Identification of sequence determinants for the ABHD14 enzymes.

Over the course of evolution, enzymes have developed remarkable functional diversity in catalyzing important chemical reactions across various organisms, and understanding how new enzyme functions might have evolved remains an important question in modern enzymology. To systematically annotate functions, based on their protein sequences and available biochemical studies, enzymes with similar catalytic mechanisms have been clustered together into an enzyme superfamily. Typically, enzymes within a superfamily have similar overall three-dimensional structures, conserved catalytic residues, but large variations in substrate recognition sites and residues to accommodate the diverse biochemical reactions that are catalyzed within the superfamily. The serine hydrolases are an excellent example of such an enzyme superfamily. Based on known enzymatic activities and protein sequences, they are split almost equally into the serine proteases and metabolic serine hydrolases. Within the metabolic serine hydrolases, there are two outlying members, ABHD14A and ABHD14B, that have high sequence similarity, but their biological functions remained cryptic till recently. While ABHD14A still lacks any functional annotation to date, we recently showed that ABHD14B functions as a lysine deacetylase in mammals. Given their high sequence similarity, automated databases often wrongly assign ABHD14A and ABHD14B as the same enzyme, and therefore, annotating functions to them in various organisms has been problematic. In this article, we present a bioinformatics study coupled with biochemical experiments, which identifies key sequence determinants for both ABHD14A and ABHD14B, and enable better classification for them. In addition, we map these enzymes on an evolutionary timescale and provide a much-wanted resource for studying these interesting enzymes in different organisms.

Hypertension in Pregnancy: What We Now Know.

PURPOSE OF REVIEW: Hypertensive disorders of pregnancy remain a highly morbid condition that affects both the mother and fetus, complicate approximately 10% of pregnancies worldwide, and contribute to immediate and long-term cardiovascular outcomes. There is still much to learn regarding pathogenesis and treatment goals. RECENT FINDINGS: There is updated information on the pathogenesis of preeclampsia and treatment thresholds for HTN in pregnancy. l-Kynurenine, a metabolite of the essential amino acid l-tryptophan, has been implicated in preeclampsia as decreased levels were found in a uninephrectomized pregnant mouse model of preeclampsia, where replacement of l-kynurenine rescued the preeclamptic state. Further, data from CHIPS (The Control of HTN in Pregnancy Study) and CHAP (Chronic HTN and Pregnancy) trials demonstrate not only the safety of lowering blood pressure to either a diastolic goal of 85 mmHg (CHIPS) or less than 160/105 mmHg (CHAP) without detriment to the fetus but the CHAPS trial has also shown a decrease in the rate of preeclampsia in the treatment group. SUMMARY: We will summarize the different types of hypertensive disorders in pregnancy, updates on the pathogenesis of preeclampsia, and appropriate HTN management based on the latest evidence in order to better care for mother and child.

Design and Synthesis of BODIPY-Hetero[5]helicenes as Heavy-Atom-Free Triplet Photosensitizers for Photodynamic Therapy of Cancer.

Designing heavy-atom-free triplet photosensitizers (PSs) is a challenge for the efficient photodynamic therapy (PDT) of cancer. Helicenes are twisted polycyclic aromatic hydrocarbons (PAHs) with an efficient intersystem crossing (ISC) that is proportional to their twisting angle. But their difficult syntheses and weak absorption profile in the visible spectral region restrict their use as heavy-atom-free triplet PSs for PDT. On the other hand, boron-containing PAHs, BODIPYs are highly recognized for their outstanding optical properties. However, planar BODIPY dyes has low ISC and thus they are not very effective as PDT agents. We have designed and synthesized fused compounds containing both BODIPY and hetero[5]helicene structures to develop red-shifted chromophores with efficient ISC. One of the pyrrole units of the BODIPY core was also replaced by a thiazole unit to further enhance the triplet conversion. All the fused compounds have helical structure, and their twisting angles are also increased by substitutions at the boron centre. The helical structures of the BODIPY-hetero[5]helicenes were confirmed by X-ray crystallography and DFT structure optimization. The designed BODIPY-hetero[5]helicenes showed superior optical properties and high ISC with respect to [5]helicene. Interestingly their ISC efficiencies increase proportionally with their twisting angles. This is the first report on the relationship between the twisting angle and the ISC efficiency in twisted BODIPY-based compounds. Theoretical calculations showed that energy gap of the S1 and T1 states decreases in BODIPY-hetero[5]helicene as compared to planar BODIPY. This enhances the ISC rate in BODIPY-hetero[5]helicene, which is responsible for their high generation of singlet oxygen. Finally, their potential applications as PDT agents were investigated, and one BODIPY-hetero[5]helicene showed efficient cancer cell killing upon photo-exposure. This new design strategy will be very useful for the future development of heavy-atom-free PDT agents.

