LTA1 is a safe, intranasal enterotoxin-based adjuvant that improves vaccine protection against influenza in young, old and B-cell-depleted (µMT) mice.

Enterotoxin-based adjuvants including cholera toxin and heat-labile toxin (LT) are powerful manipulators of mucosal immunity; however, past clinical trials identified unacceptable neurological toxicity when LT or mutant AB5 adjuvant proteins were added to intranasal vaccines. Here, we examined the isolated enzymatic A1 domain of LT (LTA1) for intranasal safety and efficacy in combination with influenza (flu) vaccination. LTA1-treated mice exhibited no neurotoxicity, as measured by olfactory system testing and H&E staining of nasal tissue in contrast with cholera toxin. In vaccination studies, intranasal LTA1 enhanced immune responses to inactivated virus antigen and subsequent protection against H1N1 flu challenge in mice (8-week or 24-months). In addition, lung H1N1 viral titers post-challenge correlated to serum antibody responses; however, enhanced protection was also observed in µMT mice lacking B-cells while activation and recruitment of CD4 T-cells into the lung was apparent. Thus, we report that LTA1 protein is a novel, safe and effective enterotoxin adjuvant that improves protection of an intranasal flu vaccination by a mechanism that does not appear to require B-cells.

The thyroid status reprograms T cell lymphoma growth and modulates immune cell frequencies.

UNLABELLED: In spite of considerable evidence on the regulation of immunity by thyroid hormones, the impact of the thyroid status in tumor immunity is poorly understood. Here, we evaluated the antitumor immune responses evoked in mice with different thyroid status (euthyroid, hyperthyroid, and hypothyroid) that developed solid tumors or metastases after inoculation of syngeneic T lymphoma cells. Hyperthyroid mice showed increased tumor growth along with increased expression of cell cycle regulators compared to hypothyroid and control tumor-bearing mice. However, hypothyroid mice showed a higher frequency of metastases than the other groups. Hyperthyroid mice bearing tumors displayed a lower number of tumor-infiltrating T lymphocytes, lower percentage of functional IFN-γ-producing CD8(+) T cells, and higher percentage of CD19(+) B cells than euthyroid tumor-bearing mice. However, no differences were found in the distribution of lymphocyte subpopulations in tumor-draining lymph nodes (TDLNs) or spleens among different experimental groups. Interestingly, hypothyroid TDLN showed an increased percentage of regulatory T (Treg) cells, while hyperthyroid mice displayed increased number and activity of splenic NK cells, which frequency declined in spleens from hypothyroid mice. Moreover, a decreased number of splenic myeloid-derived suppressor cells (MDSCs) were found in tumor-bearing hyperthyroid mice as compared to hypothyroid or euthyroid mice. Additionally, hyperthyroid mice showed increased cytotoxic activity, which declined in hypothyroid mice. Thus, low levels of intratumoral cytotoxic activity would favor tumor local growth in hyperthyroid mice, while regional and systemic antitumor response may contribute to tumor dissemination in hypothyroid animals. Our results highlight the importance of monitoring the thyroid status in patients with T cell lymphomas. KEY MESSAGES: T cell lymphoma phenotype is paradoxically influenced by thyroid status. Hyperthyroidism favors tumor growth and hypothyroidism rises tumor dissemination. Thyroid status affects the distribution of immune cell types in the tumor milieu. Thyroid status also modifies the nature of local and systemic immune responses.

Thyroid status modulates T lymphoma growth via cell cycle regulatory proteins and angiogenesis.

We have shown in vitro that thyroid hormones (THs) regulate the balance between proliferation and apoptosis of T lymphoma cells. The effects of THs on tumor development have been studied, but the results are still controversial. Herein, we show the modulatory action of thyroid status on the in vivo growth of T lymphoma cells. For this purpose, euthyroid, hypothyroid, and hyperthyroid mice received inoculations of EL4 cells to allow the development of solid tumors. Tumors in the hyperthyroid animals exhibited a higher growth rate, as evidenced by the early appearance of palpable solid tumors and the increased tumor volume. These results are consistent with the rate of cell division determined by staining tumor cells with carboxyfluorescein succinimidyl ester. Additionally, hyperthyroid mice exhibited reduced survival. Hypothyroid mice did not differ significantly from the euthyroid controls with respect to these parameters. Additionally, only tumors from hyperthyroid animals had increased expression levels of proliferating cell nuclear antigen and active caspase 3. Differential expression of cell cycle regulatory proteins was also observed. The levels of cyclins D1 and D3 were augmented in the tumors of the hyperthyroid animals, whereas the cell cycle inhibitors p16/INK4A (CDKN2A) and p27/Kip1 (CDKN1B) and the tumor suppressor p53 (TRP53) were increased in hypothyroid mice. Intratumoral and peritumoral vasculogenesis was increased only in hyperthyroid mice. Therefore, we propose that the thyroid status modulates the in vivo growth of EL4 T lymphoma through the regulation of cyclin, cyclin-dependent kinase inhibitor, and tumor suppressor gene expression, as well as the stimulation of angiogenesis.

