Weeks Of Withdrawal Research Articles

Post delivery withdrawal time of tenofovir for blocking mother to child transmission of HBV in women with high HBV load during late pregnancy

Purpose: To study the withdrawal time of tenofovir disoproxil fumarate (TDF) after delivery in parturient women having high hepatitis B virus (HBV) load who took TDF during late pregnancy to block fetal contact with HBV.
 Methods: This is a retrospective analysis of clinical data of 77 pregnant women close to parturition with high HBV density in Southern Central Hospital of Yunan Province from February 2020 to February 2021. Based on time of drug discontinuation, they were assigned to group A (immediately after delivery, n = 24); group B (1 month after delivery, n = 26), and group C (3 months after delivery, n = 27). All three groups took TDF before delivery. Liver function, with alanine aminotransferase (ALT) was assessed using Roche detection system. Deoxyribonucleic acid of hepatitis B virus (HBV-DNA) was assayed by fluorescence quantitative method. Hepatitis B surface antigen (HBsAg) and HB e antigen (HBeAg) were measured using chemiluminescence microparticle immunoassay (CMIA). All assays were done after 2, 4, 8, 12, 18 and 24 weeks of drug withdrawal.
 Results: All parameters in the three groups of patients returned to normal levels after 2, 4, 8, 12, 18 and 24 weeks of drug withdrawal. In 83 neonates, tests for hepatitis B DNA and surface antigen were negative 1 month after birth, with 100 % block of maternal neonatal transmission, and no HBV infection. The incidence of adverse reactions in pregnant women were 16.67, 19.24 and 18.52 % in groups A, B and C, respectively, with no significant difference (p > 0.05).
 Conclusion: The therapeutic effect of TDF for parturient women with high HBV density is not related to drug withdrawal time after delivery. It is necessary to increase the number of samples in subsequent studies in order to validate the research results.

Fluoroquinolone-Associated Movement Disorder: A Literature Review.

Fluoroquinolones (FQNs) are related to several central nervous system side effects. This review aims to evaluate the clinical-epidemiological profile, pathophysiological mechanisms, and management of FQNs-associated movement disorders (MDs). Two reviewers identified and assessed relevant reports in six databases without language restriction between 1988 and 2022. A total of 45 reports containing 51 cases who developed MDs secondary to FQNs were reported. The MDs included 25 myoclonus, 13 dyskinesias, 7 dystonias, 2 cerebellar syndromes, 1 ataxia, 1 tic, and 2 undefined cases. The FQNs reported were ciprofloxacin, ofloxacin, gatifloxacin, moxifloxacin, levofloxacin, gemifloxacin, and pefloxacin. The mean and median age were 64.54 (SD: 15.45) and 67 years (range: 25-87 years). The predominant sex was male (54.16%). The mean and median time of MD onset were 6.02 (SD: 10.87) and 3 days (range: 1-68 days). The mean and median recovery time after MD treatment was 5.71 (SD: 9.01) and 3 days (range: 1-56 days). A complete recovery was achieved within one week of drug withdrawal in 80.95% of the patients. Overall, 95.83% of the individuals fully recovered after management. Future cases need to describe the long-term follow-up of the individuals. Additionally, FQN-induced myoclonus should include electrodiagnostic studies.

Protective effect of anti-idiopathic pulmonary fibrosis drug Pirfenidone and Sufenidone (SC1011) on pulmonary injury induced by tuberculosis in a mouse tuberculosis model

Objective: To evaluate the protective effect of anti-idiopathic pulmonary fibrosis (IPF) marketed drug Pirfenidone and its clinical drug Sufenidone (SC1011) against lung injury in a mouse tuberculosis model. Methods: C57BL/6 mouse model of tuberculosis was established. A total of 75 C57BL/6 mice were infected with 1×107 CFU/ml H37Rv suspension by aerosol and randomly divided into untreated (n=9) group, isoniazid+rifampicin+pyrazinamide (HRZ) group (n=22), PFD+HRZ group (n=22), and SC1011+HRZ group (n=22). C57BL/6 mice were infected with H37Rv by aerosol for 6 weeks and then treated. Seven mice in each treatment group were weighed, sacrificed, dissected and observed for lung and spleen lesions at 4 and 8 weeks of treatment. HE staining and Masson staining were used to assess degree of lung injury and fibrosis, respectively. ELISA was used to assess the IFN-γ/TNF-α content in the serum of mice in each treatment group after 4 weeks of treatment. Hydroxyproline (HYP) content in lung tissue was measured by alkaline hydrolysis; meanwhile, CFU counts were used to assess the bacterial load in the lung and spleen of mice in each treatment group and the recurrence of spleen and lung tissue after 12 weeks of drug withdrawal. Results: At 8 weeks, the HYP content in the lung tissue was (630±58), (635±17), and (840±70) μg/mg in the PFD+HRZ, SC1011+HRZ, and HRZ treatment group, respectively (P<0.05).At 8 weeks, the proportion of Masson staining blue-stained area, that was, positive area, in lung tissue was 16.65%±1.82%, 10.01%±2.16%, and 21.36%±3.21%, respectively (F=27.11, P<0.001).The lung injury scores by HE staining at 8 weeks were (5.00±0.50), (5.00±0.47), and (6.89±0.99) points, respectively (F=19.81, P<0.001).The results of 4-week ELISA showed that the levels of TNF-α and IFN-γ in the serum of the SC1011+HRZ-treated group were lower than those of the HRZ-treated group (all P<0.05).The degree of lung injury and fibrosis in PFD+HRZ and SC1011+HRZ treatment groups were lower than those in HRZ treatment group (all P<0.001). The number of viable bacteria in the lung tissue of mice treated with PFD+HRZ, SC1011+HRZ, and HRZ for 4 weeks was lower than that of mice untreated [(1.82±0.10), (1.91±0.05), (1.79±0.17) vs. (5.27±0.07) lg(CFU+1)/ml, all P<0.05)]. And the aseptic transformation of the spleen of mice was achieved in each treatment group at 8 weeks of administration. After 12 weeks of drug withdrawal, the recurrence of lung infection in the SC1011+HRZ treatment group was 3/7 lower than 5/7 in the HRZ treatment group (P>0.05); the recurrence of spleen infection in the SC1011+HRZ treatment group was 1/7 lower than 5/7 in the HRZ treatment group (P>0.05).Pulmonary infection recurred more frequently in PFD+HRZ 6/7 versus HRZ 5/7 (P>0.05). Conclusions: PFD/SC1011, when combined with HRZ, reduced lung injury and reduced secondary fibrosis in pulmonary tuberculosis in C57BL/6 mice. SC1011 combined with HRZ has no significant short-term therapeutic effect on MTB, but may reduce its recurrence rate in long-term treatment, especially in reducing the recurrence rate of mouse spleen.

