Does morphology matter in 2017? An approach to morphologic clues in non-neoplastic blood and bone marrow disorders.

Numerous non-neoplastic and neoplastic conditions manifest with distinctive features in blood. Although automated complete blood count (CBC) data are essential, CBC information alone is insufficient for diagnosis. Consequently, morphologic review of blood smears is still relevant in the era of sophisticated automated analyzer systems. Pathologist interpretation of the peripheral blood smear, in conjunction with CBC and clinical information, can provide rapid diagnostic information and guide cost-effective targeted laboratory testing. Pathologist review of blood smears can be used to diagnose cases in which the clinical findings are misleading or nonspecific. Here, we discuss a selection of cases in which the pathologist examination of the blood smear can aid in rapid and accurate diagnosis and guide appropriate treatment. Exemplary non-neoplastic disorders with distinctive morphologic blood features of RBC, neutrophils, monocytes, and lymphocytes will be highlighted. The differential diagnostic considerations in blood smears with RBC destruction will be presented, expanding beyond microangiopathic hemolytic anemia. Lymphocyte morphologic assessment guides differential diagnosis including the identification of rare germline disorders. In each case presented, the integration of morphologic features guided additional testing with confirmation of the diagnosis. Peripheral blood smear review is timely and cost effective, even in an era of sophisticated automated laboratory testing.

Leukocytosis.

An increased white blood cell count, or leukocytosis, is a common laboratory finding. Appropriate specimen evaluation depends on which lineages are increased and the morphologic findings on peripheral blood smear review to guide further testing. The presence of blasts is concerning for acute leukemia and may require bone marrow biopsy. Lymphocytosis may be morphologically divided into polymorphic and monomorphic populations. Polymorphic lymphocytosis is most consistent with a reactive process, while monomorphic populations are concerning for lymphoproliferative neoplasm. The differential can be further narrowed based on morphologic findings. Myeloid leukocytosis can occur in a number of reactive conditions as well as myeloid malignancies. The types of cells present and morphology can help to guide additional workup. This study provides guidance for the appropriate evaluation and further workup of leukocytosis.

Estimating the nucleated red blood cell 'emergence time' in neonates.

OBJECTIVE: The time between onset of fetal hypoxia and first appearance of nucleated red blood cells (NRBCs) in the blood can conceptually be divided into two periods; (1) the 'erythropoietin (EPO) generation time', which previous fetal studies suggest is 4 to 5 h, and (2) the 'NRBC emergence time'. In this study, we estimated the latter as the time required for NRBC to appear in the blood after administering a dose of recombinant EPO. STUDY DESIGN: This was a retrospective analysis of data from a multihospital healthcare system (Intermountain Healthcare). Data were included only for neonates born ≥34 weeks gestation between the dates 1 January 2005 and 31 October 2012 and only if they received a dose of darbepoetin during their neonatal intensive care unit stay and had one or more complete blood cell counts (CBCs) obtained during the 3-day period before the dose was given and one or more CBCs in the 7-day period after the dose. RESULT: The study involved 31 neonates who received 34 doses of darbepoetin. Seven doses were 4 µg kg(-1) and twenty-seven doses were 10 µg kg(-1). Twenty-six CBCs were obtained during the 24-h period following the darbepoetin dose and none had NRBC identified. NRBC first appeared in the blood between 24 and 36 h after the dose. Recipients of the higher dose generally had a higher peak NRBC count but the NRBC 'emergence time' did not appear to depend on dose. CONCLUSION: Following fetal hypoxia, transcription and translation of the EPO gene result in an elevation in plasma EPO concentration. Previous fetal studies suggest this process requires 4 to 5 h. The present studies suggest that, following the increase in plasma EPO, NRBC emerge into the circulation in ≥24 h. If this model serves as a reasonable estimate, it suggests that neonates with an elevated NRBC count at birth had the onset of hypoxia at least 28 to 29 h before birth.

Whole-blood viscosity in the neonate: effects of gestational age, hematocrit, mean corpuscular volume and umbilical cord milking.

