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PubMed2001

A specific molar ratio of stabilizer to protein is required for storage stability of a lyophilized monoclonal antibody.

The selection of the appropriate excipient and the amount of excipient required to achieve a 2-year shelf-life is often done by using iso-osmotic concentrations of excipients such as sugars (e.g., 275 mM sucrose or trehalose) and salts. Excipients used for freeze-dried protein formulations are selected for their ability to prevent protein denaturation during the freeze-drying process as well as during storage. Using a model recombinant humanized monoclonal antibody (rhuMAb HER2), we assessed the impact of lyoprotectants, sucrose, and trehalose, alone or in combination with mannitol, on the storage stability at 40 degrees C. Molar ratios of sugar to protein were used, and the stability of the resulting lyophilized formulations was determined by measuring aggregation, deamidation, and oxidation of the reconstituted protein and by infrared (IR) spectroscopy (secondary structure) of the dried protein. A 360:1 molar ratio of lyoprotectant to protein was required for storage stability of the protein, and the sugar concentration was 3-4-fold below the iso-osmotic concentration typically used in formulations. Formulations with combinations of sucrose (20 mM) or trehalose (20 mM) and mannitol (40 mM) had comparable stability to those with sucrose or trehalose alone at 60 mM concentration. A formulation with 60 mM mannitol alone provided slightly less protection during storage than 60 mM sucrose or trehalose. The disaccharide/mannitol formulations also inhibited deamidation during storage to a greater extent than the lyoprotectant formulations alone. The reduction in aggregation and deamidation during storage correlated directly with inhibition of unfolding during lyophilization, as assessed by IR spectroscopy. Thus, it appears that the protein must be retained in its native-like state during freeze-drying to assure storage stability in the dried solid. Long-term studies (23-54 months) performed at 40 degrees C revealed that the appropriate molar ratio of sugar to protein stabilized against aggregation and deamidation for up to 33 months. Therefore, long-term storage at room temperature or above may be achieved by proper selection of the molar ratio and sugar mixture. Overall, a specific sugar/protein molar ratio was sufficient to provide storage stability of rhuMAb HER2.

Antibodies, Monoclonal↗
PubMed2003

Efficient synthesis of high molar mass, first- to fourth-generation distributed dendronized polymers by the macromonomer approach.

A homologous series of first- to fourth-generation (G1-G4) dendronized macromonomers, 5, 7, 10, and 12, was synthesized, and their polymerization behavior under radical conditions investigated. These conditions were thermally induced radical polymerization (TRP) and atom-transfer radical poymerization (ATRP). TRP was applied to all monomers and gave polymers PG1-PG4, whose molar masses range from several millions for PG1 to estimated several hundreds of thousands for PG2 and PG3, and to the oligomeric regime for PG4. ATRP was applied only to the G1 and G2 monomers 5 and 7. Kinetic studies on monomer 5 provide evidence that its polymerization proceeds in a controlled fashion. The highest monomer-to-initiator ratios which still gave monomodal molar mass distributions were 300:1 (for 5) and 100:1 (for 7), which correspond to achievable molar mass regime for PG1 and PG2 of approximately M(n)=100 000 (DP(calcd)(PG1)=200, DP(calcd)(PG2)=90). The polydispersities lie in the usual range (PDI=1.1-1.2). The molar masses were determined by GPC in DMF with calibration against absolute molar masses of PG1 determined by light scattering.

Journal Article↗
PubMed2005

A prospective study of ultrasound screening for molar pregnancies in missed miscarriages.

OBJECTIVE: To examine the relationship between ultrasound and histological features in the screening for molar changes in missed miscarriage. METHODS: A prospective cohort study was conducted on all missed miscarriages, with features suspicious of molar pregnancy, on transvaginal ultrasound and/or on histological examination over a 5-year period. All cases of molar pregnancy diagnosed histologically were examined and cross-referenced with cases diagnosed on ultrasound and with the supplementary report from the regional referral center. When available, maternal serum beta-human chorionic gonadotropin (hCG) levels were recorded. RESULTS: Fifty-one cases of suspected molar pregnancy were referred to the regional center for further histological opinion and follow-up, and five cases were subsequently excluded from the final analysis because of the diagnosis of hydropic abortion (HA). In 33 cases a molar pregnancy was suspected at the initial scan. Of these, 26 (78.8%) were confirmed on histology, resulting in a 56% detection rate using ultrasound alone. In 15 cases hCG results were available, of which nine were greater than two multiples of the median. CONCLUSIONS: The diagnosis of both complete (CHM) and partial (PHM) hydatidiform moles in first-trimester miscarriages is difficult. hCG is significantly higher in both CHM and PHM and, in conjunction with transvaginal ultrasound, could provide the screening test required to enable clinicians to counsel women more confidently towards non-surgical methods of management of their miscarriage, where histopathological examination is not available.

