Optimization of nanostructured lipid carriers for Zidovudine delivery using a microwave-assisted production method.

An adapted methodology for obtaining lipid nanoparticles that only uses the microwave reactor in the synthesis process was developed. The method has the following features: one-pot, one-step, fast, practical, economical, safe, readiness of scaling-up, lack of organic solvents and production of nanoparticles with low polydispersity index (PDI) (below 0.3). This new method was applied for the development of nanostructured lipid carriers (NLC) loaded with a hydrophilic drug, the antiretroviral agent zidovudine (AZT). The aim of the present work was to develop, evaluate and compare optimized NLC formulations produced by two different methods - hot ultrasonication and microwave-assisted method. The development and optimization of the NLC formulations were supported by a Quality by Design (QbD) approach. All formulations were physicochemically characterized by the same parameters. The optimized formulations presented a suitable profile for oral administration (particle size between 100 and 300 nm, PDI < 0.3 and negative zeta potential >-20 mV). Furthermore, the morphologies assessed by TEM showed spherical shape and confirmed the results obtained by DLS. Both AZT loaded formulations were physically stable for at least 45 days and non-toxic on Jurkat T cells. Drug release studies showed a controlled release of AZT under gastric and plasma-simulated conditions.

Human papillomavirus infection among sexual partners attending a Sexually Transmitted Disease Clinic in Rio de Janeiro, Brazil

Cervical cancer is a major source of illness and death among women worldwide and genital infection with oncogenic human papillomavirus (HPV) its principal cause. There is evidence of the influence of the male factor in the development of cervical neoplasia. Nevertheless, the pathogenic processes of HPV in men are still poorly understood. It has been observed that different HPV types can be found among couples. The objective of the present study was to investigate HPV infections in female patients (n = 60 females/group) as well as in their sexual partners and to identify the concordance of HPV genotypes among them. By using the polymerase chain reaction, we detected a 95% prevalence of HPV DNA in women with cervical intraepithelial neoplasia (CIN) compared to 18.3% in women with normal cervical epithelium, with a statistically significant difference (P < 0.001). The HPV DNA prevalence was 50% in male partners of women with CIN and 16.6% in partners of healthy women. In the control group (healthy women), only 9 couples were simultaneously infected with HPV, and only 22.2% of them had the same virus type, showing a weak agreement rate (kappa index = 0.2). Finally, we observed that HPV DNA was present in both partners in 30 couples if the women had CIN, and among them, 53.3% shared the same HPV type, showing moderate agreement, with a kappa index of 0.5. This finding supports the idea of circulation and recirculation of HPV among couples, perpetuating HPV in the sexually active population, rather than true recurrences of latent infections.

Human papillomavirus infection among sexual partners attending a Sexually Transmitted Disease Clinic in Rio de Janeiro, Brazil.

Cervical cancer is a major source of illness and death among women worldwide and genital infection with oncogenic human papillomavirus (HPV) its principal cause. There is evidence of the influence of the male factor in the development of cervical neoplasia. Nevertheless, the pathogenic processes of HPV in men are still poorly understood. It has been observed that different HPV types can be found among couples. The objective of the present study was to investigate HPV infections in female patients (n = 60 females/group) as well as in their sexual partners and to identify the concordance of HPV genotypes among them. By using the polymerase chain reaction, we detected a 95% prevalence of HPV DNA in women with cervical intraepithelial neoplasia (CIN) compared to 18.3% in women with normal cervical epithelium, with a statistically significant difference (P < 0.001). The HPV DNA prevalence was 50% in male partners of women with CIN and 16.6% in partners of healthy women. In the control group (healthy women), only 9 couples were simultaneously infected with HPV, and only 22.2% of them had the same virus type, showing a weak agreement rate (kappa index = 0.2). Finally, we observed that HPV DNA was present in both partners in 30 couples if the women had CIN, and among them, 53.3% shared the same HPV type, showing moderate agreement, with a kappa index of 0.5. This finding supports the idea of circulation and recirculation of HPV among couples, perpetuating HPV in the sexually active population, rather than true recurrences of latent infections.

