scholarly journals Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction

Author(s):  
Alka Shukla ◽  
Mayank Gangwar ◽  
Gaurav Sharma ◽  
Pradyot Prakash ◽  
Gopal Nath

This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen’s column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component.

Nerves to various muscles of the cat’s hind limb have been stimulated under conditions designed to give uniform distribution of current throughout the whole cross-section of the nerve, and the conducted action potentials have been recorded from the dorsal roots. Alterna­tively, stimuli have been applied to a dorsal root and the conducted action potentials recorded from the nerve to a muscle. Strong stimuli evoke a compound spike potential with a time distribution conforming with the standard fibre-calibre spectrum. Following the large group I spike there is usually a double spike attributable to group II fibres, and a very low group III spike. The stimulus strength has to be increased to about twice threshold in order to set up a maximum group I spike, but a group II spike is usually detectable when the stimulus is increased to 1.3 to 1.4 times group I threshold. Group II b fibres usually have a higher threshold than group II a , and a group III spike is not as a rule detectable until the stimulus is about three times group I threshold. The excitatory and inhibitory actions of an afferent volley from a muscle on the moto-neurones of that muscle or of a synergic muscle have been revealed by the change in mono­ synaptic reflex response to a later testing volley (maximal for group I fibres) either in that same afferent nerve (homosynaptic testing) or in the afferent nerve from a synergic muscle (heterosynaptic testing). The conditioning volley has been set up by stimuli whose strength has varied from threshold for group I fibres to m any times that value, and several standard testing intervals have been chosen. From the data so obtained it has been possible to deter­mine the reflex effects exerted by the different groups of afferent fibres. The group I afferent fibres in the nerve of a flexor or extensor muscle exert the well-known excitatory action on the motoneurones of that muscle (autogenetic excitation). Our methods have failed to reveal any autogenetic inhibitory action, though its existence is not thereby disproven. The group III impulses of extensor muscles have a powerful autogenetic inhibitory action, while the group II impulses either have no detectable action or are weak autogenetic inhibitors. On the contrary, with flexor muscles both group II and group III impulses exert an effective autogenetic excitatory action. In addition, the group I afferent impulses of both extensor and flexor muscles exert on moto­-neurones an autogenetic depressant action which is clearly distinguishable from autogenetic inhibition. It is observed at an interval of more than 10 msec, subsequent to an excitatory action which was too weak to generate a reflex discharge. Since, in contrast to inhibition, this depression is virtually restricted to a homosynaptic testing reflex, it is attributed to the subnormality associated with the positive after-potential of the activated subsynaptic areas of motoneurones, and has been called ‘subsynaptic depression’.


Author(s):  
K.K. SEKHRI ◽  
C.S. ALEXANDER ◽  
H.T. NAGASAWA

C57BL male mice (Jackson Lab., Bar Harbor, Maine) weighing about 18 gms were randomly divided into three groups: group I was fed sweetened liquid alcohol diet (modified Schenkl) in which 36% of the calories were derived from alcohol; group II was maintained on a similar diet but alcohol was isocalorically substituted by sucrose; group III was fed regular mouse chow ad lib for five months. Liver and heart tissues were fixed in 2.5% cacodylate buffered glutaraldehyde, post-fixed in 2% osmium tetroxide and embedded in Epon-araldite.


1998 ◽  
Vol 80 (09) ◽  
pp. 393-398 ◽  
Author(s):  
V. Regnault ◽  
E. Hachulla ◽  
L. Darnige ◽  
B. Roussel ◽  
J. C. Bensa ◽  
...  