Efficacy of Recombinant Human Endostatin plus Neoadjuvant Chemotherapy for Osteosarcoma and Its Influence on Serum VEGF and MMP-9 Levels.

To investigate the efficacy of recombinant human endostatin (rh-Endo) plus neoadjuvant chemotherapy (NACT) for osteosarcoma (OSA) and its influence on serum vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9). The case data of 141 OSA patients presented to the North District, Xiangyang Central Hospital Affiliated to Hubei University of Arts and Sciences from January 2018 to June 2019, were analyzed retrospectively. Patients receiving NACT (methotrexate + ifosfamide + adriamycin) were assigned into the control group (CNG; n = 65), while those treated with rh-Endo plus NACT were included in the combination group (CMG; n = 76). The following aspects were compared: clinical efficacy, serum tumor markers, serum VEGF and MMP-9 contents, inflammatory factors, incidence of adverse reactions, limb function scores at 6 months of follow-up, and prognostic quality of life (QOL). A statistically higher overall response rate (ORR) was determined in CMG versus CNG (84.2% vs. 64.6%, P < 0.05). The pretreatment serum bone alkaline phosphatase (BALP), insulin-like growth factor (IGF)-1, serum amyloid A (SAA), VEGF, MMP-9, C-reactive protein (CRP), tumor necrosis factor (TNF)-α, and interleukin (IL)-10 levels differed insignificantly between the two cohorts (P > 0.05); while except IL-10 that showed increased expression in both cohorts and was comparatively higher in CMG, the other 8 parameters reduced in both cohorts after 2 weeks of drug withdrawal, and the reduction of each parameter was more significant in CMG (P < 0.05). The total adverse reaction rate was 30.2% in CMG, which was higher than that of 36.9% in CNG, albeit without a statistical difference (P > 0.05). An evidently higher 2-year survival rate was determined in CMG (P < 0.05). rh-Endo plus NACT is more effective than NACT alone in the treatment of osteosarcoma, which can validly restore the balance of vascular endothelial cells, reduce inflammation, and is worth promoting in clinic.

Toxicological testing of Fuganlin Oral Liquid on young Sprague Dawley rats

Purpose: To further evaluate the safety of Fuganlin Oral Liquid (FGLOL) for clinical application by conducting toxicological tests in young rats.&#x0D; Methods: Based on the clinical dose for infants aged less than 1 year, 4-day-old Sprague-Dawley (SD) rats were orally administered FGLOL3.88, 11.64 and 38.75 g crude drug/kg for 18 days, followed by a 3-week withdrawal. This was followed by evaluating the effect of FGLOL on various growth and development indicators, nerve reflex function and spontaneous behavior of the rats. Based on the clinical dose for children aged 1 - 6 years, 15-day-old rats were orally administered FGLOL3.88, 11.64 and 38.75 g crude drug/kg for 31 days, followed by a 3-week withdrawal in order to evaluate the impact of the FGLOL on the development of reproductive organs and nervous systems in young rats. The effects of FGLOL on the safety of young rats were judged based on ophthalmic examination, physical examination, spontaneous behavior and other developmental effects.&#x0D; Results: FGLOL did not cause animal death, and no significant toxicological changes were observed in the body weight, growth and development, and behavior of the young rats (p &lt; 0.05). No observed adverse effect level (NOAEL) even after administering the higher dose at each stage. However, the results at each stage showed that oral administration of a large amount of FGLOL had a greater effect on normal food consumption of the rats (p &lt; 0.05).&#x0D; Conclusion: The findings indicate that Fuganlin Oral Liquid, a traditional Chinese medicine formulation, is safe in rats. However, clinical trials are required to acertain its safety in humans.

