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The World Health Organisation (WHO) defines anemia as a global public health problem. It is a medical condition in which the number of red blood cells or the hemoglobin concentration within them is below the physiological range. We present a case of a 40-year-old woman with Abnormal Uterine Bleeding (AUB) accompanied by malaise, weakness, and tachycardia. The patient reported heavy menstrual bleeding for the past 14 days. Speculum examination revealed that the bleeding was from the uterus. There were no pathological findings during a gynecological and transvaginal ultrasound examination. A complete blood count performed at the time of her arrival showed a low hemoglobin level of 24 g/L, a low hematocrit level of 7,4%, a mean corpuscular volume of 98,7 fL and a number of red blood cells 0,75 x 1012/L. Due to the severity of the anemia, she was given 6 units of red blood cell transfusion, 2 fresh frozen plasmas and tranexamic acid accompanied with calcium carbonate. The curettage was performed. The pathohistological finding was endometrium in proliferation. Afterward, the hemoglobin level increased to 90 g/L. Their past medical history revealed that she abused alcohol. On an abdominal CT scan, Alcohol-Related Liver Disease (ARLD) was confirmed. We should keep in mind that coagulopathy could be the underlying cause of abnormal uterine bleeding and that anemia must be analyzed for successful treatment. A multidisciplinary approach to anemia caused by AUB is required in cases of severe anemia.

The utilization of the Internet and wireless communication reaches its pinnacle from one side of the planet to the other. Marking the rise of criminal activity in recent years sees enormous growth in security breaches and data theft-related cases in mobile phones. To mitigate them, the implementation of security patches, safety fixes, and updates in mobile devices is of high priority for the organization. The need to foster techniques and procedures in the field to be able to extract and precisely dissect digital crime cases, providing valuable tactical data about the investigation. Mobile forensics is a developing branch assisting the investigator in criminal trials and investigations. Acquisition, Collection, and Analysis of mobile phones settle the purpose of recovering cumulative and corroborative evidence. Upgradation and innovation of mobile devices with time imposed a challenge to mobile forensic technology to extract information from such devices. The study aims at extracting comparative and statistical approaches in the analysis of Physical data acquisition utilizing significant versatile mobile criminological proprietary tools. The proposed study also introduces newly developed utility tools along with their characteristic features which help in successful data extraction from mobile devices.

Trying to understand the causes of climate change can be confusing. On the one hand, methane (CH4) emissions from cattle, and methane emissions from food wastes in landfills, are said to contribute to greenhouse gases (GHGs) that drive climate change.  People are working on feed additives for dairy cows to reduce their methane emissions. But, at the same time, cattle manure and food wastes can be fed into anaerobic digesters to convert these organic wastes to biogas; the resulting “renewable methane” or “renewable natural gas” (RNG), can be used in place of fossil natural gas and avoid extra GHG emissions and stop global warming. Can we have it both ways?  Burning gasoline in our cars and trucks generates carbon dioxide (CO2), which is said to contribute to climate change. But more than 8 billion people on planet Earth, breathe in oxygen and exhale carbon dioxide every minute of the day. And so do all the other animals who live on this planet, breathe in oxygen and exhale carbon dioxide. Is our breathing also contributing to climate change, just as the emissions from our automobile tailpipes?It is time to step back from all the hype, evaluate the various sources of CO2 and CH4 being generated and review the “mass balance” of these gases in our atmosphere. Some of these are part of the natural biogenic carbon cycle and some are simply adding to the overall mass balance. What is driving climate change - excess GHGs from the biogenic carbon cycle, excess emissions from other sources, or both? Let’s take a fresh look at the available data.  (Of course, water vapor also plays a part in the climate change story, as a “positive feedback” effect. As non-condensable GHGs rise in concentration and slightly warm the planet, slightly warming oceans add a bit more water vapor to the story and push the warming up a bit more).  During the past few decades, a great deal of pertinent data has been carefully observed and recorded, by world-class scientists and engineers. This data is available in the public domain and is now easily available on the internet. These data sources come from reputable organizations, such as NASA (National Atmospheric and Space Administration), NOAA (National Oceanic and Atmospheric Administration), USGS (United States Geological Survey) and international organizations such as the “Global Carbon Project” and “Our-World-in-Data”.  How much of the growing concentration of CO2 (and CH4) in our atmosphere can be attributed to out-of-control emissions of biogenic carbon? How much of the CO2 rise can be attributed to fossil fuel emissions: 10%, 50%, 90%, or more?  The available data appear to show that the emissions from burning fossil fuels, more than account for all of the rise of carbon dioxide in our atmosphere during the past 60 years, or longer.  In comparison, the biogenic carbon cycle data seem to be very consistent, year after year, decade after decade and have little or no impact on climate change. But, let’s have a closer look at the available data, and let the data speak for itself.