Human papillomavirus detection and p16 methylation pattern in a case of esophageal papilloma

Esophageal cancer is a prevalent cancer worldwide. Some studies have reported the possible etiology of human papillomavirus (HPV) in benign and malignant papillomas of the esophagus but the conclusions are controversial. In the present study, we investigated an esophageal papilloma from a 30-year-old male patient presenting aphasia. HPV DNA was detected by generic PCR using MY09/11 primers, and restriction fragment length polymorphism revealed the presence of HPV54, usually associated with benign genital lesions. Hypermethylation of the pINK4A gene was also investigated due to its relation to malignant transformation, but no modification was detected in the host gene. Except for an incipient reflux, no risk factors such as cigarette smoking, alcohol abuse or an infected sexual partner were recorded. Since esophageal lesions may have a malignant potential, HPV detection and typing are useful tools for patient follow-up.

Human papillomavirus detection and p16 methylation pattern in a case of esophageal papilloma.

Esophageal cancer is a prevalent cancer worldwide. Some studies have reported the possible etiology of human papillomavirus (HPV) in benign and malignant papillomas of the esophagus but the conclusions are controversial. In the present study, we investigated an esophageal papilloma from a 30-year-old male patient presenting aphasia. HPV DNA was detected by generic PCR using MY09/11 primers, and restriction fragment length polymorphism revealed the presence of HPV54, usually associated with benign genital lesions. Hypermethylation of the pINK4A gene was also investigated due to its relation to malignant transformation, but no modification was detected in the host gene. Except for an incipient reflux, no risk factors such as cigarette smoking, alcohol abuse or an infected sexual partner were recorded. Since esophageal lesions may have a malignant potential, HPV detection and typing are useful tools for patient follow-up.

Patterns of genotype distribution in multiple human papillomavirus infections.

The relationship between severe-grade cervical lesions and clusters of human papillomavirus (HPV) genotypes in a taxonomic classification was surveyed in 232 women with previous abnormal cytology. HPV co-infections were clustered according to phylogenetic criteria. Multiple infections were detected in 22.0% of the entire sample. Clade A10 (represented by HPV-6 and HPV-11) appeared more frequently in multiple infections than clade A9, which was represented by five of the most common high-risk types, including HPV-16. Although HPV-16 was the most frequent genotype, it was not more prevalent in multiple infections. Abortion and two or more sexual partners were risk-factors associated with HPV co-infections. Severe cervical dysplasia was associated with co-infections with oncogenic types from different clades, with the association being significant for the high-risk clades A7 and A9.

Human papillomavirus prevalence and predictors for cervical cancer among high-risk women from Rio de Janeiro, Brazil.

We assess the prevalence of human papillomavirus (HPV) and cofactors for cervical severe disease, as contribution for vaccine strategies at the right moment in which Brazilian health authorities have approved an anti-HPV vaccine. A case-control study was undertaken with 201 women who attended a public health service with previous abnormal cytology. The HPV status was ascertained by consensus primers My09/11 and typed by 6, 11, 16, 18, 31, 33, 35, and 58 specific primers. Patients diagnosed with high-grade squamous intraepithelial lesions (HSIL) and cervical cancer were referred as cases (n = 84). Patients with normal/inflammatory cervix or carrying benign cervical lesions were included in controls (n = 117). The overall prevalence of HPV infection was 75.6%, with 91.7% among cases. In spite of HPV 16 being the most frequent type (53.3%), 27.6% of infections were attributed to nonvaccine types. High-risk HPV were strongly associated to older women (OR = 6.7). Otherwise, age at the first intercourse (OR = 7.10), three or more parities (OR = 3.05), abortion episodes (OR = 4.80), and smoking (OR = 3.83) conferred a heavy effect in younger women. Among mediators affecting the progress from HSIL to cancer, age played the main role in easing the progression (OR = 1.09, P = 0.002) followed by education level (OR = 4.20, P = 0.066). White ethnia showed to be a protective factor (OR = 0.32, P = 0.055). Predictors from HPV exposure to malignant disease include demographic and behavioral factors. Public policies such as improvement of education and continued prevention campaigns might contribute to reduce this picture. This work also gives background, in identifying a target population, for implementing future vaccine strategies.

