SUMO: In Silico Sequence Assessment Using Multiple Optimization Parameters.

To select the most promising screening hits from antibody and VHH display campaigns for subsequent in-depth profiling and optimization, it is highly desirable to assess and select sequences on properties beyond only their binding signals from the sorting process. In addition, developability risk criteria, sequence diversity, and the anticipated complexity for sequence optimization are relevant attributes for hit selection and optimization. Here, we describe an approach for the in silico developability assessment of antibody and VHH sequences. This method not only allows for ranking and filtering multiple sequences with regard to their predicted developability properties and diversity, but also visualizes relevant sequence and structural features of potentially problematic regions and thereby provides rationales and starting points for multi-parameter sequence optimization.

Selective Wee1 Inhibitors Led to Antitumor Activity In Vitro and Correlated with Myelosuppression.

Wee1 is a tyrosine kinase that is highly expressed in several cancer types. Wee1 inhibition can lead to suppression of tumor cell proliferation and sensitization of cells to the effects of DNA-damaging agents. AZD1775 is a nonselective Wee1 inhibitor for which myelosuppression has been observed as a dose-limiting toxicity. We have applied structure-based drug design (SBDD) to rapidly generate highly selective Wee1 inhibitors that demonstrate better selectivity than AZD1775 against PLK1, which is known to cause myelosuppression (including thrombocytopenia) when inhibited. While selective Wee1 inhibitors described herein still achieved in vitro antitumor efficacy, thrombocytopenia was still observed in vitro.

Tejido adiposo epicárdico en personas que viven con VIH: ¿una ventana hacia el riesgo cardiovascular? / Epicardial adipose tissue in people living with HIV: a window towards cardiovascular risk?

Introducción: En las personas que viven con el virus de la inmunodeficiencia humana (PVVIH) se han descripto desregulaciones metabólicas que podrían vincularse a un mayor riesgo cardiovascular.Objetivo: Evaluar el espesor del tejido adiposo epicárdico (ETAE) y la relación del mismo con parámetros clínicos y bioquímicos de riesgo cardiovascular en adultos que viven con VIH, comparados con controles seronegativos.Materiales y métodos: Observacional, inclusión prospectiva. Se incluyeron PVVIH >18 años y controles seronegativos para VIH, a los cuales se les midió el espesor de TAE en dos ejes por ecocardiograma transtorácico, así como el espesor de íntima media carotídea por ecografía doppler color.Resultados: 75 pacientes, 58,7% del sexo masculino, edad de 36 años (RIQ 22). 50,7% con VIH (CD4+: 512 cél/mm3 RIQ 382; 80% indetectables). IMC de 25,2 kg/m2 (RIQ 5,3) y circunferencia de cintura de 88,5 cm (DS 12,4), sin diferencias. Las PVVIH tuvieron menor HDL, mayor proteína C reactiva, mayor dímero D y mayor glucemia en ayunas. El ETAE fue mayor en las PVVIH (4,05 vs. 3,49 mm p=0,021), y se correlacionó con la edad, glucemia en ayunas y dímero D. En las PVVIH, se correlacionó con insulinemia, índice HOMA2-IR, HDL-c y dímero D. El tratamiento con Efavirenz se asoció a un mayor ETAE.Conclusión: Las PVVIH presentaron mayor inflamación sistémica de bajo grado y un mayor espesor de TAE que los controles sanos, el cual se asoció en este grupo a insulinorresistencia

Introduction: For people living with Human Immunodeficiency Virus (PLHIV), metabolic deregulations have been described, which could be related to a higher cardiovascular risk.Objective: To assess the epicardial adipose tissue thickness (EATT), and the relationship between this value and clinical and biochemical parameters of cardiovascular risk in adults living with HIV, if compared to a healthy control group. Methods: Observational, with prospective inclusion. It included PLHIV >18 years and seronegative controls. All of them had their EAT measured in two axes by transthoracic echocardiogram, as well as the carotid intima-media thickness determined by color doppler ultrasound.Results: 75 patients, 58.7% male, age of 36 years (RIQ 22). 50.7% patients with HIV (CD4+ of 512 cells/mm3; and 80% undetectable). BMI was of 25.2 kg/m2 and waist circumference of 88.5 cm, without between-groups differences. PLHIV had lower HDL, higher C reactive protein, higher D-dimer and higher fasting blood glucose. EATT was higher in PLHIV (4.05 vs 3.49 mm, p=0.021), and this correlated with age, fasting blood glucose and D-dimer. In PLHIV, it correlated with insulinemia, HOMA2-IR index, HDL-c ; and D-dimer. Treatment with Efavirenz was associated with a higher EATT.Conclusion: PLHIV presented increased systemic inflammation of low grade and higher EATT than the seronegative control group. EATT was associated in PLHIV to insulin resistance