Зміни деяких біохімічних показників сироватки крові у разі дво- та шеститижневого застосування терапевтичних доз опіоїду та на ранніх термінах відміни препарату

An important issue today is the safety of opioids for the treatment of chronic pain syndrome, which accompanies a number of somatic diseases. Most drugs or their metabolites are excreted through the kidneys and are one of the causes of acute and chronic kidney damage. The aim of the study was to compare some serum biochemical parameters of rats at two and six weeks of therapeutic doses of nalbuphine and to compare them after early withdrawal. This study was performed on 45 white sexually mature male rats weighing 180–260 g. In the serum of experimental animals’ total protein, urea, creatinine, calcium and phosphorus were determined. Statistical calculations were performed using RStudio v. 1.1.442 and R Commander v.2.4-4. Serum total protein in the study groups had small fluctuations in control, but there was a tendency to decrease after 2 weeks of nalbuphine administration and to increase both from the second to the sixth week and with the withdrawal of nalbuphine in the early stages of the experiment. Serum creatinine increased from the second to the sixth week of the experiment both with six weeks of administration (2.23 times higher than control values) and with two weeks of nalbuphine with the subsequent four-week withdrawal (1.63 times higher than control values). Serum urea increased after 2 weeks and 6 weeks of nalbuphine administration, with mean values 6.22 ± 0.47 mmol/l and 7.63 ± 0.74 mmol/l respectively. However, after drug withdrawal the indicator decreased to the control group. Serum calcium in rats decreased after 2 weeks of the experiment to the value (2.16 ± 0.09 mmol/l). After 6 weeks of nalbuphine administration, as after drug withdrawal, serum calcium increased to the level of the control group and was 2.39 ± 0.09 mmol/l and 2.46 ± 0.05 mmol/l respectively. The serum phosphorus index increased after 2 weeks of the experiment and was 2.42 ± 0.05 mmol/l. After 6 weeks of nalbuphine administration and after drug withdrawal, the value decreased to 2.12 ± 0.12 mmol/l and 2.00 ± 0.05 mmol/l respectively. Conclusions. The use of therapeutic doses of nalbuphine for two and six weeks, as well as after drug withdrawal does not affect the concentration of serum total protein of rats. As a result of two- and six-week use of the drug, an impaired renal nitrogen excretion is observed, and hence the structure of the renal corpuscles is altered, which also takes place during four weeks of drug withdrawal after two weeks of use, as evidenced by an increase in urea and creatinine after two weeks of nalbuphine and significant increase in creatinine after drug withdrawal. Two-week administration of nalbuphine leads to hypocalcemia and hyperphosphatemia, indicating impaired renal reabsorption function, while a subsequent four-week administration and withdrawal leads to normalization of these parameters, except for phosphorus, which is reduced.

Topical Cyclosporine in Oral Lichen Planus-A Series of 21 Open-Label, Biphasic, Single-Patient Observations.

Topical cyclosporine (CSA) has been reported as an alternative treatment in steroid-refractory oral lichen planus (OLP), but evidence is limited and conflicting. An N-of-1 trial setting could be appropriate to evaluate interindividual differences in treatment response. We studied a series of 21 open-label, biphasic single-patient observations. Patients (15 women, 6 men) with OLP recalcitrant to topical steroids received four weeks of CSA mouth rinse (200 mg/twice daily) followed by four weeks of drug withdrawal. Pain (visual analogue scale (VAS) score), disease extent (physicians’ global assessment (PGA) score) and quality of life (Dermatology Life Quality Index (DLQI) score,) were assessed at baseline (T0), after four weeks of treatment (T1) and after another four weeks without treatment (T2). Median age was 58 years (interquartile range/IQR = 52–67) and median disease duration was 18 months (IQR = 12–44). Median baseline VAS score decreased significantly at T1 (p = 0.0003) and increased at T2 (p = 0.032) (T0 = 5 (IQR = 3–6.5); T1 = 2 (IQR = 0.5–3.4); T2 = 3 (IQR = 2–4.8)). Similarly, median baseline PGA score decreased significantly at T1 (p = 0.001) and increased at T2 (p = 0.007) (T0 = 2 (IQR = 1.3–2.5); T1 = 1 (IQR = 1–2); T2 = 2 (IQR = 1–2)). Median baseline DLQI score also decreased significantly at T1 (p =.027) but did not change at T2 (p = 0.5) (T0 = 2.5 (IQR = 1–5.8); T1 = 1 (IQR = 0–3); T2 = 1 (IQR = 1–4)). CSA responders (n = 16) had significantly higher median baseline VAS scores (5.2 (IQR = 5–6.5)) than nonresponders (n =5) (2 (IQR = 2–3.5) (p = 0.02). In our study, pain, disease extent and quality of life of patients with OLP improved significantly during therapy with low-dose CSA mouth rinse and exacerbated after drug withdrawal. Remarkably, patients with high initial VAS scores seemed to profit most.

