Antisense targeting of CD47 enhances human cytotoxic T-cell activity and increases survival of mice bearing B16 melanoma when combined with anti-CTLA4 and tumor irradiation.

Antibodies targeting the T-cell immune checkpoint cytotoxic T-lymphocyte antigen-4 (CTLA4) enhance the effectiveness of radiotherapy for melanoma patients, but many remain resistant. To further improve response rates, we explored combining anti-CTLA4 blockade with antisense suppression of CD47, an inhibitory receptor on T cells that limit T-cell receptor signaling and killing of irradiated target cells. Human melanoma data from The Cancer Genome Atlas revealed positive correlations between CD47 mRNA expression and expression of T-cell regulators including CTLA4 and its counter receptors CD80 and CD86. Antisense suppression of CD47 on human T cells in vitro using a translational blocking morpholino (CD47 m) alone or combined with anti-CTLA4 enhanced antigen-dependent killing of irradiated melanoma cells. Correspondingly, the treatment of locally irradiated B16F10 melanomas in C57BL/6 mice using combined blockade of CD47 and CTLA4 significantly increased the survival of mice relative to either treatment alone. CD47 m alone or in combination with anti-CTLA4 increased CD3+ T-cell infiltration in irradiated tumors. Anti-CTLA4 also increased CD3+ and CD8+ T-cell infiltration as well as markers of NK cells in non-irradiated tumors. Anti-CTLA4 combined with CD47 m resulted in the greatest increase in intratumoral granzyme B, interferon-γ, and NK-cell marker mRNA expression. These data suggest that combining CTLA4 and CD47 blockade could provide a survival benefit by enhancing adaptive T- and NK-cell immunity in irradiated tumors.

Comparative assessment of methicillin resistant Staphylococcus aureus diagnostic assays for use in resource-limited settings.

BACKGROUND: The rise of methicillin-resistant Staphylococcus aureus (MRSA) is a global health concern. Paucity of data on MRSA carriage prevalence and diagnostic methods in resource-limited settings hampers efforts to define the problem and plan an appropriate response. Additionally, high variability in cost and logistical characteristics of MRSA screening methods may impede infection control efforts. We compared the performance of locally-available chromogenic agar BD CHROMagar MRSA II and two PCR-based assays (Hain GenoQuick MRSA and Cepheid Xpert SA Complete) for the detection of asymptomatic MRSA carriage in nasal swabs. RESULTS: During 2015, we enrolled 500 patients from five hospital wards at a Ugandan regional referral hospital. We found 30% prevalence of methicillin-sensitive Staphylococcus aureus (MSSA) nasal carriage, and 5.4% MRSA nasal carriage prevalence. Compared to a composite reference standard defined as a positive test result on any one of the three assays, Hain GenoQuick MRSA demonstrated the highest sensitivity (96%) followed by direct plating on CHROMagar at (70%), with the lowest sensitivity observed with Xpert SA Complete (52%). Cepheid Xpert provided the most rapid results (< 1 h) but was the most expensive (US $45-50/test). Substantially more labor was required for the Hain GenoQuick MRSA compared to Xpert SA Complete or CHROMagar tests. CONCLUSION: MRSA nasal carriage prevalence rates were low, and high diagnostic sensitivity was achieved using Hain GenoQuick MRSA. Chromogenic media had significantly lower sensitivity, but may represent a viable local option given its lower cost compared to PCR-based assays.

Immunomodulation to enhance the efficacy of an HPV therapeutic vaccine.