Natural products are becoming increasingly important as an unlimited source for obtaining chemical substances with possible pharmacological potential. Current existing drugs for the treatment of cutaneous leishmaniosis produce major side effects; therefore the search for new drugs is justified. The stem bark of Zanthoxylum chiloperone var. Angustifolium Engl. (Rutaceae) is traditionally used in Paraguay for its antiparasitic properties. The leaf extract was evaluated for the first time to determine its leishmanicidal activity in BALB/c mice infected with amastigote forms of Leishmania amazonensis (PH8). The mice were treated orally with the extract at three concentrations (100, 50 and 10 mg/mL), intralesional (50 mg/mL), and subcutaneously using glucantime as a control (100 mg/mL). The percentage of decrease in parasite load was measured and with intralesional 50 mg/kg a reduction of 72% occurred, with the reference drug (Glucantime) a reduction of 62% was obtained with the same oral dose a reduction of 50%, while with an oral dose of 10 mg/mL the percentage of reduction was 55%. When the oral dose was increased to 100 mg/mL, the reduction percentage of the parasitic load was only 16%. These results indicated that the leaf extract of Z. chiloperone var. angustifolium Engl. at low oral concentrations (50 and 10 mg/mL) had very good activity against L. amazonensis, and it was even more efficacious intralesionally at 50 mg/mL but at the oral dose of 100 mg/kg has very reduced antiparasitic activity. This study showed the efficacy of the extract leaves of Z. chiloperone in reducing the parasite load in an in vivo test, so its use as a potential leishmanicidal could be suggested to develop and evaluate new drugs for the oral treatment of leishmaniosis disease with fewer side effects and lower cost.

Subacute sclerosing panencephalitis is a rare, slow, and insidious neurodegenerative disease caused by measles. This disease mostly has a classic course. However, sometimes it can be presented with atypical manifestations. In this paper, we aim to present a six years old male patient that was hospitalized due to seizures and ataxia. Cerebral magnetic resonance imaging was normal on the first day of admission, but within a few days, the patient started to be apathetic. On the seventh day, magnetic resonance imaging showed hyperintense lesions in the thalamic, brainstem, and periventricular areas. Periodic epileptiform discharges were detected in the repeated electroencephalogram. Investigations from the cerebrospinal fluid showed markedly elevated measles virus IgG at 230U/ml consistent with the diagnosis of SSPE which should always be ruled out when a patient comes in with uncontrollable seizures, ataxia and apathy.

Background: Having claimed lives, HIV/AIDS is still a significant global public health concern. Antiretroviral therapy (ART) is now widely available, and this rapid expansion of access is dramatically improving HIV epidemic survival rates worldwide.Objectives: The aim of this study was to identify the mortality risk factors and survival status of ART patients attending Hawassa Comprehensive Specialized Hospital in 2020.Methods: In a five-year retrospective cohort research, all patients seen between January 2015 and December 2019 were analyzed. The data were analyzed with SPSS 25.0. The Kaplan-Meier Log-rank model was employed to gauge the survival time of ART patients based on explanatory variables. Both bivariate and multivariate Cox proportional hazards regression models were employed to identify the independent causes of mortality.Results: Patients on ART had a 74% overall survival probability. With a median survival of 34 months, there are 0.135 deaths for every 100 person-years. Hemoglobin level (HR = 2.38; 95% CI = 3.3-6.3), WHO clinical stage III and IV (HR = 3; 95% CI = 2.2-9.5, p = 0.04), Age >=60 (HR = 1.6; 95% CI = 1.3-2, p = 0.04) and Functional status bed ridden (HR = 3.1; 95% CI = 1.2-9.4,p = 0.04) were all independent predictors of death among RVI patients.Conclusion: In comparison to trials conducted in wealthy countries, the survival rate of ART patients in this study was low. Patients who are anemic; WHO advanced clinical stage; old age, and functional status bedridden should be closely followed and monitored.

