for a thymoma, whose specimens were available, to compare the WHO thymoma histologic classification to the clinical behavior of the tumors. METHODS: The specimens of 69 patients, who underwent surgical treatment between 1983 and 1998, were analyzed, comparing the clinical features of the patients and the hystological typing of the neoplasm, according to the WHO classification. A survival analysis of clinical and pathological prognostic factors was carried out. RESULTS: The incidence of thymus-related syndrome was related to the histological subtype and increases progressively from A to B3, while in C subtype the incidence was nihl. With a mean follow-up of 108 months (range 54-239 months), we experienced 6 intrathoracic recurrencies, 3 of those were intrapleuric and 3 mediastinal. At the last follow-up, 52 patients were alive; 1 with disease. Five deaths were related to the tumor (2 mediastinal and 3 intrapleuric relapses). Actuarial five-year and ten-year survival was 95% and 88.9%. Because of the absence of deaths related to thymomas in most samples it was not possible to perform a comparison among different histological types and different clinical stages. CONCLUSIONS: The WHO histologic classification seems to correlate with the incidence of thymus related syndromes and the clinical stage of Masaoka. Despite the higher incidence of recurrences in type B3 and C thymoma the WHO classification did not prove to be a prognostic factor.

Viability and DNA fragmentation in differently sorted boar spermatozoa.

Sperm cell defense against DNA damage relies on two factors: the tight packaging of chromatin, based on condensation and substitution of histones with protamines, and the antioxidant agents present in seminal plasma. These defenses are extremely important as mature sperm is unable to repair DNA damage and even if a successful fertilization occurs, embryo undergoes apoptosis at the time of genomic activation. Sex-sorting exposes spermatozoa to stress sources such as high pressure, laser beam and electrical charge. The aim of this work was to determine how sorting procedures affect viability and DNA integrity in boar spermatozoa, by using the newly developed Sperm-Sus-Halomax. Four sperm populations were considered: CONTROL (no treatment), REAL (sex-sorted semen), BULK (semen sorted without sex separation) and NO LASER (semen only exposed to the high pressure, but including also cells normally discarded from sex-sorting). A significantly (P=0.019) lower viability in NO LASER (64.71%) than in CONTROL (78.6%) and REAL (80.5%) groups was found; this was accompanied by a significantly (P=0.001) higher DNA fragmentation index (DFI) in NO LASER group (6.86%) respect to CONTROL (3.30%) and REAL (3.42%) groups. BULK group did not show any difference in viability or DFI as compared to the other groups. In conclusion, we may believe that sex-sorting procedure as a whole does not affect either viability or DFI and that shear mechanical forces are a relevant source of DNA damage for sorted semen.

A strategy for thoracoscopic resection of small pulmonary nodules.

BACKGROUND: Preoperative procedures are often necessary to localize pulmonary nodules during thoracoscopic resection in order to reduce the necessity of resorting to thoracotomy. The aim of this report is to describe the strategy we developed to limit preoperative techniques without reducing the thoracoscopic success rate of localization. METHODS: Between January 2000 and December 2003, 183 patients underwent video thoracoscopic resection of small pulmonary nodules. The patients were divided into two groups on the basis of the radiological features of the nodule. The subjects in group 1 were operated on directly, and endothoracic ultrasonography was performed when necessary. The subjects in group 2 underwent preoperative radionuclide labeling of the nodule. RESULTS: In group 1, 112 out of 119 nodules (94%) were localized. Twenty-five out of 32 lesions, neither visible nor palpable, were found by endothoracic ultrasonography. In group 2, we localized 62 out of 64 nodules (97%). CONCLUSIONS: Currently, we cannot completely avoid preoperative labeling techniques for thoracoscopic resection of small pulmonary nodules. However, correct patient selection may limit this necessity, without an increased conversion rate to thoracotomy, if endothoracic ultrasonography is available.

Leptin does not seem to influence glucose uptake by bovine mammary explants.