Abortion, Missed↗
PubMed1997

Water (H2O and D2O) molar absorptivity in the 1000-4000 cm-1 range and quantitative infrared spectroscopy of aqueous solutions.

Water (H2O and D2O) molar absorptivity was measured by Fourier transform infrared transmission spectroscopy in the 1000-4000 cm-1 range at 25 degrees C. A series of assembled cells with path lengths from 1.2 to 120.5 microns was used for these measurements. The optimal path length (the path length of aqueous solution at which the IR spectrum of solute, corrected for water absorbance, has the highest signal-to-noise ratio) was calculated for all water absorbance bands. The results presented here show that the optimal path length does not depend on solute properties and is inversely proportional to the solvent (water) molar absorptivity. The maximal signal-to-noise ratio for measurements of IR spectra of aqueous solution in the 1650 cm-1 spectral region, of primary interest in biological applications, can be obtained at an optimal cell path lengths of 3-4 microns (H2O) and 40-60 microns (D2O). As an example, the signal-to-noise ratio was calculated as a function of the cell path length for the amide I (H2O) and amide I' (D2O) bands of an aqueous lysozyme solution. The molar absorptivities of water bands are several orders of magnitude weaker than those of the strongest bands of biological macromolecules in the same spectral regions. High net water absorbance in aqueous solutions is due simply to the very high molar concentration of water. A method is proposed for the quantitative measuring of the path length of the cell which exploits the molar absorptivity of the strongest water bands (stretching vibrations) or of bands which do not overlap with solute absorbance. A path length in the range from approximately 0.01 micron to approximately 1.0 mm can be determined with high precision using this technique for a samples of known concentration. Problems involved in the proper correction of strong water absorbance in IR spectra of aqueous solutions of biomolecules are discussed, including multiple reflections within the cell, the effects of pH, temperature, and perturbation of water spectral properties by polar solutes, as well as the selection of optimal spectral regions in which one may obtain the most precise absorbance corrections.

Calcium Fluoride↗
PubMed1994

Development of high-molar-mass cellobiase complex by spontaneous protein-protein interaction in the culture filtrate of Termitomyces clypeatus.

The 450 kDa cellobiase from Termitomyces clypeatus which migrates as a single band on IEF, PAGE and SDS-PAGE, was found to possess appreciable sucrase activity. The fungus produced sucrase and cellobiase constitutively in different media but with different activity ratios. The kinetics of secretion of the two enzymes was similar under in vivo and in vitro conditions. HPGPLC analysis of the culture filtrates indicated the presence of both sucrase and cellobiase in the same protein fractions of different molar mass, even in the 30-kDa protein fraction. No free sucrase or cellobiase could be detected in the culture filtrates. It was also observed that fractionation of cellobiase by (NH4)2SO4 precipitation was different with different amounts of associated sucrase activity present in the culture filtrate. The (NH4)2SO4-precipitated cellobiase fraction also contained cellobiases in proteins of widely varied molar mass ranges. However, none of the low-molar mass proteins other than the 450-kDa enzyme could be purified, as all low-molar-mass fractions spontaneously aggregated to the 450-kDa enzyme. Hydrophobic chromatography of the (NH4)2SO4-precipitated fractions followed by HPGPLC of the eluted active fraction yielded both cellobiase-free sucrase and a very low sucrase-containing cellobiase fraction. The cellobiase fraction, homogeneous in PAGE, was also a high-molar-mass protein complex dissociating into a number of protein bands on SDS-PAGE.(ABSTRACT TRUNCATED AT 250 WORDS)

Basidiomycota↗
PubMed1986

Occurrence of molar pregnancy in patients undergoing elective abortion: comparison with other clinical presentations.