BACKGROUND: While prophylactic human papillomavirus (HPV) vaccines will certainly reduce the incidence of HPV-associated cancers, these malignancies remain a major health issue. PDS0101 is a liposomal-based HPV therapeutic vaccine consisting of the immune activating cationic lipid R-DOTAP and HLA-unrestricted HPV16 peptides that has shown in vivo CD8+ T cell induction and safety in a phase I study. In this report, we have employed the PDS0101 vaccine with two immune modulators previously characterized in preclinical studies and which are currently in phase II clinical trials. Bintrafusp alfa (M7824) is a first-in-class bifunctional fusion protein composed of the extracellular domains of the transforming growth factor-ß receptor type II (TGFßRII) fused to a human IgG1 monoclonal antibody blocking programmed cell death protein-1 ligand (PDL1), designed both as a checkpoint inhibitor and to bring the TGFßRII 'trap' to the tumor microenvironment (TME). NHS-interleukin-12 (NHS-IL12) is a tumor targeting immunocytokine designed to bring IL-12 to the TME and thus enhance the inflammatory Th1 response. METHODS: We employed TC-1 carcinoma (expressing HPV16 E6 and E7 and devoid of PDL1 expression) in a syngeneic mouse model in monotherapy and combination therapy studies to analyze antitumor effects and changes in immune cell types in the spleen and the TME. RESULTS: As a monotherapy, the PDS0101 vaccine generated HPV-specific T cells and antitumor activity in mice bearing HPV-expressing mEER oropharyngeal and TC-1 lung carcinomas. When used as a monotherapy in the TC-1 model, NHS-IL12 elicited antitumor effects as well as an increase in CD8+ T cells in the TME. When used as a monotherapy, bintrafusp alfa did not elicit antitumor effects or any increase in T cells in the TME. When all three agents were used in combination, maximum antitumor effects were observed, which correlated with increases in T cells and T-cell clonality in the TME. CONCLUSION: These studies provide the rationale for the potential clinical use of combinations of agents that can (1) induce tumor-associated T-cell responses, (2) potentiate immune responses in the TME and (3) reduce immunosuppressive entities in the TME.

Factors influencing the return of inactive blood donors in a Cameroonian blood bank.

Identify factors that influence the return of donors to increase their loyalty while improving blood safety is crucial in our context. Between October 2017 and April 2018, we conducted a descriptive cross-sectional study at the Blood Bank of the Yaoundé University Teaching Hospital. The study included all former donors who had not donated blood voluntarily for over a year. Quantitative variables were described using means and standard deviations. Fisher's exact test and Chi2 test were used for association measures between qualitative variables. Statistical test results were considered significant for a P<0.05 value. We interviewed a total of 101 inactive donors. The study population was 74.3% male, donors average 30±7 years. Female gender and good staff hospitality were the factors most associated with the intention to return. The barriers to donor return were mainly lack of information on blood needs (35.60%) and time constraint for blood donation (26.73%). Pro-social motivations such as altruism (30.70%) were the main possible sources of motivation cited. To reduce blood deficiency and mortality due to lack of blood products, non-financial material compensation, good outreach and communication strategy can increase inactive donors' loyalty and consequently in improving blood safety in our context.

Prospective cohort study of the feasibility and yield of household child tuberculosis contact screening in Uganda.

SETTING: Screening and isoniazid preventive therapy (IPT) of child contacts of tuberculosis (TB) patients is poorly implemented in resource-limited countries, in part due to difficulties in TB diagnosis in children. OBJECTIVE: To assess the feasibility and yield of hospital-based screening and IPT in Uganda, and to evaluate the utility of symptom-based screening. DESIGN: Household child (age <5 years) contacts of adults with pulmonary TB were assessed for TB or latent tuberculous infection (LTBI). Children classified as 'LTBI' or 'uninfected' were prescribed IPT and followed for 9 months. Screening algorithms based on combinations of symptoms associated with TB were constructed post hoc, and their performance evaluated against a radiological-based reference standard. RESULTS: Of 281 contacts (median age 33 months), 44 (15.7%) started anti-tuberculosis treatment and 234 (83.3%) received IPT, 80.3% of whom completed a 6-month course. After adjustment for age and human immunodeficiency virus status, cough (aOR 4.10, 95%CI 1.39-12.11) and reduced playfulness (aOR 7.79, 95%CI 2.12-25.18) were associated with radiological TB. Screening based on cough or reduced playfulness had a sensitivity of 81.8% and a negative predictive value of 97.6%. CONCLUSION: Hospital-based screening appears to be feasible, and confirms the potential utility of symptom-based screening to select children for IPT and those for further investigations.

Effect of renin inhibitor, ES-8891, on renal renin secretion and storage in the marmoset: comparison with captopril.

