uti

Nucleic acid-based therapy has become an increasingly important strategy for treating a variety of human diseases. In systemic therapy, a therapeutic gene must be delivered efficiently to its target tissues without side effects. To deliver a therapeutic gene such as plasmid DNA (pDNA) or small interfering RNA (siRNA) to target tissues by systemic administration, cationic carriers such as cationic liposomes and polymers have been commonly used as a non-viral vector. However, the binary complex of therapeutic gene and cationic carrier must be stabilized in the blood circulation by avoiding agglutination with blood components, because electrostatic interactions between positively charged complexes and negatively charged erythrocytes can cause agglutination, and the agglutinates contribute to high entrapment of the therapeutic genes in the highly extended lung capillaries. One promising approach for overcoming this problem is modification of the surface of cationic complexes with anionic biodegradable polymers such as hyaluronic acid, chondroitin sulfate, or polyglutamic acid. As another approach, we recently developed a sequential injection method of anionic polymer and cationic liposome/therapeutic gene complex (cationic lipoplex) for delivery of a therapeutic gene into the liver or liver metastasis. In this review, we describe recent advances in the delivery of therapeutic genes by lipid- and polymer-based carrier systems using anionic polymers.

Since the discovery of the dystrophin gene (DMD gene) thirty years ago, several therapeutic approaches have been investigated to treat Duchenne muscular dystrophy (DMD). This includes cell therapy, exon jumping, exonic knockout, and the CinDel method. In this article, we present the challenges of developping a treatment for DMD and the advances of these various approaches. We included the new CRISPR-Cas9 system, which permits not only major progress in the development of new treatments based on genome editing but also the production of new animal models.

Köbberling-Dunnigan syndrome, also known as partial familial lipodystrophy, is a rare genetic disorder characterized by abnormal distribution of adipose tissues. Many people with Köbberling-Dunnigan syndrome develop insulin resistance, a condition in which body tissues cannot adequately respond to insulin hormone. Insulin is a hormone that helps regulate the level of your blood glucose. Köbberling-Dunnigan syndrome can be due to mutations in several different genes. However, type 2 Köbberling-Dunnigan syndrome is caused by the mutation of the LMNA gene, which is located on the long arm of chromosome 1 as 1q22.

The Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome is the most severe form of congenital malformation of the inner female reproductive tract. It is diagnosed as such when the uterus, the upper vagina and optionally the Fallopian tubes are absent. It accounts for approximately 1 in 5000 live-born females and has been classified in two subtypes: type 1 in the presence of isolated uterovaginal aplasia and type 2 when associated in various combinations with extragenital malformations of the kidneys, skeleton, heart and auditory system. Most cases of MRKH syndrome are sporadic, although a significant number of many familial cases have been reported to date. Despite numerous studies, the genetics of the syndrome remains largely unknown and appears to be heterogeneous: chromosomal abnormalities and some candidate gene variants appear to be associated with a few cases; others have been suggested but not yet confirmed. To date, mainly the GREB1L gene appears to be a serious candidate. Among the remaining hypotheses, the controversial contribution of partial duplications of the SHOX gene is still puzzling, as the deficiency of this gene is a major cause of skeletal adysplasia syndromes. We have attempted to resolve this controversy in a study of 60 MRKH cases. Our results tend to show that SHOX duplications can be the origin of a genetic mechanism responsible for MRKH syndrome.

Purpose: The purpose of the study was to examine the health status of rural adolescents and young adults in the United States through a comprehensive review of detailed health information, behavior and health awareness. The disparity in health awareness between rural and non-rural residents compared and evaluated. Methods: Rural-Urban Commuting Area (RUCA) codes were combined with respondent-level data from the Longitudinal Survey of Adolescent to Adult Health (Add Health) to classify individuals as rural/non-rural residents. Health characteristics and perceived health awareness was tested for statistically significant differences using ANOVA. Differences in weight perception accuracy was compared for systematic differences controlling for self-selection into rural areas using a two-stage logistic selection model. Findings: Analysis revealed that rural residents have a higher incidence of major health conditions including epilepsy, high cholesterol, high blood pressure and diabetes. Additionally, they have a higher prevalence of unhealthy behaviors including drinking and drug use. Rural residents are less likely to be insured, but more likely to be overweight or obese. While rural adolescents are more likely to mis-classify their body weight, this misclassification is a result of the higher incidence of overweight rather than the residential location. Conclusion: The higher prevalence of chronic conditions combined with the income and education levels suggests the rural environment is a unique and potentially challenging context for adolescent health. Improving rural adolescent health will require innovative solutions appropriate for rural environments and changes in individual health literacy. Solutions must be multisectoral, engaging education, economic development, and other community perspectives to establish key drivers for health equity.