Neuromuscular blockade characteristics of cisatracurium in patients receiving chemotherapy: A preliminary study in breast cancer patients.

Background and Aims: Cancer chemotherapeutic agents cause alteration in the response to neuromuscular blocking drugs, which can have serious perioperative implications. Magnesium, commonly found to be deficient in these patients, plays an indispensable role in neuromuscular transmission. This study aimed to understand the effect of neoadjuvant chemotherapy on the neuromuscular blocking properties of cisatracurium. Material and Methods: One hundred female patients scheduled for breast cancer surgery were divided into two groups (n = 50 each). Group B received neoadjuvant chemotherapy with taxane, adriamycin, and cyclophosphamide, and Group A did not receive neoadjuvant chemotherapy. Neuromuscular block following cisatracurium 0.15 mg/kg was measured using peripheral nerve stimulator at the ulnar nerve. Onset time, duration of intense block, clinical duration of action, time to TOF4 after the last dose of cisatracurium, along with preoperative serum magnesium concentration were measured. Correlation and multiple regression were run to analyze the relationship between history of neoadjuvant chemotherapy, preoperative magnesium, and the abovementioned time points. Mediation analysis was done to ascertain if magnesium was mediating the observed effects. Results: Onset time was prolonged by nearly 18% in Group B compared to Group A (P = 0.001). The duration of intense block was 35.27 ± 8.9 min in Group B and 42.07 ± 10.99 min in Group A (P < 0.001). The clinical duration of action of cisatracurium was significantly shorter in Group B (46.06 ± 8.68 min) compared to Group A (55.87 ± 11.04 min, P < 0.001). The time to TOF4 was 32.86 ± 5.66 min in Group B and 36.57 ± 8.49 min in Group A (P < 0.05). Preoperative serum magnesium levels were significantly lower in Group B (P < 0.001). Conclusion: Patients who had received neoadjuvant chemotherapy had a delayed onset, shorter duration of action, and faster recovery for cisatracurium. Although preoperative magnesium levels were lower in Group B, it was found to be an independent predictor rather than a mediator of these effects.

Periodontal and Dentition Status among Psychiatric Patients in Indore: A Descriptive Cross-sectional Study.

AIM: To assess and record periodontal and dentition status among psychiatric patients using modified WHO Oral Health Assessment form 1997. MATERIALS AND METHODS: A descriptive cross-sectional study was conducted at Sri Aurobindo Institute of Medical Sciences and PG Institute, Indore, among the inpatients and outpatients. Ethical approval for the study was obtained from Institutional Review Board of Sri Aurobindo College of Dentistry. Statistical analysis was done using SPSS, IBM version 20.0. Descriptive statistics was used to find the frequencies, mean, and standard deviation of variables considered in the study. RESULTS: Among all the disorders, schizophrenia subjects had shallow pockets 28 (25.2%) and deep pockets 55 (49.5%). Only 2 (2.6%) study subjects who had major depressive disorder had more than 12 mm loss of attachment (LOA). The highest mean DMFT score was recorded for schizophrenia patients (13.0 ± 7.09). CONCLUSION: The results reveal an unmet need of projecting effective planning and implementation strategies toward the improvement of periodontal and dentition status health of the psychiatric patients. CLINICAL SIGNIFICANCE: The study highlights the importance of incorporating dental health education to psychiatric rehabilitation programs.

Functional role of brain-engrafted macrophages against brain injuries.

BACKGROUND: Brain-resident microglia have a distinct origin compared to macrophages in other organs. Under physiological conditions, microglia are maintained by self-renewal from the local pool, independent of hematopoietic progenitors. Pharmacological depletion of microglia during whole-brain radiotherapy prevents synaptic loss and long-term recognition memory deficits. However, the origin or repopulated cells and the mechanisms behind these protective effects are unknown. METHODS: CD45low/int/CD11b+ cells from naïve brains, irradiated brains, PLX5622-treated brains and PLX5622 + whole-brain radiotherapy-treated brains were FACS sorted and sequenced for transcriptomic comparisons. Bone marrow chimeras were used to trace the origin and long-term morphology of repopulated cells after PLX5622 and whole-brain radiotherapy. FACS analyses of intrinsic and exotic synaptic compartments were used to measure phagocytic activities of microglia and repopulated cells. In addition, concussive brain injuries were given to PLX5622 and brain-irradiated mice to study the potential protective functions of repopulated cells after PLX5622 + whole-brain radiotherapy. RESULTS: After a combination of whole-brain radiotherapy and microglia depletion, repopulated cells are brain-engrafted macrophages that originate from circulating monocytes. Comparisons of transcriptomes reveal that brain-engrafted macrophages have an intermediate phenotype that resembles both monocytes and embryonic microglia. In addition, brain-engrafted macrophages display reduced phagocytic activity for synaptic compartments compared to microglia from normal brains in response to a secondary concussive brain injury. Importantly, replacement of microglia by brain-engrafted macrophages spare mice from whole-brain radiotherapy-induced long-term cognitive deficits, and prevent concussive injury-induced memory loss. CONCLUSIONS: Brain-engrafted macrophages prevent radiation- and concussion-induced brain injuries and cognitive deficits.

