Guiding Transient Peptide Assemblies with Structural Elements Embedded in Abiotic Phosphate Fuels.

Despite great progress in the construction of non-equilibrium systems, most approaches do not consider the structure of the fuel as a critical element to control the processes. Herein, we show that the amino acid side chains (A, F, Nal) in the structure of abiotic phosphates can direct assembly and reactivity during transient structure formation. The fuels bind covalently to substrates and subsequently influence the structures in the assembly process. We focus on the ways in which the phosphate esters guide structure formation and how structures and reactivity cross regulate when constructing assemblies. Through the chemical functionalization of energy-rich aminoacyl phosphate esters, we are able to control the yield of esters and thioesters upon adding dipeptides containing tyrosine or cysteine residues. The structural elements around the phosphate esters guide the lifetime of the structures formed and their supramolecular assemblies. These properties can be further influenced by the peptide sequence of substrates, incorporating anionic, aliphatic and aromatic residues. Furthermore, we illustrate that oligomerization of esters can be initiated from a single aminoacyl phosphate ester incorporating a tyrosine residue (Y). These findings suggest that activated amino acids with varying reactivity and energy contents can pave the way for designing and fabricating structured fuels.

Production of pharmaceutical grade [201Tl]Thallous chloride using 30 MeV cyclotron.

Multiple patient doses of [201Tl]TlCl has been produced using electrodeposited enriched 203Tl in 30 MeV cyclotron (Cyclone-30) with 28 MeV proton energy at 50 µA beam current for 8 h. Ion Exchange Column Chromatography (IECC) and liquid-liquid extraction has been employed for semi-automated radiochemical separation and purification of produced [201Tl]TlCl. The produced [201Tl]TlCl was used in coronary artery disease (CAD) patients.

Benzimidazole and piperidine containing novel 1,2,3-triazole hybrids as anti-infective agents: Design, synthesis, in silico and in vitro antimicrobial efficacy.

Cu alkyne-azide cycloaddition was used to easily synthesize a library of novel heterocycles containing benzimidazole and piperidine based 1,2,3-triazole(7a-7l) derivatives. The synthesized analogs were characterized by various spectroscopic techniques like FTIR, 1 H nuclear magnetic resonance (NMR), 13 C NMR, and mass spectrometry. All these novel bioactive compounds (7a-7l) were evaluated for in vitro antibacterial and antifungal efficacy. Compound 7k exhibited appreciable potent activity against Escherichia coli strain. Compounds 7a, 7b, 7f, and 7i showed excellent potent activity against all bacterial strains. Compound 7b, 7c, 7d, and 7g derivatives showed excellent effects when tested in vitro for antifungal activity against various fungal strains. Additionally, a molecular docking investigation revealed that compound 7k has the ability to bind to the active site of the E. coli DNA gyrase subunit protein and form hydrogen bonds with significant amino acid residues Asp73 and Asp49 in the active sites. In a 100 ns molecular dynamics simulation, the E. coli DNA gyrase protein's steady capacity to bind compound 7k was shown by the low measured root mean square deviation, which was an indication of the complex's conformational stability.

Spontaneous and Selective Peptide Elongation in Water Driven by Aminoacyl Phosphate Esters and Phase Changes.

Nature chose phosphates to activate amino acids, where reactive intermediates and complex machinery drive the construction of polyamides. Outside of biology, the pathways and mechanisms that allow spontaneous and selective peptide elongation in aqueous abiotic systems remain unclear. Herein we work to uncover those pathways by following the systems chemistry of aminoacyl phosphate esters, synthetic counterparts of aminoacyl adenylates. The phosphate esters act as solubility tags, making hydrophobic amino acids and their oligomers soluble in water and enabling selective elongation and different pathways to emerge. Thus, oligomers up to dodecamers were synthesized in one flask and on the minute time scale, where consecutive additions activated autonomous phase changes. Depending on the pathway, the resulting phases initially carry nonpolar peptides and amphiphilic oligomers containing phosphate esters. During elongation and phosphate release, shorter oligomers dominate in solution, while the aggregated phase favors the presence of longer oligomers due to their self-assembly propensity. Furthermore we demonstrated that the solution phases can be isolated and act as a new environment for continuous elongation, by adding various phosphate esters. These findings suggest that the systems chemistry of aminoacyl phosphate esters can activate a selection mechanism for peptide bond formation by merging aqueous synthesis and self-assembly.

Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid.

