Development of a construct-based risk assessment framework for genetic engineered crops.

Experience gained in the risk assessment (RA) of genetically engineered (GE) crops since their first experimental introductions in the early nineties, has increased the level of familiarity with these breeding methodologies and has motivated several agencies and expert groups worldwide to revisit the scientific criteria underlying the RA process. Along these lines, the need to engage in a scientific discussion for the case of GE crops transformed with similar constructs was recently identified in Argentina. In response to this need, the Argentine branch of the International Life Sciences Institute (ILSI Argentina) convened a tripartite working group to discuss a science-based evaluation approach for transformation events developed with genetic constructs which are identical or similar to those used in previously evaluated or approved GE crops. This discussion considered new transformation events within the same or different species and covered both environmental and food safety aspects. A construct similarity concept was defined, considering the biological function of the introduced genes. Factors like environmental and dietary exposure, familiarity with both the crop and the trait as well as the crop biology, were identified as key to inform a construct-based RA process.

A master protocol to investigate a novel therapy acetyl-L-leucine for three ultra-rare neurodegenerative diseases: Niemann-Pick type C, the GM2 gangliosidoses, and ataxia telangiectasia.

BACKGROUND: The lack of approved treatments for the majority of rare diseases is reflective of the unique challenges of orphan drug development. Novel methodologies, including new functionally relevant endpoints, are needed to render the development process more feasible and appropriate for these rare populations and thereby expedite the approval of promising treatments to address patients' high unmet medical need. Here, we describe the development of an innovative master protocol and primary outcome assessment to investigate the modified amino acid N-acetyl-L-leucine (Sponsor Code: IB1001) in three separate, multinational, phase II trials for three ultra-rare, autosomal-recessive, neurodegenerative disorders: Niemann-Pick disease type C (NPC), GM2 gangliosidoses (Tay-Sachs and Sandhoff disease; "GM2"), and ataxia telangiectasia (A-T). METHODS/DESIGN: The innovative IB1001 master protocol and novel CI-CS primary endpoints were developed through a close collaboration between the Industry Sponsor, Key Opinion Leaders, representatives of the Patient Communities, and National Regulatory Authorities. As a result, the open-label, rater-blinded study design is considerate of the practical limitations of recruitment and retention of subjects in these ultra-orphan populations. The novel primary endpoint, the Clinical Impression of Change in Severity© (CI-CS), accommodates the heterogenous clinical presentation of NPC, GM2, and A-T: at screening, the principal investigator appoints for each patient a primary anchor test (either the 8-m walk test (8MWT) or 9-hole peg test of the dominant hand (9HPT-D)) based on his/her unique clinical symptoms. The anchor tests are videoed in a standardized manner at each visit to capture all aspects related to the patient's functional performance. The CI-CS assessment is ultimately performed by independent, blinded raters who compare videos of the primary anchor test from three periods: baseline, the end of treatment, and the end of a post-treatment washout. Blinded to the time point of each video, the raters make an objective comparison scored on a 7-point Likert scale of the change in the severity of the patient's neurological signs and symptoms from video A to video B. To investigate both the symptomatic and disease-modifying effects of treatment, N-acetyl-L-leucine is assessed during two treatment sequences: a 6-week parent study and 1-year extension phase. DISCUSSION: The novel CI-CS assessment, developed through a collaboration of all stakeholders, is advantageous in that it better ensures the primary endpoint is functionally relevant for each patient, is able to capture small but meaningful clinical changes critical to the patients' quality of life (fine-motor skills; gait), and blinds the primary outcome assessment. The results of these three trials will inform whether N-acetyl-L-leucine is an effective treatment for NPC, GM2, and A-T and can also serve as a new therapeutic paradigm for the development of future treatments for other orphan diseases. TRIAL REGISTRATION: The three trials (IB1001-201 for Niemann-Pick disease type C (NPC), IB1001-202 for GM2 gangliosidoses (Tay-Sachs and Sandhoff), IB1001-203 for ataxia telangiectasia (A-T)) have been registered at www.clinicaltrials.gov (NCT03759639; NCT03759665; NCT03759678), www.clinicaltrialsregister.eu (EudraCT: 2018-004331-71; 2018-004406-25; 2018-004407-39), and https://www.germanctr.de (DR KS-ID: DRKS00016567; DRKS00017539; DRKS00020511).

Comparative studies of two-times-daily versus three-times-daily indinavir in combination with zidovudine and lamivudine.