Introduction: Sonographic scan tests are real-time procedures but the female reproductive systems are subject to continued changes beyond the mid-menstrual period. Therefore a second test may be useful after mid-menstrual ultrasound tests among women with infertility who first had normal ultrasound tests during the mid-menstrual period. Aim: The aim of this study is to find out whether physiological changes beyond the mid-menstrual period in the female reproductive system could develop into findings that could be missed at the first ultrasound test among women with infertility during mid menstrual period. Method: One hundred and forty (140) women participated in this study, over a period of 12 months. The inter-observer correlations were carried out. The ultrasound test measured the diameters of the endometrium and ovarian follicle. The study was carried out during the mid-menstrual cycle when normal changes are optimal. The subjects were women of childbearing age (18 years - 40 years) with 28 days cycles who were referred from the fertility clinic. Women who had previous pelvic surgery, women on fertility therapy, and women who were unsure of the date were excluded from the study. Only women who met the inclusion criteria were selected for the study. The selection was by convenience sampling method. The women underwent the first phase of the ultrasound test during the mid-menstrual cycle. Those who had ultrasonographic-positive infertility reports were returned to the referral clinic. Only women who had normal ultrasonographic fertility reports were included in the second phase of further ultrasonographic tests. The scans were carried out further for four days for this second phase study and the result was again compared with normal values. (Endometrium Normal range 7.4 mm - 13.5 mm and ovarian follicles normal range 17.4 mm to 23.5 mm). Results: The first phase of the scan showed 108 (77%) of the women had positive infertility results for endometrium and ovarian follicles, while 32 (23%) of the women had a normal ultrasonographic result and were rescanned over days. The findings of the second phase scan showed that 10 subjects (7.30%) showed new abnormalities, Findings were distributed in three sub-groups in this second phase of the study. Sub-group A, 4 (2.67%) had enlarged unruptured follicles, sub-group B, 6 (4.30%) had an endometrial cavity filled with fluid, sub-group C, 22 (15.70%) subjects still had normal scans who may be referred for other studies. Conclusion: Women with infertility who had a normal scan at the mid-menstrual cycle should be followed up with a second-phase ultrasound scan before other tests.

Aim: To evaluate the hemodynamic changes and side effects during endotracheal intubation with Macintosh laryngoscope and intubating laryngeal mask airway.Materials and methods: A prospective, simple randomized, comparative study on 100 patients 18 years - 60 years of age, divided into two groups: Group A comprising intubation with Macintosh laryngoscope and Group B intubation through ILMA.Results: Total intubation time (in seconds) of group A was 24.38 + 3.26 seconds and of the group, B was 42.94 + 1.24 seconds. At 2,4 and 6, a higher rise in mean heart rate was noted in group A (p < 0.05). At 2,4,6 and 8 minutes difference in mean SBP and mean DBP of the two groups was statistically significant with a p - value of < 0.05 with a significant increase of mean SBP and mean DBP in patients of group A. The difference for all complications was not significant between the two groups.Conclusion: Intubation via intubating laryngeal mask airway can be done as an alternative to direct laryngoscopy using a Macintosh blade as intubation via intubating laryngeal mask airway has shown to have lesser hemodynamic changes.

Azotobacter was selectively isolated and purified from the soil samples of Xinjiang Salt Lake Scenic spot, the fermentation technology of exopolysaccharides (EPS) by Azotobacter was optimized, and the antioxidant activity of exopolysaccharides (EPS) was studied. The bacteria were isolated and purified from the soil samples by the scribing method and the 16SrRNA gene was used for molecular identification. The carbon source, fermentation time, inoculation amount and pH of target bacteria in the exopolysaccharides (EPS) fermentation process were optimized through single-factor experiments and their antioxidant activity was measured. Eight types of Azotobacter were isolated and purified from the soil samples of Salt Lake scenic spot. Among them, As101, which showed 99.58% homology with Azotobacter salinestris, was selected as the target strain. Through single-factor experiments which used exopolysaccharides (EPS) yield and exopolysaccharides content as indexes, the optimal conditions for the As101 fermentation process were determined as follows: fermentation temperature 35, fermentation time 96h, pH 7 and mannitol as carbon source. Exopolysaccharides content from Azotobacter salinestris  was 61.35% and the yield was 6.34 g/L. The results of the exopolysaccharides (EPS) antioxidant activity experiment under optimal conditions showed that As101 EPS had excellent scavenging ability against DPPH free radical, ABTS free radical and hydroxyl free radical, with IC50 values of 6.11 mg/ml, 2.42 mg/ml and 9.57 mg/ml, respectively. As101 with high yield and high exopolysaccharides content was isolated from saline soil in a special environment of Xinjiang, and the EPS obtained showed excellent antioxidant activity. The Azotobacter found in this study would provide the material basis for further opening up the adsorption of exopolysaccharides on heavy metals and the improvement of saline-alkali soil and contribute to further understanding of the structure and other activities of exopolysaccharides derived from Azotobacter.