The U.S. Department of Energy has listed levulinic acid (LA) as one of the top 12 compounds derived from biomass. LA has gained much attention owing to its conversion into enantiopure 4-aminopentanoic acid through an amination reaction. Herein, we developed a coupled-enzyme recyclable cascade employing two transaminases (TAs) for the synthesis of (S)-4-aminopentanoic acid. TAs were first utilized to convert LA into (S)-4-aminopentanoic acid using (S)-α-Methylbenzylamine [(S)-α-MBA] as an amino donor. The deaminated (S)-α-MBA i.e., acetophenone was recycled back using a second TAs while using isopropyl amine (IPA) amino donor to generate easily removable acetone. Enzymatic reactions were carried out using different systems, with conversions ranging from 30% to 80%. Furthermore, the hybrid nanoflowers (HNF) of the fusion protein were constructed which afforded complete biocatalytic conversion of LA to the desired (S)-4-aminopentanoic acid. The created HNF demonstrated storage stability for over a month and can be reused for up to 7 sequential cycles. A preparative scale reaction (100 mL) achieved the complete conversion with an isolated yield of 62%. Furthermore, the applicability of this recycling system was tested with different ß-keto ester substrates, wherein 18%-48% of corresponding ß-amino acids were synthesized. Finally, this recycling system was applied for the biosynthesis of pharmaceutical important drug sitagliptin intermediate ((R)-3-amino-4-(2,4,5-triflurophenyl) butanoic acid) with an excellent conversion 82%.

The efficacy of paravertebral block evaluated by pain-related biomarkers and reactive oxygen species (ROS) following surgery for breast cancer: A randomized controlled study.

Background and Aims: Paravertebral block has been found to be a recommended method of analgesia for breast surgeries. We aimed to assess the efficacy of paravertebral block compared to morphine by means of the visual analogue scale (VAS), pain-related biomarkers, and reactive oxygen species (ROS) in adult patients undergoing breast cancer surgeries under general anesthesia. Methodology: Forty patients undergoing breast surgery with or without lymph outcome were randomly divided into two groups. Group M received general anesthesia with intraoperative injection morphine (0.1 mg/kg) and group P received general anesthesia with paravertebral block with 0.25% bupivacaine (0.3 ml/kg). The primary objective was to compare the postoperative pain using VAS at baseline, 2 hours, 24 hours, and 48 hours. Secondary objectives were to compare the levels of pain-related biomarkers and ROS in both the groups at baseline, 24 hours, and 48 hours and to study the correlation between the VAS and level of pain-related biomarkers and ROS. Results: There was no significant difference in the postoperative VAS scores in both the groups (P = 0.252, 0.548, 0.488 at baseline, 24 hours, and 48 hours, respectively) and no significant difference in the mean biomarkers in both the groups. The mean VAS negatively correlated with biomarker levels at 24 hours and 48 hours, but the change in VAS after 24 hours and 48 hours did not significantly correlate with the change in the biomarkers. Conclusion: Paravertebral block is equally efficacious to intraoperative morphine for breast surgeries for postoperative analgesia. The biomarker levels were not significantly different when patients received paravertebral block or morphine.

A rare case of "Brown tumor" of the axis with parathyroid adenoma and tertiary hyperparathyroidism.

"Brown tumors (BTs)" of the spine are benign rare lesions, seen in about 5%-13% of all patients with chronic hyperparathyroidism (HPT). They are not true neoplasms and are also known as osteitis fibrosa cystica or occasionally osteoclastoma. Radiological presentations are often misleading and may mimic other common lesions such as metastasis. A strong clinical suspicion is therefore necessary, especially in the background of chronic kidney disease with HPT and parathyroid adenoma. Surgical spinal fixation in case of instability due to pathological fracture may be required along with excision of the parathyroid adenoma being the treatment of choice, that maybe usually curative and carries a good prognosis. We would like to report one such rare case of BT involving the axis, or C2 vertebra, presenting with neck pain and weakness that was treated surgically. Only a few cases of spinal BTs have been reported so far in the literature. Involvement of cervical vertebrae and in particular C2 is rarer still with the one in this report only being the fourth such case.

Preparation of [68Ga]Ga-Chloride from 68Zn solid target for the synthesis of pharmaceutical grade [68Ga]Ga-PSMA-11 and [68Ga]Ga-DOTA-TATE.

68Ga is produced from enriched zinc-68 target electrodeposited on copper base material which was irradiated with 15 MeV proton energy in 30 MeV cyclotron. A modified semi-automated separation and purification module was used to obtain pharmaceutical grade [68Ga]GaCl3 in 35 ± 5 min. The quality of [68Ga]GaCl3 produced was in accordance with Pharmeuropa 30.4. The [68Ga]GaCl3 was utilized for the formulation of multiple doses of [68Ga]Ga-PSMA-11 and [68Ga]Ga-DOTATATE. The quality of [68Ga]Ga-PSMA-11 and [68Ga]Ga-DOTATATE were also in accordance with Pharmacopeia.