Leptin, a protein produced and secreted by adipocytes, is know to regulate food intake and whole-body energy metabolism, but knowledge about its possible effect in bovine mammary gland is scarce. Leptin may be involved in the regulation of glucose transport even though this effect at the tissue level remains controversial. Once uptaken by the mammary gland, glucose is utilised in several ways but the majority, about 60-70%, is drained for lactose synthesis. This study was aimed at investigating the effect of leptin on glucose regulation in bovine mammary gland. We have examined the effects of leptin on the expression of GLUT1 mRNA, pyruvate kinase (PK) as well as glucose-6-phosphate dehydrogenase (G6PDH) activity. Treatment of mammary gland explants with recombinant leptin did not influence glucose assimilation, pathway transport (GLUT1 mRNA) and glucose metabolism (PK and G6PDH) in this tissue. The results from this study seem to exclude an involvement of leptin in glucose uptake and metabolism in bovine mammary gland.

Effect of staining and sorting on boar sperm membrane integrity, mitochondrial activity and in vitro blastocyst development.

The objective of this study was to determine the effects of staining with Hoechst 33342 and of the entire sorting procedure on boar sperm membrane integrity (using Annexin-V/PI), mitochondrial activity (using JC-1/SYBR/PI) and blastocyst development in vitro; the effect of storage at 17 degrees C for 24h prior to Hoechst staining and sorting was also investigated. The Hoechst staining and the whole sorting procedure reduced the percent of live spermatozoa in both fresh (day 0) and stored (day 1) semen, as determined by both assays; nevertheless, there was no increase in live sperm cells showing signs of early damage (Annexin-V positive, propidium negative), whose percentages remained nearly zero. The majority of Annexin-V positive cells were propidium positive, therefore dead. JC-1 staining evidenced a correlation between mitochondrial activity and viability. However, a significant difference between viable sperm cells and sperm cells with active mitochondria was detected in control and stained sperm, whereas almost all viable sorted spermatozoa had active mitochondria. No significant differences in the in vitro produced blastocysts both on day 0 and 1 were observed. In conclusion, despite the damages induced by sorting procedures, semen sorted as fresh or after storage at 17 degrees C can be successfully used for in vitro production of pig embryos.

Surgical approaches to membranous tracheal wall lacerations.

BACKGROUND: Smaller postintubation tracheal tears are often misdiagnosed and, when recognized, they are effectively managed in a conservative fashion. Large membranous lacerations, especially if associated with important manifestations, require immediate surgical repair. We report our experience over the past 7 years. METHODS: From 1993 to 1999, 11 patients with a postintubation posterior tracheal wall laceration were treated in our institution. One patient was male and 10 were female, with a mean age of 68 years. Ten patients underwent orotracheal intubation under general anesthesia for elective surgery, 4 of whom were treated with a double-lumen selective tube. One patient underwent emergency intubation because of anaphylactic shock. In 9 cases the tracheal tear was promptly repaired, by way of a thoracotomy in 4 and by way of a cervicotomy and longitudinal tracheotomy in 5. In 2 cases the tear was small and was consequently managed conservatively. RESULTS: All surgical procedures proved effective in repairing the laceration, and there was no mortality or morbidity in the perioperative period. Early and late endoscopic follow-up showed no signs of tracheobronchial stenosis. CONCLUSIONS: When repair of membranous tracheal laceration is required, the surgical approach should be through a thoracotomy if the tear involves the distal trachea, a main stem, or both, and through a cervicotomy when the laceration is located in the proximal two thirds of the trachea. Performing a longitudinal tracheotomy to reach and suture the posterior tracheal wall is a reliable, quick, and safe procedure, and it avoids lateral and posterior dissection of the trachea.

Combined Nd-YAG laser/HDR brachytherapy versus Nd-YAG laser only in malignant central airway involvement: a prospective randomized study.

BACKGROUND: Laser debulking and prosthetic stents are useful modalities in the palliative treatment of initial inoperable or recurrent lung cancer. Recently, endobrochial brachytherapy was introduced to extend the duration of palliation and reduce the number of endoscopic treatments. This trial compares Nd-YAG laser alone and associated to high dose rated (HDR)-brachytherapy. PATIENTS AND METHODS: From 1995 to 1998, 29 consecutive patients, with non-small cell lung cancer (NSCLC) and central airway involvement, were randomized in two groups: group 1 (15 patients) received Nd-YAG laser only; group 2 (14 patients) underwent a combined Nd-YAG laser/ HDR brachytherapy treatment. RESULTS: There was no mortality or morbidity related to the treatment. The period free from symptoms was 2.8 months for group 1 and increased to 8.5 months in group 2 (P<0.05). The disease's progression free period grew from 2.2 months of group 1 to 7.5 months of group 2 (P<0.05) and the number of further endoscopic treatment reduced from 15 to 3 (P<0.05). CONCLUSION: The results confirm the potential of brachytherapy to prolong relief from symptoms, lessen disease progression and reduce costs of treatment. A detailed analysis is presented of both groups.