Sequential genetic testing of living-related donors for inherited renal disease to promote informed choice and enhance safety of living donation.

Living kidney donors (LKDs) with a family history of renal disease are at risk of kidney disease as compared to LKDs without such history suggesting that some LKDs may be pre-symptomatic for monogenic kidney disease. LKDs with related transplant candidates whose kidney disease was considered genetic in origin were selected for genetic testing. In each case, the transplant candidate was first tested to verify the genetic diagnosis. A genetic diagnosis was confirmed in 12 of 24 transplant candidates (ADPKD-PKD1: 6, ALPORT-COL4A3: 2, ALPORT-COL4A5: 1: nephronophthisis-SDCCAG8: 1; CAKUT-HNF1B and ADTKD-MUC1: 1 each) and 2 had variants of unknown significance (VUS) in phenotype-relevant genes. Focused genetic testing was then done in 20 of 34 LKDs. 12 LKDs screened negative for the familial variant and were permitted to donate; seven screened positive and were counseled against donation. One, the heterozygous carrier of a recessive disorder was also cleared. Six of seven LKDs with a family history of ADPKD were under 30 years and in 5, by excluding ADPKD, allowed donation to safely proceed. The inclusion of genetic testing clarified the diagnosis in recipient candidates, improving safety or informed decision-making in LKDs.

Oral Health-related Quality of Life among Psychiatric In- and Outpatients Diagnosed with Different Disorders in Indore, Central India.

AIM: To assess oral health-related quality of life among psychiatric patients. MATERIALS AND METHODS: A total of 400 inpatients and outpatients reporting to the Department of Psychiatry was taken as the final sample. Subjects diagnosed with mental illness and on medication for at least 1 year, who were able to respond to the pro forma and oral health impact profile (OHIP) questionnaire, were included. A guided, self-administered structured questionnaire was designed to include sociodemographic characteristics and short form of the OHIP-14 consisting of 14 items covering 7 domains: functional limitation, physical pain, psychological discomfort, physical disability, psychological disability, social disability and handicap. It was used to assess the impact of oral conditions on well-being and quality of life. The data collected were subjected to statistical analysis using SPSS IBM version 20.0. RESULTS: The highest mean OHIP score for the disorders was seen among the patients diagnosed with schizophrenia for functional limitation (2.73 ± 1.194), 2.91 ± 1.111 for psychological discomfort, 2.67 ± 1.203 was recorded for physical disability, 2.79 ± 1.156 was the mean score for psychological disability followed by a score of 2.87 ± 1.172 for social disability. The highest mean score for the handicap domain was recorded for schizophrenia patients (2.73 ± 1.241) whereas for physical pain, a mean score of 3.01 ± 1.261 was recorded for patients diagnosed with bipolar disorder. CONCLUSION: Based on the findings of the study, it can be stated that the psychiatrists should pay attention to the dental anxiety concerns of the patients and encourage them to visit oral health professionals. CLINICAL SIGNIFICANCE: The study highlights the importance of incorporating dental health education to psychiatric rehabilitation programs.

A Streamlined Preoperative Surgical Oncology Clinic Workflow Reduces Patient Burden.