OBJECTIVES: To compare the efficacy and safety of two-times-daily versus three-times-daily indinavir in combination with zidovudine and lamivudine. DESIGN: Two multicenter, open-label, randomized 24-week studies. METHODS: Adults HIV-1 infection, HIV-1 RNA greater than 10000 copies/ml, and no prior lamivudine or protease inhibitor therapy were eligible. In a pilot study (Study A), patients received indinavir at 800 mg every 8 h, 1000 mg every 12 h, or 1200 mg every 12 h. In a subsequent study (Study B), patients received indinavir at 800 mg every 8 h or 1200 mg every 12 h. All subjects received zidovudine (300 mg) and lamivudine (150 mg) every 12 h. An intent-to-treat analysis was used. RESULTS: In Study A, which enrolled 88 patients, neither HIV-1 RNA nor CD4 cell responses differed significantly between treatment groups at 24 weeks when corrected for multiple comparisons. Study B enrolled 433 patients, but was prematurely discontinued when interim analysis suggested greater efficacy of three-times-daily indinavir. Of the first 87 patients reaching week 24, HIV-1 RNA was less than 400 copies/ml in 91% receiving three-times-daily versus 64% receiving two-times-daily indinavir (P < 0.01). CONCLUSION: Three-times-daily indinavir appears more efficacious than two-times-daily dosing when administered with zidovudine and lamivudine. Two-times-daily indinavir dosing should only be considered in situations characterized by favorable pharmacokinetic drug-drug interactions.

Phase II study of gemcitabine, 5-fluorouracil, and leucovorin in patients with pancreatic cancer.

The primary goal of this phase II study was to determine the efficacy of gemcitabine (Gemzar; Eli Lilly and Company, Indianapolis, IN) plus 5-fluorouracil in patients with pancreatic cancer. Eligibility criteria included nonresectable locally advanced or metastatic pancreatic adenocarcinoma and measurable disease. Gemcitabine at 1,000 mg/m(2) and leucovorin at 20 mg/m(2) were administered intravenously 30 minutes before 5-fluorouracil 600 mg/m(2), weekly for 3 of every 4 weeks. Twenty nine patients were enrolled. The overall response rate was 21% (95% confidence interval: 8% to 40%), consisting of one complete response and five partial responses; 16 patients (55%) had stable disease. Median survival was 8.4 months (95% confidence interval: 2.6 to 14.2), and actuarial 1-year survival was 36%. Neutropenia (grade 3 only) was reported in 3.4% of patients, but was generally of short duration. No thrombocytopenia or evidence of cumulative myelosuppression was observed. The only significant nonhematologic events were grade 3 diarrhea and alopecia (both 3.4%). Gemcitabine plus 5-fluorouracil is active and well tolerated compared with results reported for each of these single agents. Thus, this combination justifies future comparative clinical trials. Semin Oncol 28 (suppl 10):44-49.

Paclitaxel in platinum-resistant ovarian cancer patients. Argentine Multicenter Taxol Group.

Paclitaxel (Taxol; Bristol-Myers Squibb Company; Princeton, NJ) is an antineoplastic agent that inhibits microtubular function and has shown efficacy in several solid tumors, mainly ovarian tumors, in which 20% to 40% response rates in previously treated patients were observed. We conducted a study to assess survival, response rate, and toxicity associated with paclitaxel treatment in patients with advanced ovarian cancer resistant to platinum therapy. Between September 1994 and November 1996, 38 patients were admitted for study and 37 were evaluable. All had disease progression or relapse within 1 year of receiving platinum-containing first-line chemotherapy. Mean age was 59 years (range, 30 to 75 years), all had bulky disease, and 18 showed increased carbohydrate antigen-125 at admission. They were treated every 3 weeks with paclitaxel 175 mg/m2 as a 3-hour infusion, preceded by standard premedication. Response rate was 51.3%, with a median response duration of 10.0 months and a median survival rate of 16.8 months. Mild to moderate hematologic toxicity was observed with only one episode of grade 4 neutropenia, without fever. Gastrointestinal toxicity was moderate and peripheral neuropathy was mild, except for two patients who had concomitant pathologies or previous treatment, which might have caused some neuropathy. We concluded that paclitaxel given as a 3-hour infusion was easily administered for ambulatory treatment, with mild to moderate toxicity and promising results based on rate and duration of response as well as survival.

Multicenter prospective surveillance of oral Candida dubliniensis among adult Brazilian human immunodeficiency virus-positive and AIDS patients.