First week of life respiratory management and pulmonary ventilation/perfusion matching in infants with bronchopulmonary dysplasia: a retrospective observational study.

OBJECTIVE: To investigate the association between early neonatal respiratory management in infants with bronchopulmonary dysplasia (BPD) and the degree of pulmonary ventilation perfusion-matching (V/Q) at term. METHODS: 30 preterm infants with a diagnosis of BPD who were initially treated with either controlled mechanical ventilation/continuous positive airway pressure (CMV/CPAP) (n = 14) or high-frequency oscillatory ventilation (HFOV) using a high lung-volume strategy (n = 16) were retrospectively included in this study. All infants underwent pulmonary V/Q single photon emission computed tomography at a median postmenstrual age of 37 weeks. RESULTS: Infants treated with HFOV had significantly larger proportion of the lung with matched V/Q as compared to infants treated with CMV/CPAP, median (interquartile range) 60.4% (55.5-66.0%) and 45.8% (37.8-53.1%) respectively (p = 0.01). CONCLUSIONS: In infants who needed mechanical ventilation the first week of life and later developed BPD an association was observed between treatment with a HFOV and better pulmonary V/Q matching at near-term age.

Primary hyperparathyroidism, a machine learning approach to identify multiglandular disease in patients with a single adenoma found at preoperative Sestamibi-SPECT/CT.

Objective: Successful preoperative image localisation of all parathyroid adenomas (PTA) in patients with primary hyperparathyroidism (pHPT) and multiglandular disease (MGD) remains challenging. We investigate whether a machine learning classifier (MLC) could predict the presence of overlooked PTA at preoperative localisation with 99mTc-Sestamibi-SPECT/CT in MGD patients. Design: This study is a retrospective study from a single tertiary referral hospital initially including 349 patients with biochemically confirmed pHPT and cured after surgical parathyroidectomy. Methods: A classification ensemble of decision trees with Bayesian hyperparameter optimisation and five-fold cross-validation was trained with six predictor variables: the preoperative plasma concentrations of parathyroid hormone, total calcium and thyroid-stimulating hormone, the serum concentration of ionised calcium, the 24-h urine calcium and the histopathological weight of the localised PTA at imaging. Two response classes were defined: patients with single-gland disease (SGD) correctly localised at imaging and MGD patients in whom only one PTA was localised on imaging. The data set was split into 70% for training and 30% for testing. The MLC was also tested on a subset of the original data based on CT image-derived PTA weights. Results: The MLC achieved an overall accuracy at validation of 90% with an area under the cross-validation receiver operating characteristic curve of 0.9. On test data, the MLC reached a 72% true-positive prediction rate for MGD patients and a misclassification rate of 6% for SGD patients. Similar results were obtained in the testing set with image-derived PTA weight. Conclusions: Artificial intelligence can aid in identifying patients with MGD for whom 99mTc-Sestamibi-SPECT/CT failed to visualise all PTAs.

A kinase-cGAS cascade to synthesize a therapeutic STING activator.