Histological and biochemical studies on effect of Sofosbuvir (Sovaldi) on adult male albino rat kidney

ABSTRACT Sofosbuvir (sovaldi) is the backbone of many anti‐HCV drugs. We aimed to demonstrate the effect of sofosbuvir on the adult male albino rat kidney. Sixty adult male albino rats were used. The animals were divided equally into 2 main groups (I and II), and each group was divided equally into 3 subgroups (A, B, and C). In group I (control group), each rat was gavaged 0.5 ml distilled water daily for 4 weeks. In group II (sofosbuvir treated group), each albino rat was gavaged 0.5 ml distilled water containing 7.2 mg sofosbuvir daily for 4 weeks. The rats were sacrificed at the end of the 4th week (subgroups IA and IIA), 6th week (subgroups IB and IIB), and 8th week (subgroups IC and IIC) from the start of the treatment. The kidneys were used for histological study while blood samples were used for biochemical study. The obtained data were statistically analyzed. Sofosbuvir (sovaldi) induced pathological changes that gave the criteria of acute Kidney injury in the adult male albino rats. The pathological changes were confirmed by elevation of serum level of urea and creatinine. After 2 and 4 weeks of drug withdrawal, the kidney incompletely recovered. We concluded that sofosbuvir induced criteria of acute tubular injury in the kidney of the adult male albino rats. This renal injury was proved by histological and biochemical studies. These insults were incompletely reversible after the end the treatment.

Heroin Seeking and Extinction From Seeking Activate Matrix Metalloproteinases at Synapses on Distinct Subpopulations of Accumbens Cells

BackgroundSeeking addictive drugs is regulated by synaptic plasticity in the nucleus accumbens core and involves distinct plasticity in D1 and D2 receptor–expressing medium spiny neurons (D1/2-MSNs). However, it is unknown how differential plasticity between the two cell types is coordinated. Synaptic plasticity and seeking behavior induced by drug-paired cues depends not only on plasticity in the canonical pre- and postsynapse, but also on cue-induced changes in astrocytes and the extracellular matrix adjacent to the synapse. Drug cue–induced signaling in the extracellular matrix is regulated by catalytic activity of matrix metalloproteinases MMP-2,9. We hypothesized that the cell type–specific synaptic plasticity is associated with parallel cell-specific activity of MMP-2 and MMP-9. MethodsTransgenic rats were trained on a heroin self-administration protocol in which a light/tone cue was paired with heroin delivery, followed by 2 weeks of drug withdrawal, and then reinstated to heroin-conditioned cues. Confocal microscopy was used to make morphological measurements in membrane reporter–transduced D1- and D2-MSNs and astrocytes, and MMP-2,9 gelatinase activity adjacent to cell surfaces was quantified using in vivo zymography. ResultsPresenting heroin-paired cues transiently increased MMP-9 activity around D1-MSN dendritic spines and synapse-proximal astroglial processes. Conversely, extinction training induced long-lasting increases in MMP-2 activity adjacent to D2-MSN synapses. Moreover, heroin-paired cues increased tissue inhibitor of metalloproteinases TIMP-1,2, which caused transient inhibition of MMP-2 activity around D2-MSNs during cue-induced heroin seeking. ConclusionsThe differential regulation of heroin seeking and extinguished seeking by different MMP subtypes on distinct cell populations poses MMP-2,9 activity as an important mediator and contributor in heroin-induced cell-specific synaptic plasticity.

A phase II, single-arm, open-label, multicenter clinical study to evaluate the efficacy and safety of sofosbuvir combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection

Objective: To evaluate the efficacy and safety of sofosbuvir (Nanjing Zhengda Tianqing Pharmaceutical Co., Ltd.) combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection. Methods: Treatment-naïve or treatment experienced genotype 2 chronic hepatitis C patients from sixteen research centers of China were screened. All subjects received once-daily dose of sofosbuvir (400 mg) combined with ribavirin (body weight < 75 kg, 1 000 mg/day, 400 mg in the morning and 600 mg in the evening; body weight > 75 kg, 1 200 mg/d, 600 mg in the morning and 600 mg in the evening) for 12 weeks. Patients were followed-up for a period of 12 weeks after discontinuation of treatment. Continuous variables were expressed as mean ± standard deviation. The proportion of subjects with virologic response at different follow-up time points and 95% confidence intervals were estimated by maximum likelihood ratio and Clopper-Pearson interval. Results: 132 cases with genotype 2 chronic hepatitis C virus infection from sixteen research centers of China were included, 12 cases of whom were associated with cirrhosis, and the remaining 120 cases were not associated with cirrhosis. One hundred and thirty-one cases completed the study, and one patient lost to follow-up at week 4 after the end of treatment. The sustained virological response rate was 96.2% (95% confidence interval: 92.37% - 99.16%) after 12 weeks of drug withdrawal. Virological relapse occurred in four cases. Of the 132 subjects enrolled in the study, 119 (90.2%) reported 617 adverse events during treatment, of which 359 (76.5%) were TEAE related to sofosbuvir and/or ribavirin. There were nine TEAEs of grade 3 and above, and six cases (4.5%) of them had six severe adverse events. Only one serious adverse event was associated with sofosbuvir and ribavirin (unstable angina pectoris). There were no adverse events leading to drug discontinuation or death. Conclusion: Sofosbuvir combined with ribavirin has a high SVR rate in the treatment of genotype 2 chronic hepatitis C virus infection, and most of the adverse events occurred were mild with acceptable safety profile.