The incidence of C. dubliniensis in South America has not yet been determined. In the present study, oral swab samples were taken from 108 HIV-infected/AIDS individuals attending 6 separate Brazilian HIV-treatment centers to determine the incidence of C. dubliniensis in this population. Swabs were plated onto CHROMagar Candida medium and 155 isolates, presumptively identified as C. albicans or C. dubliniensis were further investigated. In a preliminary screen for C. dubliniensis, 13 of the 155 isolates showed no or poor growth at 42 degrees C, and all them were subjected to randomly amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) analysis using C. dubliniensis-specific primers. We confirmed that 4 out of 13 isolates were C. dubliniensis, representing an incidence rate of 2.8% for the Brazilian HIV-infected population infected with yeasts exhibiting green colonies on CHROMagar Candida. This value is significantly lower than those reported in Ireland and the United States.

Prevalence of intestinal parasitic infections in patients with acquired immunodeficiency syndrome in Brazil.

OBJECTIVES: To evaluate the prevalence of intestinal parasitic infections and to investigate the possible associations of clinical status and laboratory findings with the different parasites found in stool samples. METHODS: Each patient was provided with one standard fecal collection vial containing 10% formalin for detecting ova, larvae, and cysts. To detect Cryptosporidium parvum and Isospora belli, the acid-fast Kinyoun stain and fluorescent auramine-rhodamine stain were used. RESULTS: A total of 200 patients with acquired immunodeficiency syndrome participated in this study; 40% were infected with at least one pathogenic species. The total prevalence of parasites was 16% for Giardia lamblia, 13% for Entamoeba coli, 7% for Cryptosporidium parvum, 3.5% for Endolimax nana, 2.5% for Ascaris lumbricoides, 2.5% for Strongyloides stercoralis, 2% for Isospora belli, and 0.5% for Blastocystis hominis. Results showed that diarrhea was significantly associated with cryptosporidiosis, giardiasis, and isosporiasis. However, no association was observed between the CD4+ cell counts and the manifestation of any particular parasite. CONCLUSIONS: The data support the value of standard fecal examinations in human immunodeficiency virus-infected patients, even in the absence of diarrhea, since these examinations easily can be performed, with low costs, and frequently disclose treatable conditions.

Neoadjuvant chemotherapy with cisplatin, ifosfamide and 5-fluorouracil in the treatment of locally advanced cervical cancer.

The objective of this study was to evaluate clinical and histological response, resectability, and survival in patients with cervical epidermoid carcinoma stage IB2 to IIIB with the use of neoadjuvant chemotherapy followed by radical surgery and/or radiation therapy. Between September 1989 and February 1996, 53 patients were admitted to this study. They were given three cycles of cisplatin 30 mg/m2/day, 5-fluorouracil 500 mg/m2/day, ifosfamide 2000 mg/m2/day i.v., and mesna 400 mg/m2/day i.v. at hour 0 and 400 mg/m2 at hours 4 and 8 during three days every 21-28 days. We evaluated 47 patients. Global clinical response obtained was 85% {95% (CI), 75-97%, CR in 14 patients (30%) and PR in 26 patients (55%)}. Twenty-three patients underwent surgery. Six patients (13%) had a complete histological response. Median follow-up was 42 months (5-96). In resected patients, with a median follow-up of 57 months (5-96), the estimated five-year disease-free survival was 78%. Global survival estimated to 60 months was 83% for stage IB2, 70% for IIB, and 20% for IIIB. This mode of therapy offers a new option to improve survival in locally advanced cervical cancer. Randomized trials are required in order to establish a definitive role for this therapeutic strategy.

Lactic acidosis and antiretroviral therapy: a case report and literature review.

Antiretrovirals, particularly nucleoside analogue reverse transcriptase inhibitors (RTIs) - DDI, 3TC and D4T, are widely used to effectively control human immunodeficiency virus (HIV) infection. These drugs have several adverse effects including anemia, peripheral neuropathy, pancreatitis and, on rare occasions, lactic acidosis. We describe the case of a 39 year old patient who had severe lactic acidosis after receiving stavudine (D4T) and didanosine (DDI) for an 8 month period. She had never manifested an opportunistic infection and presented a CD4 count of 378 cells/mm3 and an undetectable viral load (< 400 copies/ml). The purpose of the following report is to alert clinicians and infectious diseases specialists to the occurrence of lactic acidosis in asymptomatic HIV patients receiving antiretrovirals for long periods of time.

Human carcinogens: an evaluation study via the COMPACT and HazardExpert procedures.