Human papillomavirus infection in Rio de Janeiro, Brazil: a retrospective study

There is considerable data to support a central role for human papillomavirus (HPV) in the etiology of cervical cancer. More than a 100 HPV types have been described, and 40 have been isolated from benign and malignant genital lesions. Consequently, there is strong motivation to evaluate HPV testing for cervical cancer screening. Few studies concerning the natural history of HPV infection have been conducted in the state of Rio de Janeiro. We determined the prevalence of HPV types in female genital lesions by using Hybrid Capture Assay (HCA) and we retrospectively analyzed the course of HPV infection. Our sample included 788 women attended at Laboratórios Sérgio Franco. The average age of the participants was 29.6 years. HPV prevalence and cytological diagnosis were determined. The overall prevalence of HPV DNA in the study group was 50.1 percent (395/788), ranging from 25 percent (NORMAL) to 100 percent in high-grade intraepithelial lesions (HSIL). High risk HPV was found in 12 percent inflammatory, 58.3 percent HPV, 63.2 percent LSIL and 100 percent HSIL. A retrospective analysis of 78 patients showed that 22 presented persistent lesions, 2 had progressive lesions, 4 had regressive lesions, 13 showed latent infections, 18 were transiently infected and 19 were submitted to curative treatment. No cases of cancer were registered in this population, which can afford private medical care and regular follow-up exams. We suggest that HCA be used in specific cases involving persistent and recurrent lesions.

Human papillomavirus infection in Rio de Janeiro, Brazil: a retrospective study.

There is considerable data to support a central role for human papillomavirus (HPV) in the etiology of cervical cancer. More than a 100 HPV types have been described, and 40 have been isolated from benign and malignant genital lesions. Consequently, there is strong motivation to evaluate HPV testing for cervical cancer screening. Few studies concerning the natural history of HPV infection have been conducted in the state of Rio de Janeiro. We determined the prevalence of HPV types in female genital lesions by using Hybrid Capture Assay (HCA) and we retrospectively analyzed the course of HPV infection. Our sample included 788 women attended at Laboratórios Sérgio Franco. The average age of the participants was 29.6 years. HPV prevalence and cytological diagnosis were determined. The overall prevalence of HPV DNA in the study group was 50.1% (395/788), ranging from 25% (NORMAL) to 100% in high-grade intraepithelial lesions (HSIL). High risk HPV was found in 12% inflammatory, 58.3% HPV, 63.2% LSIL and 100% HSIL. A retrospective analysis of 78 patients showed that 22 presented persistent lesions, 2 had progressive lesions, 4 had regressive lesions, 13 showed latent infections, 18 were transiently infected and 19 were submitted to curative treatment. No cases of cancer were registered in this population, which can afford private medical care and regular follow-up exams. We suggest that HCA be used in specific cases involving persistent and recurrent lesions.

Hepatitis B infection among patients attending a sexually transmitted diseases clinic in Rio de Janeiro, Brazil.

Hepatitis B virus (HBV) has a low endemicity in Rio de Janeiro, Brazil. Sexual transmission must play an important role in this virus, but the prevalence and risk factors have never been properly investigated. The aim of this paper is to determine the prevalence and risk factors for HBV infection in patients attending a Sexually Transmitted Diseases Clinic of the Universidade Federal Fluminense, from the State of Rio de Janeiro, Brazil. In a retrospective study, HBV seroprevalence was investigated in 440 patients. Serum of each patient was assayed for antibodies against hepatitis B core antigen (anti-HBc), hepatitis B surface antigen (HBsAg) and antibodies against hepatitis B surface antigen (anti-HBs). Demographic and risk factor data were extracted from clinic notes. The overall seroprevalence of exposure markers for HBV (anti-HBc, HBsAg and anti-HBs) were 13%, 3.4% and 8.5% respectively. Homo/bisexual behaviour, anal intercourse, HIV infection, positive serology for syphilis and blood transfusion were predictors of the HBV exposure. Among demographic data, age and place of birth were associated with the anti-HBc seropositivity.