Subchronic toxicity and concomitant toxicokinetics of long-term oral administration of total alkaloid extracts from seeds of Peganum harmala Linn: A 28-day study in rats

Ethnopharmacological relevanceThe seeds of Peganum harmala Linn, in which the most abundant active compounds are harmaline and harmine, have been widely used as a traditional medicine in various countries to treat a broad spectrum of diseases including asthma, cough, depression, Parkinson's and Alzheimer's diseases. However, few studies on long-term or subchronic toxicity of seeds of P. harmala were reported after overdose. Aim of the studyTo investigate the subchronic toxicity and concomitant toxicokinetics of total alkaloid extracts from seeds of P. harmala (TAEP) after oral administration for four weeks in rats. Materials and methodsThe subchronic toxicity and concomitant toxicokinetics of TAEP were evaluated after 28-day oral administration in rats at daily dose levels of 15, 45, and 150 mg/kg. The signs of toxicity and mortality were monitored and recorded daily. The body weight and average food consumption were measured weekly. The analyses of hematology, biochemistry, urine, relative organ weights and histopathology were conducted at the termination of treatment and recovery phase. For concomitant toxicokinetics study, the plasma toxicokinetic parameters, tissue distribution, and excretion of predominant ingredients harmaline and harmine in TAEP and metabolites harmalol and harmol were tested. ResultsFollowing initial repeated exposure to high-dose (150 mg/kg/day) of TAEP excitotoxic reaction, such as tremor, was observed, but tolerated on the fourth day after multiple dosing. The significant alterations in blood glucose and lipid metabolism in liver were observed, but recovered after four weeks of drug withdrawal. The no-observed-adverse-effect level (NOAEL) of TAEP was considered to be 45 mg/kg/day under the present study conditions. There were no significant gender differences in most indexes of subchronic toxicity throughout the experimental period with the exception of food consumption and body weight. In concomitant toxicokinetics study, the alterations of dynamic characteristic for harmaline, harmine and metabolite harmol after multiple oral administration at three doses had been observed. Harmaline, harmine and metabolites harmalol and harmol were widely distributed in organs and there was no accumulation in the tissues examined. The reduction of harmaline and metabolite harmalol in brain after multiple dosing at dose of 150 mg/kg might be closely related to the tremor tolerance. The main excretory pathway for metabolites harmalol and harmol was urinary excretion via kidney. ConclusionsThe results revealed that TAEP at doses of 15 and 45 mg/kg/day in rats might be safe. Excitotoxic reaction such as tremor occurred initially at dose of 150 mg/kg/day, however, the toxicity was tolerant and reversible. In addition, harmaline and harmine in TAEP had a quick absorption into blood and metabolized to harmalol and harmol, and there was no drug accumulation in the detected tissues. Further studies should be investigated to clarify the mechanisms of tremor tolerance and neurotoxicity of TAEP.

Real-world study of paritaprevir/ritonavir-ombitasvir combined with dasabuvir for the treatment of chronic hepatitis C 1b genotype infection in China

Objective: To evaluate the real-world safety and curative effect of ombitasvir combined with dasabuvir for the treatment of chronic hepatitis C 1b genotype infection in non-cirrhotic or compensated cirrhotic patients. Methods: A real-world research method was adopted, and the research was conducted at three medical centers of mainland China. Non- cirrhotic or compensated cirrhotic patients with HCV genotype 1b infection who were initially treated with IFN/PEG-IFN-alpha combined with ribavirin, and ombitasvir combined with dasabuvir for 8 or 12 weeks were taken. Sustained virological response (SVR) and the incidence of adverse events during treatment and follow-up were evaluated after 12 weeks of drug withdrawal at OBV/PTV/r 25/150/100mg once daily and DSV 250mg, twice daily. Median and range were used for description of non-normally distributed data. Results: 80 cases of GT1b were included in this study. Of these 88.8% (71/80) were newly diagnosed, 12.5% (10/80) were compensated cirrhotic, 97.5% (78/80) received 12 weeks treatment, and 2.5% (2/80) received 8 weeks treatment. The rate of HCV RNA negative at EOT (end of treatment) was 100% (64/64). A total of 67 patients completed the treatment within 12 weeks, and 43 patients returned to the hospital for further consultations, and SVR12 was 100%(43/43). No patient discontinued the drugs because of an adverse event during treatment. Conclusion: In the real world, Ombitasvir combined with dasabuvir for the treatment of chronic hepatitis C 1b genotype infection in China has 100% rates of EOT and SVR12 with well- tolerability and safety.