Induction of apoptosis in T lymphoma cells by long-term treatment with thyroxine involves PKCζ nitration by nitric oxide synthase.

Thyroid hormones are important regulators of cell physiology, inducing cell proliferation, differentiation or apoptosis, depending on the cell type. Thyroid hormones induce proliferation in short-term T lymphocyte cultures. In this study, we assessed the effect of long-term thyroxine (T4) treatment on the balance of proliferation and apoptosis and the intermediate participants in T lymphoma cells. Treatment with T4 affected this balance from the fifth day of culture, inhibiting proliferation in a time-dependent manner. This effect was associated with apoptosis induction, as characterized through nuclear morphological changes, DNA fragmentation, and Annexin V-FITC/Propidium Iodide co-staining. In addition, increased iNOS gene and protein levels, and enzyme activity were observed. The generation of reactive oxygen species, depolarization of the mitochondrial membrane, and a reduction in glutathione levels were also observed. The imbalance between oxidants and antioxidants species is typically associated with the nitration of proteins, including PKCζ, an isoenzyme essential for lymphoma cell division and survival. Consistently, evidence of PKCζ nitration via proteasome degradation was also observed in this study. Taken together, these results suggest that the long-term culture of T lymphoma cells with T4 induces apoptosis through the increased production of oxidative species resulting from both augmented iNOS activity and the loss of mitochondrial function. These species induce the nitration of proteins involved in cell viability, promoting proteasome degradation. Furthermore, we discuss the impact of these results on the modulation of T lymphoma growth and the thyroid status in vivo.

Hysteroscopic findings in 344 women with recurrent spontaneous abortion.

STUDY OBJECTIVE: To evaluate the prevalence of different anatomic factors in women with recurrent spontaneous abortion (RSA). DESIGN: Retrospective analysis over 9 years (Canadian Task Force classification II-2). SETTING: University hospital-affiliated endoscopic unit. PATIENTS: Three hundred forty-four consecutive patients with RSA and 922 controls referred for abnormal uterine bleeding. INTERVENTION: Diagnostic hysteroscopy. MEASUREMENTS AND MAIN RESULTS: Major and minor uterine mullerian abnormalities (septate, unicornuate uteri) were found significantly more often in women with RSA than in controls (32% vs 6%, p <0.001). The frequency of acquired uterine anomalies (submucous myomas, polyps) was significantly higher in controls (32% vs 9%, p <0.001). No significant differences were observed between groups in frequency of adhesions (4% vs 2%). CONCLUSION: Major mullerian uterine abnormalities are associated with RSA, and minor uterine anomalies may be correlated with an increased risk of recurrent miscarriage.

Preceding infection as a risk factor of stroke in the young.

INTRODUCTION: The cause of stroke in the young remains unknown in 20-50% of the patients. Infections preceding stroke have been recently recognised to be an independent risk factor of stroke. MATERIAL AND METHODS: Sixty consecutive patients aged 40 years or less presenting with ischaemic completed stroke are taken up for the study. Patients with neurological deficit of less than 24 hours, evidence of haemorrhage on CT scan, infection occurring after the onset of stroke were excluded. Controls consisted of age and sex matched persons residing in the same area. Both the groups were enquired about preceding fever and infections and were examined for evidence of infections. Serum was examined for antibodies against measles, herpes simplex, and Japanese B encephalitis viruses. Cultures were put up from appropriate samples and CSF examined in patients only. RESULTS: Evidence of infection was noted in 26 (43.3%) of patients and 6 controls (p < 0.001). History of fever was elicited in 23 patients and 3 controls while 15 patients were febrile on examination at admission. Signs of local infection was observed in 14 patients and one control. The commonest site of infection was respiratory tract. Cultures were positive in 11 patients, commonest being beta haemolytic streptococci in six from throat. Conventional risk factors were identical in both groups of patients with and without evidence of preceding infection. Smoking and alcoholism were significantly higher in patients with preceding infection. CONCLUSION: Preceding infection is an important risk factor of stroke in the young. Smoking and alcoholism are more frequent in patients with preceding infection. Whether they predispose the individual for infection or infection increases the stroke risk in them needs to be examined.