The introduction of molecular complexity in an atom- and step-efficient manner remains an outstanding goal in modern synthetic chemistry. Artificial biosynthetic pathways are uniquely able to address this challenge by using enzymes to carry out multiple synthetic steps simultaneously or in a one-pot sequence1-3. Conducting biosynthesis ex vivo further broadens its applicability by avoiding cross-talk with cellular metabolism and enabling the redesign of key biosynthetic pathways through the use of non-natural cofactors and synthetic reagents4,5. Here we describe the discovery and construction of an enzymatic cascade to MK-1454, a highly potent stimulator of interferon genes (STING) activator under study as an immuno-oncology therapeutic6,7 (ClinicalTrials.gov study NCT04220866 ). From two non-natural nucleotide monothiophosphates, MK-1454 is assembled diastereoselectively in a one-pot cascade, in which two thiotriphosphate nucleotides are simultaneously generated biocatalytically, followed by coupling and cyclization catalysed by an engineered animal cyclic guanosine-adenosine synthase (cGAS). For the thiotriphosphate synthesis, three kinase enzymes were engineered to develop a non-natural cofactor recycling system in which one thiotriphosphate serves as a cofactor in its own synthesis. This study demonstrates the substantial capacity that currently exists to use biosynthetic approaches to discover and manufacture complex, non-natural molecules.

Pulmonary translocation of ultrafine carbon particles in COPD and IPF patients.

OBJECTIVE: Epidemiological studies indicate association between elevated air pollution and adverse health effects. Several mechanisms have been suggested, including translocation of inhaled ultrafine carbon (UFC) particles into the bloodstream. Previous studies in healthy subjects have shown no significant pulmonary translocation of UFC-particles. This study aimed to assess if UFC-particles translocate from damaged alveolar compartment in subjects suffering from chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF). METHODS: Eleven COPD and nine IPF subjects were exposed to a 100 nm UFC-particle-aerosol labeled with Indium-111. Activity in the body was followed up for 10 days using gamma camera planar-imaging as well as in blood and urine samples. RESULTS: The pulmonary central to periphery activity ratio was significantly higher for COPD as compared to IPF subjects at exposure, 1.8 and 1.4, respectively and remained constant throughout the test period. Ten days after exposure, the estimated median pulmonary translocation of UFC particles was 22.8 and 25.8% for COPD and IPF, respectively. Bound activity was present in blood throughout the test period, peaking at 24-h postinhalation with a median concentration of 5.6 and 8.9 Bq/ml for the COPD and IPF, respectively. Median bound activity excreted in urine (% of inhaled) after 10 days was 1.4% in COPD and 0.7% in IPF. Activity accumulation in liver and spleen could not be demonstrated. CONCLUSIONS: Our results suggest that UFC particles leak through the damaged alveolar barrier to the bloodstream in COPD and IPF patients probably distributing in a wide spectrum of whole-body tissues.

The preoperative localisation of small parathyroid adenomas improves when adding Tc-99m-Sestamibi SPECT to multiphase contrast-enhanced CT.

OBJECTIVES: To investigate the incremental value of Sestamibi SPECT combined with a non-enhanced and contrast-enhanced CT, using SPECT/CT, for the preoperative localisation of small parathyroid adenomas (PTA). METHODS: Retrospectively, 147 patients surgically cured from primary hyperparathyroidism, as verified by biochemistry 6 months postoperatively, were included. All patients had preoperatively undergone a dual time 99mTechnetium-Sestamibi SPECT (S) with multiphase CT including native (N), arterial (A) and venous (V) phases. Independently, two radiologists blinded from both the surgical and the preoperative imaging reports, sequentially performed PTA localisation starting with either [A] or [V], thereafter [A + N] or [V + N] and finally with the complete [A + N + S] or [V + N + S]. PTA localisation was reported for each image-set. The readers results were combined and the diagnostic performance for each image set was determined. Sensitivity was also calculated for the different quartiles of PTA weight distribution. RESULTS: The median adenoma weight was 315 mg. No statistically significant differences in diagnostic performance between arterial and venous based image sets were found. The net effect of adding [N] was to increase specificity. Sestamibi SPECT significantly increased the overall diagnostic accuracy for arterial- and venous-based image sets, p = 0.0008 and p = 0.001, respectively. [A + N + S] was found to have the highest diagnostic performance with 86.5% sensitivity and 94.9% overall accuracy. [A + N + S] was particularly advantageous for locating PTA in the lower weight quartiles. CONCLUSIONS: Native CT-phase and dual time point Sestamibi SPECT increase specificity and sensitivity, respectively. These, in combination with a single contrast-enhanced CT-phase is the most optimal examination protocol for preoperative localisation of PTA using SPECT/CT.