BACKGROUND: Although many institutions have focused on improving patient-centered care, little is known about how preoperative workflows affect patients. We hypothesized that a streamlined clinic workflow is associated with decreased cost and time burden on patients. METHODS: A retrospective chart review was performed on surgical oncology patients within thoracic and hepatopancreaticobiliary (HPB) clinics in a tertiary care center from January to December 2016. The clinics varied in scheduling practices, with the thoracic clinic focused on minimizing patient visits. Data collected included number of visits and phone calls made to clinic. Distance traveled, travel cost, and time burden were estimated. RESULTS: We compared 70 esophageal and 60 HPB cancer patients. Thoracic surgery patients required significantly fewer preoperative appointments compared with HPB patients (2.4 versus 4.0; P < 0.00001). About 45 of 60 HPB patients had an extra laboratory work visit, whereas the thoracic clinic incorporated this into clinic visits. The mean distance traveled by patients in the thoracic versus HPB clinic was not significantly different (105.9 versus 93.5 miles; P = 0.44); however, the total cost burden was significantly lower for thoracic patients than HPB patients ($44.0 versus $73.6; P = 0.0029). There was a significant reduction in time burden for patients in the thoracic versus HPB clinic (11.3 versus 18.5 h; P < 0.00001). CONCLUSIONS: This study suggests that a more streamlined preoperative workflow can significantly reduce travel and time burden for patients. The true burden is likely far greater, given potential lost wages and unnecessary stress. Preoperative workflow examination is a promising target for future quality improvement and patient-centered care efforts.

Prevalence and Clinical Impact of Atrial Fibrillation in Patients with Pulmonary Embolism.

OBJECTIVES: Atrial fibrillation (AF) has been suggested as a cause for pulmonary embolism (PE). We aimed to explore the prevalence and clinical impact of AF in patients with PE. METHODS: Using the 2012-2014 National (Nationwide) Inpatient Sample database, we identified "adult patients with PE" as the principal discharge diagnosis. The identified admissions were stratified into two cohorts based on the presence or absence of AF. We used multivariable regression models to evaluate in-hospital mortality, length of stay, nonhome discharge, and in-hospital complications. RESULTS: The prevalence of AF among the 201,360 patients with PE was 11.62%. Patients with AF were more likely to have massive PE (odds ratio 1.59, 95% confidence interval 1.4-1.81, P < 0.001), with higher mortality (adjusted odds ratio 1.48, 95% confidence interval 1.27-1.71, P < 0.001) and a greater risk of mechanical ventilation, cardiac arrest, and nonhome discharges. The length of hospital stay in patients with PE and comorbid AF was significantly longer than those without (6.24 ± 0.10 vs 4.79 ± 0.03 days). CONCLUSIONS: AF is associated with a higher rate of massive PE, higher in-hospital mortality, a longer length of hospital stay, and a higher incidence of in-hospital complications and nonhome discharge.

Safety of Intravenous Thrombolysis for Acute Ischemic Stroke in Patients with Thrombocytopenia.

BACKGROUND: Intravenous thrombolysis with recombinant tissue plasminogen activator (rtPA) is an effective treatment of acute ischemic stroke (AIS). The safety of intravenous rtPA in patients with thrombocytopenia is unclear. This study sought to evaluate the impact of thrombocytopenia on in-hospital outcomes in patients with AIS who received intravenous thrombolysis. METHODS: This was a retrospective study using the 2012-2014 National Inpatient Sample (20% stratified sample of US hospitals). The study identified adult patients admitted with AIS who received intravenous rtPA during hospitalization. The identified admissions were stratified into 2 cohorts based on the presence or absence of thrombocytopenia. Multilevel, multivariate regression analysis and propensity matching were performed to evaluate in-hospital mortality, length of stay, and in-hospital complications. RESULTS: Of 101,527 patients admitted for AIS and received intravenous rtPA from 2012 to 2014, 3,520 (3.47%) had thrombocytopenia. In-hospital mortality was 10.8 vs. 6.9% in patients with and without thrombocytopenia in original data, p < 0.001. In-hospital length of stay was significantly higher in the thrombocytopenia group (5.9 vs. 8.2 days, p < 0.001). The differences were significant in both the multivariate regression model and the propensity score matching model. Patients with thrombocytopenia also had a statistically higher incidence of intracranial hemorrhage, postprocedural bleeding, blood transfusion, tracheotomy, and mechanical ventilation. CONCLUSION: Thrombocytopenia is associated with higher in-hospital mortality, longer length of stay, a higher incidence of intracranial hemorrhage, postprocedural bleeding, and mechanical ventilation in stroke patients who received intravenous rtPA.

Bone Causing Abdominal Groans.

Gastrointestinal complications secondary to fish bone ingestion are rare, however important to recognize in timely manner to prevent morbidity and mortality. Diagnosis is often challenging in setting of non-specific and variable symptoms and lack of history of fish bone ingestion. Diagnostic imaging particularly computed tomography is crucial for diagnosis. However, emphasis should be given on identifying underlying cause of abdominal complications because fish bone is often missed unless specifically looked for. Identification of fish bone is essential for extraction of the inciting nidus. Emergency physician should be aware of this entity to identify it and triage the patients in timely manner. We describe here cases of sub-capsular liver abscess and acute cholecystitis caused by fish bone ingestion. The fish bone as a cause of these complication was initially missed in emergency.