The results of computer-optimized molecular parametric analysis of chemical toxicity (COMPACT) and HazardExpert evaluations on 14 established human carcinogens are reported. The concordances between COMPACT and carcinogenicity (71%) and between HazardExpert and carcinogenicity (57%) are significantly improved when taken in combination, where all 14 carcinogens are correctly identified by the two systems used in conjunction. However, if a negative energy of the highest occupied molecular orbital (E(HOMO)) value is regarded as evidence of electrophilic reactivity likely to give rise to mutagenicity and carcinogenicity, then 13/14 (93%) of the carcinogens are correctly identified by combination with the COMPACT procedure alone. It is possible, therefore, to establish likely carcinogenicity arising from either P450 mediation (CYP1 and CYP2E) or compound electrophilicity via the employment of a straightforward approach to molecular and electronic structure calculation, a process that can be performed in a relatively short time frame (i.e., less than 1 hour per chemical) and at a low cost.

[Visceral leishmaniasis and acquired immunodeficiency syndrome (AIDS). Report of a case]. / Leishmaniose visceral e síndrome da imunodeficiência adquirida (SIDA). Relato de um caso.

This is a case report that describe an association of AIDS, visceral leishmaniasis and probable disseminated tuberculosis. Due to the spread of AIDS in developing areas worldwide this association would be more frequently, seen on subjects from endemic areas where this protozoonosis is prevalent. More than one opportunistic infection related with the endemic diseases of the developing regions can be associated with those immunocompromised patients.

Randomized, double-blind trial comparing indinavir alone, zidovudine alone and indinavir plus zidovudine in antiretroviral therapy-naive HIV-infected individuals with CD4 cell counts between 50 and 250/mm3.

Treatment with indinavir has been shown to result in marked decreases in viral load and increases in CD4 cell counts in HIV-infected individuals. A randomized double-blind study to evaluate the efficacy of indinavir alone (800 mg q8h), zidovidine alone (200 mg q8h) or the combination was performed to evaluate progression to AIDS. 996 antiretroviral therapy-naive patients with CD4 cell counts of 50-250/mm3 were allocated to treatment. During the trial the protocol was amended to add lamivudine to the zidovudine-containing arms. The primary endpoint was time to development of an AIDS-defining illness or death. The study was terminated after a protocol-defined interim analysis demonstrated highly significant reductions in progression to a clinical event in the indinavir-containing arms, compared to the zidovudine arm (p<0. 0001). Over a median follow-up of 52 weeks (up to 99 weeks), percent reductions in hazards for the indinavir plus zidovudine and indinavir groups compared to the zidovudine group were 70% and 61%, respectively. Significant reductions in HIV RNA and increases in CD4 cell counts were also seen in the indinavir-containing groups compared to the zidovudine group. Improvement in both CD4 cell count and HIV RNA were associated with reduced risk of disease progression. All three regimens were generally well tolerated.

[An evaluation of the relationship between intestinal parasitoses and the risk factors for HIV in AIDS patients]. / Avaliação da relação entre parasitoses intestinais e fatores de risco para o HIV em pacientes com AIDS.

We conducted a cohort survey on 200 AIDS patients in São Paulo, SP, Brazil, to verify if the presence of enteroparasites is associated with the various risk factors for HIV infection. Diarrhea was significantly more frequent in the group of patients presenting enteroparasitosis (p < 0.00001). Giardia lamblia, found in 32 (16%) cases, was the most prevalent parasite. The presence of parasites in the stool samples was not significantly associated with risk factors for HIV infection, mainly when considering Giardia lamblia and Cryptosporidium parvum (p = 0.99 and 0.69, respectively). The controversy found in the literature indicates that additional studies should be done in the different geographical regions of the country.

Enteric parasites and AIDS.

OBJECTIVE: To report on the importance of intestinal parasites in patients with AIDS, showing relevant data in the medical literature, with special emphasis on epidemiology, diagnosis and treatment of enteroparasitosis, especially cryptosporidiosis, isosporiasis, microsporidiasis and strongyloidiasis. DESIGN: Narrative review.

Regression of AIDS-related Kaposi's sarcoma following combined antiretroviral treatment.

