Scaling up the task-sharing of psychological therapies: A formative study of the PEERS smartphone application for supervision and quality assurance in rural India.

Measurement-based peer supervision is one strategy to assure the quality of psychological treatments delivered by non-mental health specialist providers. In this formative study, we aimed to 1) describe the development and 2) examine the acceptability and feasibility of PEERS (Promoting Effective mental healthcare through peER Supervision)-a novel smartphone app that aims to facilitate registering and scheduling patients, collecting patient outcomes, rating therapy quality and assessing supervision quality-among frontline treatment providers delivering behavioral activation treatment for depression. The PEERS prototype was developed and tested in 2021, and version 1 was launched in 2022. To date, 215 treatment providers (98% female; ages 30-35) in Madhya Pradesh and Goa, India, have been trained to use PEERS and 65.58% have completed the supplemental, virtual PEERS course. Focus group discussions with 98 providers were examined according to four themes-training and education, app effectiveness, user experience and adherence and data privacy and safety. This yielded commonly endorsed facilitators (e.g., collaborative learning through group supervision, the convenience of consolidated patient data), barriers (e.g., difficulties with new technologies) and suggested changes (e.g., esthetic improvements, suicide risk assessment prompt). The PEERS app has the potential to scale measurement-based peer supervision to facilitate quality assurance of psychological treatments across contexts.

Hyperplastic Anterior Choroidal Artery-A Rare Variant Detected on MR Angiography.

Introduction The anterior choroidal artery is a branch of the terminal internal carotid artery (ICA) that is often an incidentally detected anomaly. The hyperplastic variant has been linked with an increased risk of aneurysms. We explore the role of magnetic resonance (MR) angiography in the detection of this variant and its impact of presentation of patients. Patient Presentation A 62-year-old diabetic and hypertensive male came to the casualty with complaints of giddiness, left-sided weakness, and loss of coordination for the last 2 to 3 days. He was provisionally diagnosed with a posterior circulation stroke. Management and Outcome Magnetic resonance imaging of brain revealed an acute infarct in the right thalamus and the midbrain. Time-of-flight angiography sequences were done that showed hypoplasia of the A1 segment of the left anterior cerebral artery, fetal origin of the right posterior cerebral artery, narrowing of the left ICA, and a hyperplastic left anterior choroidal artery. He was managed conservatively with antiplatelets. He later underwent a digital subtraction angiography that revealed significant narrowing of the left ICA for which he was advised carotid stenting. However, the patient was unwilling for the procedure and was discharged on oral medication with stable vitals. Conclusion Anatomical variations in the intracranial vasculature impact ischemia territory and the approach to intracranial pathology. Hyperplastic anterior choroidal artery results from the abnormal persistence of fetal pattern of vascular supply. It is important to recognize the presence of a hyperplastic anterior choroidal artery as it impacts both disease presentation and management of these patients. MR angiography allows for noninvasive and reliable detection of these anomalies in patients without the risks associated with radiation or contrast exposure in conventional/computed tomography angiography.

Resolving the Conflict Between Regional Analgesia and Developing Compartment Syndrome in Below-Knee Surgeries With the High-Volume Proximal Adductor Canal (Hi-PAC) Block: A Prospective Feasibility Study.

The consideration of regional analgesia (RA) in below-knee surgeries is always a controversial topic due to the fear of masking symptoms of developing compartment syndrome (CS) in the postoperative period. Compartment syndrome (CS) has been found frequently in below-knee surgeries, particularly among tibial diaphyseal fractures. Like any other surgery, below-knee surgeries have significant postoperative pain that requires effective postoperative analgesia protocol. The analgesia quality makes a big difference when compared with or without RA. Also, the presence or absence of RA cannot prevent or promote the development of CS. Therefore, patients should not be deprived of their right to remain pain-free in the postoperative period by compromising the analgesia protocol. The pain out of proportion to the surgery or injury is a typical symptom of developing CS, which can cause increased analgesic demands postoperatively. Timely diagnosis and treatment of CS require vigilant postoperative monitoring of the warning signs by trained staff. Avoiding RA for fear of presumed masking of symptoms and delaying CS diagnosis may not be a solution instead of choosing an appropriate RA with regular postoperative monitoring for such warning symptoms. The high-volume proximal adductor canal (Hi-PAC) block has been described as a procedure-specific and motor-sparing RA technique appropriate for below-knee surgeries. In this prospective study, we evaluated the analgesic efficacy of the Hi-PAC block in below-knee surgeries. We also observed the effect of the Hi-PAC block, due to proximal and distal drug distribution, on masking the symptoms of the developing CS during postoperative monitoring. We found the Hi-PAC block to be a safer and more effective RA alternative for below-knee surgeries with an added motor-sparing benefit that facilitated early mobility and discharge. Its property of not interfering with postoperative surveillance to detect the symptoms of CS and intervene in time helps deal with the anxiety of CS in below-knee surgeries.

