Synchronous vulvar intraepithelial neoplasia (VIN) of warty type and cervical intraepithelial neoplasia (CIN): case report

Vulvar intraepithelial neoplasia is a precancerous lesion of the vulva, which has been referred to in the past with varied terminology. It can be associated with multicentricity of other neoplastic squamous lesions in the cervix and vagina. We report a case of vaginal intraepithelial neoplasia and concomitant cervical intraepithelial neoplasia in a 30 year old female. An attempt is made to put forth the recent terminology of vulvar intraepithelial neoplasia.     

Production of calmodulin-tagged proteins in Drosophila Schneider S2 cells: a novel system for antigen production and phage antibody isolation

We report the development of an expression system for the production of soluble, calmodulin (CaM)-tagged proteins in Drosophila Schneider S2 cells and the subsequent use of these proteins for the selection of phage displayed antibodies. The CaM-tag permitted the purification of recombinant protein to >90% purity in a single step at yields of >20 mg/l. Using platelet glycoprotein VI (GP6) as a model, we demonstrated that the recombinant CaM-tagged protein was post-translationally N-glycosylated and had identical ligand specificity to native protein. A novel selection strategy, exploiting the CaM tag, was then used to isolate four single chain Fv fragments (scFvs) specific for GP6 from a non-immune phage display library. In contrast to other selection methods, which can result in antibodies that do not recognise native protein, all of the scFvs we selected bound cell surface expressed GP6. In conclusion, the production of CaM-tagged proteins in Drosophila Schneider S2 cells and the selection strategy reported here offer advantages over previously published methods, including simple culture conditions, rapid protein purification, specific elution of phage antibodies and preferential selection of phage antibodies that recognise native, cell surface expressed protein.     

Lack of effect of colesevelam HCl on the single-dose pharmacokinetics of aspirin,atenolol, enalapril,phenytoin, rosiglitazone,and sitagliptin

Aims

Drug interactions with bile acid sequestrants are primarily due to the potential of these agents to bind to concomitant drugs. Six clinical studies were performed to determine the effects of colesevelam on the pharmacokinetics of aspirin, atenolol, enalapril, phenytoin, rosiglitazone, and sitagliptin.

Methods

All six studies enrolled healthy subjects aged 18–45 years. The phenytoin study used a single-dose, three-period crossover design (phenytoin alone, phenytoin simultaneously with colesevelam, and phenytoin 4 h before colesevelam). The other studies used a two-period crossover design (test drug alone and test drug simultaneously with colesevelam). Colesevelam (3750 mg once daily) was dosed throughout the pharmacokinetic sampling period. After each single dose of the test drug, serial blood samples were collected for determination of plasma drug concentrations and calculation of pharmacokinetic parameters.

Results

For all six test drugs, 90% CIs for geometric least-squares mean ratios of AUC and C_max for the measured analytes were within specified limits, indicating no interaction between the test drug and colesevelam.

Conclusions

Aspirin, atenolol, enalapril, rosiglitazone, and sitagliptin may be taken with colesevelam. Although the phenytoin study indicated no pharmacokinetic interaction, phenytoin should continue to be taken ≥4 h before colesevelam in accordance with current prescribing information.     

Dendritic cell-based immunization induces Coccidioides Ag2/PRA-specific immune response

Valley Fever, or coccidioidomycosis, is caused by a soil-borne, highly virulent fungal pathogen, Coccidioides spp. Infection with Coccidioides can be life-threatening. Since an effective treatment is not available and the T cell-mediated immune response is protective, vaccine development is of interest. In this study, a primary dendritic cell (DC)-vaccine was evaluated for its ability to stimulate Coccidioides antigen-specific immune response in an extremely susceptible BALB/c mouse model. The DC-vaccine (Ag2-DC) was prepared by non-virally transfecting the primary bone marrow-derived DCs with a plasmid DNA encoding Ag2/PRA (protective epitope of Coccidioides). Mice were intranasally immunized with Ag2-DC on days 2 and 10. Immunized mice were necropsied on days 8, 32, and 44. Major organs and blood samples were harvested. The most common indicators of injury (protein, lactate, and albumin), Ag/PRA-specific cytokine-secreting cells, and IgG and its isotypes were determined by biochemical and immunologic assays, respectively. No signs of sickness were noted. Similarly, no significant changes were observed in the levels of total lung protein, lactate, and albumin, in immunized mice compared with healthy control mice. Interferon (IFN-γ), and interleukin (IL)-4 and IL-17 cytokine-secreting cells were observed in lung and lymph nodes upon Ag2-DC immunization. Our results showed that the levels of serum IgG and its isotypes were increased in Ag2-DC-immunized mice. This report provides evidence of DC immunization-stimulated Ag2/PRA-specific immune responses.     