Clinical data of molar pregnancies found in women undergoing elective abortion (group 1, n = 39) were compared to those of molar pregnancies in women who experienced spontaneous abortions (group 2, n = 157) and women in whom molar pregnancy was discovered before symptoms of spontaneous abortion were evident (group 3, n = 209). Group 1 women were younger and experienced uterine evacuation at an earlier stage of amenorrhea than groups 2 and 3. Group 3 had larger uteri at evacuation and longer intervals of positive tests for the beta-subunit of human chorionic gonadotropin during the postmolar phase as compared with groups 1 and 2. On the basis of available provincial data for the number of elective abortions, the estimated incidence of molar pregnancies in this population was 1:2,699. The presence of malignant gestational trophoblastic neoplasia was documented in a single patient in group 1. The incidence of malignant gestational trophoblastic neoplasia in this group was not significantly different from that in groups 2 and 3. Routine pathologic examination of the products of conception in women undergoing elective abortion coupled with routine assays of the beta-subunit of human chorionic gonadotropin when molar pregnancy is found can identify both noninvasive and invasive trophoblastic disease in these women.

Abortion, Induced↗
PubMed1983

Relationship between molar refraction and n-octanol/water partition coefficient.

The n-octanol/water partition coefficient (Pow) for organic chemicals which do not form the hydrogen bond was considered on the basis of London's dispersion force, because the dispersion force is the main component of van der Waals-type attractive forces between solute and solvent molecules. The dispersion force is a function of both the ionization potential and the molar refraction. As the variation of the ionization potential is small in comparison with that of the molar refraction, the latter is the most important factor to determine the value of Pow. Thus the relationship between Pow and the molar refraction can be established. The relationship was confirmed for some halogenated alkanes, alkenes and aromatics, and polycyclic aromatic hydrocarbons by analysis of observed values of Pow and calculated values of the molar refraction. Estimation of Pow from the molar refraction is advantageous for the preliminary hazard assessment of new chemicals, because direct measurement of Pow is laborious and time consuming for lack of information about analytical methods.

1-Octanol↗
PubMed2004

A case of molar ectopic pregnancy.

OBJECTIVE: To evaluate the occurrence of molar pregnancy in tubal ectopic pregnancy. DESIGN: Case report. SETTING: Outpatient clinic. PATIENT(S): A 27-year-old woman. INTERVENTION(S): Salpingectomy. MAIN OUTCOME MEASURE(S): Molar ectopic pregnancy. RESULT(S): Ectopic partial molar pregnancy. CONCLUSION(S): Molar pregnancy can occur in ectopic pregnancy. Molar pregnancy clinically mimics normal tubal ectopic pregnancy.

Adult↗
PubMed2005

p57KIP2 immunohistochemistry in early molar pregnancies: emphasis on its complementary role in the differential diagnosis of hydropic abortuses.

Morphologic examination still forms the main diagnostic tool in the differential diagnosis of molar pregnancies. However, the criteria are subjective and show considerable interobserver variability among pathologists. Once a diagnosis of molar pregnancy is made, DNA ploidy studies help to differentiate a triploid partial mole from diploid complete mole (CM). However, with earlier diagnosis and therapeutic evacuation of molar pregnancies, the differentiation of molar pregnancies from early nonmolar placentation is becoming increasingly difficult. The p57(KIP2) gene ( CDKN1C ) is strongly paternally imprinted and expressed from the maternal allele. Because CM lacks a maternal genome, p57(KIP2) immunostaining is correspondingly absent, whereas hydropic abortuses and partial mole show positive staining. We compared the use of p57(KIP2) staining in the differential diagnosis of 68 morphologically challenging cases of early first-trimester hydropic placentas. Diagnosis based on p57(KIP2) staining was compared with the original diagnosis based on morphology and DNA ploidy analysis. Concordant results were obtained in 65 of 68 cases studied. In 2 of 3 cases with a discordant diagnosis, microsatellite DNA genotyping analysis agreed with the results of p57(KIP2) staining, confirming that positive p57(KIP2) staining is a highly sensitive and specific marker for excluding CM in this setting. In addition, p57(KIP2) staining has the advantage of differentiating hydropic abortuses from CMs, a distinction not made by ploidy analysis. p57(KIP2) staining can be used in concert with ploidy studies to refine the diagnosis of early molar pregnancies.