Effectiveness of Health Promotion Curriculum in Enhancing Non-communicable Diseases Health Literacy in Indian Youth.

Background There is a theory-praxis gap related to health literacy interventions focused on non-communicable diseases (NCDs) among young people. We designed an NCD curriculum and investigated its' effect on health literacy in non-medical, non-nursing college students in India. We deliberately selected non-medical and non-nursing college students (age 17 to 22 years) as we hypothesized, they would have a minimum baseline knowledge of NCDs. Methods We initially carried out a pilot study on 85 students in a four-day-long workshop (32 teaching hours) using empirically developed health literacy instrument. We administered the curriculum to 120 randomly selected students in four colleges, while 50 students were assigned to the comparison group. The curriculum was given over four days for a total of 32 hours. Approval was sought to give four credits for completion of the course. Each lecture comprised didactics, followed by discussion, and skills testing of measuring blood pressure and blood sugar. Health literacy was measured using a specifically designed tool at baseline and endline. The difference in health literacy scores between the two time-points (timepoint 1: before delivering the curriculum, time-point 2: at the end of four days of training) was analyzed using the t-test. Multiple linear and Poisson regression models were used to account for covariates. Results The average difference between the intervention and the control group in baseline scores was 0.05% points (unpaired t-test statistics: -1.36, degrees of freedom 103.15, p>0.05). The same at endline was 20.59% points (unpaired t-test statistics: -11.31, degrees of freedom 138.14, p<0.001). The endline to baseline difference in health literacy scores was 18.54% points higher in the intervention group versus the control group (unpaired t-test statistics: -10.88, degrees of freedom 161.32, p<0.001). The difference-in-difference scores remained significant after accounting for college setting and baseline score (Multivariable linear regression model, ß: 19.62% points, p<0.001). None of the socio-economic characteristics were significantly associated with the difference in the difference scores, independent of the intervention effect. The proportion of participants scoring 40% or above on the health literacy measure at endline was significantly higher in the intervention versus the control group (p<0.001). Conclusions We provide empirical data to support the incorporation of NCDs as a credit course in college curricula in low- and middle-income countries. Our findings showed that a theory-driven skills-focused curriculum may be a tool for enhancing NCD health literacy in Indian youth from diverse academic and socio-economic backgrounds.

Labrune Syndrome: A Very Rare Association of Leukoencephalopathy, Cerebral Calcifications, and Cysts.

Leukoencephalopathy, cerebral calcifications, and cysts (LCC) form a very rare association which is named as "Labrune syndrome" after Labrune who reported the first case in 1996. To the best of our knowledge only eight to 10 cases have been reported in literature to date. We report a case of a 26-year-old male with onset of neurological symptoms in late adolescence (at 19 years of age) and presented with complains of continued seizures for 7 years, giddiness with imbalance, and slowly progressive motor symptoms. MRI brain revealed multiple calcifications in bilateral basal ganglia, cerebral white matter, multiple cystic lesions in the supratentorial white matter, and abnormal diffused bilateral white matter T2 hyperintensity suggesting leukoencephalopathy. Histopathological evaluation revealed prominent congested blood vessels suggestive of angiomatous changes and cystic areas suggestive of secondary gliosis.

Community-level women's education and undernutrition among Indian adolescents: A multilevel analysis of a national survey.