Background: Corona virus disease is a highly infectious disease caused by the newly innovated corona virus. An emerging respiratory disease was abbreviated as COVID-19, after it has been first reported in December 2019 in Wuhan city of China. Ethiopia Ministry of health initiated multidisciplinary approach to tackle COVID-19 of which awareness creation is the main. The aim of this study is to assess knowledge, practice and associated factors towards prevention of novel corona virus among clients in Debre Tabor general hospital, Northwest Ethiopia, 2020. Methods: Institution based cross sectional study design was conducted in Debre Tabor General hospital from May 15 to May 30, 2020. A structured questionnaire was used for data collection. The data were entered into epi data version 4.4 and exported to SPSS window version 25 for analysis. Binary and multivariable logistic regression was fitted. Odds Ratios with 95% Confidence interval and p - value ≤ 0.05 were considered to assert significance. Result: A total of 345 clients were analyzed and the response rate was 96.4%. The mean age was 32.95 with S.D ± 13.18 years. Majority of the respondents were male (75.7%). Among the study participants 54.2% with (95% CI: [49.0, 59.2%]) and 49.0% with (95% CI: [43.5, 53.4%]) have good knowledge and god practice on COVID-19 preventions respectively. Sex AOR: 4.33 (2.06, 9.09), family size AOR: 2.49 (1.01, 6.15 and heard from social media AOR: 2.78 (1.21, 6.39) were significantly associated with knowledge of respondents. Knowledge AOR: 3.11 (1.59, 6.10) was significantly associated with practice of clients. Residency and those heard from TV were significant variables for both. Conclusion and recommendation: In this study the overall knowledge and preventive practices of the respondents were found to be low. Sex, family size, residency and sources of information were associated factors for knowledge. In addition to this knowledge was significant factor for practice. Health education programs aimed at mobilizing and improving COVID-19 related knowledge and practice intend to be strengthened.

This work aims to evaluate cortical porosity through a high-resolution peripheral quantitative micro-tomography in a group of 47 patients. All patients, in vivo, were subjected to the medical care protocol of the University Hospital Clementino Fraga, 020-213. Patients were women aged from 37 to 82 years old, who did not present fractures in their lower and upper limbs, all of them showing good health. During screening, they were required to have normal BMD (as determined by DXA; T-score ≥ 1.0) and no low-trauma fractures history. The exclusion criteria for all the individuals enrolled in this control study include, for example, alcoholism, chronic drug use, and chronic gastrointestinal disease. Male patients ranging from 42 to 79 years old presented the same health issues as women group. Results showed an increase in the amount of pores on the cortical bone of the evaluated patients over time; however, this increase was also observed in pore diameter, as well as a decrease in the border between the cortical and trabecular bone, indicating a deterioration in cortical bone quality over the years.