We investigated the effects of the human renin inhibitor, ES-8891, and captopril on renal renin secretion and storage in the marmoset. Either ES-8891 (30 mg/kg) or captopril (2 mg/kg) was given orally twice a day for 1 week to conscious, sodium-depleted marmosets (n = 6 for each group). The ES-8891-treated group displayed a significant reduction in mean arterial pressure (MAP), plasma renin activity (PRA) and plasma immunoreactive renin (PIR) compared with the control group. Kidney renin content was significantly increased compared with the control group and enlarged renin granules containing heterogenous internum were observed in juxtaglomerular cells after treatment with ES-8891. Treatment with captopril significantly increased PRA and PIR compared with the control day as well as increasing kidney renin content and the number of renin granules with crystalline content in juxtaglomerular cells compared with the control group. These results suggest that ES-8891 inhibits both PRA and renin secretion from the kidney, resulting in an increase in renal renin storage.

Development of Purkinje cell bodies and processes with basic fibroblast growth factor-like immunoreactivity in the rat cerebellum.

The development of basic fibroblast growth factor-like immunoreactivity was investigated in the nuclei, cell bodies and processes of Purkinje cells with attention to basic fibroblast growth factor-containing neuronal input to the deep cerebellar nuclei. Immunoblot analysis with the use of the antisera against basic fibroblast growth factor revealed that crude homogenate of the developing rat cerebellum exhibits a main band with the same molecular weight (18,000 mol. wt) as basic fibroblast growth factor in all the postnasal stages examined. Cerebellar cells were not labeled with the antisera during embryonic life. Under light microscopy, basic fibroblast growth factor-like immunoreactivity was detected initially in cortical cells located close to deep cerebellar fissures of the newborn rat but not in superficial cortical regions. It was difficult to determine whether or not they are Purkinje cells at the fusiform stage. On postnatal day 7, immunoreactive Purkinje cells were identified throughout the cerebellar cortex, and they expressed basic fibroblast growth factor-like immunoreactivity mainly in the apical cytoplasm and proximal dendrites. From postnatal day 14 to postnatal day 28, basic fibroblast growth factor-like immunoreactivity was noted not only throughout the cytoplasm of Purkinje cells but also in the nuclei of the immunopositive cells. Our statistical analysis showed that Purkinje cells with nuclear immunoreaction peaked on postnatal day 21. At these stages, nerve fibers immunoreactive for basic fibroblast growth factor were numerous in the cerebellar medulla and deep cerebellar nuclei. After postnatal day 42, Purkinje cells with intense immunoreactivity in the nuclei showed a marked decrease in number, and immunoreactive structures were distributed in the cerebellum in a fashion similar to that in adult rats. Electron microscopy demonstrated that immunoreactivity was located mainly in the apical cytoplasm of Purkinje cells on postnatal day 7 and throughout the cytoplasm and in the nuclear euchromatin from postnatal day 14 to postnatal day 28, as was expected from light-microscopic observations. Immunoreactivity, even though distributed diffusely in the cytoplasm, was absent from the lumen of endoplasmic reticulum and mitochondria. A small population of Purkinje cell axon terminals forming synapses with the soma and dendrites of deep cerebellar nucleus neurons began to express basic fibroblast growth factor on postnatal day 21. This is much later than the starting age for synaptogenesis between Purkinje cells and deep cerebellar nucleus neurons. The age-dependent changes in the localization of basic fibroblast growth factor within Purkinje cell nucleus, soma and processes suggest a complex transport system of this factor within Purkinje cells during postnatal development.

Comparative studies of neoplastic response to a single dose of nitroso compounds. 6. The effect of diethylnitrosamine in Syrian golden hamsters.

The effect of a single subcutaneous injection of different DEN doses was examined in adult Syrian hamsters observed for life. The minimal effective dose (threshold dose) for neoplastic response, reflected by induction of papillary polyps in the larynx and/or trachea, was 1.03 mg DEN/kg body weight. No tumors were found which could be related to treatment in other segments of the respiratory epithelium nor in other tissues.

Photodynamic therapy with a diode laser for implanted fibrosarcoma in mice employing mono-L-aspartyl chlorin E6.