BACKGROUND: Little research has explored the influence of social context on health of Indian adolescents. We conceptualized community-level women's education (proxy for value placed on women's wellbeing) as exerting contextual influence on adolescent hemoglobin level and body mass index (BMI). METHODS: We derived our sample of more than 62,000 adolescent aged 15 through 17 years from the Indian National Family Health Survey 2015-16. The sample consisted of a total of 62648 adolescents (54232 girls and 8416 boys) for the hemoglobin, and 62846 adolescents (54383 girls and 8463 boys) for the BMI analysis. We fitted multilevel random intercepts linear regression models to test the association of village- and urban-ward-level-women's education with hemoglobin level and BMI of adolescents, accounting for their own and their mother's education; as well as relevant covariates. FINDINGS: Our fully adjusted model estimated that if the 52% of communities with less than 20 percent of women having a tenth-grade education in our sample were to achieve 100 percent tenth-grade completion in women, hemoglobin would be 0·2 g/dl higher (p<0·001) and BMI would be 0·62 kg/m2 higher on average among all adolescents in such communities. Unexplained variance estimates at the contextual level remained statistically significant, indicating the importance of context on adolescent undernutrition. INTERPRETATIONS: Adolescents are deeply embedded in their context, influenced by contextual factors affecting health. Promoting adolescent health therefore implies altering social norms related to adolescent health and health behaviors; along with structural changes creating a health-promoting environment. Integrating our empirical findings with theoretically plausible pathways connecting community-level women's education with adolescent undernutrition, we suggest that enhancing community-level women's education beyond high school is necessary to facilitate these processes. IMPLICATIONS: Addressing contextual determinants of adolescent undernutrition might be the missing link in India's adolescent anemia and undernutrition prevention efforts, which are currently focused heavily on individual-level biomedical determinants of the problem.

Development of self-microemulsifying lipid-based formulations of trans-resveratrol by systematically constructing lipid-surfactant-water phase diagrams using long-chain lipids.

The aim of this work was to develop self-microemulsifying lipid-based formulations of trans-resveratrol in cod liver oil, a long chain lipid, to increase its solubility, dissolution rate and oral bioavailability. Ternary phase diagrams of cod liver oil with surfactant and water as well as pseudo-ternary phase diagrams of the same by mixing cod liver oil (triglyceride) with glycerol monooleate (monoglyeride) were constructed to identify regions where microemulsions were formed. Kolliphor RH 40, Tween 80 and their 1:1-mixtures were evaluated as surfactants. No organic cosolvents were added. It was observed that cod liver oil alone did not form microemulsion with any of the surfactants used, and a 1:1 mixture of cod liver oil and glycerol monooleate was necessary to enable the formation of microemulsion. Among the surfactants, Kolliphor RH 40 provided the maximum microemulsification effect. Several formulations containing 6:4, 1:1, and 4:6 w/w ratios of lipid to surfactant using the 1:1 mixture of cod liver oil and glycerol monooleate as lipid components and Kolliphor RH 40 or its mixture with Tween 80 as surfactants were identified, and trans-resveratrol solubility in these formulations were determined. Drug concentrations used in the formulations were 80% of saturation solubility, and no organic cosolvents were used in any formulations to increase drug solubility or enable emulsification. In vitro dispersion testing in 250 mL of 0.01 N HCl (pH 2) according to the USP method 2 at 50 RPM showed that the formulations rapidly dispersed in aqueous media forming microemulsions and there was no drug precipitation.

Role of MRI in Evaluation of Spectrum of Liver Lesions in Cirrhotic Patients.