Adult↗
PubMed1994

Intraoperative molar pregnancy crisis.

Molar pregnancy is a gestation in which the ovum is transformed into a fleshy tumor mass or mole. Of all gynecologic tumors, it is one of the most feared. It is characterized by first trimester bleeding, hyperemesis, and toxemia and can be diagnosed using pelvic ultrasound. Suction currettage is the treatment of choice for molar pregnancy when a patient desires to have more children; however, hysterectomy may be necessary. Abdominal hysterectomy reduces the risk of malignant sequelae. Complications associated with molar pregnancy usually are a result of suction curettage and include pulmonary insufficiency syndrome, choriocarcinoma, hyperthyroidism, theca lutein cysts, and disseminated intravascular coagulation. The perioperative nurse can be instrumental in assessing, planning, organizing, and directing intervention for potential complications associated with the management of a molar pregnancy crisis. The perioperative nurse is encouraged to review all aspects of molar pregnancy to understand the ramifications of the surgical procedures.

Female↗
PubMed2002

Molar cornual ectopic pregnancy.

BACKGROUND: Few cases of ectopic molar pregnancies have been reported. We present one such case of a molar cornual pregnancy. CASE: A woman at 12 weeks' gestation by last menstrual period presented with ultrasound evidence of a molar pregnancy and a surgical abdomen. Diagnostic laparoscopy revealed a large amount of free intraperitoneal blood. Laparotomy revealed villous-appearing tissue with numerous fluid-filled cysts erupting from the posterior aspect of the right uterine cornu. A cornual resection was performed, followed by a suction dilation and curettage (D&C). The pathologic findings from the cornual resection were consistent with a partial molar pregnancy, while the material from the suction D&C demonstrated no villi or trophoblastic tissue. CONCLUSION: A molar cornual ectopic pregnancy is presented.

Female↗
PubMed2003

Human chorionic gonadotropin follow-up in patients with molar pregnancy: a time for reevaluation.

OBJECTIVE: To determine how often patients with molar pregnancy do not complete recommended follow-up and to identify factors that may predict failure to complete human chorionic gonadotropin (hCG) monitoring. This study also sought to determine how often patients with molar pregnancy who do not complete follow-up relapse after attaining at least one undetectable hCG value. METHODS: Four hundred randomly selected patients with molar pregnancy were analyzed regarding the serum hCG levels after molar evacuation. Demographic factors were determined for each patient: age, marital status, gravidity, parity, health insurance type, and distance from patient residence to trophoblastic center. RESULTS: Recommended hCG follow-up was completed in 63% of the uncomplicated 333 cases (n = 211). Three hundred twenty patients achieved at least one undetectable serum hCG level. Among the 320 patients, 33% achieved undetectable hCG values but did not complete recommended follow-up. However, none had any evidence of relapse. A distance of greater than 20 miles from the patient's residence to our center was associated with failure to complete hCG follow-up (P =.001). CONCLUSION: Because none of the 320 patients who achieved at least one undetectable hCG level has been diagnosed with gestational trophoblastic tumor relapse, it may be appropriate to reassess the duration of hCG monitoring for patients with molar pregnancy.

Adolescent↗
PubMed1996

Autonomic and sensory innervation of cat molar gland and blood vessels in the lower lip, gingiva and cheek.