Clinicopathological study of 113 gastrointestinal stromal tumors

Background

Gastrointestinal stromal tumor (GIST) is the most common sarcoma of the gastrointestinal tract. A retrospective study was done to evaluate the clinical and pathological features and the effect of adjuvant treatment with imatinib.

Method

The case records of 113 GIST patients were retrospectively reviewed and the clinicopathological features, treatments, and outcomes were recorded.

Results

There were 82 males and 31 females, with a median age of 51 years. All patients were symptomatic (mean duration 4 months) and abdominal pain was the most common symptom. The primary sites of GIST were small intestine (38), stomach (36), and others (39). The tumor diameter on imaging varied from 1 to 26 (mean 10.9) cm. Thirty percent of patients presented with metastasis. There was no association between tumor size and presence of metastasis (p?=?0.9). Most common histology was spindle cell morphology followed by mixed spindle cell and epithelioid morphology. Seventy percent patients had high risk (HR) category as per Fletcher risk score. Fifty-three percent had curative resection, after which 34 % had adjuvant imatinib therapy. Recurrence rates were significantly lower in patients receiving adjuvant imatinib therapy (p?=?0.003). No statistically significant association was noted between HR Fletcher score, Mib score >10, tumor size >10 cm, and the risk of recurrence (p?=?0.29, 0.07, and 0.87, respectively). Liver was the most common site of metastasis. Side effects were tolerable and edema and fluid retention were the commonest.

Conclusion

Sites of GIST in Indian patients were different from those in western studies. Adjuvant imatinib therapy significantly reduced the risk of recurrence.     

Is Direct Laryngoscopy Obsolete? “Trans Nasal Oesophagoscopy” the Complete Endoscopic Solution in Head Neck Practice

Direct laryngoscopy (DL) is the standard of care for the evaluation of suspicious lesions in the larynx and hypo pharynx but requires general anaesthesia and a dedicated operation theatre. While DL offers us the ability to map the lesion adequately and take a biopsy, it requires workup for anaesthesia well as rigid oesophagoscopy for assessing the oesophagus with its associated complications. Sixty-nine patients underwent TNE under topical anaesthesia. The lesions were mapped and biopsies taken. Those patients who had an inadequate evaluation on TNE or negative biopsy underwent direct laryngoscopy. Completeness of evaluation, adequacy of biopsy, presence of synchronous oesophageal lesions and the modified Gloucester Comfort Score for patient comfort was documented. Amongst 69 cases enrolled for TNE evaluation, 97% of cases had an adequate mapping of disease extent, and 100% adequacy of biopsy material. General anaesthesia could be avoided in 92.75% of patients. TNE took a median time of 8 min. Synchronous oesophageal tumours were seen in 5.8% of patients. There were no complications and 74% patients did not experience any discomfort. TNE appears to be simple, safe, efficient office based diagnostic procedure. TNE has the potential to be the new standard of care making DL obsolete.     