MRI provides better intrinsic soft-tissue contrast with more enhanced depiction of even subtly different tissue properties making lesion evaluation easy. Faster sequences which capture arterial sequences better, lack of ionizing radiation and simultaneous evaluation of background liver parenchyma and the liver lesions are additional advantages of using MRI as the imaging technique of choice. Comprehensive liver imaging using MRI now includes T1, T2-weighted imaging and in- and opposed-phase, in addition to dynamic post-contrast imaging with proper breath holding techniques. Wider variety of liver specific contrast agents is available for use in MR imaging with the gadolinium based agents being considered the most useful and practical, particularly for lesion characterization. AIMS AND OBJECTIVES: To evaluate MRI spectrum of liver lesions in cirrhotic patients, Role of MRI in focal liver lesion evaluation and to differentiate benign versus malignant lesions. MATERIALS AND METHODS: A prospective study of OPD or IPD patients who underwent imaging tests like Ultrasonography, or CT scan for suspected chronic liver disease was done. A total 35 patients were investigated (June 2014 - November 2016) with MRI abdomen done with the patient in supine position on a Philips Achieva 3.0T MRI scanner. Standard MRI abdomen protocol, including T2W TSE in axial and coronal plane, T2W fat suppressed (SPAIR) images in axial and coronal plane, T1W TFE, in- and out-of-phase imaging and Diffusion-weighted imaging (DWI) in axial plane along with pre-contrast baseline fat-suppressed T1W imaging in at least one plane was acquired. Breath-holding was required in few sequences. 0.1 mmol/kg Gadolinium based contrast (Gadobenate) was injected at the rate of 2.5 ml/sec followed by saline flush and dynamic contrast enhanced MRI (DCE-MRI) with post-contrast fat-suppressed T1W imaging was acquired. RESULTS AND CONCLUSIONS: In cirrhosis, there is development of nodules which are initially only microscopically detectable. With progression of cirrhosis, there is development of radiologically detectable regenerative nodules, dysplastic nodules and hepatocellular carcinoma. Amongst these regenerative nodules are completely benign lesions whereas dysplastic nodules, though benign, are considered premalignant; and hepatocellular carcinoma is a malignant condition. Differentiation of benign versus malignant lesions is possible on the basis of enhancement pattern in dynamic contrast enhanced MRI. The signal characteristics of focal lesions and other findings like portal vein thrombosis are helpful, give additional clue to the diagnosis and also helpful in assigning LIRADS grade to a lesion. Also, MRI characterization after gadolinium based contrast injection was found to be similar to the previous imaging based on non-gadolinium contrast agents.

Aftermath of pulmonary tuberculosis: computed tomography assessment.

PURPOSE: Pulmonary tuberculosis (PTB) has clinically significant sequelae, even after recommended treatment completion. It is important to recognise these sequelae for accurate assessment of severity and treatment planning, if indicated. MATERIAL AND METHODS: We retrospectively analysed contrast-enhanced computed tomography (CT) scans of chest of 100 patients with previous history of treated pulmonary tuberculosis, excluding those with active pulmonary disease. CT findings were analysed based on parenchymal, airway, pleural, mediastinal, and vascular sequelae of PTB. RESULTS: Parenchymal sequelae included fibrosis with architectural distortion and volume loss (90%), cavities (21%) (with aspergillomas noted in 19% of these cases), and tuberculomas (54%). Airway involvement was noted as bronchiectasis (77%) and bronchial stenosis (4%) but none with broncholithiasis. Mediastinal sequelae included lymph node calcification (74%), fibrosing mediastinitis (1%), and pericardial tuberculosis (2%). Pleural sequelae included pleural thickening (22%), with 40.9% of these patients showing calcifications, and one patient with chronic chylous pleural effusion. Vascular sequelae included Rasmussen aneurysms (4%), enlarged bronchial arteries (3%), and systemic bronchial collaterals in 1% of our patients. CONCLUSIONS: PTB has multiple appalling sequelae, which require due attention and appropriate treatment in symptomatic cases. Radiological evaluation forms an integral part in patient assessment and decision making.

Surgical Unmet Need in a Low-Income Area of a Metropolitan City in India: A Cross-Sectional Study.