[The use of electrosurgery in the treatment of extra-cervical genital condylomatosis]. / L'impiego dell'elettrochirurgia nel trattamento della condilomatosi genitale extra-cervicale.

BACKGROUND AND AIMS: The aim of this study was to evaluate the therapeutic efficacy of electrosurgery in the treatment of HPV correlated extra-cervical genital lesions. METHODS: A group of 78 patients with clinical or colposcopic, histologically ascertained vulvar or perineal condylomatosis were enrolled consecutively between March 1994 and July 1995 at the Cervico-Vaginal and Vulvar Pathology Unit of the Obstetrics and Gynecology Clinic at Tor Vergata University, Rome. All patients underwent physical treatment with a radiofrequency electrosurgical knife using exclusively cutting current. In addition to patient compliance, the following parameters were evaluated: length of surgery and recovery times, collateral effects and possible long-term results; a colposcopic and, if necessary, bioptic follow-up was also performed at 3, 6 and 12 months after treatment. RESULTS: Seven out of the group of 68 patients with subclinical condylomatosis presented histologically documented recidivation during the course of follow-up. In the discussion the authors evaluate the positive results obtained using this method and, based on the physical principle underlying electroradiosurgery, they suggest using the term electrosurgical vaporisation for destructive electrotherapy using a cutting current, which is suited to the physical mechanism with which this current acts.

Outpatient diagnostic hysteroscopy.

STUDY OBJECTIVE: To evaluate the feasibility, validity, indications, and results of a large series of diagnostic hysteroscopies performed without anesthesia. DESIGN: Retrospective analysis of hysteroscopy charts performed between 1989 and 1996 (Canadian Task Force classification II-2). SETTING: University-affiliated endoscopy unit. PATIENTS: Four thousand consecutive women referred for different indications. INTERVENTIONS: Diagnostic hysteroscopy was performed in 91% of patients without premedication or anesthetics. In some women premedication or general or local anesthesia was required to access the uterine cavity. MEASUREMENTS AND MAIN RESULTS: The success rate, validity indication, complication rate, and number of biopsies were critically evaluated and assessed in relation to increased experience of operators. In 91% of women we accessed the uterine cavity at the first attempt without premedication, whereas 207 (5. 1%) patients required local anesthesia and 99 (2.4%) premedication. Only 1.6% required general anesthesia. In 52% intrauterine pathology was diagnosed and in 21% further surgical treatment was suggested. CONCLUSION: Hysteroscopy was feasible when performed in an outpatient setting without general or local anesthesia in more than 90% of women. The operator's experience seems a key factor both for accurate endometrial evaluation and to reduce failure and endometrial biopsy rates. The low frequency of further surgical treatment justifies performing the procedure in the office.

Transvaginal sonographic and hysteroscopic findings in postmenopausal women receiving tamoxifen.

STUDY OBJECTIVE: To evaluate the effect of tamoxifen on the endometrium. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology of the University of Rome. PATIENTS: One hundred one postmenopausal women with breast cancer receiving tamoxifen 20 to 30 mg/day for at least 1 year; of these 78 were asymptomatic and 23 had vaginal bleeding. INTERVENTIONS: All patients underwent transvaginal color Doppler sonography. Those with endometrial thickness greater than 5 mm were advised to undergo hysteroscopy and if necessary endometrial biopsy. For women with endometrial thickness less than 5 mm, hysteroscopy was recommended only if irregular endometrial echotexture was observed. MEASUREMENTS AND MAIN RESULTS: Eleven (14%) asymptomatic patients and 1 (4.3%) with vaginal bleeding had endometrial thickness less than 5 mm (p = 0.4, NS). Women with vaginal bleeding had a significantly thicker endometrium than asymptomatic patients (15.8 +/- 7.5 vs 11.1 +/- 5.7 mm, p = 0.003). In the asymptomatic group 31 polyps, 15 atrophic endometria, and 6 hyperplasias were observed. Two endometrial cancers, 13 polyps, and 3 hyperplasias were detected in patients with vaginal bleeding. Hysteroscopy did not always allow endometrial biopsy, even in the presence of increased endometrial thickness with or without irregular surface. No statistical differences were found for mean pulsatility and resistance indexes of uterine and endometrial arteries between symptomatic and asymptomatic women, but these indexes were significantly lower compared with normal postmenopausal values. CONCLUSION: Women receiving tamoxifen, especially those who are asymptomatic, should be closely monitored by transvaginal sonography and hysteroscopy to detect endometrial pathologies.

