Spinocerebellar Ataxias: Phenotypic Spectrum of PolyQ versus Non-Repeat Expansion Forms.

Spinocerebellar ataxias (SCA) are most frequently due to (CAG)n (coding for polyglutamine, polyQ) expansions and, less so, to expansion of other oligonucleotide repeats (non-polyQ) or other type of variants (non-repeat expansion SCA). In this study we compared polyQ and non-repeat expansion SCA, in a cohort of patients with hereditary ataxia followed at a tertiary hospital. From a prospective study, 88 patients (51 families) with SCA were selected, 74 (40 families) of whom genetically diagnosed. Thirty-eight patients (51.4%, 19 families) were confirmed as having a polyQ (no other repeat-expansions were identified) and 36 (48.6%, 21 families) a non-repeat expansion SCA. Median age-at-onset was 39.5 [30.0-45.5] for polyQ and 7.0 years [1.00-21.50] for non-repeat expansion SCA. PolyQ SCA were associated with cerebellar onset, and non-repeat expansion forms with non-cerebellar onset. Time to diagnosis was longer for non-repeat expansion SCA. The most common polyQ SCA were Machado-Joseph disease (MJD/SCA3) (73.7%) and SCA2 (15.8%); whereas in non-repeat expansion SCA ATX-CACNA1A (14.3%), ATP1A3-related ataxia, ATX-ITPR1, ATX/HSP-KCNA2, and ATX-PRKCG (9.5% each) predominated. Disease duration (up to inclusion) was significantly higher in non-repeat expansion SCA, but the difference in SARA score was not statistically significant. Cerebellar peduncles and pons atrophy were more common in polyQ ataxias, as was axonal neuropathy. SCA had a wide range of genetic etiology, age-at-onset and presentation. Proportion of polyQ and non-repeat expansion SCA was similar; the latter had a higher genetic heterogeneity. While polyQ ataxias were typically linked to cerebellar onset in adulthood, non-repeat expansion forms associated with early onset and non-cerebellar presentations.

Moderately hypofractionated post-operative radiation therapy for breast cancer: preferences amongst radiation oncologists from countries in Latin America and the Caribbean.

Background: The safety and effectiveness of moderately hypofractionated post-operative radiation therapy for breast cancer were demonstrated by several trials. This study aimed to evaluate the current patterns of practice and prescription preference about moderately hypofractionated post-operative radiation therapy to assess possible aspects that affect the decision-making process regarding the use of fractionation in breast cancer patients in Latin America and the Caribbean (LAC). We also aimed to identify factors that can restrain the utilization of moderately hypofractionated post-operative radiation therapy for breast cancer. Materials an methods: Radiation oncologists from LAC were invited to contribute to this study. A 38-question survey was used to evaluate their opinions. Results: A total of 173 radiation oncologists from 13 countries answered the questionnaire. The majority of respondents (84.9%) preferred moderately hypofractionated post-operative radiation therapy as their first choice in cases of whole breast irradiation. Whole breast plus regional nodal irradiation, post-mastectomy (chest wall and regional nodal irradiation) without reconstruction, and post-mastectomy (chest wall and regional node irradiation) with reconstruction hypofractionated post-operative radiation therapy was preferred by 72.2% 71.1%, and 53.7% of respondents, respectively. Breast cancer stage, and flap-based breast reconstruction were the factors associated with absolute contraindications for the use of hypofractionated schedules. Conclusion: Even though moderately hypofractionated post-operative radiation therapy for breast cancer is considered a new standard to the vast majority of the patients, its unrestricted application in clinical practice across LAC still faces reluctance.

Computer-Aided Design/Computer-Aided Manufacturing of Customized Abutment for Rehabilitating a Malpositioned Implant Using Digital Flow: A Case Report.

This study presented a rehabilitation option for malpositioned implants; this involved obtaining their position and inclination through intraoral scanning, and producing a customized abutment with CAD/CAM technology. The patient in this case report presented a root fracture in tooth 21 and was subjected to extraction, implant installation, and immediate provisional prosthesis. The implant was installed with a distal inclination due to anatomical limitations. After osseointegration, an intraoral scanning transfer provided a digital model (file extension .stl), which reproduced the implant's position and inclination. Then, the file was sent so that a customized abutment (CAD/CAM) could be manufactured, promoting the final rehabilitation of the case; this allowed for good hygiene, load distribution in the dynamic interocclusal relationship, and favorable esthetics, whereas many would otherwise recommend implant removal. The result presented lower costs, a shorter time frame, and a lower morbidity for the patient.