Effects of vigabatrin, carbamazepine or its combination on the pituitarygonadal axis in male Wistar rats

Objectives: Epilepsy is the most common non-infectious neurologic disease in developing African countries following stroke and Alzheimer's disease. Most conventional antiepileptic drugs, due to their centrally acting potentials have been implicated in the deregulation of reproductive hormones. This study assessed the effect of the single and combined administration of vigabatrin (VIG) and carbamazepine (CBZ) on the pituitary-gonadal axis of male Wistar rats.Methods: Fifty male Wistar rats were randomly divided into 5 groups (n=10). The animals were administered with distilled water (0.1 ml/kg/day), VIG (200 mg/kg/day), CBZ (200 mg/kg/day), VIGCBZ combination (100 mg/kg/day each) and VIG-CBZ combination (200 mg/kg/day each) for 8 weeks. Twenty-four hours after the last dose, 5 rats from each group were sacrificed, while the remaining 5 eventually sacrificed after another 8 week of drug withdrawal. The level of luteinizing hormone, follicle stimulating hormone and testosterone were determined from the serum. The weight of the reproductive organs and sperm indices were assayed, while the testicular tissue were examined for signs of histological alteration.Results: The results showed significant decrease in the levels of luteinizing hormone, follicle stimulating hormone, testosterone and sperm physiological indices. Morphological alteration was noticed in the testes of all the treated rats. However, there was restoration of these parameters sequelae to 8 weeks cessation of treatment.Conclusion: Single and combined administration of VIG and CBZ resulted into pituitary-gonadal axis hormonal deregulation and alterations in the sperm profile which were however reversible upon cessation of treatment.Keywords: epilepsy, vigabatrin, carbamazepine, sperm, testosterone

Establishment of a prognostic model for clinical relapse after drug withdrawal in chronic hepatitis B patients with hepatitis B e antigen seroconversion

Objective To investigate the prognostic factors for clinical relapse in chronic hepatitis B (CHB) patients with hepatitis B e antigen (HBeAg) seroconversion after drug withdrawal and to establish a prognostic model. Methods Totally 201 CHB patients with HBeAg seroconversion after the antiviral therapy were enrolled. The epidemiological variables including age, gender and family history of hepatitis B were collected. Liver function and hepatitis B virus (HBV) DNA level one week before initiation of antiviral therapy, hepatitis B surface antigen (HBsAg) level at the time of drug withdrawal and the duration of antiviral therapy after HBeAg seroconversion were analyzed. The clinical relapse after 48 weeks of drug withdrawal was followed up. The patients were divided into relapse group and non-relapse group according to clinical variables at 48 weeks after drug withdrawal. The counting data were analyzed by chi-square test and the measurement data were analyzed by t test. The Logistic regression model was used to determine the prognostic factors for clinical relapse. The receiver operating charactenstic (ROC) curve was constructed to assess the performance of the prediction model. Results The clinical relapse rate was 16.42% (33/201) after 48 weeks of drug withdrawal. By multivariate analysis, age, the duration of antiviral therapy after HBeAg seroconversion and HBsAg level at the time of drug withdrawal were independent predictors (χ2=14.546, t=3.202, t=3.286, respectively; all P<0.05). The regression model Logit (P)=1.220×age-0.040×the duration of antiviral therapy after HBeAg seroconversion + 0.004×HBsAg level at the time of drug withdrawal -5.426. The sensitivity and specificity with the cut-off value of -0.860 were 73.10% and 90.40%, respectively. Conclusions Age, the duration of antiviral therapy after HBeAg seroconversion and HBsAg level at the time of drug withdrawal are independent predictors for clinical relapse 48 weeks after drug withdrawal in CHB patients with HBeAg seroconversion after antiviral therapy. Key words: HBeAg seroconversion; Clinical relapse; Prognostic model; Logistic regression analysis

Abstract 5561: Single-cell analysis reveals an adaptive, transiently heritable, slowly-dividing, drug-resistant state inhibitable by drug combinations

Abstract Adaptation and fractional response of tumor cells to targeted inhibitors of oncogenic pathways creates a population of viable tumor cells from which fully resistant clones can ultimately arise. Thus, understanding transient drug adaptation is key for both improving the effectiveness of treatment and delaying/controlling acquired resistance. Despite the wealth of information available about feedback mechanisms associated with adaptive resistance, most of our knowledge in this area comes from studying drug response in bulk tumor cell populations. Furthermore, the phenotypic consequences of drug adaptation have been often studied at a few fixed time-points, when drug-adapted cells exhibit a high population-average activity in multiple pro-growth signaling cascades. It therefore remains unclear how the initial responses to drug relate to subsequent phenotypes such as cell death or adaptation. This is likely a key point for designing novel approaches to overcome fractional drug response in tumor cells and to achieve durable therapy. We use real-time live-cell imaging, single-cell analysis and molecular profiling to show that exposure of BRAFV600E melanoma cells to RAF/MEK inhibitors elicits a time-variable and heterogeneous response in which some cells die, some arrest and the remainder adapt to drug. Drug-adapted cells up-regulate markers of the neural crest (e.g. NGFR), a melanocyte precursor, and grow slowly. The drug-induced slowly-cycling NFGRHigh state is only transiently stable, reverting to the drug-naïve state within two weeks of drug withdrawal as measured by the restoration of RAF/MEK inhibitor sensitivity, accelerated rate of cell division and reduced expression of NGFR. Transcriptional and biochemical profiling of cell lines and human tumors implicates a role for the c-Jun/ECM/FAK/Src cascade in driving the de-differentiated resistance program. We identify multiple drugs targeting this cascade as well as BET bromodomain inhibitors that block this resistance program in cell lines and in a BRAFV600E melanoma xenograft model and increase sensitivity and maximal effect (Emax) of RAF/MEK inhibitors. Our study reveals directly how drug adaptation happens in individual tumor cells leading to emergence of heterogeneous cell sub-populations with reduced drug-sensitivity that may be targeted by drug combinations. Citation Format: Mohammad Fallahi-Sichani, Verena Becker, Benjamin Izar, Gregory J. Baker, Jia-Ren Lin, Sarah A. Boswell, Levi A. Garraway, Peter K. Sorger. Single-cell analysis reveals an adaptive, transiently heritable, slowly-dividing, drug-resistant state inhibitable by drug combinations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5561. doi:10.1158/1538-7445.AM2017-5561

Neuronal metabolomics by ion mobility mass spectrometry in cocaine self-administering rats after early and late withdrawal.