Epidemiological aspects of human papillomavirus infection and cervical cancer in Brazil.

OBJECTIVES: A study was conducted to determine the prevalence of HPV infection in cervical lesions of Brazilian women and to search for specific risk factors associated to progression to malignancy. METHODS: Five hundred and fourteen paraffin-embedded biopsies obtained from female cervical lesions were classified according to the Bethesda System in low grade squamous intraepithelial lesion (LSIL), high grade SIL (HSIL) and squamous cell carcinoma (SCC). Specimens were tested for the presence of human papillomavirus (HPV) types 6/11,16/18 and 31/33/35 DNA using non-isotopic in situ hybridization. RESULTS: HPV prevalence ranged from 85.6% in LSIL to 55.2% in SCC. Patients were analysed through an 8 year period: 27. 1% of the lesions spontaneously regressed while 43.6% persisted and 29.3% progressed to carcinoma. High risk types were present in 80.5% (Crude OR 13.8, P=0.0003) of the progressive lesions. Possible co-factors have also been evaluated: history of other sexually transmitted diseases, mainly syphilis, showed to be positively related to progression (Adjusted OR 13.0, P=0.0003) while oral contraceptive use and tobacco smoking were not significantly related to it (P>O.1). Association of two or more co-factors also proved to be related to progression. CONCLUSIONS: Oncogenic HPV types 16 and 18 and history of other concurrent sexually transmitted diseases were found to be significantly associated with progression to cancer. Smoking and the use of oral contraceptives did not show a relation to cancer establishment, but when they were associated a significant co-operative role in progression was demonstrated. Our study indicated that HPV and other risk factors for cancer can act together, corroborating the observation of a poor prognosis for Brazilian women presenting SILs.

Human papillomavirus infection and cervical cancer in Brazil: a retrospective study.

Two hundred and thirty paraffin-embedded biopsies obtained from female cervical lesions were tested for the presence of human papillomavirus (HPV) types 6/11, 16/18 and 31/33/35 DNA using non-isotopic in situ hybridization. Specimens were classified according to the Bethesda System in low grade squamous intraepithelial lesion (LSIL), high grade SIL (HSIL) and squamous cell carcinoma (SCC). HPV prevalence ranged from 92.5% in LSIL to 68.5% in SCC. Benign types were prevalent in LSILs while oncogenic types infected predominantly HSILs and SCC. HPV infection showed to be age-dependent, but no significant relation to race has been detected. Patients were analyzed through a five-year period: 20.7% of the lesions spontaneously regressed while 48.9% persisted and 30.4% progressed to carcinoma. Patients submitted to treatment showed a 19.4% recurrence rate. High risk types were present in 78.6% (CrudeOR 13.8, P = 0.0003) of the progressive lesions, and in 73.7% of the recurrent SILs (COR 19.3, P = 0.0000001). Possible co-factors have also been evaluated: history of other sexually transmitted diseases showed to be positively related either to progression (Adjusted OR 13.0, P = 0.0002) or to recurrence (AOR 17.2, P = 0.0002) while oral contraceptive use and tobacco smoking were not significantly related to them (P > 0.1). Association of two or more co-factors also proved to be related to both progression and recurrence, indicating that they may interact with HPV infection in order to increase the risk of developing malignant lesions.

PIP: 230 paraffin-embedded biopsies obtained from female cervical lesions were tested for the presence of human papillomavirus (HPV) type 6/11, 16/18, and 31/33/35 DNA using non-isotopic in situ hybridization. Specimens were classified according to the Bethesda System as low grade squamous intraepithelial lesion (LSIL), high grade SIL (HSIL), and squamous cell carcinoma (SCC). HPV prevalence ranged from 92.5% in LSIL to 68.5% in SCC. Benign types were prevalent in LSILs, while oncogenic types infected predominantly HSILs and SCC. HPV infection was found to be age-dependent, but no significant relation to race has been detected. Patients were analyzed through a 5-year period: 20.7% of the lesions spontaneously regressed, while 48.9% persisted and 30.4% progressed to carcinoma. Patients submitted to treatment showed a 19.4% recurrence rate. High risk types were present in 78.6% (crude OR (COR), 13.8; P = 0.0003) of the progressive lesions, and in 73.7% of the recurrent SILs (COR, 19.3; P = 0.0000001). Possible cofactors have also been evaluated: history of other sexually transmitted diseases was found to be positively related either to progression (adjusted OR (AOR), 13.0; P = 0.0002) or to recurrence (AOR, 17.2; P = 0.0002), while oral contraceptive use and tobacco smoking were not significantly related to them (P 0.1). Association of two or more cofactors also proved to be related to both progression and recurrence, indicating that they may interact with HPV infection in order to increase the risk of developing malignant lesions.