The authors performed photodynamic therapy (PDT), avoiding any hyperthermic effects, using a newly developed diode laser and photosensitizer, mono-L-aspartyl chlorin e6 (NPe6), of Meth-A fibrosarcoma implanted in mice and achieved tumor therapeutic benefit. The photodynamic light treatment was performed 5 h following the photosensitizer administration. With 5.0 mg/kg NPe6 and light doses of 50, 100, 150 and 200 J/cm2, the tumor cure rates were 20, 50, 70 and 90%, respectively. With 100 J/cm2 laser exposure and NPe6 doses of 1.25, 2.5, 5.0, 7.5 and 10.0 mg/kg, the tumor cure rates were 0, 20, 50, 70 and 90%, respectively. A charge-coupled device (CCD) camera system was employed to measure the NPe6 fluorescence intensity correlating with the residual amount of the photosensitizer at deferent depth from the tumor surface. The ratios of the NPe6 fluorescence intensity at 3 mm from the tumor surface following 50, 100, 150 and 200 J/cm2 laser exposure to no laser exposure were 0.73, 0.36, 0.22 and 0.16, respectively. With samples sectioned at 1 mm depth, after 50 J/cm2 and the same photosensitizer dose (5 mg/kg) this ratio was 0.19. These results suggest that a certain increase in the tumor tissue level of NPe6 and a certain increase of laser light dose reaching deeper layer of tumor caused an increase in percent cure. In addition, the effectiveness of PDT depends on the total laser dose reaching deeper layers of tumors. Furthermore, the effectiveness of PDT tends to correlate with the amount of NPe6 photobleaching by PDT.

The role of surgical excision in the management of relapsed Wilms' tumor patients with pulmonary metastases: a report from the National Wilms' Tumor Study.

To determine the effect of surgical excision of pulmonary metastases from Wilms' tumor on postrelapse survival, we retrospectively analyzed the clinical courses of 211 patients with stages I to III, favorable or unfavorable histology Wilms' tumor entered on National Wilms' Tumor Study-1, -2, or -3 whose first recurrence was limited to the lungs. There was no difference in the 4-year postrelapse survival percentage of favorable-histology patients with a solitary pulmonary metastasis who did or did not undergo surgical removal of the metastasis in addition to pulmonary irradiation and chemotherapy. Although histological confirmation of pulmonary relapse is frequently indicated, the present data suggest that therapeutic removal of pulmonary metastases from patients with relapsed Wilms' tumor does not increase the percentage of patients who survive for 4 years postrelapse, compared with treatment with whole-lung irradiation and chemotherapy.

[Sinus bradycardia induced by talipexole hydrochloride in a patient with Parkinson disease].

We report sinus bradycardia induced by talipexole hydrochloride in a 65-year-old man with Parkinson disease. Approximately four hours after he had taken 0.8 mg of talipexole hydrochloride, he acutely developed sleepiness, delusion, akinesia, and faintness associated with hypotension and sinus bradycardia. Another similar episode occurred when he had taken talipexole hydrochloride 1.2 mg/day in combination with a daily dose of 200 mg of levodopa and 20 mg of carbidopa. These symptoms persisted for 12 hours and diminished gradually without any specific treatments. Talipexole hydrochloride, a stimulator of both the D2 and alpha 2 receptors probably induced bradycardia and hypotension in the present case.

[Complex visual hallucination in the quadrantanopsic field in a patient with a medial temporal hematoma following left putaminal hemorrhage].

A 56-year-old man with a left medial temporal lobe hematoma from the left putaminal hemorrhage presented with complex visual hallucinations in the right quadrantanopsic field. In the right superior quadrantanopsic field, he saw colored and formed hallucinations of plants, bamboo thickets and a stature of bodhisattva. Most of these hallucinations were what he had actually seen before. The visual hallucinations disappeared when he gazed them with saccadic eye movements. EEG showed no epileptic discharges. MRI revealed hematoma from the lower putamen to the medial temporal lobe including amygdala on the left. 123I-IMP SPECT showed hypoperfusion in the left temporal lobe and in the left occipital and parietal lobes. We speculate that the hallucinations of the present case were produced by the dysfunctions not only of the temporooccipital and temporoparietal regions but also of the posterior inferotemporal cortex which has strong interactions with amygdala and works as the visual memory center.

Effect of renal denervation on the development of hypertension in Dahl-Iwai salt-sensitive rats.

Renal denervation has been shown to delay the onset of hypertension in spontaneously hypertensive rats and DOCA-salt sensitive rats. We investigated the contribution of the renal nerves to the development of hypertension in Dahl-Iwai salt-sensitive (DS) rats. Bilateral renal denervation or sham-operation was carried out in DS rats, and animals were then kept on a high salt diet (study I) or on a normal salt diet (study II). DS rats became severely hypertensive (207 +/- 8 mmHg) after 4 weeks on a high salt diet. They became mildly hypertensive (156 +/- 3 mmHg) after 4 weeks on a normal salt diet. In both studies, renal denervation exerted no effect on the development of hypertension in the DS rats. The urinary sodium excretion, urinary volume, heart rate and body weight were unaltered by renal denervation. These results indicate that the renal nerves do not make a major contribution to the development of hypertension in DS rats.