The neuronal metabolomes in rat striatum (STR), prefrontal cortex (PFC), and nucleus accumbens (NAC) were analyzed by Hadamard transform ion mobility mass spectrometry (HT-IMMS) in order to reveal global and specific metabolic changes induced by cocaine self-administration after 1-day or 3-week withdrawal. Metabolite features were comprehensively separated and detected using HPLC-IMMS within minutes. Global metabolic differences were observed by PCA for comparisons between cocaine and saline treatments at 1-day withdrawal time. Metabolite features that were significantly changed were selected using PCA loadings' plot and unpaired LLL test and then tentatively identified by accurate m/z, yielding a complete profile of metabolic changes induced by cocaine self-administration. The majority of these changes were found at the 1-day withdrawal time, but several of them endured even after 3-week withdrawal from cocaine, and these changes were generally brain region specific. Putatively identified metabolites associated with oxidative stress and energy metabolism were also specifically investigated. We discovered that the dysregulation of creatine/creatinine was different between the STR and NAC, demonstrating that metabolic alterations are brain region specific. Glutathione and adenosine were also changed in their abundance, and the results agreed with previous studies. In general, this study provided a high-throughput analytical platform to perform metabolomics analyses with putative identifications for altered metabolite features induced by cocaine treatment, therefore revealing additional metabolic targets of cocaine-induced changes after early and extended withdrawal times.

Antidepressant, anxiolytic and procognitive effects of rivastigmine and donepezil in the chronic mild stress model in rats

BackgroundThe treatment of depression in old age is complicated by frequent co-morbidity with cognitive impairment. Anti-dementia drugs have some efficacy to improve cognitive performance and there is an inconsistent literature regarding the effect of such drugs on depressive symptoms. Here, we have investigated whether anti-dementia drugs would have antidepressant-like and pro-cognitive effects in a well-validated animal model of depression and cognitive impairment, chronic mild stress (CMS).MethodsRats were subjected to CMS for a total of 8 weeks. After 2 weeks, subgroups of stressed and non-stressed animals were treated daily, for 5 weeks followed by 1 week of drug withdrawal, with vehicle, imipramine (10 mg/kg), rivastigmine (2 mg/kg), donepezil (0.3 mg/kg) or memantine (5 mg/kg). Sucrose intake was tested weekly, and animals were also tested in the elevated plus maze (at week 7) and in an object recognition task (at weeks 7 and 8).ResultsCMS decreased sucrose intake, had an anxiogenic effect in the elevated plus maze, and impaired performance in the object recognition test. Imipramine, rivastigmine and donepezil normalized performance in all three tests. Memantine had anxiolytic and pro-cognitive effects, but did not reverse CMS-induced anhedonia.DiscussionThe fact that all three anti-dementia drugs reversed CMS-induced cognitive impairment and that cholinesterase inhibitors, but not memantine, have antidepressant-like effects in this model suggest that different mechanisms may underlie CMS-induced anhedonia and cognitive impairment. We discuss the clinical implications of these findings.

Transient azoospermia following rosuvastatin medication for hypercholesterolemia

The authors report a case of transient azoospermia following hydroxymethylglutaryl-coenzyme A reductase (HMGCR) inhibitor rosuvastatin medication for hypercholesterolemia. While a primary infertile couple with oligoasthenospermia was preparing for an in vitro fertilization program, the male partner had been diagnosed with hypercholesterolemia in a medical check-up and prescribed fourweek oral administration of rosuvastatin. No motile spermatozoa were found in the ejaculated semen and urine on the day of follicular aspiration. Azoospermia was confirmed by re-examination in weeks 3 and 7. Spermatozoa appeared in the ejaculated semen in two weeks of drug withdrawal. In week 16, the sperm count and motility increased to the level where intracytoplasmic sperm injection was available.

Abstract 3593: Raptor upregulation contributes to maintenance of 4EBP1 phosphorylation and TORC kinase resistance in renal cancer cells