Purpose: Sacroiliac joints (SIJ) inflammation and pain is particularly common in patients with Spondyloarthritis. Intraarticular SIJs injections represent a valuable therapeutic option in this condition. In the rheumatological outpatient clinics this procedure is usually done by landmark guidance (LG) or ultrasound guidance (USG). Thus we aimed to compare the short term efficacy of USG vs. LG SIJ injections using five outcome measures: 1. Pain; 2. SIJ status (number of positive provocation tests per symptomatic SIJ on physical examination); 3. Disability; 4. Quality of the night sleep; 5. Patients’ satisfaction. Methods: We enrolled 44 consecutive spondyloarthritis patients with pain in the SIJs that did not respond to NSAIDS and that were otherwise on a stable medical treatment. All patients also had ≥ 3 positive pain provocation tests per SIJ on physical examination. Patients were randomly allocated to receive a single SIJ injection with 7 mg Betamethasone (1 ml) and 1% Lidocaine (1.5 ml) either under USG or with LG. Results: Both groups showed significant improvement in all outcome parameters. However, the USG approach performed significantly better than the LG ones in all parameters. In addition, there was a significant correlation between the improvement in all patient reported outcomes (VAS, RMDQ, JSEQ) and the reduction in the number of positive SIJ pain provocation tests per symptomatic joint. Conclusion: Both USG and LG SIJ injections proved to be an efficient treatment for SIJ pain in SpA patients. However, USG of the intervention led to statistically better results in the present study.

Maternal mortality accounts for most deaths in agrarian communities of Nigeria due to poor access to midwives services and inadequate Skilled Birth Attendants (SBAs). The Midwives Service Scheme (MSS) was established to engage more SBAs and advocate better utilization of pre and post-natal care services. Studies have focused on maternal mortality reduction, however, information on underlying factors that predispose MSS target beneficiaries to its utilization is scarce. Therefore, utilization of MSS among women farmers in southwestern Nigeria was investigated. A four-stage sampling procedure was used. Three states from southwestern states (Oyo, Ogun and Ekiti) were randomly selected. Thereafter, ten Local Government Areas (LGAs) from eighteen LGAs that adopted MSS programme in the selected states were sampled. Also, 30% of the MSS facilities in the sampled LGAs were selected, resulting in 13 MSS facilities. Proportionate sampling technique was used to select 20% of registered women farmers in the selected 13 MSS facilities to give 207 respondents. Interview schedule was used to collect data on respondents’ socioeconomic characteristics, Maternal Health Information Sources (MHIS), Maternal Health Information Seeking Behavior (MHISB) and utilization of MSS. Data were analyzed using descriptive and inferential statistics. About (55.6%) of the respondents had formal education. MHISB and effectiveness of MSS was rated low by 53.2% and 55.6% of the respondents, respectively. MSS was moderately utilized by 64.7% of the respondents. The MSS utilization was 49.24 ± 11.39 (Oyo), 45.08 ± 9.28 (Ogun) and 44.00 ± 10.71 (Ekiti). Respondents’ education (χ2 = 12.85), family size (r = 0.02), monthly income (r = 0.48) related positively and significantly (r = 0.27) to MSS utilization.

In 1992, a patient, born July 10, 1910, aged 82, with major osteoporosis [1-3], was operated on her left hip. The surgeon performs a biomaterial transplant using natural coral [4]. Follow-up is two years. She died in 1994 of acute respiratory failure. Three bone graft osteodensitometry [5] shows a regular increase in mineralization; however, on the opposite side, bone mineralization decreases. The anatomical part is examined using x-rays, scanners, photographs, histology [6]. The article reports the findings of this study. It is noted a partial resorption of the biomaterial essentially at the periphery of the graft as well as the different local connections of the preexisting bone with the newly formed bone from the grafted area. The severity of fractures of the femoral neck is no longer to be demonstrated [7]. Multiple therapeutic trials have demonstrated their effectiveness [8,9]. Twenty two patients were transplanted. There was no failure. All died without fracturing their grafted hips. 