Análise epidemiológica da patologias genitais associadas a papilomavírus / Epidemiologic analysis of genital pathologies associated to papillomavirus

Estudando-se casos de infecçoes genitais associadas a papilomavírus em um hospital universitário, verificou-se que as patologias mais frequentes eram o condiloma acuminado e câncer epidermóide. Objetivo. Analisar a prevalência das lesoes genitais associadas a papilomavírus e distribuiçao por sexo, idade e cor. Métodos. Foram estudadas 223 pacientes atendidas no Hospital Universitário Antonio Pedro, estado do Rio de Janeiro, durante os anos de 1988 a 1992. Após diagnóstico clínico, biópsias das pacientes foram submetidas a análise histológica. Resultados. Os resultados mostraram que, entre as doenças do trato genital associadas a papilomavírus, o condiloma acuminado e o carcinoma epidermóide eram as mais frequentes. Os casos de carcinoma epidermóide mantiveram prevalência estável ao longo do período, enquanto que o número de casos condiloma acuminado aumentou de três para 33 por ano. As duas patologias foram mais frequentes em indivíduos de cor branca. Nao houve diferença significativa entre sexo e presença de condiloma. A média de idade para carcinoma epidermóide nao sofreu variaçao significativa no período estudado, enquanto decaiu para o condiloma acuminado nos períodos de 1988 para 1989 e de 1991 para 1992. Conclusao. As lesoes genitais associadas a papilomavírus aumentaram significativamente no período de cinco anos. Maior atençao deve ser dada as campanhas de controle de diagnóstico precoce.

[Epidemiologic analysis of genital diseases associated to papillomavirus]. / Análise epidemiológica de patologias genitais associadas a papilomavírus.

UNLABELLED: In order to investigate genital papillomavirus infection in a university hospital, we verified that condyloma acuminata and epidermoid carcinoma were the most common diseases associated to this virus. PURPOSE: This study was undertaken to verify the prevalence of genital lesions associated to papillomavirus and sex, age and race distribution. METHODS: We have reported 223 patients who were treated at Hospital Universitario Antonio Pedro, Rio de Janeiro State, during the period 1988-1992. After clinical diagnosis, biopsies were submitted to histological analysis. RESULTS: Results showed that condyloma acuminata and epidermoid carcinoma were the most frequent lesions among genital lesions. Carcinoma cases had a stable prevalence along the period while condyloma cases had a significant increase. Both lesions were more frequently found in Caucasian (white) patients. There were no differences between sexes and condyloma evidence. The media age in the epidermoid carcinoma cases did not change in the years, but it has decreased regarding condyloma acuminata. CONCLUSION: Genital diseases associated to papillomavirus increased during five years. Attention must be paid to control and early diagnosis to prevent neoplasic evolution.

Cure of prophage in Staphylococcus aureus by furocoumarin photoadditions.

In a previous report curing of prophage in Staphylococcus aureus by treatment with 8-methoxypsoralen plus near-UV (365 nm) radiation was described. In the present work the use of angelicin and 4,5',8-trimethylpsoralen and repeated irradiation show that monoadducts and DNA interstrand cross-links formed by furocoumarin photoaddition are involved in the curing of the prophage responsible for the suppression of the lipolytic activity in a putative repair proficient strain of Staphylococcus aureus.