Abstract Advanced renal cell cancer (RCC) is incurable, but recent advances in targeted treatment have improved the prognosis. Rapalogue inhibitors of the mammalian target of rapamycin (mTOR) are licensed for treatment of advanced RCC, although clinical benefit is modest. Novel mTOR kinase inhibitors show more complete blockade of mTOR downstream targets, and greater in vitro anti proliferative activity, although resistance remains a problem. The aim of this study was to create a model of in vitro resistance to PI3K-mTOR kinase inhibitor BEZ235, in order to identify biomarkers and mediators of resistance. After 5 months continuous culture in BEZ235, resistance to 20nM BEZ235 was induced in the RCC4 cell line, generating subline RCC4B20. These cells had 14 fold higher BEZ235 growth inhibitory 50 (GI50) concentration compared to sensitive cells (99 nM vs. 7 nM). There was cross-resistance to alternative mTOR kinase inhibitor AZD2014 (GI50 1131 vs. 166 nM), but not to PI3K, AKT or allosteric mTORC1 inhibitors, suggesting that resistance was primarily directed at mTOR kinase. RCC4B20 cells were screened at the protein level by antibody array, and RNA level by microarray, and hits were validated by western blotting and qRT-PCR. Proteins and pathways found to be up-regulated in RCC4B20 cells included MET, pMET, ABL, and MEK/ERK by antibody array, and NOTCH signalling by microarray. These were validated as showing altered regulation, but BEZ235 resistance was not blocked by depletion or inhibition of these targets, suggesting that these changes were correlates, but not mediators of resistance. An hypothesis-led approach directed attention to the mTORC1 pathway. In BEZ235 resistant cells, the mTORC1 target 4E-BP1 was re-phosphorylated, despite on-going mTOR blockade as evidenced by suppressed S6 phosphorylation. Recovery of 4E-BP1 phosphorylation was associated with 4.2 ± 0.3 fold increase in levels of RAPTOR, a scaffolding protein for mTOR substrates. Phosphorylation of 4E-BP1 was suppressed by rapamycin, or by RAPTOR depletion, both interventions partially reversing BEZ235 resistance. Finally, RCC4B20 cells lost resistance after 8 weeks of drug withdrawal, suggesting that resistance may have been associated with reversible changes in gene expression due to histone modification. Indeed, histone deacetylase inhibitor panobinostat partially reversed resistance to BEZ235. In summary, the acquisition of resistance to mTOR kinase inhibition was accompanied by complex molecular changes, many of which were passengers, not drivers of resistance. Resistance was partially suppressed by interventions directed at raptor up-regulation and histone modification, suggesting that these changes do mediate resistance. The combination of BEZ235 with rapamyin or panobinostat warrant further investigation to evaluate potential to overcome resistance to mTOR kinase inhibition in RCC. Citation Format: Philip Earwaker, Frances Willenbrock, Andrew Protheroe, Valentine Macaulay. Raptor upregulation contributes to maintenance of 4EBP1 phosphorylation and TORC kinase resistance in renal cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3593. doi:10.1158/1538-7445.AM2015-3593

Abstract 2595: Efficacy of cetuximab and mutant selective EGFR inhibitor WZ4002 in EGFR T790M and non-T790M models of erlotinib resistant non-small cell lung cancer

Abstract Background: EGFR kinase inhibitors, including erlotinib, are effective treatments for patients with EGFR mutant lung cancer. However, resistance inevitably develops, most commonly (60%) mediated by an EGFR secondary mutation EGFR T790M. Mutant selective EGFR inhibitors (WZ4002, AZD9291, CO-1686) are effective in preclinical models and clinically (AZD9291 &amp; CO-1686) in EGFR T790M patients (response rates (RR) &amp;gt; 50%) but are less effective in patients with non-T790M mediated drug resistance (AZD9291 RR ∼10%). Combination of afatinib and cetuximab is effective in both T790M (32% RR) and non-T790M (25% RR) mediated resistance but is associated with significant toxicity. We evaluated whether cetuximab could add to the efficacy of a mutant selective EGFR inhibitor and whether the combination was also effective in a model of non-T790M mediated resistance. Methods: Mice harboring xenografts from PC9GR (EGFR Del 19/T790M), H1975 (EGFR L858R/T790M), HCC827GR (EGFR Del 19/MET amplification) cells or genetically engineered mouse models (GEMMS) expressing EGFR L858R/T790M were treated with vehicle, WZ4002 alone, cetuximab alone or the combination. We compared the efficacy of the treatments and evaluated for tumor cures (defined as tumor disappearance and lack of regrowth following 6 weeks of drug withdrawal). In addition, we evaluated the impact of the treatments on EGFR signaling and apoptosis. Results: In the EGFR T790M xenograft models (PC9GR and H1975), the combination of WZ4002 and cetuximab was more effective than either single agent. Only the combination led to tumor cures (5/15 in PC9GR and 3/18 in H1975) while none were observed with the single agents. In the HCC827GR xenografts, the combination of WZ4002/cetuximab was more effective than the single agents but did not lead to any cures. Evaluation of the tumors from treated mice revealed that the efficacy of the combination correlates with effective degradation of EGFR and MET (in HCC827GR). In the L858R/T790M GEMMs, the combination led to significantly greater tumor shrinkage compared to single agents. Evaluation of treated tumors revealed enhanced apoptosis in the combination treated mice. Conclusions: The addition of cetuximab enhances the efficacy of the mutant selective EGFR inhibitor WZ4002 in vivo by increasing the single agent efficacy in both EGFR T790M and non-T790M models. Clinical trials of mutant selective EGFR inhibitors and cetuximab are warranted. Citation Format: Erin M. Tricker, Chunxiao Xu, Kwok-Kin Wong, Pasi A. Janne. Efficacy of cetuximab and mutant selective EGFR inhibitor WZ4002 in EGFR T790M and non-T790M models of erlotinib resistant non-small cell lung cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2595. doi:10.1158/1538-7445.AM2015-2595