Pseudodominance in Friedreich Ataxia-Impact of High Prevalence of Carriers and Intrafamilial Clinical Variation.

Background: Friedreich ataxia (FA) is the most common form of autosomal recessive (AR) ataxia. It is a rare disease, but carriers are frequent (1/100). Pseudodominance in FA has seldomly been reported; it may pose additional challenges for diagnosis. Cases: A family with two consecutive generations affected by FA is described. The proband and two younger siblings had typical FA, characterized by infantile-onset ataxia, hyporeflexia, Babinski sign, cardiomyopathy, and loss of ambulation in the second decade of life. Another female sibling had delayed-onset (>25 years old), with mild cerebellar and sensitive ataxia since her mid-30s. Their father presented very late-onset FA (>40 years old), with sensitive axonal neuropathy. All five patients had biallelic (GAA)n expansion in FXN. The first three had larger expansions (>800 repeats), while the latter two had one shorter expanded allele (~90 repeats). Literature Review: Pseudodominant inheritance has been described in 13 neurological disorders. Seven are movement disorders, of which three were associated with high frequency of carriers (FA, Wilson's disease and PRKN-related parkinsonism). Conclusions: Clinicians should be aware of the possibility of pseudodominance when facing an apparent autosomal dominant pedigree, particularly in disorders with high frequency of carriers and variable expression. Otherwise, genetic diagnoses may be delayed.

Before Direct-Acting Antivirals for Hepatitis C Virus: Evaluation of Core Protein R70Q and L/C91M Substitutions in Chronically Infected Brazilian Patients Unresponsive to IFN and/or RBV.

Although chronic hepatitis C has been effectively treated with direct-acting antivirals (DAAs), the use of conventional therapy with peg-interferon (Peg-IFN) or (predominantly) ribavirin (RBV), remains widespread. R70Q/H and L/C91M amino acid substitutions in the hepatitis C virus (HCV) core protein may modulate responses to IFN and/or RBV, and are associated with cirrhosis, hepatocellular carcinoma (HCC), insulin resistance, and liver steatosis. We evaluated the R70Q/H and L/C91M substitutions, clinical and epidemiological profiles, and risk factors of Brazilian patients chronically infected with HCV subgenotypes 1a and 1b (HCV-GT1a and HCV-GT1b) unresponsive to IFN and/or RBV therapy. Sequencing and pyrosequencing analyses and sociodemographic and clinical predictive variables were used to assess the relationship between R70Q/H and L/C91M substitutions. Leukocyte counts, ALT levels, and ALT/AST ratios were significantly reduced in treated individuals, but more of these patients had advanced fibrosis and cirrhosis. L91M was more prevalent (19.7%), occurring only in HCV-GT1b, followed by R70Q/P (11.5%) and R70P (1.4%). R70Q/P exhibited higher mean AST, ALT, and GGT values, whereas L91M showed higher mean GGT values. Pyrosequencing of the L91M position revealed mutant subpopulations in 43.75% of samples.

Rational design of large flat nitrogen-doped graphene oxide quantum dots with green-luminescence suitable for biomedical applications.

Rational synthesis and simple methodology for the purification of large (35-45 nm in lateral size) and flat (1.0-1.5 nm of height) nitrogen-doped graphene oxide quantum dots (GOQDs) are presented. The methodology allows robust metal-free and acid-free preparation of N-GOQDs with a yield of about 100% and includes hydrothermal treatment of graphene oxide with hydrogen peroxide and ammonia. It was demonstrated that macroscopic impurities can be separated from N-GOQD suspension by their coagulation with 0.9% NaCl solution. Redispersible in water and saline solutions, particles of N-GOQDs were characterized using tip-enhanced Raman spectroscopy (TERS), photoluminescent, XPS, and UV-VIS spectroscopies. The size and morphology of N-GOQDs were studied by dynamic light scattering, AFM, SEM, and TEM. The procedure proposed allows nitrogen-doped GOQDs to be obtained, having 60-51% of carbon, 34-45% of oxygen, and up to 7.2% of nitrogen. The N-GOQD particles obtained in two hours of synthesis contain only pyrrolic defects of the graphene core. The fraction of pyridine moieties grows with the time of synthesis, while the fraction of quaternary nitrogen declines. Application of TERS allows demonstration that the N-GOQDs consist of a graphene core with an average crystallite size of 9 nm and an average distance between nearest defects smaller than 3 nm. The cytotoxicity tests reveal high viability of the monkey epithelial kidney cells Vero in the presence of N-GOQDs in a concentration below 60 mg L-1. The N-GOQDs demonstrate green luminescence with an emission maximum at 505 nm and sedimentation stability in the cell culture medium.