[Pryce type I interlobar pulmonary sequestration with anomalous return to the left inferior pulmonary vein].

A 50-year-old woman was examined because of chest discomfort. Chest X-ray films disclosed a tumorous shadow behind the heart. Chest computed tomography (CT) scans revealed a mass connected to the descending aorta, with increased blood flow in the left basal segment. The patient was admitted for further examination. Chest CT scans and cardioscintigrams were very useful as diagnostic tools, but the final diagnosis was made on the basis of angiography. An anomalous tortuous artery ran from the descending aorta into the left basal segment and returned to the left inferior pulmonary vein. The left pulmonary arterial trunk had no basal branch (A8-A10). A loop corresponding to the superior vein (V6) ran beneath the anomalous tortuous artery. To our knowledge, this is the second case of Pryce type-I interlobar pulmonary sequestration with anomalous return to the left inferior pulmonary vein to be reported in Japan. A left inferior lobectomy was performed. Histological finding from the excised tissues showed prominent interstitial fibrosis, atypical adenomatous hyperplasia, and atherosclerosis. Following surgery, the patient's PaO2 increased from 80.4 Torr to 95.8 Torr, suggesting that left inferior lobectomy was an appropriate treatment.

Las instituciones de guarda y cuidado en el ordenamiento familiar cubano. Pautas y retos

El presente trabajo versa sobre la necesidad de regular en el Código de Familia cubano aquellas instituciones jurídicas de guarda y cuidado para complementar la capacidad de aquellas personas carentes de ella, o que la poseen, pero con limitaciones, en aras de impedir su indefensión. El objetivo es fundamentar los presupuestos teóricos normativos sobre las instituciones de guarda y cuidado en el ordenamiento jurídico familiar cubano. Para ello se emplearon métodos científicos, y técnicas propias de las ciencias jurídicas, dentro de ellos el histórico - lógico, exegético - analítico, jurídico comparado; y la técnica de investigación empleada fue el análisis de documentos. Se trata de un estudio eminentemente teórico, el tipo de investigación es descriptiva y la metodología empleada, cualitativa. Este estudio arrojó como principal resultado la necesidad de regulación de las instituciones de guarda y cuidado en el Código de Familia cubano ante la existencia de sujetos del Derecho carentes de capacidad de obrar, o con capacidad restringida.

Este trabalho enfoca a necessidade de regulamentar no Código de Família cubano essas instituições de tutela e cuidados legais para complementar a capacidade dessas pessoas sem ela, ou que a possuem, mas com limitações, a fim de evitar seu desamparo. O objectivo é informar os pressupostos teóricos normativas sobre as instituições de tutela e de cuidados no direito de família cubana. Para estes métodos científicos foram utilizados, e apropriado, entre elas as técnicas ciência jurídica históricos - lógico, exegéticos -, direito comparado analítica; e técnica de pesquisa utilizada foi a análise de documentos. Este é um estudo eminentemente teórico, o tipo de pesquisa é descritiva e metodologia, qualitativa. Este estudo resultou como principal resultado a necessidade de regulamentação das instituições de tutela e cuidados na cubana Código da Família sujeita à existência da lei falta de capacidade legal ou com capacidade limitada.

The present work turns about the necessity of regulating in the classification juridical Cuban relative guard's juridical institutions and care to supplement the capacity of those lacking people of her or that they possess it, but with limitations, for the sake of impeding its indefensión. The objective is to base the normative theoretical budgets on guard's institutions and care in the classification juridical Cuban relative. For they were used it methods scientific, and technical characteristic of the juridical sciences, inside them the historical one - logical, exegético - analytic, juridical compared; and the technique of used investigation was the analysis of documents. It is an eminently theoretical study, the investigation type is descriptive and the used methodology, qualitative. This study hurtled as main result the necessity of regulation of guard's institutions and care in the Cuban Code of Family before the existence of subject of the lacking Right of capacity of working or with restricted capacity.