OBJECTIVE: The American College of Obstetrics and Gynecology Committee on Obstetric Practice recently endorsed delayed cord clamping at preterm delivery. However, the committee report expressed the concern by some practitioners that delayed clamping or cord milking might induce hyperviscosity in preterm neonates. To address this issue we: (1) established reference ranges for whole-blood viscosity among preterm neonates (viscosity reference ranges had previously been reported only in term neonates) and (2) determined the effect of umbilical cord milking at deliveries <32 weeks gestation on subsequent blood viscosity measurements. STUDY DESIGN: This was a prospective study in two Neonatal Intensive Care Units. Blood viscosity was measured using a cone and plate viscometer. Associations were sought with gestation, hematocrit/hemoglobin and mean corpuscular volume. Reference ranges were determined for preterm infants <32 weeks gestation. Then, after umbilical cord milking at deliveries <32 weeks, viscosity was measured at birth and again during the 12 h after birth. In neonates with viscosities >95th % range, we sought signs of hyperviscosity (plethora, hypotonia, hypoglycemia, hyperbilirubinemia, thrombocytopenia). RESULT: Viscosity at higher and lower sheer rates were linearly related (n=32, r=0.971). Within the range of hematocrits measured (29-63%) viscosity correlated with hematocrit (r=0.877) and hemoglobin (r=0.853) but not with erythrocyte size (r=0.179). Viscosity was related to gestational age (n=58), primarily due to the lower hematocrits at lower gestational ages. In the 12 h after cord milking viscosity ranged from 3.1 to 9.5 centipoise. Three of twenty preterm, neonates had viscosities >95th % reference range. However, all values were well below those where hyperviscosity is defined in term neonates and all lacked features of hyperviscosity. CONCLUSION: Cord blood viscosity is directly proportional to hematocrit/hemoglobin, lower at early gestation and not associated with erythrocyte size. Cord milking at preterm delivery is associated with a low risk of clinical hyperviscosity. Practioners should not refrain from cord milking at preterm delivery because of a concern that it will commonly cause neonatal hyperviscosity.

Severe neonatal anemia from fetomaternal hemorrhage: report from a multihospital health-care system.

OBJECTIVE: The incidence of fetomaternal hemorrhage that is severe enough to cause neonatal anemia is not known. Owing to its relative rarity, much of the literature describing this condition is in the form of case reports and small case series. We performed a large, muiticentered, sequential, case series to determine the incidence, antecedents and outcomes. STUDY DESIGN: From the multicentered databases of Intermountain Healthcare, we obtained records of all neonates with hematocrit (Hct) <30% or hemoglobin (Hgb) <10 g dl(-1) on the day of birth, who had Kleihauer-Betke staining or flow cytometric evidence of fetomaternal hemorrhage. RESULT: Among 219,853 live births, 24 had anemia with evidence of fetomaternal hemorrhage (incidence estimate, 1 per 9160 live births). The initial Hgb ranged from 1.4 to 10.2 g dl(-1) (Hct 29.8%). The initial Hgb was <7 g dl(-1) in 18 (67%), <5 g dl(-1) in 12 (50%) and was <3 g dl(-1) in 7 (29%). All 7 mothers in whom neonatal Hgb was <3 g dl(-1) had reported absent fetal movement, as did 13 of 18 mothers when the initial Hgb was <7 g dl(-1). Outcomes were poorer in those with the lowest initial Hgb; in the two lowest, one died on day 1, and the other developed a grade 4 intraventricular hemorrhage (IVH). The adverse outcomes of death, IVH, periventricular leukomalacia, bronchopulmonary dysplasia or hypoxic-ischemic encephalopathy were common; occurring in 71% (17 of the 24), including all with an initial Hgb <5 g dl(-1) and all born at ≤35 weeks of gestation. CONCLUSION: Fetomaternal hemorrhage is a rare but sometimes devastating condition. Those with fetomaternal hemorrhage and an initial Hgb of <5 g dl(-1) are expected to need resuscitation at birth, to receive emergent transfusion support and to be at risk for death and major morbidities. Antenatal suspicion of this diagnosis should occur when absent fetal movement is reported. Improvements in rapid diagnosis are needed to prepare first responders and transfusion services.

Expression of GABAB receptors in magnocellular neurosecretory cells of male, virgin female and lactating rats.