A probabilistic approach to the estimation of radioactive contaminant inventories at a nuclear waste disposal site.

Routine site inspections are often conducted to gather data on radiation contamination on the surface and below ground near nuclear waste disposal areas. These observations are used to calculate total radiation inventory and its spatial delineation. The statistical kriging approach is often used to spatially interpolate contamination data, and it generates predictions at unsampled sites that are then utilized to calculate the contaminated site's radiation inventory. The kriging output, however, creates a point estimate of the inventory that omits the potential uncertainties from other sources. This paper presents a method for assessing the uncertainty of radiation inventories based on the geostatistical conditional simulation method - a simulation methodology that takes into account the observations made at the sampled sites. The radiation inventories' histograms are generated by conducting many conditional simulations of the projection map using a fitted kriging model. A practical implementation of the suggested approach is shown by evaluating total beta inventories and their spatial delineation using groundwater monitoring data at a nuclear waste disposal site.

The Nose that Broke the Hip! Tumor Induced Osteomalacia Secondary to Phosphaturic Mesenchymal Nasal Tumor : A Case Report.

Tumor induced osteomalacia is a rare paraneoplastic syndrome caused by usually small mesenchymal tumors. The tumors are commonly located in the extremities whereas occurrence in the nasal cavity is quite rare. An 18 year old male presented with asymmetric weakness of both lower limbs and difficulty in walking since one year. On evaluation he was found to have open femur head physes and iliac apophyses, low serum Phosphorus, low vitamin D, normal serum calcium, elevated PTH & alkaline phosphatase levels. FGF-23 levels in serum were elevated. DOTA NOC PET CT revealed tumor in the left nasal cavity abutting the nasal septum. Surgical excision of the tumor resulted in normalisation of the serum levels of phosphorus, vitamin D and FGF-23. This case report indicates the importance of maintaining high clinical suspicion and early detection of causative tumor using appropriate imaging techniques.

Study of Operative Outcomes of Hernioplasty Using Mosquito Net Mesh at a Tertiary Hospital.

Background Synthetic mesh is the most efficient way to repair groin hernias. However, many patients in low and middle-income countries cannot afford the above-mentioned mesh. This study's goal was to describe the mosquito net mesh's (MNM) safety, viability, complications, and recurrence rate in hernioplasty done in rural populations. Material and methods This study was a single-centre, prospective, descriptive study, conducted in patients >18 years of age, of either gender, with unilateral or bilateral primary inguinal hernias (reducible/irreducible/bilateral hernia), admitted for elective or emergency open inguinal hernia mesh repair. The surgery was performed using reasonably priced (affordable to people of low socioeconomic status) polyethylene mosquito net mesh, which was cut into 8x16 cm rectangles and wrapped in two sealed plastic bags. They were sterilised using the ethylene oxide sterilisation (EtO) method. The type of hernia (unilateral or bilateral), post-operative pain, and complications were noted. Results Among 400 study participants, the incidence of inguinal hernia was highest in the 60-79 year age group (51.5%). Direct inguinal hernia (67.5 %) was higher compared to indirect inguinal hernia (32.5 %). While a majority of the participants had a hernia on the right side (50%), 164 patients (41%) had a hernia on the left side, and 36 patients (9%) had a bilateral inguinal hernia. The average operative time for unilateral inguinal hernia was 73.96 minutes and for bilateral inguinal hernia was 106.66 minutes. Out of 400 patients, 355 patients (88.75%) had no complications. Among the postoperative patients, 14 patients (3.5%) experienced surgical site infection; 9 patients (2.25%) experienced headache; 5 patients (1.25%) experienced hematoma; 12 patients (3%) experienced urinary retention; and 5 patients (1.25%) experienced testicular pain. The average hospital stay of patients was 4.25 days. Patients returned to their daily activities and employment in an average of 7.29 days. The market cost of a single standard polypropylene mesh was Rs.1,660/-. For making a single mosquito net mesh of the same size, we had an expenditure of Rs. 11.83/- including the cost of sterilization. In this study, 382 patients (95.5%) gave us good feedback, 11 patients (2.75) gave satisfactory feedback, and 7 patients (1.75%) gave excellent feedback. Conclusion In environments with limited resources, using mosquito net mesh for hernioplasty is reasonable, acceptable, doable, and economical.

Prevalence, distribution and correlates of pain in patients with mucormycosis-A cross-sectional study.