Background: In India, the genetic disease is a disregarded service element in the community health- protection system. This study aims to gauge the accessibility of services for treating genetic disorders and also to evaluate the practices on deterrence and management services in the district health system. Methods: A cross-sectional survey of selected health amenities from 454 medical officers (MO’s), 94 accredited social health activist (ASHAs) workers, 86 multipurpose health assistant-female (MPHA-F), 34 multipurpose health assistant-male (MPHA-M), 14 multipurpose health supervisors-female (MPHS-F), 10 multipurpose health supervisors-male (MPHS-M), 6 multipurpose health extension officer/ community health officer (MPHEO/CHO), 10 public health nurse (PHN), 45 lab technicians (LT’s) working in the government health sector and 254 in the private health sector, 409 nursing staff working in the government health sector and 995 in the private health sector, 15 primary health centers (PHC’s), 4 community health centers (CHC’s), 1 district government hospital (DGH), 3 referral hospitals (RH’s). From the side of private health institutions 25 corporate hospitals (CH’s), 3 medical colleges (MC’s), and 25 diagnostic laboratories (DL’s) were conducted. Results: The findings show that adequate staff was in place at more than 70% of health centers, but none of the staff have obtained any operative training on genetic disease management. The largest part of the DH’s had rudimentary infrastructural and diagnostic facilities. However, the greater part of the CHC’s and PHC’s had inadequate diagnostic facilities related to genetic disease management. Biochemical, molecular, and cytogenetic services were not available at PHC’s and CHC’s. DH’s, RH’s, and all selected medical colleges were found to have offered the basic Biochemical genetics units during the survey. In 24% of CH’s, the basic biochemical units are available and 32% (8 out of 25) of DL’s have the advanced biochemical genetics units by study. Molecular genetics units were found to be available in 28% (7 out of 25) of DL’s during the study. About 6 (24%) diagnostic centers of cytogenetic laboratories were located in the Visakhapatnam district under the private sector. Conclusion: The district health care infrastructure in India has a shortage of basic services to be provided for the genetic disorder. With some policy resolutions and facility strengthening, it is possible to provide advanced services for a genetic disorder in the district health system.

In natural convection, the fluid motion occurs by natural means such as buoyancy. Heat transfer by natural convection happens in many physical problems and engineering applications such as geothermal systems, heat exchangers, petroleum reservoirs and nuclear waste repositories. These problems and phenomena are modeled by ordinary or partial differential equations. In most cases, experimental solutions cannot be applied to these problems, so these equations should be solved using special techniques. In this paper, natural convection of a non-Newtonian fluid flow between two vertical flat plates is investigated analytically and numerically. Collocation Method (CM) and fourth-order Runge -Kutta numerical method (NUM) are used to solve the present problem. These methods are powerful and convenient algorithms in finding the solutions for the equations. While these are capable of reducing the size of calculations. In order to compare with exact solution, velocity and temperature profiles are shown graphically. The obtained results are valid with significant accuracy.

Nanotechnology is a smart technology in the field of biomedical engineering used for the diagnosis and treatment of diseases. Nanodrugs provide better encapsulation of drug and efficiency at low dosage to kill the targeted tissue/cells. However, the chances of chronic toxicity and high cost of treatment limits its applicability [1]. To overcome these problems still, the experts of the scientific community have been working on it, to design the best one and cost-effective treatment for the human welfare.

The present report highlights our results on synthesis of NaYF4:Yb,Er@SiO2@Ag core–shell nanoparticles (CSNPs) for plasmon-enhanced upconversion luminescence (UCL). Hydrophilic surface UCL nanoparticles (UCLNPs) as cores were obtained by precipitation of Rare Earth Elements (REE) chlorides from water-alcohol solutions. The formation of a hydrophobic surface of α-NaYF4:Yb,Er NPs was achieved by thermolysis method at 280 °C and β-NaYF4:Yb,Er by precipitation method in nonpolar medium at 320 °C. Silica shell was formed by the modified Stöber method on the surfaces of UCLNPs with different polarity and phase composition. A mixture of hexane-cyclohexane-isopropyl alcohol was used as a medium for the formation of mononuclear CSNPs on hydrophobic surfaces of cores with different thicknesses of the silica shell: 5 nm and 14 nm. Formation of a predetermined thickness of silica shell was carried out by introducing a precise quantity of TEOS taking into account the size of core NPs with molar ratio TEOS: H2O equal to 1:6. The morphology and phase composition of cores and CSNPs were examined by transmission electron microscopy and selected area electron diffraction, respectively. The insertion of Ag NPs into the structure of NaYF4:Yb,Er@SiO2 was carried out in parallel at the stage of shell formation, which made this synthesis a one-step process. The control of the size of Ag NPs was implemented through the use of a colloidal solution of NPs of the cluster structure by changing the polarity of the medium. The highest intensity enhancement of 85-fold with 5 nm and 29-fold with 14 nm shell thickness was recorded, respectively. For the first time, tests on bioimaging of neutrophil cells by those CSNPs are demonstrated.