Background: Liver Transplantation (LT) is the definitive treatment for Autoimmune Hepatitis (AIH) in patients with decompensated cirrhosis, liver failure and hepatocellular carcinoma. Outcomes of LT in AIH among black-Africans are not well-defined. We performed a single-center retrospective-review of adult LT patients. The study period was from 1st August 2004-31st August 2019. The primary aim was to document 1- & 5- year patient and graft survival. A secondary aim was to compare the survival of black-Africans to Caucasians. Data was analyzed using survival-analysis. Results: A total of 56 LT were performed for AIH. Sixty-seven percent (n = 38/56) had confirmed AIH on explant histology. Of these, the majority i.e., 79% (30/38) were female and 21% (8/38) were male. There were equal numbers of black-African 42% (n = 16/38) and Caucasian 42% (n = 16/38) patients. Rejection was four-times higher in black-Africans as compared to Caucasians. Forty-four percent (n = 17/38) had an acute rejection episode and 13% (5/38) had chronic rejection. Recurrence was found in four black-African females. Post-LT patient survival at 1- and 5- years was 86.5% and 80.7%, and graft survival was 94% and 70.8% respectively. The 5- year patient survival was insignificantly lower for black-Africans (73.9%) as compared to Caucasians (83.7%) (p - value 0.26, CI 6.3 - 12.2). Five-year graft survival was significantly lower among black-Africans (55%) as compared to Caucasians (84.8%) (p - value 0.003 CI 3.8 - 8.1)Conclusion: Black-Africans had a four-fold higher rate of rejection compared to Caucasians. Recurrent AIH was only found in patients of black ethnicity. Similar 1- & 5- year patient survival rates were observed between the two ethnicities. The 5-year graft survival among black-Africans was significantly lower than Caucasians.

Aim: The current paper presents a subjective symptom survey regarding postpartum discomfort (Study 1) and a case study on postpartum care using the program developed based on the survey results (Study 2). Thereafter, health care during the postpartum period is discussed.Methods: Study 1 analyzed 1638 postpartum women who completed the Subjective Fatigue Symptom Scale (SFSS) over the period from June 2012 to December 2019. Study 2 detailed the case of a 33-year-old primiparous woman who answered questions regarding the rehabilitation care program.Results: The 1638 subjects included in Study 1 had a mean age of 32.4 ± 8.2 years and a mean postpartum duration of 4.3 ± 2.3 months. Subjective symptoms included lower back pain, shoulder stiffness, sleepiness, wanting to lie down, yawing, and eye strain. The case included in Study 2 showed certain psychological and physical changes following the exercise program. The results of Study 1 showed that motor system discomfort, such as stiff shoulders and lower back pain, occurred in women across all postpartum stages. Our results demonstrated that care and exercise geared toward improving motor system function are imperative after childbirth. Meanwhile, the results of Study 2 imply that our rehabilitation program based on postpartum physical conditions had positive psychological and physical effects.Conclusion: Taken together, our results suggest that continuing rehabilitative care based on the physical condition during each postpartum stage facilitates improvement in mothers’ physical and psychological discomfort.

Polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls are persistent organic pollutants (POPs), which in recent years received huge attention due to their extreme stability, high potential toxicity and bioaccumulation in food chains. The main source of human exposure to these compounds is discovered in foods of animal origin, especially foods rich in fat. The target of the present study was to set up an analytical method for the determination of PCDDs/PCDFs and DL-PCB in vegetable oils, sunflower meals, sunflower seeds, rapeseeds and milk powder. The first step consisted of a semi-automatic Soxhlet extraction for 3 hours, by using a mixture of Hexane: Acetone – 80:20, followed by acid digestion with 55% acid silica and filtration. After concentration, the extract is purified on a multilayer column (silica gel, silica-KOH, silica-H2SO4 anhydrous Na2SO4) followed by an alumina column separation in two fractions (first fraction containing PCDDs/PCDFs and second containing only PCBs). The purified extract was then analyzed by GC/MS/MS. The newly developed approach in our lab was capable to reduce the overall time of sample preparation to seven hours/ per sample. Since the method shows good mean recoveries for all labeled congeners spiked in the samples (for PCDDs/PCDFs – 80% - 110%, for DL-PCBs – 70% - 85%), we assumed the absence of overestimation or underestimation in the analyzed samples.

A growing number of synthetic plastics derived from fossil fuels are produced, and improper plastic waste management has caused a lot of pollution problems. There are many microplastics in the environment, and they disintegrate slowly in soil and water. The properties of microplastics (MPs) include long residence times, high stability, high fragmentation potential, and the ability to adsorb other contaminants. Invertebrates and planktonic organisms are easily able to accumulate microplastics in aquatic species. Therefore, microplastics (MPs) must be removed from the water and other media. This paper aims to review the occurrence, raw polymers and additives, and remediation methods for removing microplastics from the environment. Several methods are available for removing contaminants, including sorption, filtration and chemical treatments. Various removal methods are discussed along with their methods, efficiency and advantages.

The main focus of this study concentrates on evaluating the quality of groundwater in and around the zone of pattamadai where the production of the mat in this region is a landmark industry, which is using dyes in a larger volume. The primary work starts with the collection of water samples from 13 boreholes and 13 hand-dug wells in different time intervals in the month of March 2021, which is a dry season. As per standards the water quality parameters have been evaluated and observed to be within the range. All the physico-chemical parameters for the samples have been studied for boreholes and hand-dug wells as all the values are observed to be within the range and are highlighted in the results.

Introduction: In an earlier study, we described and validated a VKA dose-finding algorithm (B2A), based on a novel bidirectional factor (BF). We designed a prospective study to evaluate the B2A in a daily care setting. Methods: In this open-label prospective study, we compared the outcomes of the B2A over the year 2020 with the outcomes of the previous year (2019), using regular algorithms. The outcomes were the duration of Time in the Therapeutic Range (TTR), the percentage of automated dose proposals (PAuP) and the percentage of accepted dose proposals (PAcP). The data were obtained from three anticoagulation centers in the Netherlands, in four locations. The outcomes of this study were based on a non-inferiority level.Results: The TTR over the year 2020 was at least non-inferior compared with the standard of care treatment. The percentage of automated proposals increased in all centers to approximately 96% of all dosages. Conclusion: The B2A performs non-inferior compared with the existing algorithms and in some aspects even better.

Oncological diseases are one of the most significant medical and social diseases in most countries of the world. Over the past decades, the search and development of new drugs, treatment regimens and methods of molecular diagnostics of malignant neoplasms remains relevant. In turn, an important goal of molecular genetic research is to suppress the expression of genes responsible for the development of tumors. The key targets taken into account in the development of antitumor drugs are proteins involved in carcinogenic changes in the cell. One of the promising molecular targets for the development of medicinal compounds in targeted therapy of tumor diseases is poly(ADP-ribose)polymerase 1 (PARP1). A potential way to inhibit PARP1 even at the stage of protein translation is RNA interference due to small interfering RNAs (siRNAs). For the penetration of siRNAs into the target cell, it is necessary to develop a method of their transportation controlled in space and time. An actual direction for solving this problem is the use of highly stable porous silicon-based nanoparticles. In the current study, in order to increase the functionality of nanoparticles, their surface was modified with various agents (functionalization), providing increased efficiency of drug loading and more uniform release. 

Every society organizes itself to reduce the complexity of living together in order to ensure relative peace of mind for its members. Major technological accidents such as the Chernobyl or Fukushima nuclear accidents are violent disruptions that affect the quality of life of tens of thousands of people and generate increased distrust and anxiety among them as well as the questioning of the state system’s ability to restore some peace of mind. Past experience has shown that the measurement of radiation and the development of local projects with the support of professionals and experts prove to be effective levers for those affected to regain the ability to make decisions for their protection and to assess the protective actions implemented collectively. It also showed that restoring confidence and restoring the dignity of these people, seriously impaired by the accident, takes time.