Context: COVID-19 has led to a spate of rhino-orbital-cerebral mucormycosis cases in India, the epidemiology of which was least understood before. Only a few case series and case reports discuss the symptomatology of mucormycosis. Aims: The primary objective of our study was to estimate the prevalence of pain in patients with mucormycosis. The secondary objectives include the type, regional distribution, characteristics and determinants of pain in patients with mucormycosis. Settings and Design: A cross-sectional study was conducted on consecutive adult patients with mucormycosis in our hospital. Materials and Methods: Following recruitment, a preplanned written questionnaire that was tested for validity with peers, with closed-ended queries was filled on a sole visit by an anesthesia postgraduate based on the response by the patient. Statistical analysis used: Categorical variables were summarized as proportion and percentage. To compare quantitative variables, Chi-square test was used. Results: A total of 69 out of the 80 patients recruited complained of pain (P value = 0.468). A total of 76.8% of patients had pain in the supraorbital region with 84% of the patients complaining of throbbing pain and 98.6% needing analgesics. Gender, pre-existing diabetes mellitus, organs affected due to mucormycosis, prior steroid usage, prior COVID illness, surgical intervention, and previous experience of pain in the same region had no influence on the presence or severity of pain. Conclusions: Pain is one of the presenting symptoms in patients with mucormycosis which is usually in the supraorbital and maxillary region, nociceptive type and throbbing in nature, and moderate to severe in intensity usually managed with simple analgesics.

Esophageal Perforation After Anterior Cervical Surgery: A Case Report and Literature Review.

SUMMARY AND BACKGROUND: Esophageal perforation (EP) after anterior cervical surgery is a rare but potentially life-threatening condition. EP caused by malpositioned implants in cervical spine injury with multiple comorbidities is challenging to treat simultaneously. STUDY: This was a case report study. PURPOSE OF STUDY: The aim of this study was to present successful treatment of EP in a subluxated C5-C6 level with implant failure, infection, septicemia, and comorbidities. The aim was to emphasize the need for a multispecialty approach while treating serious complications. CASE: A 72-year-old woman presented to the ER with a history of operated cervical spine a week ago and having breathlessness, fever, wound infection, and tracheostomy in situ. After primary investigations, the patient was initially treated in the intensive care unit, where bleeding from the tracheostomy site was noticed. Upon endoscopy, EP was diagnosed due to implant failure. She was operated for revision cervical spine surgery (drainage of pus with anterior and posterior cervical fixation) and percutaneous endoscopic gastrostomy tube insertion (esophageal diversion). On exploration of EP, a decision was made to perform conservative treatment as initial tag sutures did not hold due to infection. Postoperatively, the patient developed rectal bleed 3 times, which was ultimately treated with cecal bleed embolization. The infected cervical wound was managed with an open dressing. The patient was managed with intermittent assisted ventilation through tracheostomy postoperatively. Barium swallow at 10 weeks confirmed healing of EP and oral feed was started. Tracheostomy closure was performed once the wound had healed, and the patient was discharged with improved neurology at 12 weeks. CONCLUSIONS: Perioperative problems after cervical surgery such as breathing difficulty, wound discharge, and worsening of neurology may lead to suspicion of underlying EP due to implant failure. Upper gastrointestinal endoscopy needs to be considered for a prompt diagnosis. Revision spine surgery with treatment of perforation simultaneously and maintenance of enteral nutrition through a percutaneous endoscopic gastrostomy tube with a multispecialty approach is recommended for this potentially life-threatening condition.

Clinical doses production of pharmaceutical grade Sodium[18F]Fluoride using modified integrated fluidic processor in a SYNTHERA® module.

A fully automated large-scale production of sodium [18F]fluoride ([18F]NaF) using SYNTHERA module with a modification in integrated fluidic processor (IFP) is reported. This modified IFP module is used to prepare [18F]NaF with more than 98% non-decay corrected radiochemical yield (RCY) within 5 min with specifications in accordance with United State Pharmacopeia (USP) monograph. The graphical user interface (GUI) is designed to perform the synthesis steps either manually or automatically and give information to the operator during the course of production. The desired clinical results add support to indigenously produced [18F]NaF as a pharmaceutical grade diagnostic radiopharmaceutical.

Addressing Iron and Zinc Micronutrient Malnutrition Through Nutrigenomics in Pearl Millet: Advances and Prospects.