[Vitamin D deficiency/insufficiency on healthy infants receiving standard supplementation]. / Deficiencia e insuficiencia de vitamina D en lactantes sanos recibiendo suplementación estándar.

Vitamin D (VD) is essential for calcium and phosphorus metabolism. Its deficiency can cause rickets. In Chile, newborns receive 400UI/day supplementation from the first day of age until the first year. OBJECTIVE: To describe the VD plasma levels in healthy infants who received supplementation and secondarily to correlate this with seasonality and nutritional status. SUBJECTS AND METHOD: Cross sectional study. Infants on exclusive or mixed breastfeeding, with monthly pediatric checkups recei ving 400 UI VD supplementation were evaluated, measuring VD plasma levels at 6 months of age, weight, and length, and their nutritional status was classified according to the WHO growth referen ces (weight/age and weight/length). The VD cut-off concentration values were < 20 ng/ml, 21- 29 ng/ ml, and ≥ 30 ng/ml considered as deficiency, insufficiency, and sufficiency, respectively. RESULTS: 40 infants were studied, 40% had insufficient levels and 40% presented deficiency. Season and nutritio nal status were variables significantly related to lower VD values (Winter-Spring p = 0.007; at risk of malnutrition p = 0.038). CONCLUSIONS: The population who received supplementation presented a high frequency of VD deficiency and insufficiency which increases during winter and spring and in subjects at risk of malnutrition.

Hospitalizations for pulmonary embolism in Brazil (2008-2019): an ecological and time series study.

OBJECTIVE: To assess the temporal trends of hospitalizations for pulmonary embolism (PE) in Brazil, its regions, and states between 2008 and 2019. METHODS: An ecological and time series study was conducted. Data were obtained from the Hospital Information System (SIH) of the Brazilian Ministry of Health. The inflection point regression model was applied for temporal trend analyses. Trends were classified as increasing, decreasing, or stationary according to the slope of the regression line. The Annual Percent Charge (APC) and the Average Annual Percent Change (AAPC) were calculated considering a confidence interval of 95% and p-value <0.05. Furthermore, spatial distribution maps of epidemiological indicators related to PE in Brazil were elaborated. RESULTS: There was an increasing trend in the hospitalization rate for PE in Brazil, ranging from 2.57 in 2008 to 4.44/100,000 in 2019 (AAPC=5.6%; p<0.001). Total and average hospitalizations costs also showed increasing trend in the country (AAPC=9.2% and 3.0%, respectively). Still, there was a decrease in the in-hospital mortality rate (from 21.21% to 17.11%; AAPC=-1.9%; p<0.001). Similar trends were observed in most regions. The average hospitalization time in Brazil showed a stationary trend. The hospitalization rate has also increased in 18 states (66.67%). Seven states showed a decrease in the mortality rate (25.93%), except for Roraima, which showed an increasing trend. CONCLUSION: Hospitalizations for PE represent a serious public health problem in Brazil and the temporal patterns observed herein demonstrate an increasing trend in all regions and states of the country.

Molecular Characterization of Portuguese Patients with Hereditary Cerebellar Ataxia.

