Antimicrobial properties of the medicinal plant Cardiospermum halicacabum L: new evidence and future perspectives.

The emergence and rapid spread of multidrug-resistance in human pathogenic microorganisms urgently require the development of novel therapeutic strategies for the treatment of infectious diseases. From this perspective, the antimicrobial properties of the natural plant-derived products may represent an important alternative therapeutic option to synthetic drugs. Among medicinal plants, the Cardiospermum halicacabum L. (C. halicacabum), belonging to Sapindaceae family, could be a very promising candidate for its antimicrobial activity against a wide range of microorganisms, including both Gram-positive and Gram-negative bacteria, as well as fungal pathogens. Although the antimicrobial properties of C. halicacabum have been intensively studied, the mechanism/s by which it exerts the inhibitory activity towards the pathogenic microbes have not yet been completely understood. This review focuses on the main antimicrobial activities displayed in vitro by the plant extract, with particular attention on our recent advances. We demonstrated that C. halicacabum is able to exert in vitro a dose-dependent fungistatic effect against Trychophyton rubrum (T. rubrum) through molecular interaction with the fungal heat shock protein (Hsp)-90 chaperone. These findings are supported by a growing body of research indicating the crucial role played by the Hsp90 in the virulence of the pathogenic microorganisms, including fungal pathogens. The possible future use of C. halicacabum for treating a wide range of infectious diseases is also discussed.

Whole exome sequencing approach to childhood onset familial erythrodermic psoriasis unravels a novel mutation of CARD14 requiring unusual high doses of ustekinumab.

BACKGROUND: Autosomal dominant gain of function mutations in caspase recruitment domain family member 14 (CARD14) is a rare condition associated with plaque-type psoriasis, generalized pustular psoriasis, palmoplantar pustular psoriasis and pityriasis rubra pilaris. Recently, a new CARD14 -associated phenotype defined as CAPE (CARD14-associated papulosquamous eruption) with clinical features of both psoriasis and pityriasis rubra pilaris was reported. We describe a family carrying a novel heterozygous mutation in CARD14 gene, with childhood-onset erythrodermic psoriasis requiring an unusual extremely high dose (up to 2 mg/kg every 8 weeks) of ustekinumab to achieve disease remission. CASE PRESENTATION: We describe a large family with three pairs of twins presenting a clinical phenotype characterized by childhood-onset erythrodermic psoriasis; in some family members is also reported psoriatic arthritis. The two probands presented poor clinical response to topic and systemic therapy with antihistamine, steroid, retinoids, cyclosporine and etanercept. After exclusion of the most common genes associated to autoinflammatory diseases (IL36RN, IL1RN, MVK, TNFRSF1A, NLRP3, NLRP12, MEFV, NOD2, PSMB8, PSTPIP1, LPIN2) we approached a new gene search by subjecting to Whole Exome Sequencing (WES) analysis five members of the family. A novel heterozygous mutation (c.446 T > G, leading to the missense amino acid substitution p.L149R) in the exon 4 of the CARD14 gene was identified in all affected members. Increasing dosages (up to 2 mg/kg every 8 weeks) of ustekinumab, a human monoclonal antibody targeting interleukin-12 (IL-12) and interleukin-23 (IL-23), allowed the complete control of the clinical manifestations, with an evident reduction of circulating Th17 and Th22 CD4+ T cell subsets. CONCLUSIONS: We describe the association of mutations of the CARD14 gene with an erythrodermic psoriasis pedigree, underlying the necessity to investigate CARD14 mutations in childhood-onset psoriasis cases and confirming the presence of CARD14 causative mutations also in erythrodermic psoriasis form, as recently reported. Also in pediatric age, ustekinumab represents a powerful therapeutic option for this rare condition, that is usually refractory to other treatments. In young children, high and frequent dosages allowed a complete control of the clinical manifestations without any severe side effects, with a long-term follow-up.

In vitro and in vivo antitumor efficacy of docetaxel and sorafenib combination in human pancreatic cancer cells.