Background: The clinical manifestations of Corona Virus Disease of 2019 (COVID-19) varied from patient to patient with evidence of multi-organ involvement. Many patients continue to have a wide range of symptoms for variable periods of time. The long-term effects of COVID-19 infection (post-COVID-19 illness or syndrome) are not yet been fully explored.This study aims to highlight the clinical manifestations of the acute COVID-19 infection and the longer-term manifestations of the disease among the attendants to a private GP clinic in the Urban Amman Area, Jordan.Methods: A clinical case-series study was conducted on a sample of 300 COVID-19-positive cases among patients attending my private GP clinic in the Urban Amman Area, Jordan. We used the structured questionnaire based mainly on World Health Organization (WHO) Case Report Form (CRF) verified tool for post-COVID-19. All patients with COVID-19 were included in the study. Data collection was conducted through phone calls and analyzed using the Statistical Package for Social Sciences (SPSS) software. Results: The incidence of COVID-19 among patients attending the clinic during the period of this study was 25%. Females were 135 (45%) of the study population and males were 165 (55%). The mean (SD) of age was 34.2 (5.0) years. Most of the infected staff (90%) were symptomatic and developed acute COVID-19 symptoms. Fever, Cough, Fatigue, joint pain and loss of smell and taste were the most common symptoms. 72% of the study population had fully recovered from the infection, while 28% of them continued to suffer from many long symptoms. Fatigue (28%), shortness of breath on activity (18%), Social withdrawal (18%), anxiety (17%), forgetfulness (16%), trouble concentrating (15) and depressed mood (14%) were the most frequently reported long symptoms.Conclusion: The prevalence of Post COVID-19 illness was 28% with a high public health burden calling for the public health system to address the medical and psychological needs of affected persons. Mental health and psychosocial support are recommended elements for the management of patients.

Background: Hemp seed (Cannabis sativa L.) is an annual herbaceous plant of the Cannabis genus that contains a large amount of protein, iron, and fatty acids, including linoleic, α-linolenic, and γ-linolenic acid. These compounds are involved in a number of biological activities, including immunity enhancement, hyperlipidemia, and inflammation reduction. Here, we investigated the antioxidant effects of hemp seed on human cognitive function.Methods: The test was administered to 34 healthy volunteers aged ≥ 20 years. Participants were selected according to age and sex and were administered 10 g of hemp seed three times daily (30 g/day) for 45 days. The outcome measurements were recorded using a survey, computerized neurocognitive tests, and artificial intelligence.Results: Survey analysis determined that both the Beck Anxiety Inventory and Beck Depression Inventory measurements decreased significantly after hemp seed consumption when compared to measurements taken before consumption (p < 0.05). Additionally, significant results were observed in the Stroop and Tower of London tasks (p < 0.05). The prediction performance for the antidepressant effect was 0.83 for the area under the curve in the random forest algorithm, which was superior to that of other machine learning methods. Conclusion: These results suggest that hemp seeds have a beneficial effect on cognitive impairment.

Hereditary spherocytosis is a common inherited type of hemolytic anemia that results from abnormal morphology of erythrocytes. It has a high occurrence in North Americans and northern Europeans with a prevalence of 1/2000. There is a wide range in age and symptoms at presentation with some individuals being asymptomatic and others having severe diseases requiring blood transfusions. Based on the severity of symptoms, management may vary from simple observation to frequent blood transfusions, cholecystectomy for gallstones, and splenectomy. Timely diagnosis may be critical to minimize complications. Diagnostic tests have been available with varying degrees of accuracy. However new diagnostic tests with greater specificity and sensitivity are now available for more accurate diagnosis of Hereditary Spherocytosis in individuals of all ages including newborns. Illustrative cases are presented that show the variability in presentation, symptoms, complications, and care. Information is presented updating diagnostic testing that allows earlier diagnosis of children with hereditary spherocytosis. Additionally, the hematologic findings suspicious and consistent for this diagnosis are presented, serving as a guide when testing should be initiated.