The morphological evolution kinetics and instabilities of alpha helical peptide 3.613, which involves large amount of stored torsional elastic deformation energy (3-40 eV/molecule), is formulated by the variational method based on the connection between the rates of internal entropy production and the changes in the global Gibbs free energy, assuming that one has isobaric irreversible processes under the isothermal conditions. The present mesoscopic nonequilibrium thermodynamic approach relies on the fact that the global Gibbs free energy of helical conformation involves not only the bulk Gibbs free energy of the amino-acid back bone structure but also the interfacial Gibbs free energy of the enclosing cylindrical shell or the cage associated with the side-wall molecular branches, and their interactions with the immediate surroundings. The proposed variational analysis applied directly on the proposed macro-model has furnished a nonlinear integral equation in terms of the normalized and scaled internal and external variables. This allows us to track down the motion of the total pitch height of the alpha polypeptide along the well-defined trajectories in the displacement-time space, dictated not only by the initial configuration of the helix but also through the gradients of the global Gibbs free energy of the strained helical conformation as the main driving force. In the negative manifold, there is a well-defined region below the dynamic instability regime, in which the helical conformation can evolve towards the nonequilibrium stationary states by expanding, or contracting, depending upon whether the interfacial free energy and/or the applied stress system are below or above the well-defined thresholds level dictated by the initial pitch height. The highest life time may be realized along that trajectory, which follows up the threshold level of the interfacial specific Gibbs free energy, which is gs = -315 erg/cm2. In the upper region of the negative manifold, the helical conformations are driven by the very large applied uniaxial tension or the negative pressure induced by the thermal expansion, in the range of p > 1GPa and/or the strong negative interfacial free energies [3-4 pH] or their combinations, they show strong kinematic instabilities, which can cause not only the accelerated unfolding phenomenon but also cause large extensions that end up with the catastrophic decimations by ruptures and fragmentations. In the positive manifold, the aging behavior of the polypeptide follows up a S-shape path having rather speedy aging behavior compared to the negative manifold, which is separated from by a well-defined boundary, which represents the isochoric path having longest relaxation times, which can be achieved with great stability. Finally, one could attempt to estimate the upper limit of the relaxation time of aging for the modern hominin, from samples of exceptional preservations, relying on the present nonequilibrium theory as well as on the very limited knowledge on the post-mortem DNA and the present pitch heights of the modern hominin, which is found to be about 25,840 yrs, with a life expectation of 451,800 yrs. These figures are very close to those calculated for Neanderthals (SH), which are found to be 31,820 yrs and 499,100 yrs, respectively.