The response of pancreatic cancer to treatments remains unsatisfactory, highlighting the need for more effective therapeutic regimens. Sorafenib, an orally available multikinase inhibitor, is active against different tumors, including pancreatic cancer. We studied the antitumor efficacy of sorafenib in combination with different antitumor drugs currently used in clinical practice in in vitro and in vivo experimental models of human pancreatic cancer. The cytotoxic effect of sorafenib and conventional antitumor drug combinations was evaluated in vitro in human pancreatic cancer cell lines and the efficacy of the most active combination was tested on tumor-bearing mice. Flow cytometric, Western blot and immunohistochemistry analyses were performed to investigate the mechanisms involved in the activity of single drugs and in their interaction when used in combination. Sorafenib showed a strong sequence-dependent synergistic interaction in vitro with docetaxel, which was highly dependent on the drug sequence employed. In vivo, human pancreatic cancer-xenografted mice treated with docetaxel followed by sorafenib reduced and delayed tumor growth, with complete tumor regression observed in half of the mice. This marked antitumor effect resulted in an overall increase in mouse survival of about 70% and in a complete cure in 3 of the 8 treated mice. The strong activity was also accompanied by marked apoptosis induction, inhibition of tumor angiogenesis and downregulation of ERK signalling. Our results show that the docetaxel and sorafenib combination exerts high therapeutic efficacy in experimental models of human pancreatic cancer, indicating a promising antitumor strategy for clinical use.

Cellular retinol-binding protein-1 expression in endometrial stromal cells: physiopathological and diagnostic implications.

AIMS: Cellular retinol-binding protein-1 (CRBP-1) contributes to the maintenance of the differentiated state of the endometrium through retinol bioavailability regulation. The aim was to analyse CRBP-1 expression in endometrial stromal cells at eutopic and ectopic sites in different physiopathological conditions. METHODS AND RESULTS: Antibodies to CRBP-1, CD10 and alpha-smooth muscle actin were applied to proliferative (n = 10), secretory (n = 9) and atrophic (n = 7) endometrium, decidua (n = 4), adenomyosis (n = 5), endometriosis (n = 10), endometrial polyps (n = 9), simple endometrial hyperplasia (n = 6), well-differentiated endometrioid carcinoma (n = 6) and submucosal leiomyomas (n = 5). In some cases, Western blotting and reverse transcription-polymerase chain reaction were also applied. CRBP-1 was expressed by eutopic and ectopic endometrial stromal cells more markedly during the late secretory phase and in decidua of pregnancy. CRBP-1 expression was low in the stroma of atrophic endometrium and absent in myometrium, leiomyomas and cervical stroma. CD10 immunoreactivity was weak in atrophic endometrium and in decidua. CONCLUSIONS: CRBP-1 expression characterizes endometrial stromal cells at eutopic and ectopic sites and appears to be more specific than CD10. The level of CRBP-1 varies in intensity according to hormonal variations, reaching its maximum in predecidua and decidua. Thus, immunodetection of CRBP-1 may help to elucidate the physiopathological changes which occur in endometrial stroma and can also be applied as an adjuvant stromal marker.

The recombinant major allergen of Parietaria judaica and its hypoallergenic variant: in vivo evaluation in a murine model of allergic sensitization.

BACKGROUND: Par j 1 represents the major allergenic component of Parietaria judaica pollen. Its three-dimensional structure is stabilized by four disulphide bridges. A family of three-dimensional mutants of the recombinant Par j 1 (rPar j 1) allergen, showing reduced allergenicity and retained T cell recognition has been recently developed by site-directed mutagenesis. OBJECTIVE: To develop and characterize a murine model of IgE sensitization to rPar j 1. To evaluate similarities between the murine model and the human IgE response. To investigate in this model the recognition of a hypoallergenic mutant of Par j 1, and to study the immune responses elicited in mice by the mutant itself. METHODS: BALB/c mice were sensitized by two intraperitoneal immunizations with rPar j 1 in alum on days 0 and 21. Allergen-specific serum IgE and IgG responses were studied by direct ELISA and immunoblotting, ELISA inhibition and competitive ELISA. Cell proliferation was evaluated in splenocyte cultures. RESULTS: Sensitization with rPar j 1 induced high levels of IgE and IgG1 vs. low levels of IgG2a. Mouse antibodies specific to rPar j 1 were able to compete with human IgE for recognition of rPar j 1. IgE from mice immunized with rPar j 1 showed a significantly reduced binding activity towards the hypoallergenic variant rPjC, which lacks three disulphide bridges. On the contrary, rPjC was recognized by IgG1 and IgG2a antibodies as well as rPar j 1. The proliferative response to rPjC by splenocytes from mice immunized with rPar j 1 was comparable to that stimulated by rPar j 1. Immunization with rPjC induced low levels of IgE antibodies to the rPjC itself, while IgG and proliferative responses were similar to those induced by rPar j 1. CONCLUSION: Conformational variants of allergens, displaying reduced allergenicity accompanied by retained IgG and T cell recognition, offer a safe, specific and flexible approach to immunotherapy of type I allergy. Our mouse model of IgE sensitization to a recombinant allergen, mimicking the human response to its native counterpart, could provide valuable information for pre-clinical testing of such hypoallergenic molecules.