SummaryMost anticardiolipin antibodies (ACA) associated with antiphospholipid syndrome (APS) are directed against epitopes expressed on β2-glycoprotein I (β2GPI). Despite a good correlation between standard ACA assays and those using purified human β2GPI as the sole antigen, some sera from APS patients only react in the latter. This is indicative of heterogeneity in anti-β2GPI antibodies. To characterize their reactivity profiles, human and bovine β2GPI were immobilized on γ-irradiated plates (β2GPI-ELISA), plain polystyrene precoated with increasing cardiolipin concentrations (CL/β2GPI-ELISA), and affinity columns. Fluid-phase inhibition experiments were also carried out with both proteins. Of 56 selected sera, restricted recognition of bovine or human β2GPI occurred respectively in 10/29 IgA-positive and 9/22 IgM-positive samples, and most of the latter (8/9) were missed by the standard ACA assay, as expected from a previous study. Based on species specificity and ACA results, IgG-positive samples (53/56) were categorized into three groups: antibodies reactive to bovine β2GPI only (group I) or to bovine and human β2GPI, group II being ACA-negative, and group III being ACA-positive. The most important group, group III (n = 33) was characterized by (i) binding when β2GPI was immobilized on γ-irradiated polystyrene or cardiolipin at sufficient concentration (regardless of β2GPI density, as assessed using 125I-β2GPI); (ii) and low avidity binding to fluid-phase β2GPI (Kd in the range 10–5 M). In contrast, all six group II samples showed (i) ability to bind human and bovine β2GPI immobilized on non-irradiated plates; (ii) concentration-dependent blockade of binding by cardiolipin, suggesting epitope location in the vicinity of the phospholipid binding site on native β2GPI; (iii) and relative avidities approximately 100-fold higher than in group III. Group I patients were heterogeneous with respect to CL/β2GPI-ELISA and ACA results (6/14 scored negative), possibly reflecting antibody differences in terms of avidity and epitope specificity. Affinity fractionation of 23 sera showed the existence, in individual patients, of various combinations of antibody subsets solely reactive to human or bovine β2GPI, together with cross-species reactive subsets present in all samples with dual reactivity namely groups III and II, although the latter antibodies were poorly purified on either column. Therefore, the mode of presentation of β2GPI greatly influences its recognition by anti-β2GPI antibodies with marked inter-individual heterogeneity, in relation to ACA quantitation and, possibly, disease presentation and pathogenesis.


2015 ◽  
Vol 18 (3) ◽  
pp. 098
Author(s):  
Cem Arıtürk ◽  
Serpil Ustalar Özgen ◽  
Behiç Danışan ◽  
Hasan Karabulut ◽  
Fevzi Toraman

<p class="p1"><span class="s1"><strong>Background:</strong> The inspiratory oxygen fraction (FiO<sub>2</sub>) is usually set between 60% and 100% during conventional extracorporeal circulation (ECC). However, this strategy causes partial oxygen pressure (PaO<sub>2</sub>) to reach hyperoxemic levels (&gt;180 mmHg). During anesthetic management of cardiothoracic surgery it is important to keep PaO<sub>2</sub> levels between 80-180 mmHg. The aim of this study was to assess whether adjusting FiO<sub>2</sub> levels in accordance with body temperature and body surface area (BSA) during ECC is an effective method for maintaining normoxemic PaO<sub>2</sub> during cardiac surgery.</span></p><p class="p1"><span class="s1"><strong>Methods:</strong> After approval from the Ethics Committee of the University of Acıbadem, informed consent was given from 60 patients. FiO<sub>2</sub> adjustment strategies applied to the patients in the groups were as follows: FiO<sub>2</sub> levels were set as 0.21 × BSA during hypothermia and 0.21 × BSA + 10 during rewarming in Group I; 0.18 × BSA during hypothermia and 0.18 × BSA + 15 during rewarming in Group II; and 0.18 × BSA during hypothermia and variable with body temperature during rewarming in Group III. Arterial blood gas values and hemodynamic parameters were recorded before ECC (T1); at the 10th minute of cross clamp (T2); when the esophageal temperature (OT) reached 34°C (T3); when OT reached 36°C (T4); and just before the cessation of ECC (T5).</span></p><p class="p1"><span class="s1"><strong>Results:</strong> Mean PaO<sub>2</sub> was significantly higher in Group I than in Group II at T2 and T3 (<em>P</em> = .0001 and <em>P</em> = .0001, respectively); in Group I than in Group III at T1 (<em>P</em> = .02); and in Group II than in Group III at T2, T3, and T4 <br /> (<em>P</em> = .0001 for all). </span></p><p class="p1"><span class="s1"><strong>Conclusion: </strong>Adjustment of FiO<sub>2</sub> according to BSA rather than keeping it at a constant level is more appropriate for keeping PaO<sub>2</sub> between safe level limits. However, since oxygen consumption of cells vary with body temperature, it would be appropriate to set FiO<sub>2</sub> levels in concordance with the body temperature in the <br /> rewarming period.</span></p>