INTRODUCTION: We investigated the burden of surgical conditions, level of unmet needs and reasons for non-utilization of surgical services in a slum of Ahmedabad, India. METHODS: A community-based cross-sectional study was carried out from August to December 2019. Inclusion criteria was age > 14 years; any type of injury/condition that requires surgery; subject has had surgery in last 1 year, and death information of family members. Data were stored and coded in Microsoft excel and exported to IBM SPSS statistics version 25 software for data analysis. Frequencies and proportions (categorical variable) are used to summarize utilization of surgical services and understanding surgical need. The Surgeons Overseas Assessment of Surgical was used to identify surgical met and unmet needs translated into local language. Open Data Kit software was used to install questionnaire in the "Tablet" to collect information and stress-free workflow in field. RESULTS: Out of 10,330 population in 2066 households, 7914 were more than 14 years of age. 3.46% (n = 274) people needed surgery; 116 did not avail surgery and were categorized in "unmet need." Fifty percent of individuals with surgical needs had abdominal- or extremities-related problems followed by eyes surgery need (14%); back, chest and breast surgical need was 13.5%. Seventeen percent of participants with surgical needs had wounds related to injury or accident while 63% had wounds that were not related to injury. Almost all participants had gone to a physician to seek healthcare, however 42% did not avail surgical care needed for a variety of reasons. Forty-six percent of participants needing surgical care underwent major surgical procedure, while 11% had minor procedures. Financial reasons (34.5%) and lack of trust (35.3%) were major reasons for not availing surgical care. CONCLUSIONS AND RELEVANCE: Ahmedabad is a relatively high income metropolitan city, has universally free health care and multiple healthcare facilities. Despite this, we have shown that there is significant unmet need for surgical procedures in the low-income population. A unique finding was that most patients sought a consultation but approximately 50% did not avail of the free surgical procedures under the universally free health care system in this city. We propose creation of community healthcare workers focused on surgical conditions.

Development of 3D Printed Tablets by Fused Deposition Modeling Using Polyvinyl Alcohol as Polymeric Matrix for Rapid Drug Release.

In this study, the processability of polyvinyl alcohol (PVA), a water-soluble polymer, into melt-extruded filaments and then into 3D printed tablets by fused deposition modeling was studied. PVA is semicrystalline with Tg and m.p. of ~45°C and ~190°C, respectively. After screening several plasticizers, sorbitol was selected to enhance melt extrudability of PVA. Carvedilol and haloperidol, 2 basic compounds with pH-dependent solubility, were used as model drugs. Miscibility of the drugs with PVA, with and without added sorbitol as plasticizer, was also tested to determine whether any amorphous solid dispersion was formed that would facilitate rapid and pH-independent dissolution. Finally, the drug release from physical mixtures, crushed extrudates, and printed tablets were determined. Owing to high m.p. and high melt viscosity of PVA, filaments containing 10% and 20% drug required 180°C-190°C for extrusion, which could be reduced to ~150°C by adding 10% sorbitol. The printing temperature of 210°C was, however, required. Miscibility of carvedilol and haloperidol with PVA were, respectively, ~20% and <10%. PVA provided complete drug release from 3D printed tablets with 10% and 20% carvedilol and 60% infill in ~45 min at both pH 2 and 6.8. However, despite relatively rapid dissolution rate, high processing temperature and limited drug-polymer miscibility could be potential development issues with PVA.

Evaluation of Cytotoxicity of Self-Emulsifying Formulations Containing Long-Chain Lipids Using Caco-2 Cell Model: Superior Safety Profile Compared to Medium-Chain Lipids.

Medium-chain (MC) and long-chain (LC) lipids are used for development of self-emulsifying drug delivery systems (SEDDS). MC lipids are often preferred because of their ability to form stable microemulsions with relatively high drug solubilization capacity. On the other hand, LC lipids could be more biocompatible as most endogenous and dietary lipids are LC glycerides. They also maintain high drug solubilization capacity after digestion. The present study was undertaken to determine the cytotoxicity of LC lipids and their formulations on Caco-2 cells of 1-day, 5-day, and 21-day maturity. The results were compared with the cytotoxicity profiles of MC lipids reported previously from our laboratory. The cell viability and cell membrane integrity were, respectively, determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and the lactate dehydrogenase assay. The cytotoxicity was partially due to lipid surfactant-induced membrane rupture, and it was influenced by cell maturity and formulation composition. The lipid-surfactant combinations showed greater tolerance than surfactants alone, and LC-SEDDS were well-tolerated at almost 10-fold higher concentration than corresponding MC-SEDDS. Furthermore, the cytotoxicity of digestion end products of both LC and MC triglycerides in the presence of 3 mM sodium taurocholate was compared on 21-day Caco-2 cultures by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The LC lipid formulations showed better tolerance than MC lipid formulations after digestion. Thus, although MC and LC lipids are well-tolerated at doses normally administered to humans, LC lipids show much better safety than MC lipids in a cell-culture model.