The global virome: The viruses have a global distribution, phylogenetic diversity and host specificity. They are obligate intracellular parasites with single- or double-stranded DNA or RNA genomes, and afflict bacteria, plants, animals and human population. The viral infection begins when surface proteins bind to receptor proteins on the host cell surface, followed by internalisation, replication and lysis. Further, trans-species interactions of viruses with bacteria, small eukaryotes and host are associated with various zoonotic viral diseases and disease progression. Virome interface and transmission: The cross-species transmission from their natural reservoir, usually mammalian or avian, hosts to infect human-being is a rare probability, but occurs leading to the zoonotic human viral infection. The factors like increased human settlements and encroachments, expanded travel and trade networks, altered wildlife and livestock practices, modernised and mass-farming practices, compromised ecosystems and habitat destruction, and global climate change have impact on the interactions between virome and its hosts and other species and act as drivers of trans-species viral spill-over and human transmission. Zoonotic viral diseases and epidemics: The zoonotic viruses have caused various deadly pandemics in human history. They can be further characterized as either newly emerging or re-emerging infectious diseases, caused by pathogens that historically have infected the same host species, but continue to appear in new locations or in drug-resistant forms, or reappear after apparent control or elimination. The prevalence of zoonoses underlines importance of the animal–human–ecosystem interface in disease transmission. The present COVID-19 infection has certain distinct features which suppress the host immune response and promote the disease potential. Treatment for epidemics like covid-19: It appears that certain nutraceuticals may provide relief in clinical symptoms to patients infected with encapsulated RNA viruses such as influenza and coronavirus. These nutraceuticals appear to reduce the inflammation in the lungs and help to boost type 1 interferon response to these viral infections. The human intestinal microbiota acting in tandem with the host’s defence and immune system, is vital for homeostasis and preservation of health. The integrity and balanced activity of the gut microbes is responsible for the protection from disease states including viral infections. Certain probiotics may help in improving the sensitivity and effectivity of immune system against viral infections. Currently, antiviral therapy is available only for a limited number of zoonotic viral infections. Because viruses are intracellular parasites, antiviral drugs are not able to deactivate or destroy the virus but can reduce the viral load by inhibiting replication and facilitating the host’s innate immune mechanisms to neutralize the virus. Conclusion: Lessons from recent viral epidemics - Considering that certain nutraceuticals have demonstrated antiviral effects in both clinical and animal studies, further studies are required to establish their therapeutic efficacy. The components of nutraceuticals such as luteolin, apigenin, quercetin and chlorogenic acid may be useful for developing a combo-therapy. The use of probiotics to enhance immunity and immune response against viral infections is a novel possibility. The available antiviral therapy is inefficient in deactivating or destroying the infecting viruses, may help in reducing the viral load by inhibiting replication. The novel efficient antiviral agents are being explored.

Due to the urgent need for water in all parts of industrial or developing societies, water supply, and transmission facilities are suitable targets for biological risks. Given that even a short interruption in water supply and water supply operations has a great impact on daily activities in the community, the deliberate contamination of urban water resources has irreparable consequences in the field of public health, and the economy of society will follow. Unfortunately, most officials in the public health control departments in our country have received limited training in detecting accidental or intentional contamination of water resources and dealing with the spread of waterborne diseases both naturally and intentionally. For this reason, there is low preparedness in the responsible agencies to deal with waterborne diseases during biological risks. In the first step of this research, a review study has been conducted on water biological risks and operational strategies to deal with them. In the following, it has studied how Escherichia coli (E. coli) bacteria spread in aqueous media. In this regard, the kinetic model of the studied microorganism was analyzed based on the implementation of (Fick Law) in polar coordinates and the combination of (Dirac Distribution) with (Legendre polynomial) distribution. Finally, after studying the factors affecting the microbial pollutant emission coefficient, the effects of all three factors of linear velocity, linear motion time period, and angle of motion on the pollutant emission flux and biofilm diffusion time in the water supply network environment were investigated. Studies have shown that the linear velocity parameter of Escherichia coli with a nonlinear relationship has the greatest effects on the release of microbial contaminants.

Background: This study aims to retrospectively investigate the comorbidity of ADHD multiple symptoms (behavioral) with alcohol addiction in a sample of adult alcohol-dependent patients and to test their current attentional skills (behavioral and cognitive). Methods: Thirty-two adult alcohol-dependent patients were examined for ADHD using a semi-structured interview and the Mini Mental State Examination to evaluate attention and inhibition functions. Brown ADD Scales were used to specifically examine ADHD syndrome. Patients were compared with thirty matched control participants selected from healthy population in few measures of attentional control and working memory. Results: 50% of patients showed evidence of primary ADHD symptoms: specifically, 28.12% showed criteria for ADHD highly probable, 12.50% for ADHD probable but not certain and 9.38% for ADHD possible but not likely. Patients also revealed several deficits in the selective visual attention, interference control and verbal working memory compared to the control group. Conclusions: These results revealed that adult alcohol-dependent patients had retrospectively high comorbidity with ADHD and significant current deficits of the executive functions. These findings suggest the importance of early diagnosis and treatment of ADHD in order to prevent the development of alcohol dependence.