Discovery of novel N-1 substituted pyrazolopyrimidinones as potent, selective PDE2 inhibitors.

A focused SAR study was conducted on a series of N1-substituted pyrazolopyrimidinone PDE2 inhibitors to reveal compounds with excellent potency and selectivity. The series was derived from previously identified internal leads and designed to enhance steric interactions with key amino acids in the PDE2 binding pocket. Compound 26 was identified as a lead compound with excellent PDE2 selectivity and good physicochemical properties.

Multiphase Iodine Contrast-Enhanced SPECT/CT Outperforms Nonenhanced SPECT/CT for Preoperative Localization of Small Parathyroid Adenomas.

PURPOSE: The aim of this study was to assess the value of intravenously contrast-enhanced CT in conjunction with Tc-MIBI SPECT for preoperative localization of parathyroid adenoma. METHODS: One hundred ninety-two patients with primary hyperparathyroidism were enrolled in the study between May 2015 and May 2017. The patients underwent a preoperative "one-stop shop" examination with Tc-MIBI SPECT/CT by using dual time-point (10 and 90 minutes) protocol and both nonenhanced CT and contrast-enhanced CT acquisition in the arterial and venous phase, 35 and 75 seconds, respectively, after contrast medium injection start. For 149 patients, the imaging results could be correlated to those at surgery and histopathology. RESULTS: The median adenoma weight was 330 mg. The addition of contrast-enhanced CT increased the sensitivity from 81.1% to 89.9% (P = 0.003). The specificity of nonenhanced SPECT/CT was similar to contrast-enhanced CT (96.1% vs 97.9%; P = 0.077). For patients with uniglandular disease (n = 140, 94.0%), the sensitivity increased from 86.4% to 93.6% (P = 0.021) and the specificity from 96.2% to 97.9% (P = 0.118) by adding contrast-enhanced CT. In patients with multiglandular disease (n = 9, 6.0%), adding contrast-enhanced CT improved detection sensitivity from 42.1% to 63.2%. However, these patients were few and significance was not reached (P = 0.125). CONCLUSIONS: In this cohort, with generally small parathyroid adenomas, the sensitivity in preoperative localization was greatly improved by adding contrast-enhanced CT to Tc-MIBI SPECT/CT.

Comparison of acquisition protocols for ventilation/perfusion SPECT-a Monte Carlo study.

One of the most commonly used imaging techniques for diagnosing pulmonary embolism (PE) is ventilation/perfusion (V/P) scintigraphy. The aim of this study was to evaluate the performance of the currently used imaging protocols for V/P single photon emission computed tomography (V/P SPECT) at two nuclear medicine department sites and to investigate the effect of altering important protocol parameters. The Monte Carlo technique was used to simulate 4D digital phantoms with perfusion defects. Six imaging protocols were included in the study and a total of 72 digital patients were simulated. Six dually trained radiologists/nuclear medicine physicians reviewed the images and reported all perfusion mismatch findings. The radiologists also visually graded the image quality. No statistically significant differences in diagnostic performance were found between the studied protocols, but visual grading analysis pointed out one protocol as significantly superior to four of the other protocols. Considering the study results, we have decided to harmonize our clinical protocols for imaging patients with suspected PE. The administered Technegas and macro aggregated albumin activities have been altered, a low energy all purpose collimator is used instead of a low energy high resolution collimator and the acquisition times have been lowered.

Pharmaceutical intervention after evaluation of the risk of ictus in elderly patients institutionalized with atrial fibrillation.