Transvaginal Sonographic and Hysteroscopic Findings in Postmenopausal Women Receiving Tamoxifen

The effect of tamoxifen on the endometrium was assessed by transvaginal color Doppler sonography, hyteroscopy, and histology in 90 postmenopausal women, of whom 71 were asymptomatic and 19 had vaginal bleeding. All were treated with tamoxifen 20 to 30 mg/day for breast cancer for at least 1 year and all underwent transvaginal color Doppler sonography. Women with endometrial thickness greater than 5 mm and with vaginal bleeding were advised to undergo hysteroscopy and endometrial biopsy if necessary. In asymptomatic women with endometrial thickness of 5 mm or less, hysteroscopy was recommended if irregular endometrial echotexture was observed. Nine (12.7%) asymptomatic patients and 1 (5.3%) with vaginal bleeding had an endometrial width of 5 mm or less (p = 0.6, NS). Those with vaginal bleeding receiving tamoxifen had a significantly thicker endometrium than asymptomatic women (16.1 ± 7.7 vs 11.1 ± 5.7 mm, p = 0.003). A typical sonographic endometrial pattern with small hypoechoic cystic areas was reported in 83% of patients. In the asymptomatic group, 25 polyps, and 7 atrophic and 13 thickened endometria were observed at hysteroscopy. Two endometrial cancers, 11 polyps, and 1 atrophic and 4 thickened endometria were present in women with vaginal bleeding. Hysteroscopy did not always allow endometrial biopsy even if endometrial thickness with or without an irregular surface was observed. No statistical differences were revealed in the two tamoxifen groups for the mean pulsatility and resistance indexes of the uterine and endometrial arteries, which were otherwise significantly lower compared with normal postmenopausal values. Women receiving tamoxifen, especially asymptomatic women, should be closely monitored by transvaginal color Doppler ultrasonography and hysteroscopy to detect endometrial pathologies.

Vaginal Ultrasonography and Diagnostic Hysteroscopy for Women with Abnormal Uterine Bleeding after Menopause

Three hundred twenty-five postmenopausal women with abnormal uterine bleeding had transvaginal ultrasound examinations. In 158 the endometrial thickness was greater than 5 mm, in 20 less than 5 mm but irregular, and in 147 less than 5 mm and regular. Diagnostic hysteroscopy was performed in the first two groups and showed mild endometrial abnormalities in 23 women, severe in 23, endometrial polyps in 55, myoma in 10, and normal endometrium (atrophic or proliferative) in 22, with synechiae in 5. Forty-six (32%) endometrial biopsies were performed and showed 20 adenocarcinomas, 3 atypical hyperplasias, 10 simple hyperplasias, 5 atrophies, and 8 proliferative. In 18 patients the examination was not possible because of cervical stenosis and was performed under general anesthesia; polyps were removed by operative hysteroscopy. In patients with endometrial thickness greater than 5 mm, hysteroscopy revealed only two cases of mild endometrial abnormalities (cystic atrophy), two polyps, and two myomas. The frequency of endometrial cancer was 7%, similar to that reported by others. Ultrasonography is sensitive in evaluating abnormal uterine bleeding with or without endometrial pathology. Hysteroscopy must be the second procedure because it can exclude pathology and allow a targeted biopsy to confirm the diagnosis.

Costs and Benefits of Office Diagnostic Hysteroscopy

In Italy, the main location for performing diagnostic hysteroscopy is the office. The success of office hysteroscopy is related to savings of time and money and the convenience for the physician and patient. We evaluated 4000 diagnostic hysteroscopies performed between January 2, 1989, and March 1, 1996. In 17.8% of these patients we subsequently performed operative hysteroscopies (metroplasty, synechiolysis, myomectomy, polypectomy for polyps >2 cm), and in 5.2% a total abdominal hysterectomy for abnormal uterine bleeding or endometrial cancer. The low percentage of operations underlines the efficacy of office diagnostic hysteroscopy, especially if we evaluate costs and accuracy of this procedure in detecting uterine pathologies.

Hysteroscopically detected asymptomatic müllerian anomalies. Prevalence and reproductive implications.