Innervation of the molar gland and blood vessels in the lower lip, gingiva and cheek mucous membrane was investigated in the cat with the aid of whole mount acetylthiocholinesterase (WATChE) histochemistry and retrograde neuronal tracing methods with horseradish peroxidase (HRP) and HRP-conjugated wheat germ agglutinin (WGA-HRP). The molar gland was found to be supplied from the buccal nerve and branches of the mylohyoid nerve on the basis of microdissection of WATChE-stained mandibular preparations under a dissecting microscope. The rostral half of the lower lip-gingiva was innervated by mental branches from the inferior alveolar nerve. The caudal half of the lower lip-gingiva and cheek mucous membrane were observed to be supplied from the buccal nerve. Following injections of HRP/WGA-HRP into the molar gland, lower lip-gingiva and cheek, many retrogradely labeled ganglion neurons were observed in the ipsilateral main and accessory otic ganglia, superior cervical ganglion and mandibular division of the trigeminal ganglion. In the pterygopalatine ganglion, a small number of positive neurons were found, but in a few cases in which the injected tracer was restricted to the lower lip-gingiva and anterior half of the molar gland, labeled neurons were not detected in the main ganglion nor in its accessory microganglia. These findings indicate that the cat molar gland receives a postganglionic parasympathetic supply from the otic ganglia, postganglionic sympathetic input from the superior cervical ganglion and sensory innervation from the trigeminal ganglion by way of the buccal nerve and mylohyoid nerve. Vessels in the rostral half of the lower lip-gingiva receive the same inputs from the inferior alveolar nerve, and vessels in the caudal half receive inputs from the buccal nerve. The vessels in the cheek mucous membrane receive dual parasympathetic supplies from the otic ganglia and the pterygopalatine ganglion by way of the buccal nerve.

Acetylcholinesterase↗
PubMed2002

Real time monitoring of lipoplex molar mass, size and density.

Time-resolved multiangle laser light scattering (TR-MALLS) is used to monitor the temporal variation of DNA/cationic liposome lipoplex molar masses and geometric sizes throughout the complexation process. The measured molar masses and geometric sizes are in turn used to estimate lipoplex density. The DNA/cationic lipid charge ratio is found to be the primary factor governing lipoplex formation kinetics and the final lipoplex molar mass, geometric size and density. Charge ratios near unity lead to a growing kinetic regime in which initially formed primary lipoplexes undergo further aggregation eventually forming large molar mass lipoplexes of high density, while charge ratios very far from unity yield low molar mass lipoplexes of lower density. It is also noted that solvent composition can play a significant role in the lipoplex formation process with lipoplexes formed in ion-containing media being larger and denser than those formed in dextrose solution.

Cations↗
PubMed2001

Ultrahigh molar mass component detected in ethylhydroxyethyl cellulose by asymmetrical flow field-flow fractionation coupled to multiangle light scattering.

Asymmetrical flow field-flow fractionation (flow FFF) was connected to multiangle light scattering (MALS) and refractive index (RI) detectors for characterization of the molar mass distribution and molecular radius of a cellulose derivative, ethylhydroxyethyl cellulose (EHEC). Experimental conditions were optimized to allow study of a wide range of molar mass including even ultrahigh molar mass (UHM) components. The weight-average molar mass was 3.1 x 10(5) g x mol(-1) representing a very broad range (of molar mass) from 4.0 x 10(4) to 10(7) g x mol(-1), which corresponds to from <20 to 200 nm rms radius. The light scattering signal showed the presence of an UHM component, possibly an aggregate of extreme size, i.e., approximately 10(8) g x mol(-1) with a hydrodynamic diameter of 0.35 microm. Careful choice of the pore size in in-line filters is necessary in order to minimize MALS detector noise without removing the UHM component. Flow FFF-MALS-RI was demonstrated to be uniquely suited to detect the presence of UHM components.

Cellulose↗
PubMed2000

The influence of poly(acrylic acid) molar mass on the properties of polyalkenoate cements formed from zinc oxide/apatite mixtures.

The influence of poly(acrylic acid) molar mass was investigated on cements formed from zinc oxide-apatite mixtures at three aging times; one, seven and 28 days. Cements based on both hydroxyapatite and fluorapatite were investigated. The compressive strength, un-notched fracture strength and fracture toughness increased markedly with poly(acrylic acid) molar mass. The fracture toughness and un-notched fracture strength increased with aging time for the two highest molar mass cements, but decreased with time for the two lowest molar mass cements. The greater chain entanglement density present in the higher molar mass cements is thought to contribute significantly to the cement stability in addition to the crosslinking of the polyacrylate chains by calcium and zinc ions. Substitution of hydroxyapatite by fluorapatite had no significant influence on the mechanical properties of the cements at aging times longer than one day.

Journal Article↗
PubMed2000

Monitoring serum insulin-like growth factor-I (IGF-I), IGF binding protein-3 (IGFBP-3), IGF-I/IGFBP-3 molar ratio and leptin during growth hormone treatment for disordered growth.