OBJECTIVES: To evaluate the risk of having a stroke and the risk of bleeding in institutionalized patients with atrial fibrillation. Atrial fibrillation is a common cardiac arrhythmia associated with increased morbidity and mortality. It is necessary to develop pharmacotherapy plans to minimize the risk. DESIGN: A prospective study. SETTING: Institutionalized patients. PARTICIPANTS: Inclusion criteria were: patients diagnosed with atrial fibrillation, with or without treatment for the prevention of stroke. MAIN OUTCOME MEASURES: The evaluation of the CHA2DS2-VASc criteria was performed. The risk of hemorrhage was assessed using the HAS-BLED scale, based on the risk factors associated with the probability of bleeding. RESULTS: We included 53 patients (86.4 ± 6.4 years, 30.2% men). Of these, 37 (69.8%) were correctly anticoagulated. Of the remaining, 5 patients (31.2%) did not have any type of anticoagulant or antiaggregant treatment and 11 (68.7%) were treated with antiaggregant alone. The pharmaceutical intervention was performed in patients who did not meet stroke criteria. Of the untreated patients: two died before the intervention, two were recommended to be referred to cardiology and in one there was no intervention because of very advanced age. In the antiaggregant patients, it was decided not to modify the treatment. The reasons were: high risk of bleeding, very advanced age, advanced dementia or terminal illness, moderate risk of stroke, and clotting factor deficiency. CONCLUSIONS: The risk of stroke in elderly patients with atrial fibrillation is high, so it is important to control the risk factors.

Development of indazole mineralocorticoid receptor antagonists and investigation into their selective late-stage functionalization.

The derivatization of pharmaceuticals is a core activity in the discovery and development of new medicines. Late-stage functionalization via modern CH functionalization chemistry has emerged as a powerful technique with which to diversify advanced pharmaceutical intermediates. We report herein a case study in late-stage functionalization towards the development of a new class of indazole-based mineralocorticoid receptor antagonists (MRA). An effort to modify the electronics of the core indazole heterocycle inspired the use of modern CH borylation chemistry. New reactivity patterns were revealed and studied computationally. Ultimately, a de novo synthesis delivered a key 6-fluoroindazole compound 26, a potent MRA with excellent metabolic stability.

Ten-year-old children with a history of bronchopulmonary dysplasia have regional abnormalities in ventilation perfusion matching.

AIM: The ratio of ventilation to blood flow is an important determinant for regional gas exchange in the lung and hypoxemia is one of the clinical hallmarks in infants with bronchopulmonary dysplasia (BPD). We have previously demonstrated ventilation/perfusion ratio (V/Q) abnormalities in infants with BPD at 36 weeks postconceptional age. The status of V/Q matching in older children with a history of BPD in infancy is unknown. In this study, we examined if 10-year-old children with a history of BPD had V/Q impairments. METHODS: Three-dimensional V/Q-scintigraphy (SPECT) was performed in 26 children. RESULTS: In the BPD group, lung volume with mismatch, (V>Q) was larger compared to areas with reverse mismatch (Q>V), 26.2% and 11.8%, respectively, implying that perfusion defects contribute more than ventilation defects in the V/Q mismatch. Also, the mean fractional distribution of V and Q to V/Q in children with BPD was reduced compared to healthy children, 31% and 51% compared to 64% and 89%, respectively (P < 0.01). CONCLUSION: At 10 years of age children with a history of BPD had ventilation/perfusion abnormalities, with prominent perfusion defects. These V/Q abnormalities suggest the presence of residual alveolar-capillary impairment.

Role of age and comorbidities in mortality of patients with infective endocarditis.

PURPOSE: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. METHODS: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. RESULTS: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32-3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39-1.88),and non-performed surgery (HR:1.64;95% CI:11.16-1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. CONCLUSION: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group.

18F FDG-PET/CT evaluation of histological response after neoadjuvant treatment in patients with cancer of the esophagus or gastroesophageal junction.