Postintubation tracheal tear repair by cervicotomy and longitudinal tracheotomy.

BACKGROUND: Membranous tracheal lacerations are a serious complication of endotracheal intubation. Smaller tears are often better managed with a conservative treatment. Larger ruptures, especially when associated with important manifestations, need an early surgical repair. METHODS: In the last 3 years, three female patients with a posterior tracheal wall laceration, related to endotracheal intubation, underwent surgical procedure in our institution. All tracheal tears were repaired with a running suture through a small cervical collar incision and longitudinal tracheotomy. RESULTS: All surgical procedures were effective and lasted less than 1 hour. Patients were discharged on average after 5 days. Endoscopic follow-up showed a perfect repair of the tear without signs of tracheal stenosis. CONCLUSIONS: This is a reliable, quick, and safe approach to a rare but insidious complication of general anesthesia. It avoids lateral and posterior dissection of the trachea, reducing the risk of a recurrent laryngeal nerve injury.

Thymic carcinoma: a report of 13 cases.

OBJECTIVE: Thymic carcinoma is a rare thymic neoplasm. It is more invasive and has a poorer prognosis than thymoma. We report our experience in the treatment of 13 thymic carcinomas. METHODS: Thirteen patients with histologically confirmed thymic carcinoma were treated from June 1989. Six patients underwent surgery, followed by adjuvant therapy. Seven patients underwent neoadjuvant chemotherapy, followed by surgery and post-operative radiotherapy. RESULTS: The diagnosis of thymic carcinoma was achieved in six cases by a mediastinotomy, in three cases by a ultrasound-guided or a CT-guided fine needle aspiration and in three cases the pre-operative diagnosis was thymoma. In one case we did not have the histological diagnosis. All seven patients treated with neoadjuvant chemotherapy responded. The surgical resection was complete in seven cases. Eight patients are still alive 8-142 months from the diagnosis, and six are disease-free. CONCLUSIONS: Our experience supports the role of surgery and post-operative radiotherapy in thymic carcinomas. Pre-operative treatment of such neoplasms by multi-drug chemotherapy may improve the resectability and the survival rate.

Primary DNA damage in chrome-plating workers.

In order to evaluate the primary DNA damage due to occupational exposure to chromium (VI), DNA strand-breaks and apoptosis in peripheral lymphocytes were measured in a group of 19 chrome-plating workers. DNA strand-breaks was assessed by alkaline (pH>13) single-cell microgel electrophoresis ('comet') assay, while apoptosis was measured by flow-cytometry after propidium iodide staining of the cells. Concentrations of chromium in urine, erythrocytes and lymphocytes were investigated as biological indicators of exposure. A group of 18 hospital workers (control group I) and another 20 university personnel (control group II) without exposure to chromium were also studied as controls. The results of the study show that chrome-plating workers have higher levels of chromium in urine, erythrocytes and lymphocytes than unexposed workers. Comet tail moment values, assumed as index of DNA damage, are increased in chromium-exposed workers and results are significantly correlated to chromium lymphocyte concentrations. No difference emerged in the percentage of apoptotic nuclei in exposed and unexposed workers. The study confirms that measurements of chromium in erythrocytes and lymphocytes may provide useful information about recent and past exposure to hexavalent chromium at the workplace. The increase in DNA strand-breaks measured by comet assay suggests this test is valid for the biological monitoring of workers exposed to genotoxic compounds such as chromium (VI).

Posterior wall laceration of the thoracic trachea: the transcervical-transtracheal approach.

A large membranous wall laceration of the thoracic trachea was surgically treated. The surgical approach consists on a low collar incision followed by a longitudinal tracheotomy. The membranous tear was repaired with a running suture and tracheotomy sutured with interrupted crossed stitches. The procedure was effective and endoscopic follow-up showed a perfect healing process with no signs of tracheal stenosis. This new technique proved to be a reliable, quick and safe procedure, which allows to repair membranous lacerations as far as the carina, avoiding thoracotomy.