In primary rat hippocampal neuron–astrocyte co-cultures, sulbactam pre-incubation (5–125 μM, 48 h) reduced cell death and preserved metabolic viability after oxygen–glucose deprivation. A separate astrocyte-culture experiment found increased GLT-1 protein expression after the same pretreatment and challenge. Saline controls did not reproduce these effects. These pretreatment findings concern cultured cells; they do not establish human neuroprotection. The paper reports sulbactam without specifying its salt form. [2]
Sulbactam-derived intermediates in SHV-1 crystals
Biochemical assay · Single crystals of purified wild-type SHV-1 β-lactamase, with deacylation-deficient SHV E166A as a mechanistic comparator. [1]
Sulbactam yielded a mixture of cis-enamine, trans-enamine and imine acyl-enzyme species in wild-type SHV-1. The E166A comparator showed a more prominent trans-enamine population; its intermediate distribution does not faithfully stand in for the wild-type enzyme.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5 mM final inhibitor concentration
- Exposure duration
- Steady-state Figure 3 spectrum; exact elapsed time for that figure is not specified.
- Method
- Raman crystallography at room temperature; difference spectra relative to apo-enzyme.
- Vehicle
- 2 mM HEPES; inhibitor stocks prepared at pH 7.0.
- Reported limits
- Sulbactam salt and lot are not specified. No IC50 or cellular treatment dose is established by this experiment; biological replicate n is not specified in the selected methods/caption.
Methods and source locations
Compound reported in the paper: Sulbactam
Biochemical mechanism evidence for reported sulbactam; no free-acid-specific potency or Adooq-lot claim.
Materials and Methods: Inhibitors S3, Raman Crystallography S6; Results and Discussion S11; Figure 3 and caption
Steady-state Raman bands assigned to sulbactam-derived acyl-enzyme intermediates in wild-type SHV-1 crystals: Raman difference spectrum with apo-enzyme spectrum subtracted; assignments supported by isotope experiments and calculations in the same article. — Figure 3 top trace and caption; Results and Discussion, S11, sulbactam assignments
Protection of hippocampal neuron–astrocyte co-cultures against OGD
Cell-based study · Primary hippocampal neuron–astrocyte co-cultures from postnatal 0–1-day Wistar rats. [2]
Sulbactam pre-incubation reduced the HO/PI-defined death fraction and preserved MTT activity after OGD. Normal saline did not protect, and 125 μM sulbactam without OGD did not measurably change basal death or viability in the reported comparisons. This is a pretreatment cell-culture result.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5, 25 or 125 μM sulbactam
- Exposure duration
- 48 h pre-incubation, then 2 h OGD without sulbactam; readouts 24 h after reoxygenation.
- Method
- Pre-incubation of primary hippocampal neuron–astrocyte co-cultures before oxygen–glucose deprivation.
- Vehicle
- Normal saline.
- Co-treatment
- No antibiotic partner.
- Reported limits
- Supplier, salt, CAS, lot and vehicle volume are not specified in the read methods. Exact bar values are not tabulated in the read locations and were not estimated from the image.
Methods and source locations
Compound reported in the paper: Sulbactam
Cell-culture results with reported sulbactam; no animal efficacy, therapeutic dose or human neuroprotection claim.
Methods: Animals, Part 1, Neuron-Astrocyte Co-culture, OGD, Neuronal Death Assay, Cell Viability Assay, Data Analysis; Results Neuronal Survival and Viability; Figure 2
Cell-death fraction after OGD in primary neuron–astrocyte co-cultures: Hoechst/propidium iodide staining; randomized fields counted by a blinded observer. n = 5 independent cultures per group; reported data mean ± SD. — Results: Neuronal Survival and Viability; Figure 2a–g and caption; Methods Part 1, Neuronal Death Assay, Data Analysis
Co-culture metabolic viability after OGD, normalized to untreated control: MTT optical-density ratio to control; this is a mixed-culture metabolic assay, not a neuron-specific cell count. n = 5 independent cultures per design; six wells per culture preparation described for MTT; mean ± SD. — Figure 2h and caption; Results: Neuronal Survival and Viability; Methods Cell Viability Assay, Part 1, Data Analysis
GLT-1 expression in OGD-treated astrocytes
Cell-based study · Primary hippocampal astrocyte cultures from neonatal Wistar rats; third or fourth passage, reported >98% purity by GFAP immunocytochemistry. [2]
Sulbactam pretreatment increased astrocytic GLT-1 expression after OGD relative to OGD-only and saline-vehicle controls. The experiment measures protein expression; it does not itself establish glutamate transport function or prove that GLT-1 mediates the separate co-culture survival result.
Literature-Reported Treatment Conditions
- Treatment concentration
- 5, 25 or 125 μM sulbactam
- Exposure duration
- 48 h pre-incubation, then 2 h OGD without sulbactam; GLT-1 assessed 12 h and 24 h after reoxygenation.
- Method
- Pretreatment of astrocyte cultures; immunocytochemistry and western blot after OGD.
- Vehicle
- Normal saline.
- Co-treatment
- No antibiotic partner.
- Reported limits
- Supplier, salt, CAS and lot are not specified in the read methods. No exact fold change was digitized from the figures.
Methods and source locations
Compound reported in the paper: Sulbactam
Astrocytic protein-expression response only; no claim of direct GLT-1 binding, transporter activity or clinical benefit.
Methods: Animals, Part 1, Astrocyte Culture, OGD, Western Blot Analysis, Immunocytochemistry Analysis, Data Analysis; Results GLT-1 Expression; Figure 3A–B
Astrocytic GLT-1 expression after OGD: GLT-1 staining integrated optical density and GLT-1/β-actin immunoblot quantification; n = 5 independent cultures per group; mean ± SD. — Figure 3A–B, panels g–h and caption; Results GLT-1 Expression; Methods Part 1, Western Blot Analysis, Immunocytochemistry Analysis, Data Analysis