2016 ◽  
pp. 108-111
Author(s):  
T.F. Tatarchuk ◽  
◽  
D.G. German ◽  

The article presents the comparative analysis of the state of the cervix in women with endometrial polyps and micropolyps. Patients and methods. The study involved 130 patients aged 18-35 years: 70 patients with endometrial polyps (group I), 30 patients with micropolyps (group II) and 30 patients of the control group (group III). Results. According to the anamnesis of women in the I group were significantly more frequent diseases of the cervix, which corrected physical surgery methods, in particular cryodestruction. In group II, the representatives of these indicators were similar to healthy. Normal colposcopic picture met significantly less frequently in patients and I, and II group. The differences in the incidence of HPV high oncogenic risk in all groups were not statistically significant. Conclusion. Destructive methods used in the detection of any changes in the cervix are often overly aggressive, form scars and contributing to inflamaciones process. In the chain of events leading to the formation of PE, cervical pathology and its correction can take the basic place. Key words: endometrial polyp, micropolyps, chronic endometritis, uterine cervix, colposcopy.


Author(s):  
Г.В. ШИРЯЕВ ◽  
Ю.Г. ТУРЛОВА ◽  
Г.С. НИКИТИН

Изучено влияние кормовой добавки в форме индивидуальных болюсов Метраболь на возможность снижения негативного влияния субклинического кетоза (СКК) и результативность искусственного осеменения высокопродуктивных молочных коров в посттранзитный период. Животные были подобраны по принципу условных аналогов и разделены на 3 группы в зависимости от концентрации β-оксимасляной кислоты в их крови. В I группе (n=18) у коров не было СКК, концентрация кислоты составляла <1,0 ммоль/л, во II группе (n=21) у животных зарегистрирован СКК, концентрация кислоты — 1,0…1,4 ммоль/л. С целью поиска решения по снижению негативного влияния СКК на репродуктивные показатели была сформирована III группа коров, переболевших СКК (n=24) с концентрацией β-оксимасляной кислоты в крови 1,0…1,4 ммоль/л, но с использованием болюсов, применяемых для ускоренного восстановления репродуктивной функции коров после отела. Важной составляющей опыта явилось то, что к моменту гормональной синхронизации у коров всех 3 групп фиксировалось отсутствие СКК (снижение β-оксимасляной кислоты до уровня <1,0 ммоль/л). Для синхронизации полового цикла выбрана схема Пресинх-овсинх. Установлено, что гормональная синхронизация животных, перенесших СКК, в сравнении со здоровыми, не позволяет снизить негативное влияние данного нарушения обмена веществ. Во II группе у животных зафиксированы самые низкие репродуктивные показатели (количество использованных доз; индекс осеменения, сервис-период; время от первого до успешного осеменения; результат первичного осеменения; количество животных, осемененных 3 и более раз). В III группе применение болюсов позволило, в сравнении с животными II группы, не только избежать отрицательного воздействия СКК, но и превзойти по изучаемым параметрам животных из I группы. The effect of the feed additive in the form of individual boluses Metrabol on the possibility of reducing the negative effect of subclinical ketosis (CCK) and the effectiveness of artificial insemination of highly productive dairy cows in the post-transit period was studied. The animals were selected according to the principle of conditional analogs and divided into 3 groups depending on the concentration of β-hydroxybutyric acid in their blood. In group I (n=18), the cows did not have SSC, the acid concentration was <1.0 mmol/L, in group II (n=21), SSC was registered in animals — 1.0...1.4 mmol / L. In order to find a solution to reduce the negative effect of CCB on reproductive indices, a group III of cows was formed who had CCB (n = 24) with a concentration of β-hydroxybutyric acid in their blood of 1.0...1.4 mmol/L, but using boluses, used to accelerate the restoration of the reproductive function of cows after calving. An important component of the experiment was that by the time of hormonal synchronization, cows of all 3 groups had no SSC (decrease in β-hydroxybutyric acid to a level of <1.0 mmol/L). To synchronize the sexual cycle, the Pressinh-ovsinh scheme was chosen. It was found that hormonal synchronization of animals that underwent SSC, in comparison with healthy ones, does not allow reducing the negative effect of this metabolic disorder. In group II, animals had the lowest reproductive indices (number of doses used; insemination index, service period; time from first to successful insemination; result of primary insemination; number of animals inseminated 3 or more times). In group III, the use of boluses made it possible, in comparison with animals of group II, not only to avoid the negative effect of SSC, but also to surpass the animals from group I in terms of the studied parameters.