The broad spectrum of heterozygous versus homozygous JAK2V617F mutated MPN consists ET, ET with early features of PV (prodromal PV), classical PV, masked PV, advanced PV and post-PV myelofibrosis. Combined use of bone marrow histology and increased erythrocyte counts above 5.8x1012/L can replace increased red cell mass at time of presentation as the pathognomonic clue for the correct diagnosis of hetero/homozygous or homozygous mutated PV. Erythrocyte counts are in the normal range below 5.8x1012/L in heterozygous JAK2V617F mutated ET and prodromal PV but above 5.8x1012/L in heterozygous-homozygous or homozygous mutated PV. The bone marrow cellularity and morphology in pre-fibrotic ET, prodromal PV and PV carrying the JAK2V617F mutation are overlapping showing clustered increase of large mature pleomorphic megakaryocytes (M) with no increase of cellularity (<60%) in ET. The bone marrow is hypercellular (60%-80%) due to increased erythropoiesis megakaryopoiesis (EM) in prodromal and classical PV and trilinear hypercellular (80%-100% due increased megakaryopoiesis, erythropoiesis and granulopoiesis (EMG) in advanced PV and masked PV. Bone marrow cellularity ranging from normal (<60%) in ET to increased erythropoiesis (EM) in prodromal PV to hypercellular (80-100%) in advanced PV and masked PV largely depends on increasing JAK2V617F mutation load from low to high on top of other biological MPN variables like constitutional symptoms during long-term follow-up. MPL515 mutated ET is featured by an increase of clustered small and giant megakaryocytes with hyper-lobulated staghorn-like nuclei in a normal cellular bone marrow. The third entity of pronounced JAK2/MPL wild type ET associated with primary megakaryocytic granulocytic myeloproliferation (PMGM) without PV features proved to be caused by calreticulin (CALR) mutation. CALR mutated thrombocythemia is characterized by dual proliferation of megakaryocytic and granulocytic bone marrow proliferation of dense clustered large to giant immature dysmorphic megakaryocytes with bulky (bulbous) hyperchromatic nuclei, which are not seen in MPL515-mutated Thrombocythemia and JAK2V617F-Thrombocythemia, prodromal PV and classical PV. 

Primary myelofibrosis (PMF) is a distinct clinicopathological myeloproliferatve disease (MPD) not preceded by any other MPD ET, PV, CML,... Combined use of bone marrow histology and increased erythrocyte counts above 5.8x1012/L can replace increased red cell mass at time of presentation as the pathognomonic clue for the correct diagnosis of hetero/homozygous or homozygous mutated PV. Erythrocyte counts are in the normal range below 5.8x1012/L in heterozygous JAK2V617F mutated ET and prodromal PV but above 5.8x1012/L in heterozygous-homozygous or homozygous mutated PV. The bone marrow cellularity and morphology in pre-fibrotic ET, prodromal PV and PV carrying the JAK2V617F mutation are overlapping showing clustered increase of large mature pleomorphic megakaryocytes (M) with no increase of cellularity (<60%) in ET. The bone marrow is hypercellular (60%-80%) due to increased erythropoiesis megakaryopoiesis (EM) in prodromal and classical PV and trilinear hypercellular (80%-100% due increased megakaryopoiesis, erythropoiesis and granulopoiesis (EMG) in advanced PV and masked PV. Bone marrow cellularity ranging from normal (<60%) in ET to increased erythropoiesis (EM) in prodromal PV to hypercellular (80-100%) in advanced PV and masked PV largely depends on increasing JAK2V617F mutation load from low to high on top of other biological MPN variables like constitutional symptoms during long-term follow-up. MPL515 mutated ET is featured by an increase of clustered small and giant megakaryocytes with hyper-lobulated staghorn-like nuclei in a normal cellular bone marrow. The third entity of pronounced JAK2/MPL wild type ET associated with primary megakaryocytic granulocytic myeloproliferation (PMGM) without PV features proved to be caused by calreticulin (CALR) mutation. CALR mutated thrombocythemia is characterized by dual proliferation of megakaryocytic and granulocytic bone marrow proliferation of dense clustered large to giant immature dysmorphic megakaryocytes with bulky (bulbous) hyperchromatic nuclei, which are not seen in MPL515-mutated Thrombocythemia and JAK2V617F-Thrombocythemia, prodromal PV and classical PV.