BACKGROUND: In most parts of the world, curatively intended treatment for esophageal cancer includes neoadjuvant therapy, either with chemoradiotherapy or chemotherapy alone, followed by esophagectomy. Currently 18F-FDG positron emission tomography/computed tomography (PET/CT) is used for preoperative disease staging, but is not well established in the evaluation of neoadjuvant treatment. PURPOSE: To evaluate changes in PET parameters in relation to the histological primary tumor response in the surgical specimen in patients randomized to neoadjuvant chemoradiotherapy or chemotherapy. MATERIAL AND METHODS: Patients were randomized between either neoadjuvant chemotherapy or chemoradiotherapy followed by esophagectomy.18F-FDG PET/CT exams were conducted at baseline and following neoadjuvant treatment. Standardized uptake ratio (SUR) values were measured in the primary tumor and compared as regards histological responders and non-responders as well as different treatment arms. RESULTS: Seventy-nine patients were enrolled and 51 were available for analysis. A significant rate of SUR reduction was observed ( P = 0.02) in the primary tumor in histological responders compared to non-responders. Changes in SUR were significantly greater in responders following chemoradiotherapy ( P = 0.02), but not following chemotherapy alone ( P = 0.49). There was no statistically significant difference in SUR in patients with a complete histological response compared to those with a subtotal response. CONCLUSION: Our results are similar to those of previous studies and show that changes in the rate of SUR can be used reliably to differentiate histological responders from non-responders after neoadjuvant treatment with either chemoradiotherapy or chemotherapy. Limitations of current PET technology are likely to restrict the possibility of accurately ruling out limited residual disease.

Lung Ventilation/Perfusion Single Photon Emission Computed Tomography (SPECT) in Infants and Children with Nonembolic Chronic Pulmonary Disorders.

Single photon emission computed tomography (SPECT) provides high contrast three dimensional images of the regional distribution of a radiotracer. SPECT is a widely used technique in pulmonary investigations of the ventilation (V) and perfusion (Q) in the adult patient, mainly in the diagnosis of pulmonary embolism. However, safety concerns among practitioners due to radiation exposure and the use of macroaggregate albumin for the perfusion scan have historically precluded the use of SPECT in pediatric patients with nonembolic pulmonary disorders. Additionally, patient cooperation at ventilation tracer administration and image artifacts from patient movements due the long acquisition times, have further limited the application of SPECT in pediatric patients. With the introduction of technegas aerosol for ventilation studies and the use of high sensitive multihead gamma cameras, both the total amount of administered activity and acquisition time have drastically been reduced allowing the application of SPECT in pediatric patients. Modern hybrid gamma camera/computed tomography systems (SPECT/CT) also brings the possibility of adding a fully diagnostic CT to the SPECT images, incrementing the clinical value of the investigation. Besides pulmonary embolism, there is now some clinical evidence that lung SPECT has diagnostic value in several pulmonary pathologies causing V/Q mismatching, which are specific to the pediatric age group. In this work, we will exemplify and briefly discuss some of these applications based on the literature and our routine clinical experience. Consideration to the risks and safety aspects associated to performing pediatric V/Q SPECT are also discussed.

Lung Scintigraphy in the Assessment of Aerosol Deposition and Clearance.

Environmental and occupational exposure to particulate aerosols is known to have negative health effects. However little is known about how these aerosols trigger the development of pathophysiological mechanisms in the body or the fate of ultrafine particles in the lungs after inhalation. The development of aerosols of different origin that can be labeled to a large variety with radionuclides compatible with clinical gamma camera systems opens the possibility of using lung scintigraphy imaging to study these causalities in detail. Lung scintigraphy (planar or SPECT) allows regional mapping of the deposition of the aerosol in the lungs and the dynamic assessment of particle clearance and translocation from the healthy and affected human lungs. In this paper, we will review the unique features of lung scintigraphy applied to aerosol clearance studies in humans.

Role of simple descriptors and applicability domain in predicting change in protein thermostability.

The melting temperature (Tm) of a protein is the temperature at which half of the protein population is in a folded state. Therefore, Tm is a measure of the thermostability of a protein. Increasing the Tm of a protein is a critical goal in biotechnology and biomedicine. However, predicting the change in melting temperature (dTm) due to mutations at a single residue is difficult because it depends on an intricate balance of forces. Existing methods for predicting dTm have had similar levels of success using generally complex models. We find that training a machine learning model with a simple set of easy to calculate physicochemical descriptors describing the local environment of the mutation performed as well as more complicated machine learning models and is 2-6 orders of magnitude faster. Importantly, unlike in most previous publications, we perform a blind prospective test on our simple model by designing 96 variants of a protein not in the training set. Results from retrospective and prospective predictions reveal the limited applicability domain of each model. This study highlights the current deficiencies in the available dTm dataset and is a call to the community to systematically design a larger and more diverse experimental dataset of mutants to prospectively predict dTm with greater certainty.