The aim of the study was to assess regression of Kaposi's sarcoma (KS) in AIDS patients in Argentina. Eighteen male AIDS patients with human immunodeficiency virus (HIV)-associated Kaposi's sarcoma at different clinical stages received KS specific treatment and/or anti-retroviral therapy. Triple anti-retroviral therapy was given to most of the patients with the exception of four who received zidovudine (ZDV) in combination with another nucleoside analogue but no protease inhibitors. Plasma viral load and CD4+ T lymphocyte number were measured in two blood samples (before and after treatment). Complete remission was found in all patients (five) at KS stage I, three out of eight patients at stage II but in none at stages III and IV. Two out of three patients at KS stage IV did not respond to treatments at all. Three patients at KS stages I and II showed complete remission of sarcoma with only anti-retroviral therapy suggesting that anti-retroviral therapy and non-KS specific chemotherapy can successfully control KS.

[Stimulus of the hypophyseal-adrenocortical axis with corticotropin releasing hormone (CRH) in acquired immunodeficiency syndrome. Evidence for activation of the immune-neuroendocrine system]. / Estímulo do eixo hipófise-adrenocortical com o hormônio liberador de corticotrofina (CRH) na síndrome de imunodeficiência adquirida. Evidência de ativação do sistema imune-neuroendócrino.

Ten-20% of patients with AIDS may present clinical evidence of primary or secondary adrenal insufficiency. PURPOSE--To evaluate the hypothalamic-pituitary-adrenocortical axis (HPAA) with CRH in patients with AIDS. METHODS--We studied 20 patients with AIDS and 17 normal subjects (NS) with exogenous ACTH (cosyntropin, 250 micrograms IV bolus) followed one week later by ovine corticotropin releasing hormone (oCRH 1 microgram/kg BW IV bolus). Basal and 60' cortisol (micrograms/dL) were determined in the former whereas ACTH (pg/mL) and cortisol were measured every 15-30' for 2 hours in the latter. RESULTS--Basal and peak values (mean +/- SD) of ACTH and cortisol for both tests were: cosyntropin test (AIDS x NS): basal cortisol 22.5 +/- 7.1 x 10.6 +/- 3.6 (p < 0.01), peak 36.0 +/- 12.8 x 28.3 +/- 7.6 (p < 0.05); oCRH test: basal ACTH 42.2 +/- 33.5 x 28.9 +/- 12.7 (NS), peak 104.7 +/- 62.2 x 59.3 +/- 17.6 (p < 0.05); basal cortisol 19.7 +/- 9.0 x 10.1 +/- 3.4 (p < 0.01), peak 27.5 +/- 8.9 x 18.3 +/- 5.1 (p < 0.05). CONCLUSION--AIDS patients had elevated basal and CRH stimulated ACTH levels and an intact glucocorticoid pathway with elevated basal and peak cortisol levels to both stimulation tests. This situation is probably due to the stressful disease condition, where lymphokines may play a role activating the hypothalamic-pituitary axis.

Gliosarcoma: an unusual cause of cerebral mass lesion in a patient with AIDS. A case report and review of the literature.

Malignant glioma is the most common primary brain neoplasm. Generally, gliomas are not included in the differential diagnosis of enhancing lesions of the central nervous system in patients infected by the human immunodeficiency virus. We report a case of gliosarcoma in a patient with AIDS presenting as a single cerebral lesion. Stereotactic brain biopsy was obtained and definitive histopathological diagnosis of gliosarcoma was made. A decline in the incidence of opportunistic infections associated with highly active antiretroviral therapy suggest the importance of early stereotactic biopsy to confirm the diagnosis of these neoplasms.

Mycobacteraemia among HIV-1-infected patients in São Paulo, Brazil: 1995 to 1998.

From July 1995 to August 1998, mycobacterial blood cultures were obtained from 1032 HIV-infected patients seen at the Centro de Referência e Treinamento de AIDS (CRTA), Hospital São Paulo (HSP), and Centro de Referência de AIDS de Santos (CRAS). Overall, 179 episodes of mycobacteraemia were detected: 111 (62.0%) at CRTA, 50 (27.9%) at HSP, and 18 (10.1%) at CRAS. The frequency of positive cultures declined sharply from 22.6% in 1995 to 6.9% in 1998, consistent with the decrease in opportunistic infections following the publicly funded distribution of highly active antiretroviral therapy. In 1995, mycobacteraemia was more frequently due to Mycobacterium avium complex (59.2%) than Mycobacterium tuberculosis (28.6%), whereas in 1998 the relative frequencies were reversed (28.6 vs. 64.3% respectively), probably justified by the increased virulence of M. tuberculosis and the greater risk of invasive infection in less-immunocompromised patients, including patients unaware they are infected with HIV.