Molecular Modeling of Cloned Bacillus subtilis Keratinase and Its Insinuation in Psoriasis Treatment Using Docking Studies.

Present study demonstrated the expression of cloned Bacillus subtilis RSE163 keratinase gene and in silico binding affinities of deduced protein with psoriasis topical drugs for systemic absorption and permeation through skin. The ker gene expressed in E. coli showed significantly higher keratinase activity 450 ± 10.43 U representing 1342 bp nucleotides encoding 447 amino acids with molecular weight of 46 kDa. The modeled structure was validated using ramachandran's plot showing 305 residues (84.3%) in most favoured region. Docking studies using extra precision (XP) method of Glide showed optimum binding affinities with the drugs Acitretin (- 39.62 kcal/mol), Clobetasol propionate (- 37.90 kcal/mol), Fluticasone (- 38.53 kcal/mol), Desonide (- 32.23 kcal/mol), Anthralin (- 38.04 kcal/mol), Calcipotreine (- 21.55 kcal/mol) and Mometasone (- 28.40 kcal/mol) in comparison to other psoriasis drugs. The results can further be correlated with in vitro enzymatic experiments using keratinase as an effective drug mediator through skin to serve the unmet need of industries.

Poland Syndrome with Ipsilateral Hypoplasia of Gluteal Muscles and Contralateral Hidradenitis Suppurativa.

BACKGROUND: Poland syndrome (PS) is a rare congenital anomaly associated with absent or hypoplastic pectoralis major muscle and a wide spectrum of ipsilateral thoracic and upper extremity deformities. Hidradenitis suppurativa (HS) is a recurrent inflammatory follicular disease that commonly affects the apocrine-bearing skin and involves follicular occlusion and hyperkeratosis. CASE REPORT: We report a case of a 46-year-old male with a history of chronic recurrent hidradenitis suppurativa who was incidentally found to have a simple type of Poland syndrome with incidental hypoplasia of ipsilateral gluteal muscles. CONCLUSIONS: Not only can PS present with HS, which we describe for the first time, but it is also associated with a wide variability of symptoms such as previously unknown co-existence of gluteal muscles hypoplasia.

A novel translational model of percutaneous fetoscopic endoluminal tracheal occlusion - baboons (Papio spp.).

INTRODUCTION: Percutaneous fetoscopic endoluminal reversible tracheal occlusion (FETO) was developed to prevent the pulmonary complications of fetal congenital diaphragmatic herniation. There is an urgent need to establish the closest to human translational model of FETO in order to improve fetal outcomes and to determine new clinical approaches and applications. MATERIAL AND METHODS: Seven non-human primates underwent two subsequent surgeries: the first, the FETO in the experimental group (n = 3) or sham operation in the control animals (S-FETO, n = 4) at 132-142 days of gestation (dGA); the second, the reversal of occlusion or sham operation at 162 ± 5 dGA. Maternal stress axis, complete blood count, and biochemical parameters were evaluated and newborn tracheal radiography was performed. RESULTS: The average pregnancy duration and neonatal weights in the FETO group did not differ from the animals in the S-FETO group. There was no bleeding or premature fetal membrane rupture during the procedures in any of the baboons. The maximal tracheal width was 7.02 ± 0.6 mm in the FETO versus 5.46 ± 0.6 mm in S-FETO group. DISCUSSION: This is the very first report of a successful FETO model in non-human primates. Similarities to human tracheomegaly were for the first time documented in any model studied.

Hydrolyzing proficiency of keratinases in feather degradation.

The keratinase degrade highly rigid, cross linked structural polypeptides with different efficiency depending on the type of source. Two newly isolated strains of Bacillus subtilis (RSE163 and RSE165; NCBI Accession no JQ887983 and JQ887982) were found to be efficient keratinase producers with unusual catalytic activity result in different morphological changes in degradation pattern of feather, confirmed by their scanned electron micrographs. Maximum keratinolytic activity of both the strains B. subtilis RSE163 and RSE165 were found to be 366 ± 15.79 and 194 ± 7.26 U after 72 h of incubation. While the disulphide reductase activity of RSE163 and RSE165 estimated 0.24 ± 0.05 and 0.15 ± 0.03 U/ml of enzyme after 24 h of incubation. A total of 16 free amino acids of variable concentration were also analyzed in the cell free supernatant of hydrolyzed feather from two strains. Present study demonstrates the action of two different keratinases in feather degradation.