Hereditary cerebellar ataxia (HCA) comprises a clinical and genetic heterogeneous group of neurodegenerative disorders characterized by incoordination of movement, speech, and unsteady gait. In this study, we performed whole-exome sequencing (WES) in 19 families with HCA and presumed autosomal recessive (AR) inheritance, to identify the causal genes. A phenotypic classification was performed, considering the main clinical syndromes: spastic ataxia, ataxia and neuropathy, ataxia and oculomotor apraxia (AOA), ataxia and dystonia, and ataxia with cognitive impairment. The most frequent causal genes were associated with spastic ataxia (SACS and KIF1C) and with ataxia and neuropathy or AOA (PNKP). We also identified three families with autosomal dominant (AD) forms arising from de novo variants in KIF1A, CACNA1A, or ATP1A3, reinforcing the importance of differential diagnosis (AR vs. AD forms) in families with only one affected member. Moreover, 10 novel causal-variants were identified, and the detrimental effect of two splice-site variants confirmed through functional assays. Finally, by reviewing the molecular mechanisms, we speculated that regulation of cytoskeleton function might be impaired in spastic ataxia, whereas DNA repair is clearly associated with AOA. In conclusion, our study provided a genetic diagnosis for HCA families and proposed common molecular pathways underlying cerebellar neurodegeneration.

Porcine circovirus-2 in Africa: Identification of continent-specific clusters and evidence of independent viral introductions from Europe, North America and Asia.

Porcine circovirus-2 (PCV-2) is associated with several disease syndromes in domestic pigs that have a significant impact on global pig production and health. Currently, little is known about the status of PCV-2 in Africa. In this study, a total of 408 archived DNA samples collected from pigs in Burkina Faso, Cameroon, Cape Verde, Ethiopia, the Democratic Republic of the Congo, Mozambique, Nigeria, Senegal, Tanzania and Zambia between 2000 and 2018 were screened by PCR for the presence of PCV-2. Positive amplicons of the gene encoding the viral capsid protein (ORF2) were sequenced to determine the genotypes circulating in each country. Four of the nine currently known genotypes of PCV-2 were identified (i.e. PCV-2a, PCV-2b, PCV-2d and PCV-2 g) with more than one genotype being identified in Burkina Faso, Ethiopia, Nigeria, Mozambique, Senegal and Zambia. Additionally, a phylogeographic analysis which included 38 additional ORF2 gene sequences of PCV-2s previously identified in Mozambique, Namibia and South Africa from 2014 to 2016 and 2019 to 2020 and available in public databases, demonstrated the existence of several African-specific clusters and estimated the approximate time of introduction of PCV-2s into Africa from other continents. This is the first in-depth study of PCV-2 in Africa and it has important implications for pig production at both the small-holder and commercial farm level on the continent.

Hospitalizations for pulmonary embolism in Brazil (2008-2019): an ecological and time series study / Hospitalizações por embolia pulmonar no Brasil (2008-2019): um estudo ecológico e de séries temporais

ABSTRACT Objective To assess the temporal trends of hospitalizations for pulmonary embolism (PE) in Brazil, its regions, and states between 2008 and 2019. Methods An ecological and time series study was conducted. Data were obtained from the Hospital Information System (SIH) of the Brazilian Ministry of Health. The inflection point regression model was applied for temporal trend analyses. Trends were classified as increasing, decreasing, or stationary according to the slope of the regression line. The Annual Percent Charge (APC) and the Average Annual Percent Change (AAPC) were calculated considering a confidence interval of 95% and p-value <0.05. Furthermore, spatial distribution maps of epidemiological indicators related to PE in Brazil were elaborated. Results There was an increasing trend in the hospitalization rate for PE in Brazil, ranging from 2.57 in 2008 to 4.44/100,000 in 2019 (AAPC=5.6%; p<0.001). Total and average hospitalizations costs also showed increasing trend in the country (AAPC=9.2% and 3.0%, respectively). Still, there was a decrease in the in-hospital mortality rate (from 21.21% to 17.11%; AAPC=-1.9%; p<0.001). Similar trends were observed in most regions. The average hospitalization time in Brazil showed a stationary trend. The hospitalization rate has also increased in 18 states (66.67%). Seven states showed a decrease in the mortality rate (25.93%), except for Roraima, which showed an increasing trend. Conclusion Hospitalizations for PE represent a serious public health problem in Brazil and the temporal patterns observed herein demonstrate an increasing trend in all regions and states of the country.