Survival analysis of haematopoietic cell transplantation for childhood cerebral X-linked adrenoleukodystrophy: a comparison study

BACKGROUND: Favourable outcomes have been reported for patients with childhood cerebral adrenoleukodystrophy (CCALD) who had received haematopoietic cell transplantation (HCT) at the early stage of cerebral involvement. However, comparative data for non-transplanted CCALD patients are limited. We analysed survival of CCALD patients who had not received HCT and, in a subgroup with early cerebral disease, compared survival in those who underwent HCT with those who did not. METHODS: Retrospective survival analyses were done on 283 CCALD patients identified at the Kennedy Krieger Institute who had not received HCT, focusing on a 30-member early stage cerebral subgroup whose neurological disability and MRI severity scores matched those in a 19-member transplanted subgroup previously reported. A Kaplan-Meier survival curve and log-rank test were used for survival analysis and to estimate the difference between the survival probabilities of the groups with statistical significance set at alpha=0.05. FINDINGS: Mean age at onset of symptoms in the entire 283 non-transplanted group was 7 years (SD 2 years). 131 (46%) patients died during the mean follow-up period of 5.9 years (5.3) at a mean age of 12.3 years (4.9). 5-year survival was 66%. The 5-year survival probability of 54% in the early stage group was significantly poorer (chi(2)=7.47, p=0.006) than the 5-year survival of 95% in the transplanted group with early stage cerebral disease. INTERPRETATION: HCT done in the early and progressive stages of CCALD is beneficial, and our data support the recommendation that transplantation be offered to patients in the early stages of CCALD.     

Comparison of survival outcomes among older adults with major trauma after trauma center versus non-trauma center care in the United States

Objective

To compare level 1 and 2 trauma centers with similarly sized non-trauma centers on survival after major trauma among older adults.

Data Sources and Study Setting

We used claims of 100% of 2012–2017 Medicare fee-for-service beneficiaries who received hospital care after major trauma.

Study Design

Survival differences were estimated after applying propensity-score-based overlap weights. Subgroup analyses were performed for ambulance-transported patients and by external cause. We assessed the roles of prehospital care, hospital quality, and volume.

Data Collection

Data were obtained from the Centers for Medicare and Medicaid Services.

Principal Findings

Thirty-day mortality was higher overall at level 1 versus non-trauma centers by 2.2 (95% confidence interval [CI]: 1.8, 2.6) percentage points (pp). Thirty-day mortality was higher at level 1 versus non-trauma centers by 2.3 (95% CI: 1.9, 2.8) pp for falls and 2.3 (95% CI: 0.2, 4.4) pp for motor vehicle crashes. Differences persisted at 1 year. Level 1 and 2 trauma centers had similar outcomes. Hospital quality and volume did not explain these differences. In the ambulance-transported subgroup, after adjusting for prehospital variables, no statistically significant differences remained.

Conclusions

Trauma centers may not provide longer survival than similarly sized non-trauma hospitals for severely injured older adults.     

Pathogenic effects of novel mutations in the P-type ATPase ATP13A2 (PARK9) causing Kufor-Rakeb syndrome, a form of early-onset parkinsonism

Kufor-Rakeb syndrome (KRS) is a rare form of autosomal recessive juvenile or early-onset, levodopa responsive parkinsonism and has been associated with mutations in ATP13A2(also known as PARK9), a lysosomal type 5 P-type ATPase. Recently, we identified novel compound heterozygous mutations, c.3176T>G (p.L1059R) and c.3253delC (p.L1085WfsX1088) in ATP13A2 of two siblings affected with KRS. When overexpressed, wild-type ATP13A2 localized to Lysotracker-positive and LAMP2-positive lysosomes while both truncating and missense mutated ATP13A2 were retained in the endoplasmic reticulum (ER). Both mutant proteins were degraded by the proteasomal but not the lysosomal pathways. In addition, ATP13A2 mRNA with c.3253delC was degraded by nonsense-mediated mRNA decay (NMD), which was protected by cycloheximide treatment. To validate our findings in a biologically relevant setting, we used patient-derived human olfactory neurosphere cultures and fibroblasts and demonstrated persistent ER stress by detecting upregulation of unfolded protein response-related genes in the patient-derived cells. We also confirmed NMD degraded ATP13A2 c.3253delC mRNA in the cells. These findings indicate that these novel ATP13A2 mutations are indeed pathogenic and support the notion that mislocalization of the mutant ATP13A2, resultant ER stress, alterations in the proteasomal pathways and premature degradation of mutant ATP13A2 mRNA contribute to the aetiology of KRS.