GABA is one of the key neurotransmitters that regulate the firing activity of neurones in the supraoptic (SON) and paraventricular (PVN) nuclei. In the present study, we used immunohistochemical techniques to study the distribution and subcellular localisation of metabotropic GABA(B) receptors in magnocellular neurones in the SON and PVN. Robust GABA(B) receptor immunoreactivity (GABA(B)R; both subunit 1 and subunit 2 of the heterodimer), was observed in the SON and PVN. At the light microcope level, GABA(B)R immonoreactivity displayed a clustered pattern localised both intracytoplasmically and at the plasma membrane. Densitometry analysis indicated that GABA(B)R immunoreactivity was significantly more intense in vasopressin cells than in oxytocin cells, both in male, virgin female and lactating rats, and was denser in males than in virgin females. Light and electron microscope studies indicated that cytoplasmic GABA(B)R was localised in various organelles, including the Golgi, early endosomes and lysosomes, suggesting the cycling of the receptor within the endocytic and trafficking pathways. Some smaller clusters at the level of the cell plasma membrane were apposed to glutamic acid decarboxylase 67 immunoreactive boutons, and appeared to be colocalised with gephyrin, a constituent protein of the postsynaptic density at inhibitory synapses. The presence of GABA(B)R immunoreactivity at synaptic and extrasynaptic sites was supported by electron microscopy. These results provide anatomical evidence for the expression of postsynaptic GABA(B) receptors in magnocellular neurosecretory cells.

Preinduction cervical assessment.

Systems of quantifying and scoring cervical factors have been sought for years to predict the duration of labor and to determine which patients may successfully and safely undergo induction of labor. Early methods of cervical assessment generally attempted to classify patients as having favorable or unfavorable cervices based on dichotomous variables. Since then, obstetricians have begun to appreciate the usefulness of having a single score derived from the degree of ripeness of several cervical characteristics. The scoring system that has become most prevalent is the Bishop score. This system and its modifications take into account the dilation, effacement, consistency, and position of the cervix in addition to the station of the presenting part. Many have evaluated and confirmed the validity of the Bishop score. Among the factors considered in assigning the score, the strongest association with successful labor seems to be with cervical dilation. The Bishop score has been criticized for not attributing more significance to cervical dilation. However, despite this criticism, none of the modifications to the original scoring system have been shown to improve predictability. More recently, the use of ultrasound assessment of the cervix has been suggested to improve prediction of the success of labor induction. However, convincing evidence that this technique provides significant additional information when compared to digital examination is lacking. The Bishop score would seem to be the best and most cost-effective method currently available to assess the cervix and predict the likelihood of success of labor induction and the duration of such an induction.

Patient acceptance of endovaginal ultrasound.

OBJECTIVE: To assess women's experiences and attitudes regarding endovaginal ultrasound. METHODS: Women attending the obstetrics and gynecology clinics at the University of Florida were asked to complete an anonymous questionnaire. RESULTS: Of respondents who had had a prior endovaginal ultrasound examination, 26% reported that 'it hurt a lot' and 50% reported that 'it hurt a little'. In contrast, only 23% of patients had experienced any pain with a prior abdominal ultrasound (P < 0.001). Multivariate analysis showed that discomfort with a prior vaginal ultrasound was related to patient age category, with less discomfort being experienced with increasing ages (P = 0.001). A history of unwanted sex, sexual or physical abuse, or dyspareunia was not associated with more painful examinations. Most patients were willing to undergo a future vaginal ultrasound investigation if it were recommended by their doctor. Multivariate analysis showed that more willingness was associated with older age category (P = 0.004), a history of dyspareunia (P = 0.03) and bleeding in a current pregnancy (P = 0.005). Of the patients (less than half) who had a preference, most preferred a female sonographer for endovaginal sonography (P < 0.001) and most preferred that a doctor, rather than a nurse or technician, perform the examination (P < 0.001). CONCLUSIONS: The majority of women who had experienced a vaginal ultrasound examination found it somewhat uncomfortable, but almost all women who returned the questionnaire expressed a willingness to undergo endovaginal ultrasound if it were recommended.

Congenital ischemic forearm necrosis associated with a compound presentation.

An infant born in a compound presentation had ischemic forearm necrosis requiring amputation. Bilateral renal vein thrombosis predating labor and delivery was also diagnosed. The possibility of congenital ischemic necrosis in an extremity should not affect the obstetrical management of compound presentation, but if this complication occurs a search for an underlying coagulopathy is important.

Nonimmune hydrops fetalis: fetal and neonatal outcome during 1983-1992.

Prognostic factors for survival of 62 fetuses and neonates with nonimmune hydrops fetalis (NIHF) were studied retrospectively. Twenty-eight infants survived >/=28 days which is 45% for all fetuses and newborns diagnosed with NIHF and 61% for liveborns with unresolved NIHF. Univariate analysis identified that mortality was associated with the presence of >/=2 serous cavity effusions and a need for chest compressions at birth. Multivariate logistic regression analysis confirmed that the presence of >/=2 serous cavity effusions was significantly associated with mortality from NIHF <28 days after birth [OR = 48.2 (CI 3.6, 662.9) (p < 0.004)]. We conclude that, compared to published cases from the 1970s and early 1980s, survival of liveborns with NIHF seems improved. The decrease in stillbirths is more notable. The severity of hydrops at birth is the key determinant for survival.