RESUMO Objetivo Avaliar as tendências temporais das hospitalizações por Embolia Pulmonar (EP) no Brasil, assim como suas regiões e estados no período entre 2008 e 2019. Métodos Foi realizado um estudo ecológico e de série temporal. Os dados foram obtidos do Sistema de Informação Hospitalar (SIH) do Ministério da Saúde (MS) do Brasil. O modelo de regressão de pontos de inflexão foi aplicado para análises de tendências temporais. As tendências foram classificadas como crescentes, decrescentes ou estacionárias de acordo com a inclinação da linha de regressão. O percentual de variação anual (APC) e Percentual de Variação Médio do Período (AAPC) foram calculados considerando Intervalo de Confiança de 95% (IC 95%) e significância de 5%. Além disso, foram elaborados mapas de distribuição espacial dos indicadores epidemiológicos relacionados à EP no Brasil. Resultados Houve uma tendência crescente na taxa de hospitalização de EP no Brasil, variando de 2,57 em 2008 a 4,44/100.000 em 2019 (AAPC=5,6%; p<0,001). Os custos totais e médios de hospitalização também mostraram uma tendência crescente no país (AAPC=9,2% e 3,0%, respectivamente). Ainda assim, houve uma diminuição na taxa de mortalidade hospitalar (de 21,21% para 17,11%; AAPC=-1,9%; p<0,001). Tendências similares foram observadas na maioria das regiões. O tempo médio de hospitalização no Brasil mostrou uma tendência estacionária. A taxa de hospitalização também aumentou em 18 estados (66,67%). Sete estados mostraram uma diminuição na taxa de mortalidade (25,93%), exceto Roraima, que mostrou uma tendência crescente. Conclusão As hospitalizações de EP representam um grave problema de saúde pública no Brasil, e os padrões temporais aqui observados demonstraram uma tendência crescente em todas as regiões e estados do país.

Novel KMT2B mutation causes cerebellar ataxia: Expanding the clinical phenotype.

Hereditary cerebellar ataxias comprise a heterogeneous group of neurodegenerative disorders affecting the cerebellum and/or cerebellar pathways. Next-generation sequencing techniques have contributed substantially to the expansion of ataxia-causing genes, including genes classically described in alternative phenotypes. Herein, we describe a patient with adult-onset cerebellar ataxia, minor dystonia, neuropathy, seizure and ophthalmological pathology, who bears a novel variant in KMT2B (NM_014727.2:c.3334 + 1G > A). Bioinformatic analysis suggested this variant completely abolished the splice-site at exon 8/intron 8, which was confirmed through analysis of mRNA extracted from fibroblasts. Exon 8 skipping would ultimately translate as an in-frame deletion at the protein level, corresponding to the loss of 91 aminoacids [p.(Gly1020_Asn1111del)]. So far, KMT2B disease causing variants have been described in patients with dystonia or neurodevelopmental delay, with no reports of a cerebellar predominant phenotype. Our findings highlight the possible role of KMT2B as a gene involved in hereditary cerebellar ataxias.

Rare occurrence of severe blindness and deafness in Friedreich ataxia: a case report.

BACKGROUND: Friedreich ataxia is the most frequent hereditary ataxia worldwide. Subclinical visual and auditory involvement has been recognized in these patients, with co-occurrence of severe blindness and deafness being rare. CASE REPORT: We describe a patient, homozygous for a 873 GAA expansion in the FXN gene, whose first symptoms appeared by the age of 8. At 22 years-old he developed sensorineural deafness, and at 26 visual impairment. Deafness had a progressive course over 11 years, until a stage of extreme severity which hindered communication. Visual acuity had a catastrophic deterioration, with blindness 3 years after visual impairment was first noticed. Audiograms documented progressive sensorineural deafness, most striking for low frequencies. Visual evoked potentials disclosed bilaterally increased P100 latency. He passed away at the age of 41 years old, at a stage of extreme disability, blind and deaf, in addition to the complete phenotype of a patient with Friedreich ataxia of more than 30 years duration. DISCUSSION: Severe vision loss and extreme deafness has been described in very few patients with Friedreich ataxia. Long duration, severe disease and large expanded alleles may account for such an extreme phenotype; nonetheless, the role of factors as modifying genes warrants further investigation in this subset of patients.

Revision of the deep-sea Eulimidae (Gastropoda, Caenogastropoda) from off Northeast Brazil.