Iron (Fe) and zinc (Zn) micronutrient deficiencies are significant health concerns, particularly among the underprivileged and resource-poor people in the semi-arid tropics globally. Pearl millet is regarded as a climate-smart crop with low water and energy footprints. It thrives well under adverse agro-ecologies such as high temperatures and limited rainfall. Pearl millet is regarded as a nutri-cereal owing to health-promoting traits such as high grain Fe and Zn content, metabolizable energy, high antioxidant and polyphenols, high proportion of slowly digestible starches, dietary fibers, and favorable essential amino acid profile compared to many cereals. Higher genetic variability for grain Fe and Zn content has facilitated considerable progress in mapping and mining QTLs, alleles and genes underlying micronutrient metabolism. This has been made possible by developing efficient genetic and genomic resources in pearl millet over the last decade. These include genetic stocks such as bi-parental RIL mapping populations, association mapping panels, chromosome segment substitution lines (CSSLs) and TILLING populations. On the genomics side, considerable progress has been made in generating genomic markers, such as SSR marker repository development. This was followed by the development of a next-generation sequencing-based genome-wide SNP repository. The circa 1,000 genomes re-sequencing project played a significant role. A high-quality reference genome was made available by re-sequencing of world diversity panel, mapping population parents and hybrid parental lines. This mini-review attempts to provide information on the current developments on mapping Fe and Zn content in pearl millet and future outlook.

Chemical modification of enzymes to improve biocatalytic performance.

Improvement in intrinsic enzymatic features is in many instances a prerequisite for the scalable applicability of many industrially important biocatalysts. To this end, various strategies of chemical modification of enzymes are maturing and now considered as a distinct way to improve biocatalytic properties. Traditional chemical modification methods utilize reactivities of amine, carboxylic, thiol and other side chains originating from canonical amino acids. On the other hand, noncanonical amino acid- mediated 'click' (bioorthogoal) chemistry and dehydroalanine (Dha)-mediated modifications have emerged as an alternate and promising ways to modify enzymes for functional enhancement. This review discusses the applications of various chemical modification tools that have been directed towards the improvement of functional properties and/or stability of diverse array of biocatalysts.

Annulation of Enals with Carbamoylpropiolates via NHC-Catalyzed Enolate Pathway: Access to Functionalized Maleimides/Iso-maleimides and Synthesis of Aspergillus FH-X-213.

Herein we report the N-heterocyclic carbene (NHC)-catalyzed [3 + 2] annulation of α,ß-unsaturated aldehydes with carbamoylpropiolates via an unusual enolate pathway leading to the construction of highly functionalized maleimides or isomaleimides. The electronic effect imposed by the alkyl/aryl group present on the amide nitrogen of carbamoylpropiolates plays a crucial role in the selective formation of these important five-membered heterocyclic building blocks. The developed protocol is mild and tolerates a wide range of substituents on both substrates. The application of this protocol in the synthesis of the antibacterial natural product Aspergillus FH-X-213 has also been demonstrated.

Promoter engineering-mediated Tuning of esterase and transaminase expression for the chemoenzymatic synthesis of sitagliptin phosphate at the kilogram-scale.

Here, we report a bienzymatic cascade to produce ß-amino acids as an intermediate for the synthesis of the leading oral antidiabetic drug, sitagliptin. A whole-cell biotransformation using recombinant Escherichia coli coexpressing a esterase and transaminase were developed, wherein the desired expression level of each enzyme was achieved by promotor engineering. The small-scale reactions (30 ml) performed under optimized conditions at varying amounts of substrate (100-300 mM) resulted in excellent conversions of 82%-95% for the desired product. Finally, a kilogram-scale enzymatic reaction (250 mM substrate, 220 L) was carried out to produce ß-amino acid (229 mM). Sitagliptin phosphate was chemically synthesized from ß-amino acids with 82% yield and > 99% purity.

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet.

The two main strategies for enzyme engineering, directed evolution and rational design, have found widespread applications in improving the intrinsic activities of proteins. Although numerous advances have been achieved using these ground-breaking methods, the limited chemical diversity of the biopolymers, restricted to the 20 canonical amino acids, hampers creation of novel enzymes that Nature has never made thus far. To address this, much research has been devoted to expanding the protein sequence space via chemical modifications and/or incorporation of noncanonical amino acids (ncAAs). This review provides a balanced discussion and critical evaluation of the applications, recent advances, and technical breakthroughs in biocatalysis for three approaches: (i) chemical modification of cAAs, (ii) incorporation of ncAAs, and (iii) chemical modification of incorporated ncAAs. Furthermore, the applications of these approaches and the result on the functional properties and mechanistic study of the enzymes are extensively reviewed. We also discuss the design of artificial enzymes and directed evolution strategies for enzymes with ncAAs incorporated. Finally, we discuss the current challenges and future perspectives for biocatalysis using the expanded amino acid alphabet.