Prenatal ultrasound diagnosis of massive subchorionic thrombohematoma.

Massive subchorionic thrombohematoma is a rare condition in which a large maternal blood clot separates the chorionic plate from the villous chorion. This condition is usually complicated by intrauterine growth restriction, and is often associated with fetal distress and perinatal death. We present a case in which the diagnosis of massive subchorionic thrombohematoma was made at 24 weeks' gestation. Doppler ultrasound helped to confirm the diagnosis and demonstrated severely abnormal umbilical blood flow. Two days after the diagnosis, fetal distress prompted emergency Cesarean delivery of a growth-restricted infant.

Second-trimester vaginal bleeding: correlation of ultrasonographic findings with perinatal outcome.

OBJECTIVE: Our purpose was to determine the relationship between ultrasonographic findings and perinatal outcome in patients with second-trimester vaginal bleeding. STUDY DESIGN: A retrospective case-control study was performed. One hundred sixty-seven patients with ultrasonographic examinations performed for bleeding between 13 and 26 weeks' gestation were identified through a comprehensive ultrasonography database. The main ultrasonographic findings of interest were the presence of an intrauterine clot, membrane separation, and placenta previa. A control group of 167 patients was obtained by selecting the next consecutive patient from the comprehensive perinatal database. Perinatal outcome measures for both groups were recorded. RESULTS: Multiparity was more common in patients with bleeding than in controls (69% vs 58%, p = 0.036), as was history of two or more previous preterm deliveries (6% vs none, p = 0.005). Second-trimester vaginal bleeding was associated with increased risk of preterm delivery (relative risk 1.9, 95% confidence interval 1.4 to 2.8), fetal death (relative risk 6.3, 95% confidence interval 1.9 to 2.1), and perinatal death (relative risk 5.4, 95% confidence interval 2.1 to 13.7). The perinatal mortality rate was 162:1000 in these patients versus 30:1000 in controls. To assess the impact of ultrasonographic abnormalities, the study group was divided into two groups. Among the patients with second-trimester vaginal bleeding those with abnormal ultrasonographic findings had an increased risk of preterm delivery (relative risk 2.0, 95% confidence interval 1.4 to 2.8), fetal death (relative risk 2.6, 95% confidence interval 1.1 to 6.3), perinatal death (relative risk 2.6, 95% confidence interval 1.3 to 5.3), and neonatal intensive care unit admissions (relative risk 3.2, 95% confidence interval 1.6 to 6.1). The perinatal mortality rate was 258:1000 for patients with abnormal ultrasonographic examinations. CONCLUSION: Second-trimester vaginal bleeding is more common in multiparous women and in women with a history of a previous preterm delivery. Perinatal morbidity and mortality is increased in patients with bleeding during the second trimester. The risk is compounded when abnormalities are detected by ultrasonography.

On-line liquid chromatography coupled with high field NMR and mass spectrometry (LC-NMR-MS): a new technique for drug metabolite structure elucidation.

High performance liquid chromatography has been coupled simultaneously to high field NMR and MS detectors, giving UV, NMR and mass spectra for each component in a mixture, after on-line separation. This powerful new tool for the structure elucidation of components in mixtures without isolation has been successfully applied to the analysis of the metabolites of paracetamol in human urine.

The etiology of fetal acidosis as determined by umbilical cord acid-base studies.

OBJECTIVE: Our purpose was to determine whether umbilical cord acid-base values might be used to differentiate abruptio placentae from cord prolapse. STUDY DESIGN: By use of a retrospective study design, umbilical cord blood acid-base values of 32 cases of severely acidotic acute abruptio placentae were statistically compared with those of 19 cases of severely acidotic acute cord prolapse with Fisher's exact test, Mann-Whitney U tests, and receiver-operator curve characteristic analyses. RESULTS: No significant differences in electronic fetal heart rate record interpretations were detected. Highly significant differences in umbilical arterial and umbilical venous blood gas values were noted between the two groups. Most notable were the differences between umbilical venous and arterial blood gas values in the cord prolapse group. Receiver-operator characteristic curve analysis demonstrated that an umbilical venous-arterial pH difference of 0.15 was an effective cutoff value in differentiating cord prolapse from abruptio placentae (accuracy 92%). CONCLUSION: Umbilical arterial blood gas values combined with umbilical venous blood gas values can assist in determining the pathogenesis of marked fetal acidosis.