Author(s):  
M. M. Chaudhary ◽  
C. T. Khasatiya ◽  
S. B. Patel ◽  
S. S. Chaudhary ◽  
V. B. Atara ◽  
...  

The serum progesterone and estradiol profiles during synchronization of estrus by buck effect and PGF2α treatments were monitored in Surti does. Total eighteen non-pregnant does selected were evenly divided into 3 groups, 6 does in each group. The does of Group I were teased with a sexuallyactive- apronized buck; and those of Group II were treated with PGF2α, i.e., Inj. Lutalyse® @ 7.5 mg/doe IM twice 11 days apart, while the Group III served as untreated control. Blood samples were collected from all the animals on day 0 (before 1st PGF2α injection), 3rd day (during treatment), 11th day (before 2nd PGF2α injection), 14th day (after treatment) and 40th day (post-service) by jugular vein puncture. The serum separated was stored at -20°C till further analysis. In all the three groups, 83.33% does, conceived at first service in the sampling cycle. The overall mean serum progesterone concentration of Group I does (5.82±0.72 ng/ml) was significantly higher (p less than 0.01) as compared to Group II (2.93±0.38 ng/ml) and III (2.88±0.30 ng/ml). Similarly, the overall mean serum progesterone concentration of Surti does on day 0 (2.65±0.46 ng/ml), 3rd (2.56±0.80 ng/ml), 11th (4.45±0.84 ng/ml) and 14th (3.40±0.63 ng/ml) did not differ significantly, but the overall mean level at day 40 (6.31±0.45 ng/ml) was significantly (p less than 0.01) higher, because most of animals became pregnant at that time. The overall mean serum oestradiol-17β levels of Group I (24.40±2.98 pg/ ml) was significantly higher (p less than 0.01) than in Group II (15.77±1.77 pg/ml) and III (12.21±1.45 pg/ ml). On the other hand, the overall mean serum oestradiol-17β levels of Surti does on day 0 (12.89±1.21 pg/ml), 3rd (15.84±1.74 pg/ml), 11th (14.81±1.96 pg/ml), 14th (22.15±2.97 pg/ml) and 40th (21.64±5.16 pg/ml) did not differ significantly (p>0.05) and the slightly higher overall mean level found at 40th day might be the influence of the non-pregnant does at first service in the cumulative animals. The hormonal profile reflected the initiation of cyclicity and establishment of pregnancy in treated and control animals.


2020 ◽  
Author(s):  
Jan Niclas Mumm ◽  
Lucas Bohn ◽  
Lennert Eismann ◽  
Alexander Buchner ◽  
Theresa Vilsmaier ◽  
...  