OBJECTIVE: Serum IGF-I levels are monitored during GH replacement treatment in adults with GH deficiency (GHD) to guide GH dose adjustment and to minimize occurrence of GH-related side-effects. This is not routine practice in children treated with GH. The aim of this study was to evaluate changes in (1) serum IGF-I, IGFBP-3 and IGF-I/IGFBP-3 molar ratio, and (2) serum leptin, an indirect marker of GH response, during the first year of GH treatment in children with disordered growth. DESIGN: An observational prospective longitudinal study with serial measurements at five time points during the first year of GH treatment was carried out. Each patient served as his/her own control. PATIENTS: The study included 31 patients, grouped as (1) GHD (n = 20) and (2) non-GHD (Turner syndrome n = 7; Noonan syndrome n = 4), who had not previously received GH treatment. MEASUREMENTS: Serum IGF-I, IGFBP-3 and leptin levels were measured before treatment and after 6 weeks, 3 months, 6 months and 12 months of GH treatment, with a mean dose of 0.5 IU/kg/wk in GHD and 0.7 IU/kg/wk in non-GHD groups. IGF-I, IGFBP-3 and the calculated IGF-I/IGFBP-3 molar ratio were expressed as SD scores using reference values from the local population. RESULTS: In the GHD group, IGF-I SDS before treatment was lower compared with the non-GHD (-5.4+/-2.5 vs. -1.8+/-1.0; P<0.001). IGF-I (-1.8 SDS +/- 2.2) and IGFBP-3 (-1.1 SDS +/- 0.6) levels and their molar ratios were highest at 6 weeks and remained relatively constant thereafter. In the non-GHD group, IGF-I levels increased throughout the year and were maximum at 12 months (0.3 SDS +/- 1.4) while IGFBP-3 (1.1 SDS +/- 0.9) and IGF-I/IGFBP-3 molar ratio peaked at 6 months. In both groups, IGF-I SDS and IGF-I/IGFBP-3 during treatment correlated with the dose of GH expressed as IU/m2/week (r-values 0. 77 to 0.89; P = 0.005) but not as IU/kg/week. Serum leptin levels decreased significantly during GH treatment in the GHD (median before treatment 4.0 microg/l; median after 12 months treatment 2.4 microg/l; P = 0.02) but not the non-GHD (median before treatment 3.0 microg/l; median after 12 months treatment 2.6 microg/l). In the GHD group, serum leptin before treatment correlated with 12 month change in height SDS (r = 0.70, P = 0.02). CONCLUSIONS: The pattern of IGF-I, IGFBP-3 and their molar ratio during the first year of GH treatment differed between the GHD and non-GHD groups. Calculation of GH dose by surface area may be preferable to calculating by body weight. As a GH dose-dependent increase in serum IGF-I and IGF-I/IGFBP-3 may be associated with adverse effects, serum IGF-I and IGFBP-3 should be monitored routinely during long-term GH treatment. Serum leptin was the only variable that correlated with first year growth response in GHD.

Adolescent↗
PubMed1997

Early ultrasound diagnosis and follow-up of molar pregnancies.

The objective of this study was to investigate the role of ultrasound in the differential diagnosis and management of early pregnancies presenting with placental molar changes. Placental features were recorded over a 10-month period in women undergoing ultrasound examination at 10-14 weeks of gestation. In cases of a molar pregnancy, the fetal karyotype was obtained in utero and, if the pregnancy continued, the maternal concentration of human chorionic gonadotropin (hCG) and uterine artery resistance to flow were measured serially. A histopathological examination of the placenta was performed in all cases after delivery. During the study period, 9425 women had an early scan and 11 molar pregnancies were identified including one classical mole, four hydatidiform moles coexisting with a normal pregnancy, three partial triploid moles and three partial moles associated in one case with a fetus presenting congenitial anomalies diagnostic of Beckwith-Wiedemann syndrome. The hCG levels were high in all cases except one case of triploidy and remained high during the rest of the pregnancy in cases of hydatidiform moles coexisting with a fetus. In these cases, the uterine artery resistance was normal. The present data indicate that placental ultrasound examination can correctly identify molar changes in early pregnancy and together with hCG level and uterine Doppler measurements can establish the differential diagnosis in utero of the various forms of placental molar transformations.

Adult↗