Transcriptional dysregulation in developing trigeminal sensory neurons in the LgDel mouse model of DiGeorge 22q11.2 deletion syndrome.

LgDel mice, which model the heterozygous deletion of genes at human chromosome 22q11.2 associated with DiGeorge/22q11.2 deletion syndrome (22q11DS), have cranial nerve and craniofacial dysfunction as well as disrupted suckling, feeding and swallowing, similar to key 22q11DS phenotypes. Divergent trigeminal nerve (CN V) differentiation and altered trigeminal ganglion (CNgV) cellular composition prefigure these disruptions in LgDel embryos. We therefore asked whether a distinct transcriptional state in a specific population of early differentiating LgDel cranial sensory neurons, those in CNgV, a major source of innervation for appropriate oropharyngeal function, underlies this departure from typical development. LgDel versus wild-type (WT) CNgV transcriptomes differ significantly at E10.5 just after the ganglion has coalesced. Some changes parallel altered proportions of cranial placode versus cranial neural crest-derived CNgV cells. Others are consistent with a shift in anterior-posterior patterning associated with divergent LgDel cranial nerve differentiation. The most robust quantitative distinction, however, is statistically verifiable increased variability of expression levels for most of the over 17 000 genes expressed in common in LgDel versus WT CNgV. Thus, quantitative expression changes of functionally relevant genes and increased stochastic variation across the entire CNgV transcriptome at the onset of CN V differentiation prefigure subsequent disruption of cranial nerve differentiation and oropharyngeal function in LgDel mice.

Six1 proteins with human branchio-oto-renal mutations differentially affect cranial gene expression and otic development.

Single-nucleotide mutations in human SIX1 result in amino acid substitutions in either the protein-protein interaction domain or the homeodomain, and cause ∼4% of branchio-otic (BOS) and branchio-oto-renal (BOR) cases. The phenotypic variation between patients with the same mutation, even within affected members of the same family, make it difficult to functionally distinguish between the different SIX1 mutations. We made four of the BOS/BOR substitutions in the Xenopus Six1 protein (V17E, R110W, W122R, Y129C), which is 100% identical to human in both the protein-protein interaction domain and the homeodomain, and expressed them in embryos to determine whether they cause differential changes in early craniofacial gene expression, otic gene expression or otic morphology. We confirmed that, similar to the human mutants, all four mutant Xenopus Six1 proteins access the nucleus but are transcriptionally deficient. Analysis of craniofacial gene expression showed that each mutant causes specific, often different and highly variable disruptions in the size of the domains of neural border zone, neural crest and pre-placodal ectoderm genes. Each mutant also had differential effects on genes that pattern the otic vesicle. Assessment of the tadpole inner ear demonstrated that while the auditory and vestibular structures formed, the volume of the otic cartilaginous capsule, otoliths, lumen and a subset of the hair cell-containing sensory patches were reduced. This detailed description of the effects of BOS/BOR-associated SIX1 mutations in the embryo indicates that each causes subtle changes in gene expression in the embryonic ectoderm and otocyst, leading to inner ear morphological anomalies.

Effects of Surfactants on Itraconazole-Hydroxypropyl Methylcellulose Acetate Succinate Solid Dispersion Prepared by Hot Melt Extrusion. II: Rheological Analysis and Extrudability Testing.

Although hydroxypropyl methylcellulose acetate succinate (HPMCAS) has been widely used as a carrier for amorphous solid dispersion of poorly water-soluble drugs, its application has mostly been limited to spray drying, and the solvent-free method of hot melt extrusion has rarely been used. This is on account of the high temperature (≥170°C) required for extrusion where the polymer and even a drug may degrade. In part 1 of this series of papers, we demonstrated that HPMCAS is miscible with surfactants such as, poloxamer 188, poloxamer 407 and d-alpha tocopheryl polyethylene glycol 1000 succinate, which may also serve as plasticizers (Solanki et al., J Pharm Sci. 2019; 108 (4):1453-1465). The present investigation was undertaken to determine plasticization effects of the surfactants and a model drug, itraconazole, in reducing melt extrusion temperatures of HPMCAS. The determination of complex viscosity as functions of temperature and also as functions of angular frequency at certain fixed temperatures showed that the surfactants and the drug greatly reduce viscosity of HPMCAS by their plasticization effects. Surfactants and drug also had synergistic effects in reducing viscosity. The torque analysis during melt extrusion demonstrated that these additives greatly enhanced extrudability of HPMCAS. Surfactant-drug-polymer mixtures were successfully extruded as stable amorphous solid dispersions at 130°C, which is much lower than the minimum extrusion temperature of 170°C for neat HPMCAS.