The present study reviews the taxonomy of the marine gastropods belonging to the family Eulimidae Philippi, 1853 from the continental slope off Northeast Brazil. The genera Abyssoaclis Barros, Mello, Barros, Lima, Santos, Cabral Padovan, 2003 and Aclis Lovén, 1846 were not treated here. A total of 20 taxa were identified in this region, excluding Eulima hebes Watson, 1883, a species with a doubtful classification in Eulimidae. The species were assigned to the following genera: Costaclis Bartsch, 1947, Eulima Risso, 1826, Fusceulima Laseron, 1955, Melanella Bowdich, 1822, Ophieulima Warén Sibuet, 1981, Sticteulima Laseron, 1955, Thaleia Warén, 1979, and Umbilibalcis Bouchet Warén, 1986. The genera Sticteulima, Ophieulima and Umbilibalcis are reported for the first time in the southwestern Atlantic. Four species represent new records for the southwestern Atlantic. A redescription of the shell morphology is provided for: Costaclis egregia (Dall, 1889b), Melanella doederleini (Brusina, 1886), and Umbilibalcis lata (Dall, 1889b). Additional information to the original description are reported for other species. Melanella sarissa is considered as a synonym of Melanella cinca Dall, 1927. Six new species are described: Eulima cracentis sp. nov., Melanella paraabida sp. nov., M. adiastalta sp. nov., M. anapetes sp. nov., Sticteulima cabrali sp. nov. and S. canopusensis sp. nov. Lectotypes are designated for: Costaclis hyalina, C. egregia, Eulima ephamilla (Watson, 1883), E. psila and E. (?) hebes. Eulima sp. 1, Melanella sp. 1 and Melanella sp. 2 are potential new species, but the scarcity of material precludes a formal description at this moment.

Congenital ataxia due to novel variant in ATP8A2.

Congenital ataxias are a heterogeneous group of disorders characterized by congenital or early-onset ataxia. Here, we describe two siblings with congenital ataxia, who acquired independent gait by age 4 years. After 16 years of follow-up they presented near normal cognition, cerebellar ataxia, mild pyramidal signs, and dystonia. On exome sequencing, a novel homozygous variant (c.1580-18C > G - intron 17) in ATP8A2 was identified. A new acceptor splice site was predicted by bioinformatics tools, and functionally characterized through a minigene assay. Minigene constructs were generated by PCR-amplification of genomic sequences surrounding the variant of interest and cloning into the pCMVdi vector. Altered splicing was evaluated by expressing these constructs in HEK293T cells. The construct with the c.1580-18C > G homozygous variant produced an aberrant transcript, leading to retention of 17 bp of intron 17, by the use of an alternative acceptor splice site, resulting in a premature stop codon by insertion of four amino acids. These results allowed us to establish this as a disease-causing variant and expand ATP8A2-related disorders to include less severe forms of congenital ataxia.

Is there a place for optimizing thoracic radiotherapy in limited-stage small cell lung cancer after twenty years?

Thoracic radiotherapy (TRT) is one of the main treatments in limited-stage small cell lung cancer (LS-SCLC). Hyperfractionated TRT (45 Gy, 1.5 Gy twice daily) has been the standard of care (SOC) since Turrisi and colleagues published the results of their clinical trial in 1999. Two meta-analyses have demonstrated the benefits of concurrent chemotherapy and TRT in terms of intrathoracic disease control at 2 years and 3-year overall survival (OS). The phase 2 trial by Grønberg et al (2016) comparing once-daily hypofractionated TRT to twice-daily hyperfractionated TRT in LS-SCLC found similar outcomes in both groups in terms of response rate, progression-free survival (PFS), grade 3-4 adverse effects, and OS. The CONVERT trial, published in 2017, failed to demonstrate the superiority of the conventional scheme (once-daily TRT) vs twice-daily radiotherapy, despite the application of modern radiotherapy techniques and a quality assurance programme, thus confirming the twice-daily hyperfractionated regimen as the SOC. At the 2020 American Society of Clinical Oncology (ASCO) annual meeting, Grønberg et al reported preliminary findings from a phase 2 trial comparing two different TRT dose regimens (45 Gy vs 60 Gy), both administered twice daily. Those data demonstrated a marked improvement in 2-year survival rates in the high dose arm (70.2% vs 46.1%, P = 0.002), despite similar objective response rates and PFS outcomes. Those findings provide a new treatment alternative to consider: Hyperfractionated, high-dose TRT. However, the results of that trial will need to be validated in a large, randomized phase 3 study. The results of the phase 2 CALCG 30610 trial will help to clarify the optimal dose and regimen. The potential role of upfront immunotherapy, which early data suggest may improve OS, also needs to be determined.