Evolucion de prolactinomas en adolescentes / The course of prolactinomas in adolescents

Se presentan los resultados del tratamiento y seguimiento a mediano y largo plazo en 13 pacientes (8 niñas y 5 varones) de 14.2 a 20 años (x:16,5) portadores de prolactinoma. En el momento de la consulta los signos clínicos, más frecuentes fueron: cefaleas, detención o retraso de la pubertad, galactorrea y alteraciones visuales. La evaluación pretratamiento mostró respsuestas variables y discordantes de LH y FSH al ensayo de LH-RH, mientras que la prueba de TRH (TSH) resultó normal en 8/9 casos (una joven mostró un hipotiroidismo primario asociado). Los niveles de prolactina (PRL) basales estaban elevados en todos los casos entre 77 y 1150 ng/ml (x ñ SD : 378 ñ 285), sin incremento luego del TRH (x : 25%). En 8 casos, la cirugía constituyó el tratamiento inicial; 7 de ellos persistieron hiperprolactinémicos y requirieron bromocriptina (BEC) postquirúrgica. Cinco enfermos recibieron BEC como tratamiento primario; la TC mostró desaparición o importante reducción tumoral en 4 y ausencia de modificaciones en 1 niña por lo que se indicó cirugía secundariamente. La evolución pudo seguirse en 11 pacientes entre 2 y 12 años. De los operados inicialmente, 1 recuperó espontáneamente la función gonadal y 5 casos requirieron tratamiento complementario. De los 5 casos tratados primariamente con BEC, una recuperó ciclos espontáneamente logrado embarazo a los 10 meses, una completó el desarrollo puberal y presenta sangrados periódicos, una requirió tratamiento complementario con progesterona y el varón recuperó la función gonadal. La niña que fue secundariamente operada persiste normoprolactinémia sin recuperar sus ciclos. Concluimos que: 1) la PRL basal, junto con la evaluación radiológica, constituyen los elementos fundamentales en el diagnóstico de prolactinoma; 2) las evidencias clínicas y radiológicas abonarían en favor de una mayor rapidez evolutiva en los varones; 3) la BEC constituye un recurso valioso en el tratamiento primario o complementario de los prolactinomas en pacientes infantojuveniles (AU)

Evolucion de prolactinomas en adolescentes / The course of prolactinomas in adolescents

Se presentan los resultados del tratamiento y seguimiento a mediano y largo plazo en 13 pacientes (8 niñas y 5 varones) de 14.2 a 20 años (x:16,5) portadores de prolactinoma. En el momento de la consulta los signos clínicos, más frecuentes fueron: cefaleas, detención o retraso de la pubertad, galactorrea y alteraciones visuales. La evaluación pretratamiento mostró respsuestas variables y discordantes de LH y FSH al ensayo de LH-RH, mientras que la prueba de TRH (TSH) resultó normal en 8/9 casos (una joven mostró un hipotiroidismo primario asociado). Los niveles de prolactina (PRL) basales estaban elevados en todos los casos entre 77 y 1150 ng/ml (x ñ SD : 378 ñ 285), sin incremento luego del TRH (x : 25%). En 8 casos, la cirugía constituyó el tratamiento inicial; 7 de ellos persistieron hiperprolactinémicos y requirieron bromocriptina (BEC) postquirúrgica. Cinco enfermos recibieron BEC como tratamiento primario; la TC mostró desaparición o importante reducción tumoral en 4 y ausencia de modificaciones en 1 niña por lo que se indicó cirugía secundariamente. La evolución pudo seguirse en 11 pacientes entre 2 y 12 años. De los operados inicialmente, 1 recuperó espontáneamente la función gonadal y 5 casos requirieron tratamiento complementario. De los 5 casos tratados primariamente con BEC, una recuperó ciclos espontáneamente logrado embarazo a los 10 meses, una completó el desarrollo puberal y presenta sangrados periódicos, una requirió tratamiento complementario con progesterona y el varón recuperó la función gonadal. La niña que fue secundariamente operada persiste normoprolactinémia sin recuperar sus ciclos. Concluimos que: 1) la PRL basal, junto con la evaluación radiológica, constituyen los elementos fundamentales en el diagnóstico de prolactinoma; 2) las evidencias clínicas y radiológicas abonarían en favor de una mayor rapidez evolutiva en los varones; 3) la BEC constituye un recurso valioso en el tratamiento primario o complementario de los prolactinomas en pacientes infantojuveniles