DNA damage response activates respiration and thereby enlarges dNTP pools to promote cell survival in budding yeast.

The DNA damage response (DDR) is an evolutionarily conserved process essential for cell survival. Previously, we found that decreased histone expression induces mitochondrial respiration, raising the question whether the DDR also stimulates respiration. Here, using oxygen consumption and ATP assays, RT-qPCR and ChIP-qPCR methods, and dNTP analyses, we show that DDR activation in the budding yeast Saccharomyces cerevisiae, either by genetic manipulation or by growth in the presence of genotoxic chemicals, induces respiration. We observed that this induction is conferred by reduced transcription of histone genes and globally decreased DNA nucleosome occupancy. This globally altered chromatin structure increased the expression of genes encoding enzymes of tricarboxylic acid cycle, electron transport chain, oxidative phosphorylation, elevated oxygen consumption, and ATP synthesis. The elevated ATP levels resulting from DDR-stimulated respiration drove enlargement of dNTP pools; cells with a defect in respiration failed to increase dNTP synthesis and exhibited reduced fitness in the presence of DNA damage. Together, our results reveal an unexpected connection between respiration and the DDR and indicate that the benefit of increased dNTP synthesis in the face of DNA damage outweighs possible cellular damage due to increased oxygen metabolism.

Solubility-pH profile of desipramine hydrochloride in saline phosphate buffer: Enhanced solubility due to drug-buffer aggregates.

Although solubility-pH data for desipramine hydrochloride (DsHCl) have been reported previously, the aim of the present study was to critically examine the aqueous solubility-pH behavior of DsHCl in buffer-free and buffered solutions, in the presence of physiologically-relevant chloride concentration, using experimental practices recommended in the recently-published "white paper" (Avdeef et al., 2016). The computer program pDISOL-X was used to design the structured experiments (pH-RSF method), to process the data, and to refine the equilibrium constants. Low-to-high and high-to-low pH assays (using HCl, H3PO4, or NaOH to adjust pH) were performed on phosphate-buffered (0.12­0.15 M) saturated solutions of DsHCl in the pH 1.3-11.6 range. After equilibration (stirring 6 h, followed by 18 h stir-free sedimentation), filtration or centrifugation was used for phase separation. Concentration was measured using HPLC with UV/VIS detection. The 2:1 drug-phosphate solubility product (Ksp2:1 = [DsH+]2[HPO42-]) was determined from data in the pH 4-9 region. The free base of desipramine was prepared and used to determine the Ksp1:1 ([DsH+][H2PO4-]) in chloride-free acidified suspension. In addition, phosphate-free titrations were conducted to determine the intrinsic solubility, S0, and the 1:1 drug-chloride solubility product, KspDsHCl = [DsH+][Cl-]. Under the assay conditions, only the phosphate-free solutions showed some supersaturation near pHmax 8.0. In phosphate-containing solutions, pHmax was indicated at higher pH (8.8-9.6). Oils mixed with solids were observed to form in alkaline solutions (pH > 11). Notably, soluble drug-phosphate complexes appeared to form below pH 3.9 and above pHmax in saturated phosphate­containing saline solutions. This was indicated by the systematic pH shift to higher values in the log S-pH curve in alkaline solution than expected from the Henderson-Hasselbalch equation. For pH < 3.9, saturated phosphate-containing saline solutions exhibited elevated solubility, with drug-hydrochloride as the sole precipitate. Salt solubility products, intrinsic solubility, and complexation constants, which rationalized the data, were determined. Elemental, thermogravimetric (TGA), differential scanning calorimetric (DSC), and powder X-ray diffraction (PXRD) analyses were used to characterize the precipitates isolated from suspensions at different pH.