OBJECTIVE: To assess the prevalence of uterine anomalies and relative reproductive function in 322 women with abnormal uterine bleeding (AUB) evaluated by diagnostic hysteroscopy. STUDY DESIGN: Uterine contours were classified as septate/bicornuate, arcuate or normal on the basis of hysteroscopy. A complete medical history was obtained from all the subjects; it included a questionnaire on menstrual characteristics, reproductive history and pregnancy outcome. Cumulative birth rates, frequency of spontaneous abortion, preterm delivery, malpresentation and mode of delivery in patients with normal and abnormal uteri were compared using life table analysis, the log-rank test and chi 2 analysis. RESULTS: Arcuate, septate/bicornuate and unicornuate uteri were observed in 6.5%, 3.7% and 0.3% of women, respectively. Although 24-month pregnancy rates and monthly fecundability rates were similar in women with and without müllerian anomalies, the 36-month cumulative live birth rate was significantly lower in women with a septate/bicornuate uterus. Overall, women with uterine malformations showed a significantly higher miscarriage rate (P < .05) and a significantly lower term delivery rate (P < .05) than women with a normal-shaped uterus. CONCLUSION: Diagnostic hysteroscopy in women with AUB detected a 10% prevalence of uterine anomalies, which were associated with a significantly higher incidence of spontaneous abortion and lower cumulative live birth rates.

Disseminated Burkitt's lymphoma presenting as multiple cranial nerve palsies.

A case of disseminated Burkitt's lymphoma with nervous system involvement in a HIV negative 35 year old lady is described. She primarily presented with multiple cranial nerve palsies. At autopsy, diffuse involvement of parenchymatous organs and lymphomatous meningitis with conspicuous sparing of gastrointestinal system was observed. In addition, there was an unusual feature of paraneoplastic demyelinating peripheral neuropathy. Incidentally, a large hydatid cyst was also seen in the left lobe in addition to the lymphomatous involvement of the liver.

A new hysteroscopic classification of and nomenclature for endometrial lesions.

STUDY OBJECTIVE: To establish a hysteroscopic classification of endometrial lesions based on a combined score: DESIGN: A prospective study to grade four features of endometrial lesions (thickness, surface, color, vascularization) and subdivide them as mild (low risk) and severe (high risk). SETTING: Gynecologic endoscopy unit of an obstetric and gynecology clinic. PATIENTS: Two hundred twenty-nine women with hysteroscopically diagnosed endometrial lesions. INTERVENTIONS: Hysteroscopic inspection of endometriotic lesions, with grading performed on the four features. MEASUREMENTS AND MAIN RESULTS: The sensitivity and specificity of our system were 86.9% and 87.4% for mild pathology, and 96% and 92.9% for severe pathology. CONCLUSIONS: This hysteroscopic classification of endometrial pathology can be useful for a better definition of endometrial lesions. The method has good sensitivity and specificity.

The use of topical anesthesia in diagnostic hysteroscopy and endometrial biopsy.

OBJECTIVE: To show that intrauterine anesthesia is a reliable method for reducing pain associated with endocavitary procedures. DESIGN: A prospective, randomized, double-blind study. SETTING: The Department of Obstetrics and Gynecology, Tor Vergata University of Rome, Rome, Italy. PATIENTS: Forty-five patients undergoing diagnostic hysteroscopy (n = 27) or hysteroscopy and endometrial biopsy (n = 18). INTERVENTIONS: Five milliliters of 2% mepivacaine or saline solution were injected transcervically into the uterine cavity before performing the procedures. MAIN OUTCOME MEASURES: Evaluation of pain reduction on a visual analogue scale. RESULTS: Pain expectation and pain reported were reduced during and after the procedures. CONCLUSION: Topical anesthesia effectively reduces pain during hysteroscopy and endometrial biopsy.

Hysteroscopic Appearance of Benign and Malignant Endometrial Polyps. What are the Differences?

The objective of this study is to evaluate the differences between benign and malignant endometrial polyps at hysteroscopy. In our collected experience of 1793 diagnostic hysteroscopies performed from December 1989 to December 1993, we found 3 malignant polyps in 151 cases of hysteroscopically diagnosed endometrial polyps. The differences in appearance of the malignant polyps are mainly represented by surface irregularities such as necrosis, vascular irregularities and whitish thickened areas. We conclude that when these patterns are present it is better to perform a targeted biopsy to confirm the hysteroscopic diagnosis. This approach allows selection of patients that can be treated by hysteroscopic polypectomy from those who require more radical treatment.