BACKGROUND Pelvic floor training (PFT) is the gold standard for conservative treatment of male stress urinary incontinence. OBJECTIVE To evaluate patients´ perspective at risk of incontinence on PFT and application of digital technologies for PFT. METHODS Patients undergoing transurethral surgery of the prostate (group I), radical prostatectomy (group II) or treatment at a specialized incontinence outpatient clinic (group III) were surveyed anonymously. Chi-Square test and Kruskal-Wallis-analysis were used for statistical analysis. RESULTS 180 patients were included in the final analysis. In group I (n=35) no patient underwent PFT prior to transurethral surgery. 23.5% of patients in group II (n=51) and 95.7% of patients in group III (n=94) performed PFT. 11.4% in group I, 80.4% in group II and 91.5% in group III have been advised to perform PFT by their urologist. Regarding the information level on PFT, patients from group I (median 1, range 0-5) are less satisfied than patients from group II (median 3, 0-9) or group III (median 5, range 0-10, p<0.001). 88.6% of patients from group I are willing to perform PFT as preventive treatment or to avoid incontinence surgery, 100% from group II and 68.4% from group III (p<0.001). The likelihood to use digital PFT is higher in group I (median: 9, range 0-10) and II (median: 9, range 0-10) than in group III (median: 4, range 0-10, p<0.001). CONCLUSIONS Patients at risk of incontinence currently have limited access to PFT, although they are willing to perform PFT. Digital PFT is highly accepted by patients preoperatively and might be a valuable tool to increase PFT participation.


Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 471
Author(s):  
Jongmin Lee ◽  
Sung-Soo Park ◽  
Tong Yoon Kim ◽  
Dong-Gun Lee ◽  
Dong-Wook Kim

We aimed to identify whether lymphopenia is a reliable prognostic marker for COVID-19. Using data derived from a Korean nationwide longitudinal cohort of 5628 COVID-19 patients, we identified propensity-matched cohorts (n = 770) with group I of severe lymphopenia (absolute lymphocyte counts [ALC]: <500/mm3, n = 110), group II of mild-to-moderate lymphopenia (ALC: ≥500–<1000/mm3, n = 330), and group III, no lymphopenia (ALC: ≥1000/mm3, n = 330). A significantly higher mortality rate was associated with lymphopenia severity: 40% in group I, 22.7% in group II, and 13.0% in group III (p < 0.001). At 28 days, the estimated inferior overall survival associated with intensified lymphopenia: 62.7% in group I, 79.9% in group II, and 89.0% in group III (p < 0.001). Lymphopenia contributed significantly toward a greater need for interventions in all groups but at varying degrees: requirements of invasive ventilation, intensive oxygen supply, or adequate oxygen supply, respectively (p < 0.001). The lymphopenia intensity was independently associated with higher COVID-19 mortality in multivariable analysis; adjusted odds ratios of 5.63 (95% CI, 3.0–10.72), and 2.47 (95% CI, 1.5–4.13) for group I and group II, respectively. Lymphopenia and its severity levels may serve as reliable predictive factors for COVID-19 clinical outcomes; thus, lymphopenia may provide the prognostic granularity required for clinical use in the management of patients with COVID-19.


Author(s):  
Mahir Tıraş ◽  
Emrah Can ◽  
Şahin Hamilçıkan

Objective This study aimed to assess whether cord blood carboxyhemoglobin (COHb) levels in jaundiced term neonates with and without a positive direct Coombs test (DCT) and in healthy controls could be used as a predictor of severe hyperbilirubinemia. The percentage of cord blood COHb should be higher among neonates with Coombs-positive ABO hemolytic disease than among those with Coombs-negative ABO incompatibility and higher than that of ABO-compatible control neonates. Study Design This cross-sectional descriptive study of 198 term neonates comprised three subgroups: group I featured 68 DCT-positive ABO-incompatible neonates (ABO + DCT), group II featured 60 DCT-negative ABO-incompatible neonates with hyperbilirubinemia (ABO–DCT), and group III featured 70 healthy controls. COHb was determined by an OSM3 hemoximeter. Results Group I differed from groups II and III for cord blood bilirubin, cord blood hemoglobin, and cord blood hematocrit. Groups I and II had higher mean total serum bilirubin (TSB) levels than group III, while there was no difference in the mean TSB levels between groups I and II. There was no significant difference between the COHb group means for groups I, II, and III (p = 0.98). The area under the receiver operating characteristic curve calculated for group I/group III and group II/group III were found to be 0.62 and 0.54, respectively. Conclusion COHb levels did not prove to be superior to the DCT for predicting the risk of developing severe hyperbilirubinemia in term neonates. Key Points


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