Prevalence of human papillomavirus DNA in female cervical lesions from Rio de Janeiro, Brazil

A hundred-sixty paraffin-embedded specimens from female cervical lesions were examined for human papillomavirus (HPV) types 6, 11, 16 and 18 infections by non-isotopic in situ hybridization. The data were compared with histologic diagnosis. Eighty-eight (55) biopsies contained HPV DNA sequences. In low grade cervical intraepithelial neoplasias (CIN I), HPV infection was detected in 78.7 of the cases, the benign HPV 6 was the most prevalent type. HPV DNA was detected in 58 of CIN II and CIN III cases and in 41.8 of squamous cell carcinomas (SCC). Histologically normal women presented 20 of HPV infection. Oncogenic HPV was found in 10 of these cases, what may indicate a higher risk of developing CINs and cancer. Twenty-five percent of the infected tissues contained mixed infections. HPV 16 was the most common type infecting the cervix and its prevalence raised significantly with the severity of the lesions, pointing its role in cancer pathogenesis. White women presented twice the cervical lesions of mulatto and African origin women, although HPV infection rates were nearly the same for the three groups (approximately 50). Our results showed that HPV typing by in situ hybridization is a useful tool for distinguishing between low and high risk cervical lesions. Further studies are required to elucidate risk factors associated with HPV infection and progression to malignancy in Brazilian population.

Prevalence of human papillomavirus DNA in female cervical lesions from Rio de Janeiro, Brazil.

A hundred-sixty paraffin-embedded specimens from female cervical lesions were examined for human papillomavirus (HPV) types 6, 11, 16 and 18 infections by non-isotopic in situ hybridization. The data were compared with histologic diagnosis. Eighty-eight (55%) biopsies contained HPV DNA sequences. In low grade cervical intraepithelial neoplasias (CIN I), HPV infection was detected in 78.7% of the cases, the benign HPV 6 was the most prevalent type. HPV DNA was detected in 58% of CIN II and CIN III cases and in 41.8% of squamous cell carcinomas (SCC). Histologically normal women presented 20% of HPV infection. Oncogenic HPV was found in 10% of these cases, what may indicate a higher risk of developing CINs and cancer. Twenty-five percent of the infected tissues contained mixed infections. HPV 16 was the most common type infecting the cervix and its prevalence raised significantly with the severity of the lesions, pointing its role in cancer pathogenesis. White women presented twice the cervical lesions of mulatto and African origin women, although HPV infection rates were nearly the same for the three groups (approximately 50%). Our results showed that HPV typing by in situ hybridization is a useful tool for distinguishing between low and high risk cervical lesions. Further studies are required to elucidate risk factors associated with HPV infection and progression to malignancy in Brazilian population.

Human papillomavirus detection in genital lesions by in situ hybridization and ultrastructural observations

Detection of papillomavirus DNA in sity hybridization technique was perfomed in 29 symptomatic patients (6 males and 23 females) during the period of 1989-1991 at the Clinic for Sexually Transmitted Diseases, Universidade Federal Fluminense, State of rio de Janeiro. All the male patients had condyloma acuminata. Only HPV 6/11 were found in these lesions. Clinical features inthe female patients included vulvar condyloma acuminata, bowenoid populosis, flat cervical condyloma, cervical condyloma acuminatum and cervical intraepithelialneoplasia grade II (CIN II). We also found cases of condyloma acuminata associated to vulvar intraepithelial neoplasia grade III (VIN III), as well as to vaginal invasive carcinoma. HPV 6/11 and 16/18 were found in vulvar condyloma acuminata. Mixed infection by 6/11-16/18 HPV were also seen in these lesions as well as in the patient who had cervical condyloma acuminatum. HPV 16/18 were found in the condyloma acuminatum plus VIN III and in the CIN II lesions. We have found HPV31/33/51 in the specimen of condyloma acuminatum plus invasive carcinoma. In order to investigate the ultrastructural aspects of HPV infection in genital tissue, the biopsies of three female patients were observed under electron microscope.Mature virus particles were found in the cells of a condyloma acuminatum as wellas in the condyloma acuminatum plus invasive carcinoma case. In another sample, chromosome breakages were found in the nuclei of the infected cells although no viral particles were observed