The SARS‐CoV‐2 pandemic has created an urgent need for diagnostic tests to detect viral RNA. Commercial RNA extraction kits are often expensive, in limited supply, and do not always fully inactivate ...the virus. Together, this calls for the development of safer methods for SARS‐CoV‐2 extraction that utilize readily available reagents and equipment present in most standard laboratories. We optimized and simplified a RNA extraction method combining a high molar acidic guanidinium isothiocyanate (GITC) solution, phenol and chloroform. First, we determined the GITC/RNA dilution thresholds compatible with an efficient two‐step RT‐qPCR for B2M mRNA in nasopharyngeal (NP) or oropharyngeal (OP) swab samples. Second, we optimized a one‐step RT‐qPCR against SARS‐CoV‐2 using NP and OP samples. We furthermore tested a SARS‐CoV‐2 dilution series to determine the detection threshold. The method enables downstream detection of SARS‐CoV‐2 by RT‐qPCR with high sensitivity (~4 viral RNA copies per RT‐qPCR). The protocol is simple, safe, and expands analysis capacity as the inactivated samples can be used in RT‐qPCR detection tests at laboratories not otherwise classified for viral work. The method takes about 30 min from swab to PCR‐ready viral RNA and circumvents the need for commercial RNA purification kits.
Kohlschütter–Tönz syndrome (KTS) is a rare, autosomal recessive syndrome characterized by a triad of epilepsy, amelogenesis imperfecta and severe global developmental delay. It was first described in ...a Swiss family in 1974 by Alfried Kohlschütter and Otmar Tönz. It is caused by pathogenic variants in the ROGDI gene. To the best of our knowledge, there are currently 43 patients with a confirmed ROGDI gene pathogenic variant reported. Here, we review in detail the clinical manifestations of KTS, provide an overview of all reported genetically confirmed patients, and document an additional case of KTS—a 6‐year‐old Latvian girl—with a confirmed ROGDI gene pathogenic variant. In contrast to previous reports, we detected idiopathic bilateral nephrocalcinosis in this newly identified KTS patient. Perampanel proved an effective treatment for our patient with prolonged super‐refractory status epilepticus. In order to better characterize this rare syndrome and its clinical course, it is important to report any additional symptoms and also the effectiveness of used therapies. Future research should focus on elucidating the mechanisms by which the absence/insufficiency of ROGDI‐encoded protein causes the clinical manifestations of KTS. This knowledge could shape possible ways of influencing the disease's natural history with more effective therapies.
Vitamin D insufficiency is associated with poor prognosis in COVID-19 patients. Vitamin D supplementation is related to improved clinical outcomes in terms of intensive care unit admission and death, ...particularly in individuals with moderate-to-severe forms of COVID-19. The placebo-controlled five-day study was performed on 99 hospitalised COVID-19 patients with vitamin D insufficiency randomised into two groups. Vitamin D in the form of a sublingual sprayable microemulsion was given three times daily (daily dose 12,000 IU) to 51 patients with blood 25(OH)D levels below 30 ng/ml. Forty-eight patients in the control group received a placebo spray in the same daily regimen. Intention-to-treat (ITT) analysis and pre-protocol analysis were used to verify the impact of 25(OH)D level elevation on inflammatory markers. There was a statistically significant increase by 8.7 ± 7.6 ng/ml in 25(OH)D level from the baseline level of 15.6 ± 6.5 ng/ml in the case group. Individuals with moderately severe disease showed negative correlation between changes in 25(OH)D and C-reactive protein (CRP) levels in both ITT and pre-protocol analysis (
< 0.05). Mild and severe cases showed no statistical significance in CRP levels. There were no statistically significant changes in ferritin and IL-6 levels in ITT and pre-protocol analysis. In conclusion, high-dose vitamin D therapy was accompanied by significant decrease in CRP levels in COVID-19 patients with a moderate to severe illness.
The complexity of quantum query algorithms computing Boolean functions is strongly related to the degree of the algebraic polynomial representing this Boolean function. There are two related ...difficult open problems. First, Boolean functions are sought for which the complexity of exact quantum query algorithms is essentially less than the complexity of deterministic query algorithms for the same function. Second, Boolean functions are sought for which the degree of the representing polynomial is essentially less than the complexity of deterministic query algorithms. We present in this paper new techniques to solve the second problem.
PDF
